AU2005305765A1 - Method and kit for detecting a risk of essential arterial hypertension - Google Patents

Description

WO 2006/053955 PCT/F12005/050429 1 Method and kit for detecting a risk of essential arterial hypertension BACKGROUND OF THE INVENTION 5 Field of the Invention The present invention relates generally to the field of diagnosis of cardiovascular diseases (CVD) such as arterial hypertension (HT). More particularly, it provides a method of 10 diagnosing or detecting a predisposition or propensity or susceptibility for HT. Specifically, the invention focuses on a method that comprises the steps of providing a biological sample from the subject to be tested and detecting the presence or absence of one or several genomic single nucleotide polymorphism (SNP) markers in the biological sample. Furthermore, the invention utilizes both genetic and phenotypic information as well as information obtained by 15 questionnaires to construct a score that provides the probability of developing HT. In addition, the invention provides a kit to perform the method. The kit can be used to set an etiology based diagnosis of HT for targeting of treatment and preventive interventions such as dietary advice, as well as stratification of the subject in clinical trials testing drugs and other interventions. 20 Description of Related Art Public health significance of CVD and HT 25 Cardiovascular Diseases (CVD) (ICD/10 codes 100-199, Q20-Q28) include ischemic (coronary) heart disease (IHD, CHD), hypertensive diseases, cerebrovascular disease (stroke) and rheumatic fever/rheumatic heart disease, among others (AHA, 2004). HT (ICD/10 110 115) is defined as systolic pressure of 140 mm Hg or higher, or diastolic pressure of 90 mm Hg or higher, or taking antihypertensive medicine (AHA, 2004). Apart from being a CVD 30 itself, HT is a risk factor for other CVD, such as IHD, stroke and congestive heart failure (CHF). About half of those people who have a first heart attack and two thirds of those who have a first stroke, have blood pressure (BP) higher than 160/95 mm Hg. HT precedes the development of CHF in 91% of cases (AHA, 2004). 35 Of patients with HT, 90-95% have essential HT in which the underlying cause remains unknown. Essential HT refers to a lasting increase in BP with heterogeneous genetic and environmental causes. Its prevalence rises with age irrespective of the type of BP measurement and the operational thresholds used for diagnosis. HT aggregates with other cardiovascular risk factors such as abdominal obesity, dyslipidaemia, glucose intolerance, 40 hyperinsulinaemia and hyperuricaemia, possibly because of a common underlying cause (Salonen JT et al, 1981, 1998, Staessen JA et al, 2003). In 2001 an estimated 16.6 million - or one-third of total global deaths - resulted from the various forms of CVD (7.2 million due to HT, 5.5 million to cerebrovascular disease, and an 45 additional 3.9 million to hypertensive and other heart conditions). At least 20 million people survive heart attacks and strokes every year, a significant proportion of them requiring costly clinical care, putting a huge burden on long-term care resources. It is necessary to recognize that CVDs are devastating to men, women and children (AHA, 2004).

WO 2006/053955 PCT/F12005/050429 2 Around 80% of all CVD deaths worldwide took place in developing, low and middle-income countries. It is estimated that by 2010, CVD will be the leading cause of death in both developed and developing countries. The rise in CVDs reflects a significant change in dietary habits, physical activity levels, and tobacco consumption worldwide as a result of 5 industrialization, urbanization, economic development and food market globalization (WIO, 2004). This emphasizes the role of relatively modem environmental or behavioral risk factors. However, ethnic differences in the incidence and prevalence of CVD and the enrichment of CVD in families suggest that heritable risk factors play a major role. 10 In terms of disability measured in disability-adjusted life years (DALYs) CVD caused 9.7% of global DALYs, 20.4% of DALYs in developed countries and 8.3% of DALYs in the developing countries. HT caused 1.4% of global DALYs, 4.7% of DALYs in developed countries and 0.9% of DALYs in the developing countries (Murray CJL and Lopez AD, 1997). 15 On the basis of data from the NHANES III study (1988-1994), it is estimated that in 2001, 64.4 million Americans were affected by some form of CVD, which corresponds to a prevalence of 22.6% (21.5% for males, 22.4% for females). Of these, 50 million had HT (20% prevalence). Of those with HT, 30% do not know they have HT; 34% are on medication and 20 have HT controlled; 25% are on medication but do not have their HT under control; and 11% are not on medication (AHA, 2004). HT is also a public health problem in developing countries where prevalences of 10% or higher are common and it is frequently associated with low levels of awareness, treatment and control (Fuentes RM et al, 2000). 25 The cost of CVD in the United States in 2004 was estimated at $368.4 billion ($133.2 billion for HT, $53.6 billion for stroke, $55.5 billion for hypertensive disease). This figure includes health expenditures (direct costs) and lost productivity resulting from morbidity and mortality (indirect costs) (AHA, 2004). 30 Pathophysiology of essential HT The pressure required to move blood through the circulatory bed is provided by the pumping action of the heart [cardiac output (CO)] and the tone of the arteries [peripheral resistance (PR)]. Each of these primary determinants of BP is, in turn, determined by the interaction of a complex series of factors. 35 Factors affecting cardiac output An increased CO has been found in some young, borderline hypertensives who may display a hyperkinetic circulation. If it is responsible for HT, the increase in CO could logically arise in two ways: either from an increase in fluid volume (preload) or from an increase in 40 contractility from neural stimulation of the heart. However, even if it is involved in the initiation of HT, the increased CO probably does not persist. The typical hemodynamic finding in established HT is an elevated PR and normal CO (Cowley AW, 1992). Although an increased heart rate may not simply be a reflection of a hyperdynamic circulation 45 or an indicator of increased sympathetic activity, multiple epidemiologic surveys have shown that an elevated heart rate is an independent predictor of the development of HT (Palatini P and Julius S, 1999). Left ventricular hypertrophy has generally been considered a compensatory mechanism to an 50 increased vascular resistance. However, it could also reflect a primary response to repeated WO 2006/053955 PCT/F12005/050429 3 neural stimulation and, thereby, could be an initiating mechanism for HT (Julius S et al., 1991c) as well as an amplifier of CO that reinforces the elevation of BP from arterial stiffening (Segers P et al., 2000). 5 Another mechanism that could induce HT by increasing CO would be an increased circulating fluid volume (preload). However, in most studies, subjects with high BP have a lower blood volume and total exchangeable sodium than normal subjects (Harrap SB et al., 2000). Even without an expanded total volume, blood may be redistributed so that more is in the central or cardiopulmonary section because of greater peripheral vasoconstriction (Schobel HP et al., 10 1993). Venous return to the heart would thereby be increased and could mediate an increased CO. Excess sodium intake induces HT by increasing fluid volume and preload, thereby increasing CO (Chobanian AV and Hill M, 2000). Both experimental data (Tobian L, 1991) and 15 epidemiologic evidence (Stamler J et al., 1997) support a close association between HT and a high sodium-potassium ratio in humans. Because almost everyone in industrialized societies ingests a high-sodium diet, the fact that only about half will develop HT suggests a variable degree of BP sensitivity to sodium (Weinberger MH, 1996). 20 In healthy people, when BP increases, renal excretion of sodium and water increases, shrinking fluid volume and returning the BP to normal - this phenomenon is pressure natriuresis. On the basis of animal experiments and computer models, the regulation of body fluid volume by the kidneys is considered to be the dominant mechanism for the long-term control of BP (Guyton AC 1961, 1992). Therefore, if HT develops, something must be wrong 25 with the pressure-natriuresis control mechanism; otherwise the BP would return to normal (Cowley AW and Roman RJ, 1996). In patients with primary HT a resetting of the pressure sodium excretion curve prevents the return of BP to normal (Palmer BF, 2001). The shift in pressure-natriuresis requires increased BP to maintain fluid balance. The pressure-natriuresis relationship can be modified by neural and humoral factors including the renin-angiotensin 30 system (RAS), sympathetic nervous activity, atrial natriuretic factor, metabolites of arachidonic acid, and intrarenal nitric oxide (Moreno C et al., 2001; Majid DS et al., 2001). The major modifier is likely to be theRAS (Hall JE et al., 1999; van Paassen P et al., 2000), with an increase in renal sodium reabsorption occurring at concentrations of Angiotensin II much below those needed for peripheral vasoconstriction. Angiotensin II acts not only on 35 vascular smooth muscle and the adrenal cortex but also within the heart, kidneys, and central and autonomic nervous systems. These actions amplify its volume-retaining and vasoconstrictive effects on the peripheral vascular system, thus affecting both CO and PR. Furthermore, Angiotensin II induces cell growth and hypertrophy independent of its effect on BP (Su EJ et al., 1998). Moreover, Angiotensin II appears to induce an inflammatory response 40 in vascular smooth muscle cells (Kranzh6fer R et al., 1999), with activation of nuclear factor k-B (Luft FC, 2001) and adhesion molecule-I expression (Tummala PE et al., 1999), which may serve as direct links to atherosclerosis. Stress may activate the sympathetic nervous system (SNS) directly; and SNS overactivity, in 45 turn, may interact with high sodium intake, the RAS, and insulin resistance, among other possible mechanisms. Considerable evidence supports increased SNS activity in early HT (Esler M et al., 2001) and, even more impressively, in the still-normotensive offspring of hypertensive parents, of whom a large number are likely to develop HT. Whatever the specific role of SNS activity in the pathogenesis of HT, it appears to be involved in the 50 increased cardiovascular morbidity and mortality that afflicts hypertensive patients during the WO 2006/053955 PCT/F12005/050429 4 early morning hours. Epinephrine levels begin to increase after awakening and norepinephrine rises sharply on standing (Dodt C et al., 1997). As a consequence of the increased SNS activity, BP rises suddenly and markedly, and this rise is at least partly responsible for the increase in sudden death, heart attack, and stroke during the early morning hours. Increased 5 sympathetic activity is probably also responsible for the increased heart rate present in many hypertensives that was previously noted to be associated with increased cardiovascular mortality. Factors affecting peripheral resistance 10 HT is maintained by increased PR, largely due to decreased arterial lumen size or radius. According to Poiseuille's law, vascular resistance is positively related to both the viscosity of blood and the length of the arterial system, and negatively related to the third power of the luminal radius. Because neither viscosity nor length is altered much if at all, and because small changes in the luminal radius can have a major effect, it is apparent that the increased 15 vascular resistance seen in established HT must reflect changes in the calibre of the small resistance arteries and arterioles (Folkow B et al., 1970). Because of the increased wall thickness-lumen diameter ratio, higher wall stress and intraluminal pressure develop when resistance vessels are stimulated. 20 In HT, small arteries undergo functional, structural and mechanical changes, resulting in reduced lumen size and increased peripheral resistance (Mulvany MJ, 2002; Intengan HD and Schiffrin EL. 2001). Functional alterations include enhanced reactivity or impaired relaxation, and reflect changes in excitation-contraction coupling, altered electrical properties of vascular smooth muscle cells, or endothelial dysfunction (Johns DG et al, 2000; Feldman RD and Gros 25 R, 1998). Major structural changes include remodelling due to increased cell growth, extracellular matrix deposition and inflammation (Mulvany MJ, 2002; Intengan H-D and Schiffrin EL, 2001; Brasier AR, 2002). Vascular smooth muscle cells are central to these events and play a fundamental role in the dynamic processes underlying the alterations that occur in HT. 30 Vascular changes in HT are associated with humoral and mechanical factors that modulate signalling events, resulting in abnormal function and growth of cellular components of the media (Touyz RM, 2000; Koller A, 2002). The humoral factors that regulate arteries in HT include vasoconstrictor agents such as angiotensin II, endothelin-1, catecholamines and 35 vasopressin; vasodilator agents such as nitric oxide, endothelium-derived hyperpolarizing factor and natriuretic peptides; growth factors such as insulin-like growth factor-1, platelet derived growth factor (PDGF), epidermal growth factor (EGF) and basic fibroblast growth factor; and cytokines such as transforming growth factor-[beta], tumour necrosis factor and interleukins (Touyz RM, 2000). Mechanical factors that influence the vasculature in HT 40 include shear stress, wall stress and the direct actions of pressure itself (Touyz RM, 2000; Koller A, 2002). In addition to these factors, there is growing evidence that reactive oxygen species (ROS) that act as intercellular and intracellular signalling molecules, regulate vascular tone and structure (Wilcox CS, 2002; Berry C et al, 2001). A recent advance in the field of angiotensin II signalling was the demonstration that, in 45 addition to its vasoconstrictor properties, angiotensin II has potent mitogenic-like and proinflammatory-like characteristics. These actions are mediated through phosphorylation of both nonreceptor tyrosine kinases and receptor tyrosine kinases (Touyz RM, 2003). It is also becoming increasingly apparent that many signalling events that underlie abnormal vascular function in HT are influenced by changes in intracellular redox status. In particular, increased 50 bioavailability of ROS stimulates growth-signalling pathways, induces expression of WO 2006/053955 PCT/F12005/050429 5 proinflammatory genes, alters contraction-excitation coupling and impairs endothelial function (Touyz RM, 2003). In concert with the various functional and structural changes that are responsible for HT, the arteries become stiffer or less elastic. Vascular stiffness progressively increases with age 5 (Slotwiner DJ et al., 2001) and is responsible for the progressive increase in systolic as compared to diastolic pressure, leading to the typical increase of pulse pressure that is now recognized to be the major determinant of cardiovascular risk (Beltran A et al., 2001). Measures of stiffness and elasticity have been shown to be an independent predictor of the development of HT (Liao D et al., 1999) and a marker of cardiovascular risk in those with HT 10 (Blacher J et al., 1999). Changes in the physical characteristics of the large arteries reflected in the BP pulse contour alter not only BP and pulse pressure, but also cardiac work and performance. The complexity of pathophysiologic mechanisms that lead to BP elevation is such that 15 selective, mechanistically based antihypertensive treatment is rarely possible in any hypertensive patient. HT is highly prevalent among middle-aged and elderly persons, and the success rate in controlling BP in these individuals is poor. Current treatment guidelines generally recommend a generic approach to treating HT, with little emphasis on selecting therapy on the basis of the underlying pathophysiology of the elevated BP (Chobanian AV et 20 al, 2003; ESH/ESC, 2003). With increased recognition of specific causes, it may be possible to develop therapies selective for distinct pathophysiologic mechanisms with fewer adverse effects, resulting in more effective BP reduction. The use of powerful new techniques of genetics, genomics, and proteomics, integrated with systems physiology and population studies, will make more selective and effective approaches to treating and even preventing HT 25 possible in the coming decades (Oparil S et al, 2003). Essential HT: a polygenic disease Nuclear family studies show greater similarity in BP within families than between families, with heritability estimates ranging between 0.20 and 0.46 (Fuentes RM, 2003). Twin studies 30 document greater concordance of BP in monozygotic than dizygotic twins, giving the highest heritability estimates between 0.48 and 0.64 (Fuentes RM, 2003). Adoption studies demonstrate greater concordance of BP among biological siblings than adoptive siblings living in the same household, estimating heritability between 0.45 and 0.61 (Fuentes RM, 2003). 35 Single genes can have major effects on BP, accounting for the rare Mendelian forms of high and low BP (Lifton RP et al, 2001). Although identifiable single-gene mutations account for only a small percentage of HT cases, studying these rare disorders may elucidate pathophysiologic mechanisms that predispose to more common forms of HT and may suggest 40 novel therapeutic approaches (Lifton RP et al, 2001). Mutations in 10 genes that cause Mendelian forms of human HT and 9 genes that cause hypotension have been described to date (Lifton RP et al, 2001; Wilson FH et al, 2001). These mutations affect BP by altering renal salt handling, reinforcing the hypothesis that the development of HT depends on genetically determined renal dysfunction with resultant salt and water retention (Guyton AC, 45 1991). Importantly, all the monogenic HT syndromes identified to date are caused by defects resulting in renal salt retention, whereas all the low BP syndromes share a common mechanism of excess renal sodium loss (Hopkins PN and Hunt SC, 2003). The best studied monogenic cause of HT is the Liddle syndrome, a rare but clinically 50 important disorder in which constitutive activation of the epithelial sodium channel WO 2006/053955 PCT/F12005/050429 6 predisposes to severe, treatment-resistant HT (Shimkets RA et al, 1994). Epithelial sodium channel activation has been traced to mutations in the beta or gamma subunits of the channel, resulting in inappropriate sodium retention at the renal collecting duct level. Patients with the Liddle syndrome typically display volume-dependent, low-renin, and low-aldosterone HT. 5 In most cases, HT results from a complex interaction of genetic, environmental, and demographic factors. Improved techniques of genetic analysis, especially candidate gene association studies and genome wide linkage analysis (genome wide scan, GWS), have enabled a search for genes that contribute to the development of primary HT in the 10 population. Thus far, the candidate gene approach has provided more examples than the linkage approach of gene variants that appear to affect BP. Reasonable candidate genes to consider include genes related to physiological systems known to be involved in the control of BP and genes 15 known to affect BP in mouse models. To date more than 80 candidate genes have been evaluated for HT (Fuentes RM, 2004, unpublished review). However, the association with HT of only three genes have been widely replicated: angiotensinogen precursor (AGT), adducin 1 (ADD1) and guanine nucleotide-binding protein, beta-3 subunit (GNB3) (Hopkins PN and Hunt SC, 2003). Gene-environment interactions affecting HT treatment have been shown 20 between AGT, ADDI and salt intake reduction (Hunt SC et al, 1998; Hunt SC et al, 1999; Cusi D et al, 1997), and between ADDI, GNB3 and diuretic treatment (Cusi D et al, 1997; Turner ST et al, 2001). Gene-gene interactions affecting HT risk development have been shown between ADDI and the ACE gene I/D polymorphisms (Staessen JA et al, 2001). Lessons learned from the studies of candidate genes to date include the shortcomings that 25 result from the limited statistical power of many studies, expected variation from one population to another, the need for better phenotyping of study subjects, the relatively small effect of the genes studied on population prevalence of HT, and the lack of sufficient certainty of consequences of any genes studied thus far to make treatment recommendations based on genotype (Hopkins PN and Hunt SC, 2003). 30 To date more than 30 GWS studies have been reported to identify loci for BP/HT (Fuentes RM, 2004, unpublished review).Some studies utilized families, others affected or dissimilar sibling pairs. Linked loci with at least indicative LOD scores to BP/HT have been observed on every chromosome. Perhaps most striking is the lack of consistently linked loci. 35 Koivukoski L et al, 2004 found evidence of susceptibility regions for BP/HT on chromosomes 2p12-q22.1 and 3p14.1-ql2.3 that had modest or non-significant linkage in each individual study when applying the genome-search meta-analysis method (GSMA) to nine published genome-wide scans of BP (n = 5) and HT (n = 4) from Caucasian populations. This may serve to illustrate the heterogeneity of human HT as well as the potential shortcomings of 40 attempting to compare studies using different methodologies. Opportunity for population genetics 45 Previous medical research concerning the genetic etiology of HT has been based to a large extent on retrospective case-control and family studies in humans and studies in genetically modified animals. As recognized only recently, retrospective case-control studies are prone to survival and selection biases, and they have produced a myriad of biased findings concerning a large number of candidate genes. A commonly used approach is to compare gene expression 50 between affected and unaffected persons. Gene expression studies, which are mostly cross- WO 2006/053955 PCT/F12005/050429 7 sectional, cannot however separate cause and consequence. Findings from animal models concerning HT cannot be generalised to humans, as the pathophysiology in humans is unique. The unsuitability of the animal studies is the main reason why genetic epidemiologic studies are the most important means in the clarification of genetic etiologies of human diseases. 5 Prospective cohort studies in humans overcome these problems. Developments in GWS and sequencing technology and methods of data analysis render possible the attempt to identify liability genes in complex, multifactorial traits, and to dissect the role of genetic predisposition and environment/life style factors in these disorders with new precision. 10 Genetic and environmental effects vary over the life span, and only longitudinal studies in genetically informative data sets permit the study of such effects. A major advantage of population genetics approaches in disease gene discovery over other methodologies is that it will yield diagnostic markers that are valid in humans. 15 The identification of genes causing major public health problems such as HT is now enabled by the following recent advances in molecular biology, population genetics and bioinformatics: 1. the availability of new genotyping platforms that will dramatically lower operating costs and increase throughputs; 2. the application of genome scans using dense marker maps; 3. data analysis using new powerful statistical methods testing for linkage 20 disequilibrium using haplotype sharing analysis, and 4. the recognition that a smaller number of genetic markers than previously thought is sufficient for genome scans in genetically homogeneous populations. Traditional GWS using microsatellite markers with linkage analyses have not been successful 25 in finding genes causing common diseases. The failure has in part been due to too small a number of genetic markers used in GWS, and in part due to too heterogeneous study populations. With the advancements of the human genome project and genotyping technology, the first dense marker maps have recently become available for mapping the entire human genome. The microarrays used by Jurilab include probes for over 100,000 single 30 nucleotide polymorphism (SNP) markers. These SNPs form a marker map covering, for the first time, the entire genome tightly enough for the discovery of most disease genes causing HT. Genetic homogeneity of the East Finland founder population 35 Finns descend from two human immigration waves occurring about 4,000 and 2,000 years ago. Both Y-chromosomal haplotypes and mitochondrial sequences show low genetic diversity among Finns compared with other European populations and confirm the long standing isolation of Finland (Sajantila A et al, 1996). During King Gustavus of Vasa (1523 40 1560) over 400 years ago, internal migrations created regional subisolates, the late settlements (Peltonen L et al, 1999). The most isolated of these are the East Finns. The East Finnish population is the most genetically-homogenous population isolate known that is large enough for effective gene discovery program. The reasons for homogeneity are: 45 the young age of the population (fewer generations); the small number of founders; long-term geographical isolation; and population bottlenecks because of wars, famine and fatal disease epidemics. Owing to the genetic homogeneity of the East Finland population, there are fewer mutations 50 in important disease predisposing genes and the affected individuals share a similar genetic WO 2006/053955 PCT/F12005/050429 8 background. Because of the stronger linkage disequilibrium (LD), fewer SNPs and fewer subjects are needed for GWS than in other populations. 5 SUMMARY OF THE INVENTION The present invention relates to single nucleotide polymorphism (SNP) markers, combinations of such markers and haplotypes associated with altered risk of HT, and genes associated with HT within or close to which said markers or haplotypes are located. Said SNP 10 markers may be associated either with increased HT risk or reduced HT risk i.e. protective of HT. The "prediction" or risk implies here that the risk is either increased or reduced. Thus, the present invention provides individual SNP markers associated with HT and combinations of SNP markers and haplotypes in genetic regions associated with HT, genes 15 previously known in the art, but not known to be associated with HT, methods of estimating susceptibility or predisposition of an individual to HT, as well as methods for prediction of clinical course and efficacy of treatments for HT using polymorphisms in the HT risk genes. Accordingly, the present invention provides novel methods and compositions based on the disclosed HT associated SNP markers, combinations of SNP markers, haplotypes and genes. 20 The invention further relates to a method for estimating susceptibility or predisposition of an individual to HT comprising the detection of the presence of SNP markers and haplotypes, or an alteration in expression of an HT risk gene set forth in tables 2 through 8, as well as alterations in the polypeptides encoded by the said HT risk genes. The alterations may be 25 quantitative, qualitative, or both. The invention yet further relates to a method for estimating susceptibility or predisposition of an individual to HT. The method for estimating susceptibility or predisposition of an individual to HT is comprised of detecting the presence of at-risk haplotypes in an 30 individual's nucleic acid. The invention further relates to a kit for estimating susceptibility to HT in an individual comprising wholly or in part: amplification reagents for amplifying nucleic acid fragments containing SNP markers, detection reagents for genotyping SNP markers and interpretation 35 software for data analysis and risk assessment. In one aspect, the invention relates to methods of diagnosing a predisposition to HT. The methods of diagnosing a predisposition to HT in an individual include detecting the presence of SNP markers predicting HT, as well as detecting alterations in expression of genes which 40 are associated with said markers. The alterations in expression can be quantitative, qualitative, or both. A further object of the present invention is a method of identifying the risk of HT by detecting SNP markers in a biological sample of the subject. The information obtained from this 45 method can be combined with other information concerning an individual, e.g. results from blood measurements, clinical examination and questionnaires. The blood measurements include but are not restricted to the determination of plasma or serum cholesterol and high density lipoprotein cholesterol. The information to be collected by questionnaire includes information concerning gender, age, family and medical history such as the family history of 50 HT and diabetes. Clinical information collected by examination includes e.g. information WO 2006/053955 PCT/F12005/050429 9 concerning height, weight, hip and waist circumference, systolic and diastolic BP, and heart rate. The methods of the invention allow the accurate diagnosis of HT at or before disease onset, 5 thus reducing or minimizing the debilitating effects of HT. The method can be applied in persons who are free of clinical symptoms and signs of HT, in those who already have clinical HT, in those who have a family history of HT, or in those who have an elevated level or levels of risk factors of HT. 10 The invention further provides a method of diagnosing susceptibility to HT in an individual. This method comprises screening for at-risk haplotypes that predict HT that are more frequently present in an individual susceptible to HT, compared to the frequency of its presence in the general population, wherein the presence of an at-risk haplotype is indicative of a susceptibility to HT. The "at-risk haplotype" may also be associated with a reduced rather 15 than increased risk of HT. An "at-risk haplotype" is intended to embrace one or a combination of haplotypes described herein over the markers that show high correlation to HT. Kits for diagnosing susceptibility to HT in an individual are also disclosed. Those skilled in the art will readily recognize that the analysis of the nucleotides present in 20 one or several of the SNP markers of this invention in an individual's nucleic acid can be done by any method or technique capable of determining nucleotides present in a polymorphic site. As it is obvious in the art the nucleotides present in SNP markers can be determined from either nucleic acid strand or from both strands. 25 The major application of the current invention involves prediction of those at higher risk of developing HT. Diagnostic tests that define genetic factors contributing to HT might be used together with or independent of the known clinical risk factors to define an individual's risk relative to the general population. Better means for identifying those individuals at risk of HT should lead to better preventive and treatment regimens, including more aggressive 30 management of the current clinical risk factors for sequelae of HT such as cigarette smoking, hypercholesterolemia, elevated LDL cholesterol, low HiDL cholesterol, HT and elevated BP, diabetes mellitus, glucose intolerance, insulin resistance and the metabolic syndrome, obesity, lack of physical activity, and inflammatory components as reflected by increased C-reactive protein levels or other inflammatory markers. Information on genetic risk may be used by 35 physicians to help convince particular patients to adjust life style (e.g. to stop smoking, reduce caloric intake, to increase exercise). Finally, preventive measures aimed at lowering blood pressure such as reduction of weight, intake of salt and alcohol, can be both better motivated to the patients and selected on the basis of the molecular subdiagnosis of HT. 40 A further object of the invention is to provide a method for the selection of human subjects for studies testing antihypertensive effects of drugs. Another object of the invention is a method for the selection of subjects for clinical trials testing antihypertensive drugs. 45 Still another object of the invention is to provide a method for prediction of clinical course and efficacy of treatments for HT using polymorphisms in the HT risk genes. The genes, gene products and agents of the invention are also useful for treating HT, for monitoring the effectiveness of the treatment, and for drug development. Kits are also provided for the 50 diagnosis, treatment and prognosis of HT.

WO 2006/053955 PCT/F12005/050429 10 DETAILED DESCRIPTION OF THE INVENTION Representative Target Population 5 An individual at risk of HT is an individual who has at least one risk factor of HT, such as family history of HT, central or other type of obesity, lack of physical activity, high sodium intake, high intake of saturated fats, low intake of potassium and/or magnesium, low H-DL cholesterol, diabetes mellitus, glucose intolerance, insulin resistance and the metabolic 10 syndrome, elevated inflammatory marker, and an at-risk allele or haplotype with one or several HT risk SNP markers. In another embodiment of the invention, an individual who is at risk of HT is an individual who has a risk-increasing allele in an HT risk gene, in which the presence of the 15 polymorphism is indicative of a susceptibility to HT. The term "gene," as used herein, refers to an entirety containing all regulatory elements located both upstream and downstream as well as within of a polypeptide encoding sequence, 5' and 3' untranslated regions of mRNA and the entire polypeptide encoding sequence including all exon and intron sequences (also alternatively spliced exons and introns) of a gene. 20 Assessment for At-Risk Alleles and At-Risk Haplotypes The genetic markers are particular "alleles" at "polymorphic sites" associated with HT. A 25 nucleotide position at which more than one sequence is possible in a population, is referred to herein as a "polymorphic site". Where a polymorphic site is a single nucleotide in length, the site is referred to as a SNP. For example, if at a particular chromosomal location, one member of a population has an adenine and another member of the population has a thymine at the same position, then this position is a polymorphic site, and, more specifically, the 30 polymorphic site is a SNP. Polymorphic sites may be several nucleotides in length due to insertions, deletions, conversions or translocations. Each version of the sequence with respect to the polymorphic site is referred to herein as an "allele" of the polymorphic site. Thus, in the previous example, the SNP allows for both an adenine allele and a thymine allele. 35 Typically, a reference nucleotide sequence is referred to for a particular gene. Alleles that differ from the reference are referred to as "variant" alleles. The polypeptide encoded by the reference nucleotide sequence is the "reference" polypeptide with a particular reference amino acid sequence, and polypeptides encoded by variant alleles are referred to as "variant" polypeptides with variant amino acid sequences. 40 Nucleotide sequence variants can result in changes affecting the properties of a polypeptide. These sequence differences, when compared to a reference nucleotide sequence, include insertions, deletions, conversions and substitutions: e.g. an insertion, a deletion or a conversion may result in a frame shift generating an altered polypeptide; a substitution of at 45 least one nucleotide may result in a premature stop codon, aminoacid change or abnormal mRNA splicing; the deletion of several nucleotides, resulting in a deletion of one or more amino acids encoded by the nucleotides; the insertion of several nucleotides, such as by unequal recombination or gene conversion, resulting in an interruption of the coding sequence of a reading frame; duplication of all or a part of a sequence; transposition; or a rearrangement WO 2006/053955 PCT/F12005/050429 11 of a nucleotide sequence, as described in detail above. Such sequence changes alter the polypeptide encoded by an HT susceptibility gene. For example, a nucleotide change in a gene resulting in a change in corresponding polypeptide aminoacid sequence can alter the physiological properties of a polypeptide resulting in a polypeptide having altered biological 5 activity/function, distribution or stability. Alternatively, nucleotide sequence variants can result in changes affecting transcription of a gene or translation of it's mRNA. A polymorphic site located in a regulatory region of a gene may result in altered transcription of a gene e.g. due to altered tissue specifity, altered 10 transcription rate or altered response to transcription factors. A polymorphic site located in a region corresponding to the mRNA of a gene may result in altered translation of the mRNA e.g. by inducing stable secondary structures to the mRNA and affecting the stability of the mRNA. Such sequence changes may alter the expression of an HT susceptibility gene. 15 A "haplotype," as described herein, refers to any combination of genetic markers ("alleles"), such as those set forth in tables 3, 4, 5, 7 and 8. A haplotype can comprise two or more alleles. As it is recognized by those skilled in the art, the same haplotype can be described differently by determining the haplotype defining alleles from different strands e.g. the haplotype 20 rs222151 1, rs4940595, rs1522723, rs1395266 (A T C C) described in this invention is the same as haplotype rs222151 1, rs4940595, rs1522723, rs1395266 (T A G G) in which the alleles are determined from the other strand or haplotype rs222151 1, rs4940595, rs1522723, rs1395266 (T T C C), in which the first allele is determined from the other strand. 25 The haplotypes described herein, e.g. having markers such as those shown in tables 3, 4, 5, 7 and 8, are found more frequently in individuals with HT than in individuals without HT. Therefore, these haplotypes have predictive value for detecting HT or a susceptibility to HT in an individual. Therefore, detecting haplotypes can be accomplished by methods known in the art for detecting sequences at polymorphic sites. 30 It is understood that the HT associated at-risk alleles and at-risk haplotypes described in this invention may be associated with other "polymorphic sites" located in HT associated genes of this invention. These other HT associated polymorphic sites may be either equally useful as genetic markers or even more useful as causative variations explaining the observed 35 association of the at-risk alleles and at-risk haplotypes of this invention to HT. In certain methods described herein, an individual who is at risk of HT is an individual in whom an at-risk allele or an at-risk haplotype is identified. In one embodiment, the at-risk allele or the at-risk haplotype is one that confers a significant risk of HT. In one embodiment, 40 significance associated with an allele or a haplotype is measured by an odds ratio. In a further embodiment, the significance is measured by a percentage. In one embodiment, a significant risk is measured as an odds ratio of at least about 1.2, including but not limited to: 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.5, 3.0, 4.0, 5.0, 10.0, 15.0, 20.0, 25.0, 30.0 and 40.0. In a further embodiment, a significant increase or reduction in 45 risk is at least about 20%, including but not limited to about 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% and 98%. In a further embodiment, a significant increase in risk is at least about 50%. It is understood however, that identifying whether a risk is medically significant may also depend on a variety of factors, including the specific disease, the allele or the haplotype, and often, environmental factors.

WO 2006/053955 PCT/F12005/050429 12 An at-risk haplotype in, or comprising portions of, the HT risk gene, is one where the haplotype is more frequently present in an individual at risk of HT (affected), compared to the frequency of its presence in a healthy individual (control), and wherein the presence of the 5 haplotype is indicative of HT or susceptibility to HT. In a preferred embodiment, the method comprises assessing in an individual the presence or frequency of SNPs in, comprising portions of, an HT risk gene, wherein an excess or higher frequency of the SNPs compared to healthy control individuals is indicative that the 10 individual has HT, or is susceptible to HT. See, for example, tables 3, 4, 5, 7 and 8 for SNPs that can form haplotypes that can be used as screening tools. These SNP markers can be identified in at-risk haploptypes. For example, an at-risk haplotype can include microsatellite markers and/or SNPs such as those set forth in tables 3, 4, 5, 7 and 8. The presence of the haplotype is indicative of HT, or a susceptibility to HT, and therefore is indicative of an 15 individual who falls within a target population for the treatment methods described herein. Consequently, the method of the invention is particularly directed to the detection of one or several of the SNP markers defining the following at-risk haplotypes indicative of HT: 20 1) rs4845303 (A/T) (SEQ ID NO: 980), rs6428195 (C/G) (SEQ ID NO: 1030) and rs1935659 (A/G) (SEQ ID NO: 637) defining the haplotype ACG; 2) rs1997454 (A/G) (SEQ ID NO: 656), rs2139502 (A/G) (SEQ ID NO: 709) and rs1519991 (A/C) (SEQ ID NO: 542) defining the haplotype AGC; 3) rs1521409 (A/G) (SEQ ID NO: 544), rs10511365 (C/T) (SEQ ID NO: 316) and 25 rs10511366 (C/T) (SEQ ID NO: 317) defining the haplotype ACT; 4) rs7679959 (C/G) (SEQ ID NO: 1178), rs10517338 (C/G) (SEQ ID NO: 381) and rs959297 (A/T) (SEQ ID NO: 1338) defining the haplotype CGA; 5) rs2278677 (A/G) (SEQ ID NO: 749), rs3886091 (C/G) (SEQ ID NO: 899), rs1998167 (A/G) (SEQ ID NO: 657), rs1998168 (A/G) (SEQ ID NO: 658) and rs2235280 (A/G) (SEQ 30 ID NO: 740) defining the haplotype GCAGG; 6) rs10521062 (A/C) (SEQ ID NO: 404), rsl10512296 (A/G) (SEQ ID NO: 331), rs1924001 (C/G) (SEQ ID NO: 633) and rs2417359 (A/G) (SEQ ID NO: 784) defining the haplotype AACG; 7) rs10508933 (C/G) (SEQ ID NO: 289), rs10509071 (A/G) (SEQ ID NO: 295) and 35 rs10490967 (A/G) (SEQ ID NO: 94) defining the haplotype GGA; 8) rs10508771 (A/T) (SEQ ID NO: 286), rs3006608 (C/T) (SEQ ID NO: 854), rs10508773 (C/T) (SEQ ID NO: 287) and rs950132 (C/T) (SEQ ID NO: 1325) defining the haplotype TCCC; 9) rs1386486 (C/T) (SEQ ID NO: 472), rs1386485 (A/C) (SEQ ID NO: 471), rs1386483 40 (A/G) (SEQ ID NO: 470) and rs7977245 (C/T) (SEQ ID NO: 1212) defining the haplotype CAGT; 10) rs276002 (A/G) (SEQ ID NO: 814) and rs274460 (A/G) (SEQ ID NO: 810) defining the haplotype AA; 11) rs1245383 (A/G) (SEQ ID NO: 430), rs2133829 (C/T) (SEQ ID NO: 707), rs2173738 45 (C/T) (SEQ ID NO: 722), rs2050528 (C/T) (SEQ ID NO: 677) and rs202970 (C/T) (SEQ ID NO: 671) defining the haplotype GCTTC; 12) rs1395266 (C/T) (SEQ ID NO: 476), rs931850 (A/G) (SEQ ID NO: 1303) and rs1522722 (C/T) (SEQ ID NO: 547) defining the haplotype TAC; WO 2006/053955 PCT/F12005/050429 13 13) rs2221511 (A/G) (SEQ ID NO: 733), rs4940595 (G/T) (SEQ ID NO: 986), rs1522723 (C/T) (SEQ ID NO: 548) and rs1395266 (C/T) (SEQ ID NO: 476) defining the haplotype ATCC; 14) rs2825555 (A/G) (SEQ ID NO: 819), rs2825583 (C/T) (SEQ ID NO: 820), rs2825601 5 (A/G) (SEQ ID NO: 821), rs2825610 (G/T) (SEQ ID NO: 822) and rs1489734 (A/G) (SEQ ID NO: 532) defining the haplotype ATGGA Monitoring Progress of Treatment 10 The current invention also pertains to methods of monitoring the effectiveness of a treatment of HT on the expression (e.g. relative or absolute expression) of one or more HT risk genes. The HT susceptibility gene mRNA, the polypeptide it is encoding, or the biological activity of the encoded polypeptide can be measured in a sample of peripheral blood or cells derived 15 therefrom. An assessment of the levels of expression or biological activity of the polypeptide can be made before and during treatment with HT therapeutic agents. Alternatively the effectiveness of a treatment of HT can be followed by monitoring biological networks and/or metabolic pathways related to one or several polypeptides encoded by HT 20 risk genes listed in table 6. Monitoring biological networks and/or metabolic pathways can be done e.g. by measuring one or several polypeptides from plasma proteome and/or by measuring one or several metabolites from plasma metabolome before and during treatment. Effectiveness of a treatment is evaluated by comparing observed changes in biological networks and or metabolic pathways following treatment with HT therapeutic agents to the 25 data available from healthy subjects. For example, in one embodiment of the invention, an individual who is a member of the target population can be assessed for response to treatment with an HT inhibitor, by examining the HT risk gene encoding polypeptide biological activity or absolute and/or relative levels of HT 30 risk gene encoding polypeptide or mRNA in peripheral blood in general or specific cell subfractions or combination of cell subfractions. In addition, variations such as haplotypes or mutations within or near (within one to hundreds of kb) the HT risk gene may be used to identify individuals who are at higher risk for HT to 35 increase the power and efficiency of clinical trials for pharmaceutical agents to prevent or treat HT or its complications. The haplotypes and other variations may be used to exclude or fractionate patients in a clinical trial who are likely to have involvement of another pathway in their HT in order to enrich patients who have pathways involved that are relevant regarding to the treatment tested and boost the power and sensitivity of the clinical trial. Such variations 40 may be used as a pharmacogenetic test to guide selection of pharmaceutical agents for individuals. Primers, probes and nucleic acid molecules 45 "Probes" or "primers" are oligonucleotides that hybridize in a base-specific manner to a complementary strand of nucleic acid molecules. "Base specific manner" means that the two sequences must have a degree of nucleotide complementarity sufficient for the primer or probe to hybridize. Accordingly, the primer or probe sequence is not required to be perfectly complementary to the sequence of the template. Non-complementary bases or modified bases 50 can be interspersed into the primer or probe, provided that base substitutions do not inhibit WO 2006/053955 PCT/F12005/050429 14 hybridization. The nucleic acid template may also include "nonspecific priming sequences" or nonspecificc sequences" to which the primer or probe has varying degrees of complementarity. Such probes and primers include polypeptide nucleic acids (Nielsen PE et al, 1991). 5 A probe or primer comprises a region of nucleic acid that hybridizes to at least about 15, for example about 20-25, and in certain embodiments about 40, 50, or 75 consecutive nucleotides of a nucleic acid of the invention, such as a nucleic acid comprising a contiguous nucleic acid sequence. 10 In preferred embodiments, a probe or primer comprises 100 or fewer nucleotides, in certain embodiments, from 6 to 50 nucleotides, for example, from 12 to 30 nucleotides. In other embodiments, the probe or primer is at least 70% identical to the contiguous nucleic acid sequence or to the complement of the contiguous nucleotide sequence, for example, at least 15 80% identical, in certain embodiments at least 90% identical, and in other embodiments at least 95% identical, or even capable of selectively hybridizing to the contiguous nucleic acid sequence or to the complement of the contiguous nucleotide sequence. Often, the probe or primer further comprises a label, e.g. radioisotope, fluorescent compound, enzyme, or enzyme co-factor. 20 Antisense nucleic acid molecules of the invention can be designed using the nucleotide sequences available e.g. in GenBank database for HT associated genes of table 6 as well as nucleotide sequences containing polymorphic sites listed in tables 2 to 5 and 7 to 11. Antisense oligonucleotides can be constructed using chemical synthesis and enzymatic 25 ligation reactions using procedures known in the art. For example, an antisense nucleic acid molecule (e.g. an antisense oligonucleotide) can be chemically synthesized using naturally occurring nucleotides or variously modified nucleotides designed to increase the biological stability of the molecules or to increase the physical stability of the duplex formed between the antisense and sense nucleic acids, e.g. phosphorothioate derivatives and acridine 30 substituted nucleotides can be used. Alternatively, the antisense nucleic acid molecule can be produced biologically using an expression vector into which a nucleic acid molecule has been subcloned in an antisense orientation (i.e. RNA transcribed from the inserted nucleic acid molecule will be of an antisense orientation to a target nucleic acid of interest). 35 The nucleic acid sequences of the HT associated genes of table 6 described in this invention can also be used to compare with endogenous DNA sequences in patients to identify genetic disorders (e.g. a predisposition for, or susceptibility to HT), and as probes, such as to hybridize and discover related DNA sequences or to extract known sequences from a sample. The nucleic acid sequences can further be used to derive primers for genetic fingerprinting, to 40 raise anti-polypeptide antibodies using DNA immunization techniques, and as an antigen to raise anti-DNA antibodies or elicit immune responses. Portions or fragments of the nucleotide sequences identified herein (and the corresponding complete gene sequences) can be used in numerous ways as polynucleotide reagents. For example, these sequences can be used to: (i) map their respective genes on a chromosome; and thus locate gene regions associated with 45 genetic disease; (ii) identify an individual from a minute biological sample (tissue typing); and (iii) aid in forensic identification of a biological sample. Additionally, the nucleotide sequences of the invention can be used to identify and express recombinant polypeptides for analysis, characterization or therapeutic use, or as markers for tissues in which the corresponding polypeptide is expressed, either constitutively, during tissue differentiation, or 50 in diseased states. The nucleic acid sequences can additionally be used as reagents in the WO 2006/053955 PCT/F12005/050429 15 screening and/or diagnostic assays described herein, and can also be included as components of kits (e.g. reagent kits) for use in the screening and/or diagnostic assays described herein. 5 Polyclonal and monoclonal antibodies Polyclonal and/or monoclonal antibodies that specifically bind one form of the gene product but not to the other form of the gene product are also provided. Antibodies are also provided that bind a portion of either the variant or the reference gene product that contains the 10 polymorphic site or sites. The term "antibody" as used herein refers to immunoglobulin molecules and immunologically active portions of immunoglobulin molecules, i.e. molecules that contain an antigen binding site that specifically binds an antigen. A molecule that specifically binds to a polypeptide of the invention is a molecule that binds to that polypeptide or a fragment thereof, but does not substantially bind other molecules in a sample, e.g. a 15 biological sample, which naturally contains the polypeptide. Examples of immunologically active portions of immunoglobulin molecules include F(ab) and F(ab') fragments which can be generated by treating the antibody with an enzyme such as pepsin. The invention provides polyclonal and monoclonal antibodies that bind to a polypeptide of the invention. The term "monoclonal antibody" or "monoclonal antibody composition" as used herein, refers to a 20 population of antibody molecules that contain only one species of an antigen binding site capable of immunoreacting with a particular epitope of a polypeptide of the invention. A monoclonal antibody composition thus typically displays a single binding affinity for a particular polypeptide of the invention with which it immunoreacts. 25 Polyclonal antibodies can be prepared as known by those skilled in the art by immunizing a suitable subject with a desired immunogen, e.g. a polypeptide of the invention or fragment thereof. The antibody titer in the immunized subject can be monitored over time by standard techniques such as with an enzyme linked immunosorbent assay (ELISA) using immobilized polypeptide. If desired, the antibody molecules directed against the polypeptide can be 30 isolated from the mammal (e.g. from blood) and further purified by well-known techniques, such as protein A chromatography to obtain the IgG fraction. At an appropriate time after immunization, e.g. when the antibody titers are highest, antibody-producing cells can be obtained from the subject and used to prepare monoclonal antibodies by standard techniques, such as the hybridoma technique (Kohler G and Milstein C, 1975), the human B cell 35 hybridoma technique (Kozbor D et al, 1982), the EBV-hybridoma technique (Cole SP et al, 1994), or trioma techniques (Hering S et al, 1988). To produce a hybridoma an immortal cell line (typically a myeloma) is fused to lymphocytes (typically splenocytes) from a mammal immunized with an immunogen as described above, and the culture supernatants of the resulting hybridoma cells are screened to identify a hybridoma producing a monoclonal 40 antibody that binds a polypeptide of the invention. Any of the many well known protocols used for fusing lymphocytes and immortalized cell lines can be applied for the purpose of generating a monoclonal antibody to a polypeptide of the invention (Bierer B et al, 2002). Moreover, the ordinarily skilled worker will appreciate 45 that there are many variations of such methods that would also be useful. As an alternative to preparing monoclonal antibody-secreting hybridomas, a monoclonal antibody to a polypeptide of the invention can be identified and isolated by screening a recombinant combinatorial immunoglobulin library (e.g. an antibody phage display library) 50 with the polypeptide to thereby isolate immunoglobulin library members that bind the WO 2006/053955 PCT/F12005/050429 16 polypeptide (Hayashi N et al, 1995; Hay BN et al, 1992; Huse WD et al, 1989; Griffiths AD et al, 1993). Kits for generating and screening phage display libraries are commercially available. 5 Additionally, recombinant antibodies, such as chimeric and humanized monoclonal antibodies comprising both human and nonhuman portions, which can be made using standard recombinant DNA techniques, are within the scope of the invention. Such chimeric and humanized monoclonal antibodies can be produced by recombinant DNA techniques known in the art. 10 In general, antibodies of the invention (e.g. a monoclonal antibody) can be used to isolate a polypeptide of the invention by standard techniques such as affinity chromatography or immunoprecipitation. A polypeptide-specific antibody can facilitate the purification of natural polypeptide from cells and of recombinantly produced polypeptide expressed in host cells. 15 Moreover, an antibody specific for a polypeptide of the invention can be used to detect the polypeptide (e.g. in a cellular lysate, cell supernatant, or tissue sample) in order to evaluate the abundance and pattern of expression of the polypeptide. Antibodies can be used diagnostically to monitor protein levels in tissue such as blood as part of a test predicting the susceptibility to HT or as part of a clinical testing procedure, e.g. to determine the efficacy of 20 a given treatment regimen. Detection can be facilitated by coupling the antibody to a detectable substance. Examples of detectable substances include various enzymes, prosthetic groups, fluorescent materials, luminescent materials, bioluminescent materials, and radioactive materials. Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, beta-galactosidase, and acetylcholinesterase; examples of suitable prosthetic 25 group complexes include streptavidin/biotin and avidin/biotin; examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin; an example of a luminescent material includes luminol; examples of bioluminescent materials include luciferase, luciferin, and aequorin, and examples of suitable radioactive material include 1251 30 131, 3 S and 3 H. Diagnostic Assays 35 The probes, primers and antibodies described herein can be used in methods of diagnosis of HT or diagnosis of a susceptibility to HT, as well as in kits useful for the diagnosis of HT or susceptibility to HT, or to a disease or condition associated with HT. In one embodiment of the invention, diagnosis of HT or susceptibility to HT (or diagnosis of 40 or susceptibility to a disease or condition associated with HT), is made by detecting one or several of at-risk alleles or at-risk haplotypes or a combination of at-risk alleles and at-risk haplotypes described in this invention in the subject's nucleic acid as described herein. In one embodiment of the invention, diagnosis of HT or susceptibility to HT (or diagnosis of 45 or susceptibility to a disease or condition associated with HT) is made by detecting one or several polymorphic sites that are associated with at-risk alleles and/or at-risk haplotypes described in this invention, in the subject's nucleic acid. Diagnostically, the most useful polymorphic sites are those altering the polypeptide structure of an HT associated gene due to a frame shift; due to a premature stop codon, due to an aminoacid change or due to abnormal 50 mRNA splicing. Nucleotide changes in a gene resulting in a change in corresponding WO 2006/053955 PCT/F12005/050429 17 polypeptide aminoacid sequence in many case alter the physiological properties of a polypeptide by resulting in a polypeptide having altered biological activity/function, distribution or stability. Other diagnostically useful polymorphic sites are those affecting transcription of an HT associated gene or translation of it's mRNA due to altered tissue 5 specifity, altered transcription rate, altered response to physiological status, altered translation efficiency of the mRNA and altered stability of the mRNA. The presence of nucleotide sequence variants altering the polypeptide structure of HT associated genes or altering the expression of HT associated genes is diagnostic for susceptibility to HT. 10 For diagnostic applications, there may be informative polymorphisms for prediction of disease risk that are in linkage disequilibrium with the functional polymorphism. Such a functional polymorphism may alter splicing sites, affect the stability or transport of mRNA, or otherwise affect the transcription or translation of the nucleic acid. The presence of nucleotide sequence variants associated with functional polymorphism is diagnostic for susceptibility to 15 HT. While we have genotyped and included a limited number of example SNP markers in the experimental section, any functional, regulatory or other mutation or alteration described above in any of the HT risk genes identified herein is expected to predict the risk of HT. 20 In diagnostic assays determination of the nucleotides present in one or several of the HT associated SNP markers of this invention, as well as polymorphic sites associated with HT associated SNP markers of this invention, in an individual's nucleic acid can be done by any method or technique which can accurately determine nucleotides present in a polymorphic 25 site. Numerous suitable methods have been described in the art (Kwok P-Y, 2001; Syvinen A-C, 2001). These methods include, but are not limited to, hybridization assays, ligation assays, primer extension assays, enzymatic cleavage assays, chemical cleavage assays and any combinations of these assays. The assays may or may not include PCR, solid phase step, modified oligonucleotides, labeled probes or labeled nucleotides, and the assay may be 30 multiplex or singleplex. As it is obvious in the art the nucleotides present in polymorphic site can be determined from one nucleic acid strand or from both strands. In another embodiment of the invention, diagnosis of a susceptibility to HT can also be made by examining transcription of one or several HT associated genes. Alterations in transcription 35 can be analyzed by a variety of methods as described in the art, including e.g. hybridization methods, enzymatic cleavage assays, RT-PCR assays and microarrays. A test sample from an individual is collected and the alterations in the transcription of HT associated genes are assessed from the RNA present in the sample. Altered transcription is diagnostic for a susceptibility to HT. 40 In another embodiment of the invention, diagnosis of a susceptibility to HT can also be made by examining expression and/or structure and/or function of an HT susceptibility polypeptide. A test sample from an individual is assessed for the presence of an alteration in the expression and/or an alteration in structure and/or function of the polypeptide encoded by an HT risk 45 gene, or for the presence of a particular polypeptide variant (e.g. an isoform) encoded by an HT risk gene. An alteration in expression of a polypeptide encoded by an HT risk gene can be for example, an alteration in the quantitative polypeptide expression (i.e. the amount of polypeptide produced); an alteration in the structure and/or function of a polypeptide encoded by an HT risk gene is an alteration in the qualitative polypeptide expression (e.g. expression 50 of a mutant HT susceptibility polypeptide or of a different splicing variant or isoform). In a WO 2006/053955 PCT/F12005/050429 18 preferred embodiment, detection of a particular splicing variant encoded by an HT risk gene, or a particular pattern of splicing variants makes diagnosis of the disease or condition associated with HT or a susceptibility to a disease or condition associated with HT possible. 5 Alterations in expression and/or structure and/or function of an HT susceptibility polypeptide can be determined by various methods known in the art e.g. by assays based on chromatography, spectroscopy, colorimetry, electrophoresis, isoelectric focusing, specific cleavage, immunologic techniques and measurement of biological activity as well as combinations of different assays. An "alteration" in the polypeptide expression or 10 composition, as used herein, refers to an alteration in expression or composition in a test sample, as compared with the expression or composition of polypeptide by an HT risk gene in a control sample. A control sample is a sample that corresponds to the test sample (i.e. is from the same type of cells), and is from an individual who is not affected by HT. An alteration in the expression or composition of the polypeptide in the test sample, as compared with the 15 control sample, is indicative of a susceptibility to HT. Western blotting analysis using an antibody as described above that specifically binds to a polypeptide encoded by a mutant HT risk gene, or an antibody that specifically binds to a polypeptide encoded by a nonmutant gene, or an antibody that specifically binds to a 20 particular splicing variant encoded by an HT risk gene, can be used to identify the presence in a test sample of a particular splicing variant or isoform, or of a polypeptide encoded by a polymorphic or mutant HT risk gene, or the absence in a test sample of a particular splicing variant or isoform, or of a polypeptide encoded by a nonpolymorphic or nonmutant gene. The presence of a polypeptide encoded by a polymorphic or mutant gene, or the absence of a 25 polypeptide encoded by a nonpolymorphic or nonmutant gene, is diagnostic for susceptibility to HT, as is the presence (or absence) of particular splicing variants encoded by an HT risk gene. In one embodiment of this method, the level or amount of polypeptide encoded by an HT risk 30 gene in a test sample is compared with the level or amount of the polypeptide encoded by an HT risk gene in a control sample. A level or amount of the polypeptide in the test sample that is higher or lower than the level or amount of the polypeptide in the control sample, such that the difference is statistically significant, is indicative of an alteration in the expression of the polypeptide encoded by an HT risk gene, and is diagnostic for susceptibility to HT. 35 Alternatively, the composition of the polypeptide encoded by an HT risk gene in a test sample is compared with the composition of the polypeptide encoded by an HT risk gene in a control sample (e.g. the presence of different splicing variants). A difference in the composition of the polypeptide in the test sample, as compared with the composition of the polypeptide in the control sample, is diagnostic for susceptibility to HT. In another embodiment, both the level 40 or amount, and the composition of the polypeptide can be assessed in the test sample and in the control sample. A difference in the amount or level of the polypeptide in the test sample compared to the control sample; a difference in composition in the test sample compared to the control sample; or both a difference in the amount or level, and a difference in the composition, is indicative of susceptibility to HT. 45 In another embodiment, assessment of the splicing variant or isoform(s) of a polypeptide encoded by a polymorphic or mutant HT risk gene can be performed. The assessment can be performed directly (e.g. by examining the polypeptide itself), or indirectly (e.g. by examining the mRNA encoding the polypeptide, e.g. by mRNA profiling). For example, probes or 50 primers as described herein can be used to determine which splicing variants or isoforms are WO 2006/053955 PCT/F12005/050429 19 encoded by an HT risk gene mRNA, using standard methods. The presence in a test sample of a particular splicing variant(s) or isoform(s) associated with HT or risk of HT, or the absence in a test sample of a particular splicing variant(s) or 5 isoform(s) not associated with HT or risk of HT, is diagnostic for a disease or condition associated with an HT risk gene or susceptibility to a disease or condition associated with an HT risk gene. Similarly, the absence in a test sample of a particular splicing variant(s) or isoform(s) associated with HT or risk of HT, or the presence in a test sample of a particular splicing variant(s) or isoform(s) not associated with HT or risk of HT, is diagnostic for the 10 absence of disease or condition associated with an HT risk gene or susceptibility to a disease or condition associated with an HT risk gene. The invention further pertains to a method for the diagnosis and identification of susceptibility to HT in an individual by identifying an at-risk allele or an at-risk haplotype in an HT risk 15 gene. In one embodiment, the at-risk allele or the at-risk haplotype is an allele or haplotype for which the presence of the haplotype increases the risk of HT significantly. Although it is to be understood that identifying whether a risk is significant may depend on a variety of factors, including the specific disease, the haplotype, and often, environmental factors, the significance may be measured by an odds ratio or a percentage. In a further embodiment, the 20 significance is measured by a percentage. In one embodiment, a significant risk is measured as an odds ratio of 0.8 or less or at least about 1.2, including but not limited to: 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.5, 3.0, 4.0, 5.0, 10.0, 15.0, 20.0, 25.0, 30.0 and 40.0. In a further embodiment an odds ratio of at least 1.2 is significant. In a further embodiment, an odds ratio of at least about 1.5 is significant. In a further 25 embodiment a significant increase or decrease in risk is at least about 1.7. In a further embodiment, a significant increase in risk is at least about 20%, including but not limited to about 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% and 98%. In a further embodiment a significant increase or reduction in risk is at least about 50%. It is understood, however, that identifying whether a risk is medically significant may 30 also depend on a variety of factors, including the specific disease, the allele or the haplotype, and often, environmental factors. The invention also pertains to methods of diagnosing HT or susceptibility to HT in an individual, comprising screening for an at-risk haplotype in the HT risk gene that is more 35 frequently present in an individual susceptible to HT (affected), compared to the frequency of its presence in a healthy individual (control), wherein the presence of the haplotype is indicative of HT or susceptibility to HT. See tables 3, 4, 6, 7 and 8 for SNP markers that comprise haplotypes that can be used as screening tools. SNP markers from these lists represent at-risk haplotypes and can be used to design diagnostic tests for determining 40 susceptibility to HT. Kits (e.g. reagent kits) useful in the methods of diagnosis comprise components useful in any of the methods described herein, including for example, PCR primers, hybridization probes or primers as described herein (e.g. labeled probes or primers), reagents for genotyping SNP 45 markers, reagents for detection of labeled molecules, restriction enzymes (e.g. for RFLP analysis), allele-specific oligonucleotides, DNA polymerases, RNA polymerases, marker enzymes, antibodies which bind to altered or to nonaltered (native) HT susceptibility polypeptide, means for amplification of nucleic acids comprising one or several HT risk genes, or means for analyzing the nucleic acid sequence of one or several HT risk genes or for 50 analyzing the amino acid sequence of one or several HT susceptibility polypeptides, etc. In WO 2006/053955 PCT/F12005/050429 20 one embodiment, a kit for diagnosing susceptibility to HT can comprise primers for nucleic acid amplification of a region in an HT risk gene comprising an at-risk haplotype that is more frequently present in an individual susceptible to HT. The primers can be designed using portions of the nucleic acids flanking SNPs that are indicative of HT. 5 This invention is based on the principle that one or a small number of genotypings are performed and the sequence variations to be typed are selected on the basis of their ability to predict HT. For this reason any method to genotype sequence variations in a genomic DNA sample can be used. 10 Thus, the detection method of the invention may further comprise a step of combining information concerning age, gender, the family history of HT, diabetes and hypercholesterolemia, and the medical history concerning CVD or diabetes of the subject with the results obtained from step b) of the method (see claim 1) for confirming the indication 15 obtained from the detection step. Said information may also concern hypercholesterolemia in the family, smoking status, HT in the family, history of CVD, obesity in the family, and waist-to-hip circumference ratio (cm/cm) The detection method of the invention may also further comprise a step determining blood, 20 serum or plasma cholesterol, HiDL cholesterol, LDL cholesterol, triglyceride, apolipoprotein B and Al, fibrinogen, ferritin, transferrin receptor, C-reactive protein, serum or plasma insulin concentration. The score that predicts the probability of HT may be calculated using a multivariate failure 25 time model or a logistic regression equation. The results from the further steps of the method as described above render possible a step of calculating the probability of developing HT using a logistic regression equation as follows. Probability of HT = 1/[1 + e (-(-a + E(bi*Xi))], where e is Napier's constant, Xi are variables 30 related to HT, bi are coefficients of these variables in the logistic function, and a is the constant term in the logistic function, and wherein a and bi are preferably determined in the population in which the method is to be used, and Xi are prefereably selected among the variables that have been measured in the population in which the method is to be used. Preferable values for bi are between -20 and 20; and for i between 0 (zero) and 100,000. A 35 negative coefficient bi implies that the marker is risk-reducing and a positive coefficient implies that the marker is risk-increasing. Xi are binary variables that can have values or are coded as 0 (zero) or 1 (one) such as SNP markers. The model may additionally include any interaction (product) or terms of any 40 variables Xi, e.g. biXi. An algorithm is developed for combining the information to yield a simple prediction of HT as percentage of risk in one year, two years, five years, 10 years or 20 years. Alternative statistical models are failure-time models such as the Cox's proportional hazards' 45 model, other iterative models and neural networking models. The test can be applied to test the risk of developing HT in both healthy persons, as a screening or predisposition test, and high-risk persons (who have e.g. family history of HT, central or other type of obesity, lack of physical activity, high sodium intake, high intake of 50 saturated fats, low intake of potassium and/or magnesium, low HiDL cholesterol, diabetes WO 2006/053955 PCT/F12005/050429 21 mellitus, glucose intolerance, insulin resistance and the metabolic syndrome, elevated inflammatory marker, or any combination of these or an elevated level of any other risk factor for HT). 5 The method can be used in the prediction and early diagnosis of HT in adult persons, stratification and selection of subjects in clinical trials, and/or stratification and selection of persons for intensified preventive and curative interventions. The aim is to reduce the cost of clinical drug trials and health care. 10 Pharmaceutical Compositions The present invention also pertains to pharmaceutical compositions comprising agents described herein, particularly nucleotides in HT risk genes, and/or comprising other splicing 15 variants encoded by HT risk genes; and/or an agent that alters (e.g. enhances or inhibits) HT risk gene expression or HT susceptibility gene polypeptide activity as described herein. For instance, a polypeptide, protein (e.g. a receptor), an agent that alters an HT risk gene expression, or an HT susceptibility polypeptide binding agent or binding partner, fragment, fusion protein or prodrug thereof, or a nucleotide or nucleic acid construct (vector) 20 comprising a nucleotide of the present invention, or an agent that alters HT susceptibility gene polypeptide activity, can be formulated with a physiologically acceptable carrier or excipient to prepare a pharmaceutical composition. The carrier and composition can be sterile. The formulation should suit the mode of administration. 25 In a preferred embodiment pharmaceutical compositions comprise an agent or agents reversing, at least partially, HT associated changes in biological networks and/or metabolic pathways related to the HT associated genes of this invention (Table 6). Suitable pharmaceutically acceptable carriers include but are not limited to water, salt 30 solutions (e.g. NaCl), saline, buffered saline, alcohols, glycerol, ethanol, gum arabic, vegetable oils, benzyl alcohols, polyethylene glycols, gelatin, carbohydrates such as lactose, amylose or starch, dextrose, magnesium stearate, talc, silicic acid, viscous paraffin, perfume oil, fatty acid esters, hydroxymethylcellulose, polyvinyl pyrolidone, etc., as well as combinations thereof. The pharmaceutical preparations can, if desired, be mixed with 35 auxiliary agents, e.g. lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, coloring, flavoring and/or aromatic substances and the like that do not deleteriously react with the active agents. The composition, if desired, can also contain minor amounts of wetting or emulsifying agents, 40 or pH buffering agents. The composition can be a liquid solution, suspension, emulsion, tablet, pill, capsule, sustained release formulation, or powder. The composition can be formulated as a suppository, with traditional binders and carriers such as triglycerides. Oral formulation can include standard carriers such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, polyvinyl pyrolidone, sodium saccharine, cellulose, magnesium 45 carbonate, etc. Methods of introduction of these compositions include, but are not limited to, intradermal, intramuscular, intraperitoneal, intraocular, intravenous, subcutaneous, topical, oral and intranasal. Other suitable methods of introduction can also include gene therapy (as described 50 below), rechargeable or biodegradable devices, particle acceleration devices ("gene guns") WO 2006/053955 PCT/F12005/050429 22 and slow release polymeric devices. The pharmaceutical compositions of this invention can also be administered as part of a combinatorial therapy with other agents. The composition can be formulated in accordance with the routine procedures as a 5 pharmaceutical composition adapted for administration to human beings. For example, compositions for intravenous administration are typically solutions in sterile isotonic aqueous buffer. Where necessary, the composition may also include a solubilizing agent and a local anesthetic to ease pain at the site of the injection. Generally, the ingredients are supplied either separately or mixed together in unit dosage form, for example, as a dry lyophilized 10 powder or water free concentrate in a hermetically sealed container such as an ampule or sachette indicating the quantity of active agent. Where the composition is to be administered by infusion, it can be dispensed with an infusion bottle containing sterile pharmaceutical grade water, saline or dextrose/water. Where the composition is administered by injection, an ampule of sterile water for injection or saline can be provided so that the ingredients may be 15 mixed prior to administration. For topical application, nonsprayable forms, viscous to semi-solid or solid forms comprising a carrier compatible with topical application and having a dynamic viscosity preferably greater than water, can be employed. Suitable formulations include but are not limited to solutions, 20 suspensions, emulsions, creams, ointments, powders, enemas, lotions, sols, liniments, salves, aerosols, etc., which are, if desired, sterilized or mixed with auxiliary agents, e.g. preservatives, stabilizers, wetting agents, buffers or salts for influencing osmotic pressure, etc. The agent may be incorporated into a cosmetic formulation. For topical application, sprayable aerosol preparations wherein the active ingredient, preferably in combination with a solid or 25 liquid inert carrier material, is packaged in a squeeze bottle or in admixture with a pressurized volatile, normally gaseous propellant, e.g. pressurized air, are also suitable. Agents described herein can be formulated as neutral or salt forms. Pharmaceutically acceptable salts include those formed with free amino groups such as those derived from 30 hydrochloric, phosphoric, acetic, oxalic, tartaric acids, etc., and those formed with free carboxyl groups such as those derived from sodium, potassium, ammonium, calcium, ferric hydroxides, isopropylamine, triethylamine, 2-ethylamino ethanol, histidine, procaine, etc. The agents are administered in a therapeutically effective amount. The amount of agents 35 which will be therapeutically effective in the treatment of a particular disorder or condition will depend on the nature of the disorder or condition, and can be determined by standard clinical techniques. In addition, in vitro or in vivo assays may optionally be employed to help identify optimal dosage ranges. The precise dose to be employed in the formulation will also depend on the route of administration, and the severity of the symptoms of HT, and should be 40 decided according to the judgment of a practitioner and each patient's circumstances. Effective doses may be extrapolated from dose-response curves derived from in vitro or animal model test systems. 45 Methods of Therapy The present invention encompasses methods of treatment (prophylactic and/or therapeutic) for HT or a susceptibility to HT, such as individuals in the target populations described herein, using an HT therapeutic agent. An "HT therapeutic agent" is an agent that alters (e.g. 50 enhances or inhibits) HT risk affecting polypeptide (enzymatic activity or quantity) and/or an WO 2006/053955 PCT/F12005/050429 23 HT risk gene expression, as described herein (e.g. an agonist or antagonist). HT therapeutic agents can alter an HT susceptibility polypeptide activity or nucleic acid expression by a variety of means, for example, by providing additional HT susceptibility polypeptide or by upregulating the transcription or translation of the HT risk gene; by altering posttranslational 5 processing of the HT susceptibility polypeptide; by altering transcription of an HT risk gene splicing variants; or by interfering with an HT susceptibility polypeptide activity (e.g. by binding to an HT susceptibility polypeptide); or by downregulating the transcription or translation of the HT risk gene, or by inhibiting or enhancing the elimination of an HT susceptibility polypeptide. 10 In particular, the invention relates to methods of treatment for HT or susceptibility to HT (for example, for individuals in an at-risk population such as those described herein); as well as to methods of treatment for manifestations and subtypes of HT. 15 Representative HT therapeutic agents include the following: nucleic acids or fragments or derivatives thereof described herein, particularly nucleotides encoding the polypeptides described herein and vectors comprising such nucleic acids (e.g. a gene, cDNA, and/or mRNA, double-stranded interfering RNA, a nucleic acid encoding an HT 20 susceptibility polypeptide or active fragment or derivative thereof, or an oligonucleotide; for examples see tables 2 through 8; other polypeptides (e.g. HT susceptibility receptors); HT susceptibility polypeptide binding agents; peptidomimetics; fusion proteins or prodrugs thereof, antibodies (e.g. an antibody to a 25 mutant HT susceptibility polypeptide, or an antibody to a non-mutant HT susceptibility polypeptide, or an antibody to a particular splicing variant encoded by an HT risk gene, as described above); ribozymes; other small molecules; and other agents that alter (e.g. inhibit or antagonize) an HT risk gene expression or 30 polypeptide activity or that regulate transcription of an HT risk gene splicing variants (e.g. agents that affect which splicing variants are expressed, or that affect the amount of each splicing variant that is expressed); and other reagents that alter (e.g. induce or agonize) an HT risk gene expression or 35 polypeptide activity or that regulate transcription of an HT risk gene splicing variants (e.g. agents that affect which splicing variants are expressed or that affect the amount of each splicing variant that is expressed). More than one HT therapeutic agent can be used concurrently, if desired. 40 The HT therapeutic agent that is a nucleic acid is used in the treatment of HT. The term, "treatment" as used herein, refers not only to ameliorating symptoms associated with the disease, but also preventing or delaying the onset of the disease and also lessening the severity or frequency of symptoms of the disease, preventing or delaying the occurrence of a second 45 episode of the disease or condition; and/or also lessening the severity or frequency of symptoms of the disease or condition. In the case of atherosclerosis, "treatment" also refers to a minimization or reversal of the development of plaques. The therapy is designed to alter (e.g. inhibit or enhance), replace or supplement activity of an HT polypeptide in an individual. For example, an HT therapeutic agent can be administered in order to upregulate or increase 50 the expression or availability of an HT risk gene or of specific splicing variants of an HT WO 2006/053955 PCT/F12005/050429 24 susceptibility, gene or, conversely, to downregulate or decrease the expression or availability of an HT risk gene or specific splicing variants of an HT risk gene. Upregulation or increasing expression or availability of a native HT risk gene or of a particular splicing variant could interfere with or compensate for the expression or activity of a defective gene or another 5 splicing variant; downregulation or decreasing expression or availability of a native HT risk gene or of a particular splicing variant could minimize the expression or activity of a defective gene or the particular splicing variant and thereby minimize the impact of the defective gene or the particular splicing variant. 10 The HT therapeutic agent(s) are administered in a therapeutically effective amount (i.e. an amount that is sufficient to treat the disease, e.g. by ameliorating symptoms associated with the disease, preventing or delaying the onset of the disease, and/or also lessening the severity or frequency of symptoms of the disease). The amount which will be therapeutically effective in the treatment of a particular individual's disorder or condition will depend on the symptoms 15 and severity of the disease and can be determined by standard clinical techniques. In addition, in vitro or in vivo assays may optionally be employed to help identify optimal dosage ranges. The precise dose to be employed in the formulation will also depend on the route of administration and the severity of the disease or disorder, and should be decided according to the judgment of a practitioner and each patient's circumstances. Effective doses may be 20 extrapolated from dose-response curves derived from in vitro or animal model test systems. In one embodiment, a nucleic acid of the invention (e.g. a nucleic acid encoding an HT susceptibility polypeptide set forth in table 6 optionally comprising at least one polymorphism shown in tables 2 through 11; or another nucleic acid that encodes an HT susceptibility 25 polypeptide or a splicing variant, derivative or fragment thereof, can be used, either alone or in a pharmaceutical composition as described above. For example, an HT risk gene or a cDNA encoding an HT susceptibility polypeptide, either by itself or included within a vector, can be introduced into cells (either in vitro or in vivo) such that the cells produce native HT susceptibility polypeptide. If necessary, cells that have been transformed with the gene or 30 cDNA or a vector comprising the gene or cDNA can be introduced (or re-introduced) into an individual affected with the disease. Thus, cells that in nature lack a native HT risk gene expression and activity, or have mutant HT risk gene expression and activity, or have expression of a disease-associated HT risk gene splicing variant, can be engineered to express an HT susceptibility polypeptide or an active fragment of an HT susceptibility polypeptide (or 35 a different variant of an HT susceptibility polypeptide). In a preferred embodiment, nucleic acid encoding an HT susceptibility polypeptide, or an active fragment or derivative thereof, can be introduced into an expression vector, such as a viral vector, and the vector can be introduced into appropriate cells in an animal. Other gene transfer systems including viral and nonviral transfer systems can be used. Alternatively, nonviral gene transfer methods such as 40 calcium phosphate coprecipitation, mechanical techniques (e.g. microinjection); membrane fusion-mediated transfer via liposomes; or direct DNA uptake, can also be used. Alternatively, in another embodiment of the invention, a nucleic acid of the invention; a nucleic acid complementary to a nucleic acid of the invention; or a portion of such a nucleic 45 acid (e.g. an oligonucleotide as described below) can be used in "antisense" therapy in which a nucleic acid (e.g. an oligonucleotide) that specifically hybridizes to the mRNA and/or genomic DNA of an HT risk gene is administered or generated in situ. The antisense nucleic acid that specifically hybridizes to the mRNA and/or DNA inhibits expression of the HT susceptibility polypeptide, e.g. by inhibiting translation and/or transcription. Binding of the 50 antisense nucleic acid can be by conventional base pair complementarity, or for example in WO 2006/053955 PCT/F12005/050429 25 the case of binding to DNA duplexes, through specific interaction in the major groove of the double helix. An antisense construct of the present invention can be delivered, for example, as an 5 expression plasmid as described above. When the plasmid is transcribed in the cell it produces RNA that is complementary to a portion of the mRNA and/or DNA which encodes an HT susceptibility polypeptide. Alternatively, the antisense construct can be an oligonucleotide probe that is generated ex vivo and introduced into cells; it then inhibits expression by hybridizing with the mRNA and/or genomic DNA of an HT risk gene. In one embodiment, 10 the oligonucleotide probes are modified oligonucleotides that are resistant to endogenous nucleases, e.g. exonucleases and/or endonucleases, thereby rendering them stable in vivo. Exemplary nucleic acid molecules for use as antisense oligonucleotides are phosphoramidate, phosphothioate and methylphosphonate analogs of DNA. Additionally, general approaches to constructing oligomers useful in antisense therapy are also described by van der Krol AR et 15 al, 1988 and Stein CA and Cohen JS, 1988. With respect to antisense DNA, oligodeoxyribonucleotides derived from the translation initiation site, e.g. between the -10 and +10 regions of an HT risk gene sequence, are preferred. To perform antisense therapy, oligonucleotides (mRNA, cDNA or DNA) are designed that are 20 complementary to the mRNA encoding an HT susceptibility polypeptide. The antisense oligonucleotides bind to HT susceptibility mRNA transcripts and prevent translation. Absolute complementarity, although preferred, is not required. A sequence "complementary" to a portion of an RNA, as referred to herein, indicates that a sequence has sufficient complementarity to be able to hybridize with the RNA forming a stable duplex; in the case of 25 double-stranded antisense nucleic acids, a single strand of the duplex DNA may thus be tested, or triplex formation may be assayed. The ability to hybridize will depend on both the degree of complementarity and the length of the antisense nucleic acid, as described in detail above. Generally, the longer the hybridizing nucleic acid, the more base mismatches with an RNA it may contain and still form a stable duplex (or triplex, as the case may be). One skilled 30 in the art can ascertain a tolerable degree of mismatch by use of standard procedures. The oligonucleotides used in antisense therapy can be DNA, RNA, or chimeric mixtures or derivatives or modified versions thereof, single-stranded or double-stranded. The oligonucleotides can be modified at the base moiety, sugar moiety, or phosphate backbone, 35 for example, to improve stability of the molecule, hybridization, etc. The oligonucleotides can include other appended groups such as peptides (e.g. for targeting host cell receptors in vivo), or agents facilitating transport across the cell membrane (Letsinger RL et al, 1989; Lemaitre M et al, 1987) or the blood-brain barrier (Jaeger LB and Banks WA, 2004), or hybridization triggered cleavage agents (van der Krol AR et al, 1988) or intercalating agents. (Zon G, 40 1988). To this end, the oligonucleotide may be conjugated to another molecule (e.g. a peptide, hybridization triggered cross-linking agent, transport agent, hybridization-triggered cleavage agent). The antisense molecules are delivered to cells that express an HT risk gene in vivo. A number 45 of methods can be used for delivering antisense DNA or RNA to cells; e.g. antisense molecules can be injected directly into the tissue site, or modified antisense molecules, designed to target the desired cells (e.g. antisense linked to peptides or antibodies that specifically bind receptors or antigens expressed on the target cell surface) can be administered systematically. Alternatively, in a preferred embodiment, a recombinant DNA 50 construct is utilized in which the antisense oligonucleotide is placed under the control of a WO 2006/053955 PCT/F12005/050429 26 strong promoter (e.g. pol III or pol II). The use of such a construct to transfect target cells in the patient results in the transcription of sufficient amounts of single stranded RNAs that will form complementary base pairs with the endogenous HT risk gene transcripts and thereby prevent translation of the HT susceptibility mRNA. For example, a vector can be introduced 5 in vivo such that it is taken up by a cell and directs the transcription of an antisense RNA. Such a vector can remain episomal or become chromosomally integrated, as long as it can be transcribed to produce the desired antisense RNA. Such vectors can be constructed by recombinant DNA technology methods standard in the art and described above. For example, a plasmid, cosmid, YAC or viral vector can be used to prepare the recombinant DNA 10 construct that can be introduced directly into the tissue site. Alternatively, viral vectors can be used which selectively infect the desired tissue, in which case administration may be accomplished by another route (e.g. systemically). An endogenous HT risk gene expression can be also reduced by inactivating or "knocking 15 out" an HT risk gene or its promoter using targeted homologous recombination (Smithies 0 et al, 1985; Thomas KR and Capecchi MR, 1987; Thompson S et al, 1989). For example, a mutant, non-functional HT risk gene (or a completely unrelated DNA sequence) flanked by DNA homologous to the endogenous HT risk gene (either the coding regions or regulatory regions of an HT risk gene) can be used, with or without a selectable marker and/or a negative 20 selectable marker, to transfect cells that express an HT risk gene in vivo. Insertion of the DNA construct, via targeted homologous recombination, results in inactivation of the HT risk gene. The recombinant DNA constructs can be directly administered or targeted to the required site in vivo using appropriate vectors, as described above. Alternatively, expression of nonmutant HT risk gene can be increased using a similar method: targeted homologous 25 recombination can be used to insert a DNA construct comprising a nonmutant, functional HT risk gene (e.g. any gene shown in table 6 that may optionally comprise at least one polymorphism shown in tables 2 through 11), or a portion thereof, in place of a mutant HT risk gene in the cell as described above. In another embodiment, targeted homologous recombination can be used to insert a DNA construct comprising a nucleic acid that encodes 30 an HT susceptibility polypeptide variant that differs from that present in the cell. Alternatively, an endogenous HT risk gene expression can be reduced by targeting deoxyribonucleotide sequences complementary to the regulatory region of an HT risk gene (i.e. the HT risk gene promoter and/or enhancers) to form triple helical structures that prevent 35 transcription of an HT risk gene in target cells in the body (Helene C, 1991; Helene C et al, 1992; Maher LJ, 1992). Likewise, the antisense constructs described herein can be used in the manipulation of tissue, by antagonizing the normal biological activity of one of the HT proteins, e.g. tissue differentiation both in vivo and for ex vivo tissue cultures. Furthermore, the anti-sense techniques (e.g. microinjection of antisense molecules, or transfection with 40 plasmids whose transcripts are anti-sense with regard to an HT mRNA or gene sequence) can be used to investigate the role of an HT risk gene in developmental events, as well as the normal cellular function of an HT risk gene in adult tissue. Such techniques can be utilized in cell culture, but can also be used in the creation of transgenic animals. 45 In yet another embodiment of the invention, other HT therapeutic agents as described herein can also be used in the treatment or prevention of HT. The therapeutic agents can be delivered in a composition, as described above, or by themselves. They can be administered systemically, or can be targeted to a particular tissue. The therapeutic agents can be produced by a variety of means including chemical synthesis; recombinant production; in vivo 50 production, e.g. a transgenic animal (Meade H et al, 1990) and can be isolated using standard WO 2006/053955 PCT/F12005/050429 27 means such as those described herein. A combination of any of the above methods of treatment (e.g. administration of non-mutant HT susceptibility polypeptide in conjunction with antisense therapy targeting mutant HT 5 susceptibility mRNA; administration of a first splicing variant encoded by an HT risk gene in conjunction with antisense therapy targeting a second splicing encoded by an HT risk gene), can also be used. The invention will be further described by the following nonlimiting examples. The teachings 10 of all publications cited herein are incorporated herein by reference in their entirety. EXPERIMENTAL SECTION 15 East Finnish HT Patients and Phenotype Characterization The subjects were participants of the Kuopio Ischaemic Heart Disease Risk Factor Study 20 (KIHD), which is an ongoing prospective population-based study designed to investigate risk factors for chronic diseases, including HT and CVD, in middle-aged men (Salonen JT 1988, Salonen JT et al 1998, 1999, Tuomainen T-P et al 1999). The study population was a random age-stratified sample of men living in Eastern Finland who were 42, 48, 54 or 60 years old at baseline examinations in 1984-1989. A total of 2682 men were examined during 1984-89. 25 The male cohort was complemented by a random population sample of 920 women first examined during 1998-2001, at the time of the 11-year follow up of the male cohort. The recruitment and examination of the subjects has been described previously in detail (Salonen JT, 1988). The University of Kuopio Research Ethics Committee approved the study. All participants gave their written informed consent. 30 The analyses are based on logistic modeling in a case-control set of 81 cases with HT (SBP 140 mmHg or more or DBP 90 mmHg or more or antihypertensive medication) and HT in either sibling or parent, and 82 controls who had neither HT nor family history of HT, both from the KID cohort. Three of the subjects (two cases, one control) were women, 160 were 35 men. Thirty-eight of the 81 cases had antihypertensive medication at the time of BP measurements in the KID baseline examination. HT was defined as either systolic BP (SBP) 140 mmHg or diastolic BP (DBP) 90 mmHg or antihypertensive medication. Both BPs were measured in the morning by a nurse with a 40 random-zero mercury sphygmomanometer. The measuring protocol included three measurements in supine, one in standing and two in sitting position with 5-minutes intervals. The mean of all six measurements were used as SBP and DBP (Salonen JT et al, 1998). The family history of HT was defined positive, if either father, the mother or a sibling of the study subject had reported a history or prevalent hypertension. 45 50 WO 2006/053955 PCT/F12005/050429 28 Table 1. Selected characteristics of the cases and controls Hypertensive cases (n=81) Normotensive controls (n=82) Mean Min Max Mean Min Max Age (years) 54.6 42.1 71.9 54.6 42.2 61.1 Cigarettes/day 5.3 0 40 7.4 0 40 S-Cholesterol (mmol/L) 6.2 3.8 9.1 6.0 3.2 8.7 S-HDL-Chol (mmol/L) 1.21 0.82 2.15 1.34 0.76 2.77 B-Glucose (mmol/L) 5.13 3.3 12.6 4.55 3.5 5.9 S-Insulin (U/L) 14.7 4.7 59.6 9.33 1.7 22.5 Mean SBP (mmHg) 140.0 110.0 182.33 124.5 99.0 148.33 Mean DBP (mmHg) 92.1 63.3 122.3 81.3 66.0 94.3 In table 1 selected characteristics of the cases and controls are summarized. Age and tobacco smoking were recorded on a self-administered questionnaire checked by an interviewer. 5 Fasting blood glucose was measured using a glucose dehydrogenase method after precipitation of proteins by trichloroacetic acid. Serum insulin was determined with a Novo Biolabs radioimmunoassay kit (Novo Nordisk). H-DL fractions were separated from fresh serum by combined ultracentrifugation and precipitation. The cholesterol contents of lipoprotein fractions and serum triglycerides were measured enzymatically. Fibrinogen was 10 measured based on the clotting of diluted plasma with excess thrombin. Adulthood socioeconomical status (SES) is an index comprised of measures of education, occupation, income and material living conditions. The scale is inverse, low score corresponding to high SES. These data have been collected by a self administered 15 questionnaire. Serum ferritin was assessed with a commercial double antibody radioimmunoassay (Amersham International, Amersham, UK). Lipoproteins, including high density lipoprotein (HDL) and low density lipoprotein (LDL), were separated from fresh serum samples by 20 ultracentrifugation followed by direct very low density lipoprotein (VLDL) removal and LDL precipitation (Salonen et al 1991). Cholesterol concentration was then determined enzymically. Serum C-reactive protein was measured by a commercial high-sensitive immunometric assay (Immulite High Sensitivity CR Assay, DPC, Los Angeles). 25 Genomic DNA isolation and quality testing High molecular weight genomic DNA samples were extracted from frozen venous whole blood using standard methods, and dissolved in standard TE buffer. The quantity and purity of 30 each DNA sample was evaluated by measuring the absorbance at 260 and 280 nm and integrity of isolated DNA samples was evaluated with 0.9% agarose gel electrophoresis and Ethidiumbromide staining. A sample was qualified for genome wide scan (GWS) analysis if the A260/A280 ratio was 1.7 and the average size of isolated DNA was over 20 kb in agarose gel electrophoresis. Before GWS, analysis samples were diluted to a concentration of 35 50 ng/gl in reduced EDTA TE buffer (TEKnova).

WO 2006/053955 PCT/F12005/050429 29 Genome-Wide Scan Genotyping of SNP markers was performed using the technology access version of Affymetrix GeneChip@ human mapping 100k system. The assay consisted of two arrays, Xba and Hind, which were used to genotype over 126,000 SNP markers from each DNA sample. 5 The assays were performed according to the instructions provided by the manufacturer. A total of 250 ng of genomic DNA was used for each individual assay. The DNA sample was digested with either Xba I or Hind III enzyme (New England Biolabs, NEB) in the mixture of NE Buffer 2 (1 x; NEB), bovine serum albumin (1 x; NEB), and either Xba I or Hind III (0,5 U/ l; NEB) for 2h at +37'C followed by enzyme inactivation for 20 min at +70'C. Xba I or 10 Hind III adapters were then ligated to the digested DNA samples by adding Xba or Hind III adapter (0,25 gM, Affymetrix), T4 DNA ligase buffer (1 x; NEB), and T4 DNA ligase (250 U; NEB). Ligation reactions were allowed to proceed for 2h at +16'C followed by 20 min incubation at +70'C. Each ligated DNA sample was diluted with 75 l of molecular biology grade water (BioWhittaker Molecular Applications/Cambrex). 15 Diluted ligated DNA samples were subjected to four identical 100 l volume polymerase chain reactions (PCR) by implementing a 10 l aliquot of DNA sample with Pfx Amplification Buffer (1 x; Invitrogen), PCR Enhancer (1 x; Invitrogen), MgSO 4 (1 mM; Invitrogen), dNTP (300 gM each; Takara), PCR primer (1 gM; Affymetrix), and Pfx 20 Polymerase (0,05 U/g1; Invitrogen). The PCR was allowed to proceed for 3 min at +94'C, followed by 30 cycles of 15 sec at +94'C, 30 sec at +60'C, 60 sec at +68'C, and finally for the final extension for 7 min at +68'C. The performance of the PCR was checked by standard 2% agarose gel electrophoresis in 1 x TBE buffer for lh at 120V. 25 PCR products were purified according to the Affymetrix manual using MinElute 96 UF PCR Purification kit (Qiagen) by combining all four PCR products of an individual sample into the same purification reaction. The purified PCR products were eluted with 40 l of EB buffer (Qiagen), and the yields of the products were measured at the absorbance 260 nm. A total of 40 gg of each PCR product was then subjected to fragmentation reaction consisting of 0.2 30 U/g1 fragmentation reagent (Affymetrix) in 1x Fragmentation Buffer. The fragmentation reaction was allowed to proceed for 35 min at +37'C followed by 15 min incubation at +95'C for enzyme inactivation. Completeness of fragmentation was checked by running an aliquot of each fragmented PCR product in 4% agarose 1 x TBE (BMA Reliant precast) for 30-45 min at 120V. 35 Fragmented PCR products were then labeled using 1 x Terminal Deoxinucleotidyl Transferase (TdT) buffer (Affymetrix), GeneChip DNA Labeling Reagent (0.214 mM; Affymetrix), and TdT (1,5 U/g1; Affymetrix) for 2h at +37'C followed by 15 min at +95'C. Labeled DNA samples were combined with hybridization buffer consisting of 0.056 M MES 40 solution (Sigma), 5% DMSO (Sigma), 2.5 x Denhardt's solution (Sigma), 5.77 mM EDTA (Ambion), 0.115 mg/ml Herring Sperm DNA (Promega), 1 x Oligonucleotide Control reagent (Affymetrix), 11.5 gg/ml Human Cot-i (Invitrogen), 0.0115% Tween-20 (Pierce), and 2.69 M Tetramethyl Ammonium Chloride (Sigma). DNA-hybridization buffer mix was denatured for 10 min at +95'C, cooled on ice for 10 sec and incubated for 2 min at +48'C prior to 45 hybridization onto corresponding Xba or Hind GeneChip® array. Hybridization was completed at +48'C for 16-18 h at 60 rpm in an Affymetrix GeneChip Hybridization Oven. Following hybridization, the arrays were stained and washed in GeneChip Fluidics Station 450 according to fluidics station protocol Mappingl0Kvl_450 as recommended by the WO 2006/053955 PCT/F12005/050429 30 manufacturer. Arrays were scanned with GeneChip 3000 Scanner and the genotype calls for each of the SNP markers on the array were generated using Affymetrix Genotyping Tools (GTT) software. The confidence score in SNP calling algorithm was adjusted to 0.20. 5 Initial SNP selection for statistical analysis Prior to the statistical analysis, SNP quality was assessed on the basis of three values: the call rate (CR), minor allele frequency (MAF), and Hardy-Weinberg equilibrium (H-W). The CR is 10 the proportion of samples genotyped successfully. It does not take into account whether the genotypes are correct or not. The call rate was calculated as: CR = number of samples with successful genotype call / total number of samples. The MAF is the frequency of the allele that is less frequent in the study sample. MAF was calculated as: MAF = min(p, q), where p is frequency of the SNP allele 'A' and q is frequency of the SNP allele 'B'; p = (number of 15 samples with "AA"-genotype + 0.5*number of samples with "AB"-genotype) / total number of samples with successful genotype call; q = 1 - p. SNPs that are homozygous (MAF=0) cannot be used in genetic analysis and were thus discarded. H-W equilibrium is tested for controls. The test is based on the standard Chi-square test of goodness of fit. The observed genotype distribution is compared with the expected genotype distribution under H-W 20 equilibrium. For two alleles this distribution is p 2 , 2pq, and q 2 for genotypes 'AA', 'AB' and 'BB', respectively. If the SNP is not in H-W equilibrium it can be due to genotyping error or some unknown population dynamics (e.g. random drift, selection). Only the SNPs that had CR > 50%, MAF > 1%, and were in H-W equilibrium (Chi-square 25 test statistic < 23.93) were used in the statistical analysis. A total of 107,895 SNPs fulfilled the above criteria and were included in the statistical analysis. Statistical Methods 30 Single SNP analysis Differences in allele distributions between cases and controls were screened for all 107,895 SNPs. The screening was carried out using the standard Chi-square independence test with 1 df (allele distribution, 2x2 table). SNPs that gave a P-value less than 0.005 (Chi-square 35 distribution with 1 df of 7.88 or more) were considered as statistically significant and selected for further analysis. There were 529 SNPs that fulfilled this criterium. Haplotype analysis 40 The data set was analyzed with a haplotype pattern mining algorithm either with H-PM-G software (Sevon P et al, 2004) or with 1PM software (Toivonen HT et al, 2000). For 11PM software, genotypes must be phase known to determine which alleles come from the mother and which from the father. Without family data, phases must be estimated based on population data. We used HaploRec-program (Eronen L et al, 2004) to estimate the phases. 45 1PM-G and 11PM are very fast and can handle a large number of SNPs in a single run The difference between 11PM and HPM-G is that 1PM-G can use phase unknown genotypic data and 11PM uses phase known (or estimated by HaploRec or similar program) data. HPM G finds all haplotype patterns that fit the genotype configuration. For phase-known data 11PM WO 2006/053955 PCT/F12005/050429 31 finds all haplotype patterns that are in concordance with the phase configuration. The length of the haplotype patterns can vary. As an example, if there are four SNPs and an individual has alleles A T for SNP1, C C for SNP2, C G for SNP3, and A C for SNP4, then H-PM-G considers haplotype patterns (of length 4 SNPs): ACCA, TCGC, TCCA, ACGC, ACGA, 5 TCCC, TCGA, ACCC. 1PM considers only haplotype patterns that are in concordance with the estimated phase (done by HaploRec). If the estimated phase is ACGA (from the mother/father) and TCCC (from the father/mother) then 11PM considers only two patterns (of length 4 SNPs): ACGA and TCCC. 10 A SNP is scored based on the number of times it is included in a haplotype pattern that differs between cases and controls (a threshold Chi-square value can be selected by the user). Significance of the score values is tested based on permutation tests. Several parameters can be modified in the HPM-G and HPM programs including the Chi 15 square threshold value (-x), the maximum haplotype pattern length (-1), the maximum number of wildcards that can be included in a haplotype pattern (-w), and the number of permutation tests in order to estimate the P-value (-p). Wildcards allow gaps in haplotypes. The 1PM-G program was run with the following parameter settings: haplotype analysis with 5 SNPs (-x9 -15 -wl -p10000). HaploRec+ 11PM was run with the following parameter settings: 20 haplotype analysis with 5 SNPs (-x9 -15 -wl -p10000). 1PM-G analysis was based on the order of the SNP given in dbSNP122 and HaploRec+HPM was based on the order of the SNP given in dbSNP123. Based on 10,000 replicates (-p10000) in the 1PM-G analyses 570 SNPs were significant at P-value less than 0.005 and 642 SNPs were significant in the 11PM analysis. 25 Definition of terms used in the haplotype analysis results 30 The term "haplotype genomic region" or "haplotype region" refers to a genomic region that has been found significant in the haplotype analysis (HPM, 1PMG or similar statistical method/program). The haplotype region is defined as 10OKbp up/downstream from the physical position of the first/last SNP that was included in the statistical analysis (haplotype analysis) and was found statistically significant. This region is given in base pairs based on 35 the given genome build e.g. SNP physical position (base pair position) according to NCBI Human Genome Build 35. The term "haplotype" as described herein, refers to any combination of alleles e.g. A T C C that is found in the given genetic markers e.g rs222151 1, rs4940595, rs1522723, rs1395266. 40 A defined haplotype gives the name of the genetic markers (dbSNP rs-id for the SNPs) and the alleles. As it is recognized by those skilled in the art, the same haplotype can be described differently by determining alleles from different strands e.g. the haplotype rs222151 1, rs4940595, rs1522723, rs1395266 (A T C C) is the same as haplotype rs222151 1, rs4940595, rs1522723, rs1395266 (T A G G) in which the alleles are determined from the other strand, or 45 haplotype rs222151 1, rs4940595, rs1522723, rs1395266 (T T C C), in which the first allele is determined from the other strand. The haplotypes described herein, e.g. having markers such as those shown in tables 3, 4, 5, 7 and 8, are found more frequently in individuals with HT than in individuals without HT. 50 Therefore, these haplotypes have predictive value for detecting HT or a susceptibility to HT in WO 2006/053955 PCT/F12005/050429 32 an individual. Therefore, detecting haplotypes can be accomplished by methods known in the art for detecting sequences at polymorphic sites. It is understood that the HT associated at-risk alleles and at-risk haplotypes described in this 5 invention may be associated with other "polymorphic sites" located in HT associated genes of this invention. These other HT associated polymorphic sites may be either equally useful as genetic markers or even more useful as causative variations explaining the observed association of at-risk alleles and at-risk haplotypes of this invention to HT. 10 Multivariate modeling For modeling for hypertension as a binary outcome, the 734 strongest predicting SNP markers from the individual SNP analysis and 14 strongest haplotypes from the 1PM analysis were tested for entry to the model. These were recoded as 0, if homozygote of the major allele, 1, if 15 heterozygote and 2, if homozygote of the minor allele. A multivariate binary logistic function regression analysis was used to: a) Find the SNPs that were most predictive of HT and b) Construct a multivariate model that predicted HT the strongest. A forward step-up model construction was used with p-value to enter of 0.01 and p-value to exclude from the model of 0.02. The predictivity of the models was estimated by two methods: the Nagelkerke R square 20 and the reclassification of the subjects to cases and controls on the basis of the logistic model contructed. The predicted probability used as cut-off was 0.5. A data reduction analysis was carried out by step-down and step-up logistic modeling. Multivariate least-squares linear regression modeling was used to identify the SNP markers 25 that were most strongly associated with the mean systolic and diastolic blood pressure as quantitative traits. A forward step-up model construction was used with p-value to enter of 0.001 and p-value to exclude from the model of 0.005. The statistical software used was SPSS for Windows, version 11.5. 30 Results In table 2 are summarized the characteristics of the SNP markers with the strongest association with HT in the individual marker analysis (n = 529). SNP identification numbers 35 are according to NCBI dbSNP database build 124. Physical positions of SNP markers are according to NCBI Human Genome Build 35. Gene locus as reported by NCBI dbSNP database build 124. SNP flanking sequence provided by Affymetrix "csv" commercial access Human Mapping 1OOK array annotation files. 40 In table 3 are summarized the characteristics of the haplotype genomic regions with the strongest association with HT in the HIPM-G analysis with 5 SNPs. SNP identification numbers are according to NCBI dbSNP database build 124. Physical positions of SNP markers are according to NCBI Human Genome Build 35. Associated genes are those genes positioned within 1OOKbp up/downstream from the physical position of the SNPs bordering 45 the haplotype genomic region found using NCBI MapViewer, based on NCBI Human Genome Build 35. SNP flanking sequence provided by Affymetrix "csv" commercial access Human Mapping 1OOK array annotation files. In table 4 are summarized the characteristics of the haplotype genomic regions with the 50 strongest association with HT in the HaploRec + 1PM analysis with 5 SNPs. SNP WO 2006/053955 PCT/F12005/050429 33 identification numbers are according to NCBI dbSNP database build 124. Physical positions of SNP markers are according to NCBI Human Genome Build 35. Associated genes are those genes positioned within 100Kbp up/downstream from the physical position of the SNPs bordering the haplotype genomic region found using NCBI MapViewer, based on NCBI 5 Human Genome Build 35. SNP flanking sequence provided by Affymetrix "csv" commercial access Human Mapping 1OOK array annotation files. In table 5 are listed haplotype blocks with the strongest association with HT based on HaploRec + 1PM analysis (n = 14). SNP identification numbers are according to NCBI 10 dbSNP database build 124. In table 6 are listed all genes found associated with HT according to point wise and haplotype analyses (n = 722). Names of genes are according to HUGO Gene Nomenclature Committee (HGNC). 15 In table 7 are listed the SNP-markers and haplotypes that best predicted risk of familial HT in a multivariate logistic model. SNP identification numbers are according to NCBI dbSNP database build 124. The 8-variable model predicts 91.4% of familial HT correctly. The statistics are based on 81 KIHD participants who were hypertensive in the KIHD baseline 20 examination (SBP 140 mmHg or more or DBP 90 mmHg or more or antihypertensive medication) and either sibling or parent had HT and 82 KIHD participants who neither had HT at KIHD baseline nor had family history of HT. The controls were matched according to age. 25 In table 8 are listed the SNP-markers, haplotypes and phenotypic data that best predicted risk of familial HT in a multivariate logistic model. SNP identification numbers are according to NCBI dbSNP database build 124. The 12-variable model, including two haplotypes, five SNP markers and two phenotypic variables, predicted 87.1% of familial HT correctly. The strongest loci pinpointed by the multivariate logistic models were SERPINs B3, B4, B7 and 30 B1I and EPC1, OR1J4 and LOC401406, 439953, 441550 and 441551. Table 9 presents a multivariate linear regression model of the strongest SNPs predicting the mean systolic and diastolic BP. Tables 10 and 11 show the means and standard deviations of the mean systolic (Table 10) and diastolic (Table 11) BP in the genotypes of the strongest 35 SNP markers, which predicted BP the strongest in both the univariate single-SNP, haplotype and multivariate analyses. The rank order of markers is according to the strength of association with the diastolic BP. The strongest pinpointed genes concerning BP as quantitative trait were SERPINS B3, B7 and BI 1, A100A7, S100A6, FARSI, SPOCK3, and TLL1. 40 Implications and Conclusions We have found 1365 SNP markers associated with the risk of HT and/or blood pressure in a 45 population-based set of familial cases and healthy controls without family history. Of these, 529 were identified in the analysis of individual SNPs and 1080 in haplotype pattern mining or haplotype analysis. Of the 1365 markers, 244 predicted HT in both types of statistical analysis. We further identified SNP markers, which predict in a multivariate logistic model virtually fully the development of HT. 50 WO 2006/053955 PCT/F12005/050429 34 The results of the point wise and haplotype analyses identified a total of 722 genes associated with HT, of which 330 genes had at least one of the 1365 SNP markers physically linked to the gene. 5 Thus, we have discovered a total of 722 HT genes, in which any genetic markers can be used to predict HT, and thus these markers can be used as part of molecular diagnostic tests of HT predisposition. In addition, we have disclosed a set of 1365 SNP markers which are predictive of HT. The markers can also be used as part of pharmacogenetic tests predicting the efficacy and adverse reactions of antihypertensive agents and compounds. The genes discovered are 10 also targets to new therapies of HT, such as drugs. Other therapies are molecular, including gene transfer. The new genes can also be used to develop and produce new transgenic animals for studies of antihypertensive agents and compounds. While this invention has been particularly shown and described with reference to preferred 15 embodiments thereof, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.

WO 2006/053955 PCT/F12005/050429 35 Table 2. SNP markers with the strongest association with HT in the individual marker analysis (n = 529). d | G I S I C IA IA I M I A | 0 | b | e | e I h |l |l1 i | 1 | d | S n | q | r |l |li n | 1 | d | N e u I o le le I o I e I s | P | e l m |l |li r | 1 | | | 1 | n | o le le I I e I r | r | o Ic I s I I A l I a I s c | e | o |A IB | 1 X I t | | u I I m l | | 1 | 2 | i | I | s I I e I I | e | I o | D | ID | | | 1 | | 1 | | | e | I | rs1395266 SERPINBll 476 18 C T C 20.29 0.23 rs931850 None 1303 18 A G G 18.10 0.27 rs1386483 TPH2 470 12 A G A 17.88 2.61 rs1386485 TPH2 471 12 A C C 17.80 2.84 rs10506395 None 246 12 C T T 17.04 0.29 rs1410425 None 491 13 C T T 16.89 0.11 rs10500913 None 186 11 A G A 16.68 3.02 rs1935659 None 637 1 A G G 16.53 0.36 rs1318392 OR1J4 438 9 A G G 15.69 2.50 rs6428195 None 1030 1 C G C 15.39 0.39 rs7112382 None 1105 11 A G A 15.28 2.64 rs2469828 None 792 17 C T T 14.97 4.59 rs1386486 TPH2 472 12 C T T 14.87 2.46 rs10513404 NT 008470 343 9 C T C 14.74 0.32 rs10515283 None 356 5 C T C 14.55 5.73 rs1884389 None 618 20 C T T 14.21 2.66 rs10494019 None 124 1 A T T 14.03 0.33 rs4756190 None 965 11 A G G 14.03 2.38 rs10517655 PDGFC 387 4 A G G 13.88 10.16 rs1157122 GABRAl 417 5 C T C 13.83 0.22 rs668498 FBXO18 1058 10 C T T 13.74 2.60 rs473232 COL24A1 959 1 C T C 13.72 0.34 rs1986902 None 653 1 A G A 13.72 2.83 rs627777 LOC388458 1026 18 C T T 13.70 0.41 rs9315991 None 1294 13 G T T 13.68 4.55 rs9299552 PCDH15 1267 10 C T T 13.66 0.39 rs868299 None 1233 15 C T C 13.65 2.37 rs4845303 None 980 1 A T T 13.59 2.64 rs6935462 None 1082 6 A T A 13.51 6.29 rs7536312 None 1161 1 A G A 13.26 0.30 rs7521497 None 1159 1 C T T 13.25 2.86 rs270454 None 805 4 A G A 13.20 0.36 rs1374038 None 465 5 A C C 13.03 3.56 rs3213829 ANK3 869 10 G T G 12.94 2.30 rs10494628 None 128 1 C T C 12.86 0.43 rs2885625 LOC145497 839 14 A G G 12.82 0.29 rs2476847 SEC5L1 793 6 C T T 12.77 2.26 rs1521409 None 544 3 A G G 12.77 9.50 rs274460 None 810 12 A G G 12.72 0.35 rs10483354 STRN3 21 14 A C A 12.62 0.00 rs5957594 None 1010 X G T G 12.61 0.00 rs1394380 None 475 12 A G G 12.59 0.37 rs1391130 None 473 12 A G G 12.58 0.36 rs1842328 None 609 12 C T C 12.56 0.34 WO 2006/053955 PCT/F12005/050429 36 rs387619 None 898 11 A G A 12.56 2.22 rs9291304 GABRB1 1259 4 A G A 12.53 0.06 rs10514437 WWOX 349 16 C T T 12.53 5.88 rs2209672 None 730 10 A G A 12.49 2.26 rs1912914 None 628 1 A G G 12.45 0.41 rs2416472 None 781 5 A G G 12.44 0.42 rs1837426 None 607 2 A G G 12.44 0.43 rs301748 USH2A 855 1 A G A 12.31 0.00 rs1335311 None 447 10 C G C 12.31 2.29 rs10499961 None 175 7 C T C 12.27 3.24 rs1866565 None 616 2 C T T 12.25 0.40 rs2060688 None 682 8 C T T 12.23 0.35 rs951573 UTRN 1328 6 A G G 12.20 2.22 rs572166 None 1008 18 A G G 12.17 2.32 rs10493787 COL24A1 122 1 C G C 12.17 0.32 rs10514689 None 352 3 A G G 12.15 2.86 rs10494663 None 134 1 A G A 12.14 0.29 rs1997454 None 656 2 A G G 12.12 0.31 rs10499538 DNAH11 171 7 A G A 12.00 0.06 rs474106 COL24A1 961 1 A G A 11.96 0.35 rs916853 None 1240 7 C T T 11.93 0.29 rs10492479 None 112 13 A G G 11.91 0.24 rs4680266 KCNAB1 955 3 C T C 11.83 2.18 rs4691246 None 956 4 C T C 11.82 2.36 rs9289965 KCNAB1 1257 3 C G C 11.82 2.17 rs10519989 MGC14798 397 15 A C A 11.82 0.07 rs10485822 SNPH 49 20 C G C 11.82 3.90 rs9298688 None 1266 9 C T T 11.81 0.39 rs892271 None 1236 12 A G G 11.81 0.11 rs29800 None 849 5 A C C 11.71 0.38 rs1598978 None 575 12 A G A 11.70 2.22 rs10494659 None 130 1 C T C 11.69 0.27 rs10515461 None 362 5 A G A 11.68 0.28 rs9283795 CMYA5 1248 5 C T T 11.67 3.39 rs503208 None 989 11 C G G 11.66 2.32 rs1578704 None 568 1 A G A 11.63 0.43 rs715732 None 1114 8 C G G 11.60 0.24 rs6469061 None 1038 8 C T T 11.53 0.00 rs1578705 None 569 1 A G G 11.51 0.45 rs716193 NPAS3 1117 14 G T T 11.49 2.28 rs9328130 None 1314 6 C T C 11.46 3.37 rs1487275 TPH2 531 12 A C C 11.45 2.20 rs10488914 None 76 4 A G A 11.42 0.11 rs1521770 None 545 8 C T C 11.42 0.44 rs6976996 CNTNAP2 1089 7 A G A 11.42 2.34 rs1173476 None 424 5 G T G 11.40 2.16 rs10486619 None 58 7 C T C 11.31 2.55 rs2105233 None 696 1 A G G 11.31 2.89 rs921382 None 1243 8 C G C 11.28 0.35 rs1452356 FLJ31810 513 9 C T C 11.25 2.16 rs956316 None 1336 7 C T C 11.25 0.43 rs7524749 SSBP3 1160 1 C T T 11.22 3.11 rs1913157 None 630 3 C G G 11.22 0.44 rs10499668 PKD1L1 172 7 A G G 11.21 2.13 rs1614336 None 582 5 A C A 11.18 2.17 rs477862 COL24A1 972 1 C T T 11.14 0.39 rs1912941 None 629 1 C T C 11.13 0.44 rs2198044 None 729 8 A G A 11.09 0.38 rs7156058 None 1113 14 A C A 11.09 2.90 rs10511192 LOC152225 314 3 C T T 11.09 2.57 rs10507107 EB-1 257 12 C G C 11.07 0.40 WO 2006/053955 PCT/F12005/050429 37 rsl0505140 None 238 8 A G A 11.04 0.35 rs6601146 None 1054 5 G T G 11.04 2.33 rs2861215 None 832 2 C T C 11.03 2.19 rs10482862 None 19 21 A G G 11.03 3.19 rs175741 None 592 X G T T 11.02 0.00 rs964888 None 1344 6 C T C 11.02 2.16 rs1649882 SP100 586 2 G T T 10.89 0.34 rs8047401 CDYL2 1221 16 C T T 10.84 0.41 rs2048506 None 674 4 C T T 10.84 2.12 rs1409367 None 490 1 C T C 10.79 0.00 rs8010717 LOC388001 1217 14 C T C 10.76 2.14 rs792762 None 1206 17 A C A 10.74 2.11 rs2175550 None 724 1 A G G 10.70 0.46 rs1391533 None 474 11 C T C 10.70 2.11 rs3856852 None 897 3 A G A 10.70 0.12 rs1160054 None 419 8 A G G 10.66 0.46 rs1882233 None 617 2 A T T 10.63 2.10 rs536617 FAT3 1001 11 C T C 10.63 2.09 rs10488849 None 73 4 C T C 10.62 2.49 rs666899 None 1055 11 A G G 10.61 0.39 rs1376089 None 468 4 A G A 10.60 13.87 rs475553 None 964 3 C T T 10.59 0.07 rs1408356 None 486 10 A G G 10.58 3.43 rs1113983 MTHFS 413 15 G T T 10.57 2.09 rs1512828 None 538 3 A C C 10.57 0.46 rs6746500 None 1062 2 C T T 10.56 2.33 rs1430692 None 506 2 A C C 10.55 3.09 rs10270433 None 11 7 A T A 10.51 0.24 rs10489134 None 81 1 C G C 10.47 6.23 rs1580964 None 570 12 C T T 10.47 2.08 rs2849597 PARK2 828 6 A G G 10.45 0.00 rs2279120 MATN2 751 8 C T T 10.44 5.27 rs9300909 None 1269 13 A G G 10.41 2.39 rs1281590 None 434 1 A G A 10.40 2.61 rs2417359 None 784 9 A G G 10.37 2.06 rs10484614 STX11 35 6 C T T 10.37 3.44 rs10491030 None 95 10 C T C 10.34 2.06 rs605627 None 1015 7 C T T 10.33 2.28 rs2222186 None 735 2 C T T 10.28 2.74 rs2121956 None 705 1 C T T 10.25 2.11 rs10495082 None 137 1 C T C 10.25 0.47 rs9288697 ACVR1 1256 2 A C A 10.25 0.29 rs9304252 None 1277 18 A T A 10.25 4.15 rs438418 None 936 5 A G G 10.24 0.29 rs1865290 None 615 18 A T T 10.23 4.18 rs1944404 RIT2 639 18 C T C 10.21 0.35 rs10504127 None 226 8 A G A 10.20 0.00 rs10504596 None 228 8 C T T 10.19 0.00 rs2372643 EB-1 768 12 A G A 10.19 2.07 rs10502283 LOC338661 202 11 C T T 10.18 0.32 rs9308066 None 1281 4 A G G 10.17 7.97 rs9321214 None 1306 6 C T C 10.15 0.40 rsl0501504 None 199 11 C T C 10.15 2.61 rs10513802 DGKG 346 3 A T T 10.14 0.20 rs9290835 None 1258 3 A T A 10.13 2.21 rs9287120 None 1251 1 C G C 10.13 0.26 rs10515406 None 360 5 A G G 10.12 3.40 rs2429555 None 785 17 C G C 10.11 3.81 rs10494664 None 135 1 A C C 10.09 0.34 rs6961386 HIC 1085 7 C T T 10.07 0.26 rs1978503 None 649 18 A G G 10.06 2.85 WO 2006/053955 PCT/F12005/050429 38 rs3847653 None 894 12 C G C 10.03 2.41 rs10502667 KIAA1328 209 18 C G G 10.02 0.38 rs10502582 None 206 18 A T T 10.01 2.05 rs730694 LOC401406 1149 7 C T C 10.00 2.18 rs10486935 None 59 7 A G A 10.00 0.31 rs9323707 LOC400234 1312 14 A G A 10.00 0.31 rs723107 None 1138 8 A G A 9.98 13.22 rs1281611 None 435 1 C T T 9.98 2.52 rs956274 None 1335 5 A C A 9.97 0.41 rs7777049 None 1190 7 A G A 9.96 2.13 rs10517654 PDGFC 386 4 G T T 9.96 5.09 rs10515622 LOC255187 364 5 G T G 9.96 2.65 rs10510664 LOC389105 311 3 C G G 9.95 3.44 rs2173802 NPAS3 723 14 A G A 9.95 0.34 rs1423485 ADAMTS12 503 5 A G A 9.95 5.08 rs4131804 RGS7 923 1 G T T 9.91 2.45 rs10508220 None 272 10 A G G 9.90 2.04 rs2192947 None 728 2 A G G 9.90 2.05 rs10492232 None 109 12 C T C 9.88 3.77 rs383096 None 892 5 A G A 9.86 13.08 rs701665 None 1098 6 C T C 9.85 2.30 rs256825 SGCD 799 5 C T T 9.83 2.18 rs6879659 None 1074 5 C T T 9.80 2.81 rs10496096 COMMD1 140 2 C G G 9.80 0.44 rs10508229 None 274 10 A G G 9.79 0.31 rs8089921 NETO1 1223 18 C T C 9.79 2.05 rs7081811 CACNB2 1101 10 A G G 9.78 0.43 rs420350 None 929 6 A C A 9.77 2.05 rs8007568 NPAS3 1214 14 C G C 9.75 0.28 rs2381194 None 773 9 C T T 9.74 0.34 rs9741 PSD3 1350 8 C T C 9.72 0.43 rs1514471 LOC152225 540 3 A G G 9.71 2.40 rs10512853 None 339 5 C T C 9.71 0.08 rs895800 None 1237 1 A T A 9.70 0.48 rs542873 DAB1 1002 1 A G A 9.69 0.49 rs10515314 None 357 5 C T C 9.66 0.13 rs10508223 None 273 10 C T C 9.66 2.05 rs6990997 ZFPM2 1091 8 C T C 9.64 0.30 rs6832042 None 1071 4 A G G 9.63 0.45 rs2089918 None 694 1 A G A 9.60 2.87 rs2113030 None 700 5 A G G 9.59 0.13 rs10515942 AOX2 371 2 A G A 9.59 2.68 rs1827309 None 600 21 A G G 9.58 2.14 rs741283 MYO3B 1154 2 G T T 9.58 0.29 rs10499728 None 174 7 A G A 9.58 0.46 rs1041778 None 18 21 A G G 9.57 2.18 rs1446255 None 510 9 C T T 9.57 0.23 rs10485601 TMC2 44 20 A G A 9.57 0.35 rs2077833 None 689 12 A G A 9.56 0.45 rs129752 FBLN1 437 22 A G A 9.56 0.08 rs293443 None 847 4 C T C 9.56 0.36 rs285790 None 829 8 C T C 9.55 0.08 rs10270360 LOC401406 10 7 A G G 9.55 2.04 rs10513702 None 345 3 A G G 9.50 2.03 rs10497952 ERBB4 159 2 C T T 9.49 2.29 rs10516740 None 378 4 A G G 9.49 0.00 rs1529647 None 550 7 A G G 9.48 2.95 rs1362828 None 458 3 C G C 9.48 2.06 rs6981960 None 1090 8 C T T 9.48 2.33 rs1992906 None 655 5 A G G 9.46 7.64 rs1928326 None 635 1 C T C 9.42 2.06 WO 2006/053955 PCT/F12005/050429 39 rs607872 None 1017 7 C G C 9.42 2.19 rs953160 None 1330 7 A G G 9.42 0.46 rs360559 CRISP2 877 6 G T T 9.40 2.09 rs10488631 None 68 7 C T C 9.39 0.25 rs4719652 None 958 7 C T T 9.39 0.49 rs10520170 None 398 2 C T C 9.39 2.54 rs10511824 FLJ31810 323 9 A G A 9.38 2.92 rs10501243 None 190 11 A G A 9.38 0.21 rs8092610 None 1225 18 C T T 9.38 0.21 rs1513120 LOC152225 539 3 C T C 9.38 2.20 rs9316871 None 1298 13 C T C 9.37 2.36 rs10501242 None 189 11 A G A 9.37 0.21 rs10516731 None 377 4 A G A 9.37 0.00 rs3906713 None 904 7 A T A 9.36 4.31 rs997661 KIAA1328 1362 18 C T T 9.36 0.39 rs2089273 None 693 4 A T A 9.34 0.39 rs960791 LOC389938 1339 10 A T T 9.34 2.11 rs7176436 ADAM10 1122 15 A G G 9.34 0.38 rs10512731 LOC389285 338 5 C G C 9.33 2.55 rs4775086 ADAM10 970 15 C T T 9.31 0.48 rs4086865 None 913 11 C T T 9.29 0.17 rs10506726 None 253 12 A G A 9.29 0.29 rs10490758 None 93 2 C G G 9.29 0.43 rs10488777 None 71 11 C T C 9.29 3.92 rs4884068 None 984 13 C G G 9.28 2.15 rs802296 TCBAl 1220 6 A G A 9.28 2.17 rs572241 FAT3 1009 11 C T C 9.27 2.00 rs2168792 None 721 19 C G C 9.26 0.00 rs344924 LOC391046 872 1 A G G 9.25 2.74 rs1438048 PDE11A 508 2 A C A 9.25 2.78 rs716376 None 1118 5 A G G 9.24 2.53 rs10488747 LOC260340 69 11 C G C 9.22 0.45 rs10490757 None 92 2 C T C 9.21 0.44 rs3897770 None 901 12 A C A 9.21 2.29 rs1544774 None 554 5 G T T 9.20 0.21 rs10509035 None 293 10 G T G 9.19 0.49 rs6586445 None 1052 1 A C C 9.18 2.01 rs2091169 None 695 2 A G G 9.17 1.98 rs2164857 PDE11A 719 2 A C C 9.16 2.47 rs9308440 XM 372814 1282 1 G T T 9.16 3.12 rs1550740 None 557 12 C T C 9.14 0.08 rs237112 None 767 6 C T T 9.14 2.46 rs2416863 NT 008470 782 9 C T T 9.13 0.26 rs1331772 None 445 13 A G G 9.13 0.35 rs1363522 LOC255187 460 5 C T T 9.12 2.81 rs422621 None 930 21 C T C 9.12 0.48 rs6972578 OSBPL3 1087 7 C T C 9.09 2.19 rs10503506 None 220 8 C T C 9.09 0.36 rs747334 None 1156 10 A G G 9.08 1.97 rs342818 None 871 1 C T C 9.08 2.12 rs824848 None 1230 5 A G A 9.08 2.06 rs10483100 DGCR2 20 22 C T C 9.07 3.38 rs1548348 NUMAl 555 11 C T T 9.07 12.23 rs10502666 KIAA1328 208 18 C T C 9.07 0.40 rs10502671 KIAA1328 210 18 C T T 9.07 0.40 rs2514288 None 796 11 C T C 9.07 0.46 rs9320728 None 1305 6 C T C 9.06 0.45 rs10512709 None 337 5 A G A 9.06 2.02 rs10498935 C6orfl57 167 6 A T T 9.06 2.96 rs10484484 None 33 6 A G G 9.05 2.73 rs1411270 LOC138882 492 9 C G G 9.05 0.45 WO 2006/053955 PCT/F12005/050429 40 rs8010105 LOC388001 1216 14 A G A 9.04 1.99 rs9379628 None 1319 6 C T C 9.04 5.75 rs10505797 None 240 12 A G A 9.03 0.00 rs2152634 None 714 9 C T T 9.03 0.00 rs4766492 VPS29 968 12 A G A 9.03 1.96 rs7085103 MYO3A 1102 10 C T C 9.03 2.02 rs2373717 None 771 11 A T T 9.01 2.12 rs997418 None 1361 10 A G A 9.01 0.50 rs951232 None 1327 4 C T T 9.01 0.48 rs927348 SLC35F1 1246 6 A G G 9.00 1.96 rs1497763 SCHIP1 533 3 C G G 8.99 4.83 rs10507130 DRIM 258 12 A G A 8.98 0.34 rs2871869 None 838 1 A G A 8.97 0.37 rs1370656 PDE11A 463 2 A C A 8.97 2.59 rs10512117 None 326 9 A T A 8.97 0.50 rs1346671 None 454 11 A T T 8.96 2.32 rs970604 DNAH11 1348 7 A G A 8.95 2.12 rs1959429 None 644 14 C T T 8.92 3.07 rs4814909 None 975 20 C T T 8.92 0.08 rs1405589 LOC402640 484 7 A G A 8.92 0.00 rs7069564 None 1100 10 C T T 8.92 0.31 rs891255 None 1235 2 C T T 8.91 2.04 rs4519041 NY-BR-1 943 10 A C A 8.91 12.03 rs10512241 None 329 9 C T T 8.90 2.00 rs987312 None 1354 10 A G A 8.89 2.12 rs2000112 None 660 9 C T T 8.87 0.43 rs721599 None 1136 1 C G G 8.86 2.03 rs948159 LOC390263 1322 11 C T C 8.86 0.39 rs10502774 None 213 18 G T G 8.86 3.19 rs717293 None 1120 16 C T T 8.85 0.50 rs2070513 None 686 21 G T T 8.83 2.34 rs10508731 MPP7 285 10 C T C 8.82 2.10 rs3169160 ARRDC4 864 15 A G A 8.81 3.38 rs3934674 None 906 4 A G A 8.79 0.36 rs2750452 None 811 6 C G C 8.79 2.11 rs10521300 None 406 16 C T T 8.78 11.87 rs10521301 None 407 16 G T T 8.78 11.87 rs305218 PRKCL2 858 1 C T T 8.78 11.87 rs724417 None 1141 12 G T G 8.78 3.85 rs9283603 None 1247 3 A G A 8.77 2.18 rs1181875 FLJ32825 427 1 A G G 8.77 0.28 rs4600228 None 950 11 A G A 8.77 2.10 rs1928322 None 634 1 A G A 8.76 2.01 rs3846211 None 893 3 A G A 8.76 2.01 rs9332453 None 1317 9 A G A 8.76 0.49 rs10511366 None 317 3 C T C 8.75 7.16 rs2177520 DNAH11 725 7 C G C 8.74 2.10 rs2319248 None 759 4 C T T 8.74 0.50 rs1893814 None 622 11 C T T 8.73 2.17 rs2892734 FARP1 841 13 A G G 8.72 1.96 rs6799641 None 1067 3 A T A 8.72 2.56 rs2017712 None 665 21 A G G 8.71 2.05 rs1489734 None 532 21 A G A 8.71 0.51 rs12654 LOC401255 432 6 C T C 8.70 0.30 rs1969025 LOC391858 645 5 A G G 8.70 0.47 rs382190 None 889 5 A G A 8.68 0.08 rs10511049 None 313 3 A G G 8.67 0.30 rs1581413 FLJ12604 571 3 A G G 8.67 1.96 rs10497893 CRYGEP1 156 2 A C A 8.66 0.51 rs2217110 C18orflO 732 18 A G G 8.64 0.41 rs7667905 LOC339979 1176 4 A G A 8.63 2.24 WO 2006/053955 PCT/F12005/050429 41 rs10502700 None 211 18 A G A 8.63 0.24 rs4627439 None 951 18 G T T 8.63 0.24 rs251470 FLJ10904 797 5 G T T 8.63 1.96 rs10507224 LOC390359 262 12 C T T 8.63 0.46 rs543224 None 1003 18 C T T 8.63 0.46 rs2241748 ARL4 742 7 A G A 8.62 5.48 rs2263356 None 746 9 C T T 8.62 2.33 rs935661 PRKCE 1318 2 G T T 8.61 1.97 rs2183024 None 727 9 C G C 8.61 0.38 rs10495321 SIPAlL2 138 1 A G G 8.61 4.64 rs994214 None 1357 2 C T C 8.61 1.95 rs10507505 None 265 13 C T C 8.61 2.29 rs10490387 LOC130576 90 2 C T T 8.61 3.30 rs361161 None 878 4 A G G 8.60 2.35 rs2394081 None 776 10 A G A 8.60 7.07 rs719639 C6orfl57 1128 6 G T G 8.59 2.18 rs1744510 None 591 6 A T A 8.59 0.45 rs1860752 None 612 7 C G C 8.58 0.48 rs2357761 None 763 6 C G G 8.58 0.48 rs10516708 None 376 4 C T T 8.57 1.96 rs10499381 PHF14 169 7 C T T 8.57 0.39 rs10511318 None 315 3 C G C 8.56 2.71 rs3964705 LOC390561 907 15 A G G 8.56 0.43 rs10487921 None 62 7 A G A 8.56 0.38 rs10512097 None 325 9 C T T 8.56 2.80 rs10485602 TMC2 45 20 C T C 8.54 0.37 rs3751877 RRN3 882 16 A G G 8.54 2.72 rs10485608 None 47 20 A G G 8.54 2.94 rs2060850 FLJ31810 683 9 A G G 8.54 1.94 rs1120712 None 416 7 C T C 8.53 1.95 rs6864370 None 1072 5 C T T 8.53 0.42 rs10508410 LOC389938 276 10 C G G 8.53 0.09 rs10509281 CTNNA3 298 10 C T C 8.53 0.09 rs1904641 CTNNA3 626 10 A C C 8.53 0.09 rs4102401 RRM2B 914 8 C T C 8.53 0.09 rs4404874 None 937 8 A G A 8.52 0.48 rs10496213 REGL 145 2 A C C 8.52 3.48 rs10500296 GRLF1 180 19 C G G 8.52 0.00 rs1403105 FLJ12604 482 3 A G G 8.51 1.95 rs4129499 None 919 8 A C A 8.50 0.18 rs2252646 None 745 12 A G G 8.49 0.50 rs161912 None 583 3 A G G 8.49 2.03 rs10484683 None 39 6 A G G 8.49 11.67 rs802279 TCBAl 1219 6 C G C 8.48 2.40 rs6592456 NUMAl 1053 11 C T C 8.47 5.41 rs9301083 None 1273 13 A G A 8.47 1.94 rs2241443 DGKI 741 7 C T C 8.46 2.05 rs10512918 None 340 5 A G G 8.46 2.64 rs1402319 None 481 12 C T C 8.45 2.02 rs10520415 FLJ13110 400 2 C T C 8.44 0.49 rs10503122 None 217 18 C T T 8.44 0.22 rs966118 ASAH1 1345 8 C T T 8.43 2.02 rs10491771 None 99 9 A G A 8.43 0.51 rs1037124 ARNT2 15 15 C T T 8.43 0.32 rs10513473 None 344 9 C T T 8.42 2.39 rs10508171 None 271 13 A G A 8.42 1.96 rs10518928 MEIS2 394 15 C T C 8.42 0.45 rs10508412 LOC389938 278 10 C T T 8.42 1.93 rs2439538 TBClD7 787 6 A G G 8.42 1.93 rs950171 pp9099 1326 15 C G G 8.41 1.95 rs10512629 TRAD 334 3 A T A 8.41 2.35 WO 2006/053955 PCT/F12005/050429 42 rs3731572 LTBP1 880 2 C T C 8.41 0.00 rs10516839 None 379 4 A C C 8.40 0.42 rs2888378 EB-1 840 12 A G A 8.40 1.92 rs4762559 EB-1 967 12 C T T 8.39 1.92 rs10491747 None 97 9 A G G 8.39 2.01 rs1561365 None 560 5 A T A 8.38 2.64 rs10485603 TMC2 46 20 A G G 8.38 0.36 rs10519536 KIAA0882 396 4 C T T 8.36 0.42 rs4336940 None 934 10 A G G 8.36 0.51 rs964664 BAI3 1343 6 A G A 8.36 2.06 rs7908118 None 1202 10 C T T 8.35 0.50 rs10515457 KIAA1061 361 5 C T C 8.35 2.27 rs10484858 BPAG1 40 6 C T C 8.35 0.46 rs1998570 None 659 13 A G A 8.35 2.00 rs6969674 HDAC9 1086 7 A C A 8.35 0.32 rs7763613 None 1187 6 A G G 8.35 1.91 rs10498789 None 165 6 A G A 8.34 1.99 rs10503493 SGCZ 219 8 G T T 8.34 3.44 rs10503869 RBPMS 225 8 C T C 8.33 5.36 rs10507223 LOC390359 261 12 C G G 8.32 0.46 rs9295562 LOC389370 1261 6 C T T 8.32 2.14 rs10504702 None 230 8 A G A 8.32 0.19 rs10484126 None 31 14 C T T 8.32 5.34 rs10487982 None 64 1 A C C 8.32 2.27 rs1498767 None 535 16 C T C 8.31 2.10 rs923312 None 1245 11 A G A 8.31 0.19 rs10515934 GORASP2 370 2 A G A 8.31 2.18 rs4073846 MAST2 911 1 C T C 8.31 1.92 rs10485760 None 48 20 C T C 8.31 0.00 rs1407508 None 485 9 A G G 8.30 0.00 rs1455780 SIAT8B 517 15 A G A 8.30 1.91 rs4669134 None 953 2 A G A 8.30 0.49 rs2369942 None 765 12 A G A 8.30 1.96 rs2274154 None 748 6 C T T 8.29 2.78 rs6808403 None 1068 3 A G A 8.28 2.51 rs438405 RCL1 935 9 A C C 8.27 0.22 rsl0515512 NDFIP1 363 5 C T C 8.25 3.69 rs1116163 LOC400019 414 12 A G G 8.24 0.35 rs340978 None 870 11 A C C 8.24 2.32 rs10509071 None 295 10 A G A 8.23 2.18 rs719548 TLK1 1127 2 A G A 8.21 2.28 rs6490721 None 1042 13 G T G 8.21 0.00 rs10498018 None 161 2 A G A 8.21 2.60 rs10503837 None 224 8 C G G 8.21 3.67 rs1335310 None 446 10 C T C 8.20 2.02 rs10489505 None 84 1 C G C 8.20 11.33 rs2008927 None 662 5 C T T 8.20 1.97 rs2160836 None 716 3 A C C 8.20 2.00 rs2298457 NUMAl 757 11 C T T 8.19 4.52 rs10508230 None 275 10 A G A 8.18 0.40 rs10496800 None 147 2 A G A 8.18 2.40 rs10520676 NTRK3 402 15 A G A 8.18 2.40 rs2901383 None 846 4 C T C 8.17 0.51 rs731693 GORASP2 1151 2 C T T 8.17 2.09 rs4976621 None 988 5 C T C 8.16 0.41 rs1398868 FAFl 478 1 C T C 8.16 2.44 rs159347 None 574 20 C T C 8.14 2.04 rs10500474 None 181 16 A G A 8.14 0.37 rs4131886 LHCGR 924 2 A C C 8.13 2.46 rs770163 NAV3 1179 12 C T C 8.13 1.99 rs1947032 CTNNA2 640 2 A G A 8.12 2.21 WO 2006/053955 PCT/F12005/050429 43 rs2297139 LOC400758 754 1 A G G 8.12 0.37 rs1573219 NTRK2 563 9 C T T 8.12 2.32 rs10508139 None 270 13 C T T 8.10 0.00 rs3774258 MRPL47 886 3 C T C 8.10 0.00 rs9312054 None 1284 4 C T C 8.10 0.00 rs10486445 OSBPL3 55 7 C T C 8.10 2.33 rs974059 None 1349 3 A G A 8.09 0.52 rs10484595 None 34 6 C T C 8.09 0.43 rs10491806 ECM2 100 9 C T C 8.09 2.85 rs2861427 None 834 1 G T G 8.09 0.50 rs508544 None 991 2 C T C 8.08 2.10 rs10493731 None 121 1 A G A 8.08 3.98 rs2900362 None 844 12 A C C 8.07 0.35 rs7905537 None 1201 10 G T G 8.07 0.47 rs2270861 DRIM 747 12 C T C 8.07 0.37 rs1857706 FSHR 610 2 C T T 8.07 0.40 rs6461373 None 1035 7 G T T 8.07 2.02 rs6534623 None 1045 4 G T T 8.06 0.49 rs2414768 None 780 15 A T T 8.06 0.40 rs8092873 None 1226 18 A G G 8.05 2.92 rs2981196 None 850 8 A C C 8.05 2.34 rs9312682 LOC255130 1285 4 C T T 8.04 0.47 rs1173085 None 423 9 A G G 8.04 0.43 rs1834476 UNQ739 606 7 A T T 8.03 0.32 rs7007471 None 1094 8 A G A 8.03 0.38 rs9301082 None 1272 13 C T T 8.02 1.90 rs362492 ANK2 879 4 A G G 8.02 0.38 rs2248815 ITSN1 744 21 C T C 8.02 2.82 rs2725653 None 808 8 A C C 8.01 0.09 rs3770688 LANCL1 885 2 C T T 8.01 0.38 rs7720796 None 1182 5 A G A 8.01 0.28 rs10500943 GAS2 187 11 A C A 8.00 1.99 rs7975812 None 1211 12 C G G 8.00 2.27 rs10502526 None 204 18 A G A 7.98 2.13 rs7763189 None 1186 6 A G A 7.98 2.64 rs10498017 None 160 2 A T A 7.98 2.58 rs10517087 None 380 4 C T T 7.97 0.39 rs4125962 None 915 2 C T C 7.96 2.52 rs1519956 None 541 12 C T T 7.96 0.48 rs10508904 ClOorf64 288 10 G T G 7.96 1.98 rs445607 CTNND2 940 5 A G A 7.95 0.45 rs627069 MGC29875 1024 1 C G G 7.94 2.12 rs10493583 SIAT7C 120 1 C T C 7.94 1.91 rs720884 C6orf65 1134 6 C T C 7.94 2.22 rs6972736 OSBPL3 1088 7 C T T 7.93 2.11 rs1357194 None 457 6 C T C 7.93 0.43 rs1906923 None 627 5 C G G 7.92 0.40 rs633198 PROCR 1027 20 A G G 7.92 1.89 rs997285 RASGRF1 1360 15 A C A 7.91 2.30 rs10497919 None 158 2 C T C 7.91 0.45 rs1430242 None 505 2 A G G 7.90 1.95 rs10275635 None 12 7 A G A 7.90 0.00 rs10492406 None 110 13 C T C 7.90 0.00 rs10503636 EFA6R 221 8 C T T 7.90 1.88 rs1354348 None 456 3 A C C 7.89 0.15 rs3212220 IL12B 868 5 A C A 7.89 2.34 rs9315132 None 1290 13 C T T 7.89 0.37 rs1458088 None 519 11 C G G 7.88 1.99 dbSNP rs ID: SNP identification number in NCBI dbSNP database Gene locus: Gene locus as reported by NCBI dbSNP database build 124 WO 2006/053955 PCT/F12005/050429 44 Sequence ID: Sequence identification number Allele A: Alternate SNP allele or its complementary nucleotide in the position indicated by dbSNP RS ID and basepair position Allele B: Alternate SNP allele or its complementary nucleotide in the position indicated by dbSNP RS ID and basepair position Minor Allele: SNP allele or its complementary nucleotide that is less common in the control population Allele X2: Chi-squared test based on allele frequencies WO 2006/053955 PCT/F12005/050429 45 I WD2- m0 0H~ OCN (N , 0 DWr. W G0 r. PW r.-PI r. 22 0 u 4U P ~ -U 0 0 PF z0 0 0 (DO 0 0 0 NN H -P )r.U H 0 C- zl )( (N Lr) r-(DD -N - N (N (N (N ( LC) LC) I.LO m 0 (N LO 4] r-Y LC) H1 H 0 r- LC)a ( I( N (N 00LC) ( I ~ (N r- LC) '. oH - - - - - - - - - - - -H I(N LC) H I '0 U) M.0- (N (1)Lna) ) U) r- '.0 H1 '. m (N4(~~- IN m LC a) (N LC) '00 a) -N (N() Ca a ) )( (N)( N - zlL (1 N 0 0 N 1 1 0 -r - D(D 0 (N (1) '.0 0~. N. H H) HH- H 0 r N- 1 1- 0 L n 0r zlr zlL n a -r -P ) (N zl '.0r-r zlzl L nL n ( N LCN aC) 1( W)N 1 1 (NHC ( OOL(N ( (N H- r-r - -r zl zl zl) U 0 )H-j 0 W 0 rNH 0H ~ L L'0-r- LL) O - -( N -- ( - - - -H(- - - - 4-Ja) IN. zzl-r---- a)O OL L 00N( (N - 0O1 0- 0 -P --------------------------------------- I(D ( zzl0'(N0(N0(1)HHHlH HH IWD( D(D( D ( ( D(ID D ( D(D ( D( U~) &H HHH HH HH HHH H HH( ( ( ( ( ( - - ~ - - W I (N F<~~ ~ ~ *- *- (1 (1 F< . . . . . u u .<u uF 0 F < uF < U)r HHW H -1 (D a) U Ur) -1 - 0 - D (D ( ( , -H1-0----l----------ll-0---I00-100LnL U) - - - - - - - - 0) 0 0r.I u oo z 01 0 0 00 0 00 0 00 0 00 < 0 0 (a)---------------------------- I I zzl () Lr 1 I 1) (1) (N (N (D '.0 (1 (N r a) a) (1)(N o a)zll M (N MM D(N (N zzl .0 (N Lf)LO I U) ) coU)aU)a) U) U)U) U) U) ) U) U) ) a) U) ) WO 2006/053955 PCT/F12005/050429 46 m W00 00 H) < N R4 o0 o H m o o R0 0 0 0 C) 0 < 0 0 LOL O O O O C CO L ND N L (D (D (D ( L0 L 00 (D ( 0 T H O~ H (D ~LO N 00 LO '0'0'0 LO M- LO O~ N0 00 00 HN HN MN N N HY 0a- 00 r -1 (N (Y r- a) N 1 MY -0 - LO O 00 a0 L0- n( 00 -1 r- 1 LL a) -0 MY) LO H1 LL (D 00 00 00 a) H H1 (Y O -10 -1OLOH 0 LOLO -N LN N 00 NL00 00 - LN 00 N OMT0 MQ0 OC LO r- r NY a) N 1 LO(OY0 )NL 0M O O M 0 00 M ML r- L '.L 0 LO CO O . (DOL O M OLO N L a) ( 0 L0 -O n (D - (N 00 N N Q M H O H O &T0 O r 0 0 0 L LO L t-H 0000 0 O O O O O O N 0 H '.0 CY O- 1 CN t o ooooQ 000 000 0000 00 0 0 H 0 0 0 000000 N Na)N 0 ~LOn (D, 0.0 Nn '0 0(D)00 OY () Cn O - O a ) - -1 (N Ln Ln 0 L HL 00 - - r H 00 & H L LO M Ln LC 0 ( H0 (Yo o o o C e a o o H a 1 ) C Co ooooooo Co CCo CC o C Co o0 oo a H ) H ) a) a) -0 00 00 LO ) a) O -1 00 r- -1 Ln LO LO LO LO -I -1 0 00 ) O Y N Y (N M ZI -ZI -ZI M N N N ZI N N (N (n N (n N M N M Y O Y MZI ZI ( M D CD C0 00 N 0 0 0 0 0p pFA F p p 0p p0 p p p 0 E-00 0 0F 00F 00 00 00 00 00~0 o0 00 00 Gm oa 00 LO LO -ZZI LO (N (N (N LO N ( N a) a) 00 OC 00 (N -1 00 M -1 r Lo M- M O OY Y (D (D , 1 (Y 1 1 tl o ( N 00 (D -1 N -ZI r- r- -ZI MI LO 00 (D 00 (N 00 00 a) a) a) m LO m 1 m m m m m -ZI - a) a) (D (D M O (D (D(Y 'T r-T 1 'T H- r- - 'T 'T 00 'T 'T 00 'T M 'T 'T 'T 00 'T LO H- 'T 00 LO LO 'T N LO LO r 00 -l (D (N (D LO L (D (D (N (D (D r- (D L (D (D (D (D - (D (D (D (D (D r- (N (D (D WO 2006/053955 PCT/F12005/050429 47 og~~L (D a)r o M o OD o4 mDYU RO O 4 -- 14o O ZUH0 D O U(N P0 u 0E P Dm m O 0 RE4U R4 < 0 U 2 Q o 0 0 0 W~< R4- CR4 P- DZ zm 0 u < u22 mo(D -ZI (D -Z mU r-D CU Zn(D (

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o( c o (D em oD (N (D (D ( ( (N Lo m m o o o oo om -i mm -ZI -Zm I o m (Nm (N m (o me o r- e m e m co m n o0 oc ZI o cm o 110 n om m1mo (Dc (D m em (N co (D m La (N o o m o :l o r- oc co :Zl Oo o co I ( (n LO LO 110 r -r o c co (N (n o m o on :Zl tl :Zl o m om :Zll c o m ( c m (D N co 00 mo 00 0 0 N r 0 NLO '.0 :Zl Zl m co LO Lo L- mo C( -i oN mo o oo co o :Zl o co o o o -i a o m moLO Lo .0o c o o N no O m m oo ( o0o N M N :o : N o em m :Zl co m (Nm :Zll 00 r- (D CD 1 0 c ( m( ( co n 10 ( oo 1N N Mooo o o -I :l :l o 0 EED U CC U CD U<<M CC C C << C o L : 0O O-H Lm r- m m 1 N (N m 00O D or r- mN :Zl :Zll Or-L r-Q' co m (N N t-H ~- mU :Zl 00 N 00 mZ > 000 0 0ZI tm tm 0 o t-l (N m :Zl LC CD mZZ m :Zl UU (N UUI N( ( i -N:Zl N:l CD~ CD CD CD' CD ( D( D D( D ( D D( D ( ( D ( D( CJ)I C )) C J)J C ) C JJ C J)C)J CJ)JC)J C)) C)) ( ( ( C) C)) C))C) WO 2006/053955 PCT/F12005/050429 48 H- L zzll (N - (NM < <N (D R4A ~ < pox oOO U PD H : < F< U)I U) 0 0 0 u H r (D CI0r 1IO (D 110 11 (D a) (D a (D 0 o (D m o o co co co coo a o v cn me a (D zz om (om (D ma -o on D m o s zzm (D ( oD (D o(D m(D (om m (Dov o '0 oso (D 00 r- oo -H HHHH HH LOH H H HH H H H H H '. 0 ( 00 '00m r m m aa mn m o( m 00 o zzom zzl r m .zl LO r- Lto-im( zl < UU<< U< U << < << CD<U CD << UU tl o zzl zzl (D mo zl I CCN LC NL C- C (D L 00000 000 000 0 00000 omo (D 1m me (D 1m m Ln 0< m m zzl r-m ( zo zl -L .zl o(D -oo o ac0 n m o r m (D (D o D mezll moo o(Do (om (Dm 0o o-i tom mN 0 oo m l - L mOO C 0C o Om S o -- oN ooN O o ( (D Cc 00LL ~ L---'~O ()OH LL~ L OOOL 'O 110(D N ( 0 (n~ 00H O' lo '. '-H LON- (D M CmNnLOr tCo (D o~~ Cozoa) r- 00oo~~ CooO (Do~ Co mooo~~ (DrCo- 0 ( 0 1 zl0 m L No mL i WO 2006/053955 PCT/F12005/050429 49 LO r- (N m 0 0 0 0 0 0 p -H 2 0 z2 z2 z2 F- 44 P4 44 (D (N (D -1(N r- -1 (D 0 -ZZII(D (D Ln a (D a D 1C L) (D (D(D (D( LO )(Y '.0 '.0 a) 00 0 a) LY '.N L 0 -1(D -0r ~N Q L O N Q:0 LO.00L C '.L n fLO O ~ '0 (D - ~ ( 00U 000~ N LL-L r- L(--H' (Y)C ( CNN r- L(N 00 L(N 00 LL- (Y) a) LL- a) -1- a) Lr- -1- (D 0000000 - -ZI ) N -ZI () ( 00 00 0000 00 D( 1M n() ( 0000 00 000 r- (D( 1 0M (N-zl - N 0ZIr - r~- o'(D'. 0 M ( (D -1 M-(D '.o cn a)a -1 -. m m m-ma) -1 ( LO LO O'zzl r r r ( - ( a r-L 00000 (~N (Y) -1 - - LONLO)(Y r-r-r-r-r-w ( ( () ZZC ZZl ZZO ZZLn (Y) (Y) 00 M'- M~~L~ M QD'Y Y ) )a a r- 0000 000000000 0 a) a) a) a)))a) )) )) JC) )JC)J JC WO 2006/053955 PCT/F12005/050429 50 I- LO ~-HW (-HC i -i (N WD m m 40 40 0 0 0 (D ( n-ZIC CD CD 00 CD CD co c (D (DH L00 (Dc D ( - C D a CD~L CD LO (D (D ( D ( D -ZI m~ m ~ L LO m LO r-( D 1 - co (N O m mco( LO m LO r- m mO m. LO r-r - - r m~ Ln L (D (NCN NO ~O r-0 (D (D co (D l (N (D (D r- (D (DQ' Q'Q' r-H m . (D '. -ZZI m mo o -Hi kC o CDCCU CD-EE- coUC E-cDC Un E-o mE- CD E-~ (DE-CE-C (NC (N mU U U C U m m (N Un co Ln -ZI UCn U r (D OiL -i Lnco LO m~ LO LO -H.0 m'0N -ZZI -ZI c D(D-0 D ( m LO Ln m Lmr-- N Z LO r- (D(D -zlOOM00LOL L (D LOLOLO 0 -(N 1 (D -Z LOC m~ mO L(D -HL ( OLOLO.m mo (N oooo~~~~~ C oa) ooa mC nr N( 1 1 o C L o mo~ Cn L o Co Coooo~ C ocoo WO 2006/053955 PCT/F12005/050429 51 O m 00 00 C 00 m ( (D (D N L 0 R-H M Hi M M ( a u LZ ( R. M M 2 ( R. R4 U (N u (N Hi zzl H oD R4 N zzl <: Q m E- H (D = P4 N R4 pNz. N R4 44 EF-. U) F- 4 U > (N (D(D 00 -H -1 (D 00 r-( D 00 r-Ln-( 00 .zl (D( 00 (N -((1 LO O LO LO T N ND ND N N(DNN N - N ) D a) 00 ( LO H1 00 - (D--1Ln HY a)m 00 a Yr- 00 T L T 00 00 00 (DOY Ln Y N (-00 H Ln Nr 00 (N() Nr D a) a)Q 00 ()00 (YH 00 -1 zzlr )r L a L L a) - N Ln ( ( 00 O 00 00 O L LO (D N 00 zzl (D r- ( H r- H H1 (N H (D 00 r- 00 H1 a) H H) a) (N LO LO N (Nr 00 (N (N LO - (N (N a) (Y (D (N 00 (N zzl a) r a) N a) a) (D a) M (D M 00 00 -l (D (Y LO M (D CO LO zzl (Y -1 N LO .zzl LO -1 00 -1 'zl LO M (D (D r 00 - M r- (D - r- a) - LO a) a) N 00 LO 00 - a) LO L L CO 00 - -a) (D - ( ( N (N - C0 O a) M ( ( - N (Y- CM 0 0 0 Q z - - -1 LO LO LO M M M ( H1 00 00 00 00 00 D D (D (D (D LO LO LO LO - - - L N N 00 LOO O N O a N 00 a z LO z 00 - - a 00 H1 H (O D N l N (D (D (D Ol (N (N zzll M N H H CY ( ND u 0 0 E-A 0 E- 0- 00 E00 0 0 E 0 0 0 0 p p E E- E-0 U E < F u FF< u u u F< < u F< u F< F< < F< u F< 0 u < u a) -H 00 a) -1 N Y zzl LO (D (Y 00 00 Lz a) a 00 ) ) ) zl zl )zz a)z a) (D k N LO O a N L ( (N (N ( LO a L N ( N L O N N (N N - a) zzl - LO a) - - - 00 - 00 - C N N - - - C -o-|o-| C C-T- C)C))C)C)C) C))C)C CC))C)) C)C)C) C) C)C)C) (D 0000 0 LO ) U) 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 L0 -1 LO r- a) ( LO LO 00 , , (D (N - zzl LO -l LO (D CO LO 1 (N 00 (Y (D (N -1 a) 1 -- LO r- - - a) (D M M - N -1 LO (D - - 00 -1 CO LO (D MY N - zzl (Y LO (N Y MY zzl (D ' M (Y (N (Y (D (D (Y (Y (n a) CD WO 2006/053955 PCT/F12005/050429 52 110' (D w D wN R4 (D w U U) Z r r. O H N O 0 0 0 P 0 0 H 0. > 14 LO ND Nzl ND N ND Nzl ND N N N ND LO O '. M- T H LO ( H L H ( L CO L O O- O O L~L D (D O D H '0 m zzll O H N( m m N Hm0 ( O Nm 00 a) IfN '0(D.0L N O O O O LO zzl Om .0m 0 O O0 m m M -i -NiO m OL~m H & H NNO O 00 LO ( (D 0 Om H N] H -H-H M NO M O oN mOO zOLO CNON ( Toom T- ON-MN O 0 0 H (D-N LOOO HN H0 N OHNNO zz ll zzlO 00 Q ' Q C 1Q LOOHH 00 -1 (N -i Q OO OOOO m HH HH On HHH L0 HHH -i zllm w c m r- m MM M0 0 00 LO r- ( 00 zzll DC 000 zl 0 0nr-000( 10 0 CDZZ CD 000l r 0 000 UUU 0n- 0- n Co ~~a) )~ ~))C~ m) mooo~~a m) moo))) mJC)C m) mooa m za)l zl nL L H - HHH HH HHHHH H HH HH HN HHH HHH HHHl H0mL - 0 0zl zlzl Nzl 0- 1 m NO (N mN M zzl me o (D (DT C CDC CD- i zl (N N(N zl (N T (H H (D( < uU0G < U <uU< < u F< uu < < < F< F< < < < < u< F< < < u o o H0 o 00 o- r- ( i( i mm r zl zl n ( H MNT 1 1 N N (NH H H m TH~ zzl zzl00 0m N r00a)mo m zllm Hi HH iN N HH THTH Hzl zlN NH o -- 1 0 N HHHH - - (N N Ln NTH R4 0 0 0 0 0 0 00 00 0OP 0 0 00 0 0 00 uU O (N zzltoo OO 0 ( D ( N -zl N(N m nr N N m 00D00 O m m m m o m 0zzl 0 a) N O O H H CD( i- N O M (D M - 0 0r -tozl 0 m m T aH) 000 zlzzl 0 (D L MMM (D (DMM m m D zM( ( DMD M r- (D (D M D Dr r M M (D WO 2006/053955 PCT/F12005/050429 53 rm Cn -ZI (N a)) m OLOLC) L .ZIU rU u U 40m 0 0 P-ZH U u E-uE-pE (DO (Dl ( D Ln CD (D H- > C00 ( (D~ (l~ D (D (DY r- (D m mE U lZ (DZZ (D (D (D LO( D 1 D ZI (D (D (D ( 00 D -ZII m(DL0 (D (D ( D (D m( -ZZI -ZZI LO HZI i r ZI 0r 00 ( m r - ir- 00 10 -ZI D ( LO m~ r- H. (N (N 10 00 H N C (N~ '. N c 0 m ( 00 00 H(H m r- H N 0 (D H LO N( r- ( m m r- - r- 00 m -i( ( 0 LO ( 0 QLO - H - L --Z OLO LO- r- LOOm 00-ZILO 1.0 (N 1.0 mfl1.0 (N ( 0 00 (CN N H(m ZZI-ZI) - - H LO ( rL- M CNCNN (N (D 0 (N m~ mT mO H (N (N N 0'.10 '.0 '0 (N LO LO (NHCN H ZI m (N (N -ZIILO -ZI (N ( N ( D(N m m ZI (N H- -Z (-~D D (D (D (D CDC( D( D ( D( D CD (D (DE~C (DUED(D(D~C (D (D~ FA~L H C0 0 ~ p u 0 '0 000 u pNO L'0 pO p 0 FA FA Hi (N -ZZI m ( -ZI mN HHLO Q'01 O' LOOL r- r-CNCN HZI -ZZII -ZINWWmLn Ln uU uuU r 0 0 00 u . . 0 00 0 0 CD CD CD 0 0 0 0 (Y) 1 () - 0 ) D zzl N LO LO LO -ZI .0 (N ~ H H 1 ) 1- 0-ZI 00 00LOn -ZI (Y) - H1 Lr-O a) O a) ) - (D - 0 )0 (~N 0 (N M( ) )a HC N '.0 a) Lr- H1 (Y)O Ha) ~ ~ CN H -~ HC ~ C U) U) U) U) C) C)) C))C)) C)))) C) C)CC) C))C))wCw)w)w C) ))) ) U) ) ) WO 2006/053955 PCT/F12005/050429 54 Q 00 om OOHCN~m LON H (N (CN HCN N (NH (N N a) (D WD WN R4 4 O4 4 0 0 0 H 0 44ZZ 44 FAx ) R4 Z2 Z2 Z Cl) o Lo (D ( ( - -L LO mLO ol (D (D (on ( zl( o Ln on 0) mn m DI 0 in .zl In In0 n(N zl l mn r- In( -m n ( oD mn o0 on t.0 o00 (D zzl o o0 tl '.0 I0mm n In nm me -10 m m (D (D com omm oo0 om mo (D r- LL - '0.0 D - (D om m to ooD o (D << m0 00 m> zzl o (D o o 0oo m ozl . o r- L o- m m m In In (N m m o o ooo o00o o oo ooo o 00 (D m to (D m o o ooi Lm m (D -i ( D- 00 D( DML 00 zlto zzl 00 m ~-i L- 00 0 N zl zl zl zlzl zlzl zzl In n nLn r- r- Lr- r- - In t.o'.o '.o 1- o o-o-n( (noo (n (no (n ono o onn In UU U U <<< UCD<CDC UCD<<< UU <U < 00 0 0 0 00000 000000 HH 000 0 m N zzl zzll -CNCNlO (Dm m m m ClNC N CN (DN M~. InN N CJCI C)) C)) C)))C)C)) C )))C)C) C))C))C))C))C))C))C C)C) C))C))C))( ( ( ( C) WO 2006/053955 PCT/F12005/050429 55 H HH (D 40 D r ( R4R4R (NO0m( (CN (N N m Ln H mr 4 r 0 - H H H H H HH 0 0 0 R4 W C) U) U) 44U 444 HZ Z Z 4 u > R4 a) 0 ( ( (D ( 00 (Y) - (D LO (N 00 LO 00 .zzl Y)(D (N ~-1 (Y (Y)(D00LO LO LO zzl r- (N a zl () () zl ( )LO ILn (N ()LO in 00 .0 '. in r-a - (D .zl n in '0 00 in (N .zl.0Y ( (N ~-1 in izl n (Din in in a) (N a)() Y (Y) in in 00 '.0 '.0 (D'. LnM(DM-( 0 - N DLn ( L ( (Y)( r-i (D OD , r-r- zzl ) )-1 (N (N ( -0 ( D( D ( , Q'00 Ln in in in a) -1(N Ln n i Ln '.0 '. 0 1 (D (D( 1- 1- 0 00 1zl (N (N (( Ln in o 0 '0 '. 0 1 10 wM(TMh(MTh(r iniLni Ln in 00 . '0.0. CD~ CDE~-~-UD CDE~DU-E~ CDDC E-CD CD CD (D( D(CD( D D( D( D ( D(D (D ( ( ( ( ( ( ( ( (N (N (N (N (N (N (N ( ( ( ( 00 00000 000 00 HN-HO - 1 00 0) 0n C) oo (- ' O (NY- (N) m- m m-H( WO 2006/053955 PCT/F12005/050429 56 R4 mmm N 0 R40 u < > r- D r- m o-Z -ZI (D (HD ( - ( Ln (N (D ( co D (N (n ( LO ZZI (D( Ln- mO '.0 (D0 m~ LO HZI (NLO-i m N LO n LO Hl ( LON '.o m~ (D -ZI N i co Ho H-L N ( l LO (D -i mN 1- Hi Hi H LO (N (D0 -ZI to LO Hnr- L '.0 (n- r- r- co( c m N In In In 1.0 r 00MM D( (NHCN H o OL-ZZI '. r-toto ( zl ot zl 0'0 C N N (N .0 Hi(D c m'O In m to - ZZI - ZZI c c -ZI r Ln -i Hc N In m -r ( N r- (N (N (N LO '.fm In mHH( In '0 ( 110 m (HN (N n RZI N tl -c - i (H N LOn Ln HHZI -i-i (N( ZI N m L i- ntl l l l - c oc 0 00n~ In mI ~ 110 10 110 110 coc 1 (N(N (N (N N (N (N ( N (N N N N N (N N (N (N(N (N (N N (N (N (N (N (N(N (N (D (D (D ( ( ( CD CD CD CD -E-CD (D(D(D (D ( D D D (~D (DE (DE- CD- (D( ( D D( CD CD (D (D (D( D D( D D ( ( D( ( D C CD CD CD CD (D( D ( D ( D ( D ( pC( 0. H'. 0 0u0pp 00p0u0 p~ H HH HH mm HHl l ( ZI 00000 mmN 00zl (D 0ZI U U00 00 >n( nL nL iNL D ( D m-i -i 0D ZZZZ -ZZI r- cDCCDD co r-Z ZZ a) co r '.0< F<O H r.~ -- r.C Lf-H f~(TiN H~ 0H ~~ O H . 0f'. H~i fNH In 2>>>> WO 2006/053955 PCT/F12005/050429 57 4-1 (N u~- r- -410-ZI Dr 0o m~~ = MR4D4-O 4 N (-HN LO M H 0 Z2 LocI) (DRmUM4 R4 ~ uu u u ZCD 0 P4P4U LO r C u U) U) ZN c4 ) p 0 0DCD CD02 uE- pEp pE 0 4 P (D F: P U WW 4 4 U)E P E- H Z4 R4 u c H p u x LO) If)N LNO0( 00 (N n -1 Y) LY LO LO 000 -Z0 Y (N (Y)(0 '.0 LO 1-1( '. C CN'. L(Y) ~ C '.0cL 00~- -oLZ-I I-1Q LO -H~H~ O(T O'LO ~ Q~ '~CO~ ()~ - (Y C1 1LnL ZZ- C)CCC C)CC C ) C)CC C)CC CNCC)C M l nr 0r 00 a) -zzl- (D (D -1 r- - ~ CDC0 - E0-0 1 E-- UE-N - CD-(N ~ - (N ( NCN D 00 00 00 WN (NW (N N ( N( N( (N WW N Cf(NW WN WNWW Y)() 0 pOQ pDDC 0U 000 0F A AF uF 0000p 0 000 u cfZZ uZ uZ FcZ Z ZZZ u u c F u0F c c r- ~ ~ ~ ~ - 00a Y - na (D 0 a 1 1( - 10( ()- a-cN'.c o m cn r 1 DL (D -1-1 D D ( (D -zl. D . ( -. o '. r-~a N~ ZZOO( - -H0N a)-Oa '.0 ZZI L())LO -1H LOrHL 00NL CN 0C ~- L ( 00 0)a QLO ~ O'~L~ ~H~-H CN~- '. '.0 n LLO( QOn ~-~ Co ~ ~ - -1o~~ -1CJ~ 00oa 00o) C~a o~~~ oC~a WO 2006/053955 PCT/F12005/050429 58 ZzZCI) a) u zD( m (D PU 40 4P 0 0 p- 0 F< 0 Z 4 -0 L4 2 R40 P )0U r m Ln LO( m( m ( LOm(n( (D (D0 (D - (D LO (n0 LO m. (N (D (D 1LODm D ( r- (D m- '. LO LO '.0 tl 0 -1 -ZI LO (DH m i O (N '.0 m. tO D n- Ln (D m m~~ -ZZI tl l -r l i ( '.0 LO If) m ~ m O '. mO (DH LO nL ZZI L l 00 00~HCC m~~-- t -I '.0 m~- mf O m mOO '0 a)~--~ 00m r l l D ( r- ~-'0 L OLr00 ( ( ~-I -H'.0tlo-ZZ mO -~ tO m m Z LOU LO- LL ~-Z r- -ZI l o n ooo 000 Ln 000 0N E-N- 000 (N -Z 00 0a 00 (n -ZZI -ZZZ Z -EZEZ-IZZ Z Z /) -ZZI ZZ CDI -ZI Z -ZZI -Z N NL 0 a - 0 LO m -ZIm L -( l N L '0.0( tot 00 L0. Lr ZZI o-'.o'. LN '.N (DOL M (N M DCNC m m m N m( -ZO Z M ( CD C DCD C D(D( D( (D0 H (D- - (DhL Q'(D (D (D CD.0 (D (D. '.0 (D(D(D (D (D(D (D (D(D(D( (D- (D-(D-(D- ( CDO L (D (D~- (D (D. (D' (D (DL (D (D( D ( D ( D( D D ( D ( D( D( a) a( a aC aa aC aa aC aa a) aCa aC aa a) aCa a) a C ( a( ( a C C a) C WO 2006/053955 PCT/F12005/050429 59 u LO (D WD W N Wr ( N WD WD W r z 0 fo x H r r. Z0 0 2 0 Nm 0 0 0 Z Z E- GCD Z HU Z Z Z U 2 FA '0 R4 2.u (D o o (D (D N o (D m (D (N (D r-( n N( 11 (D LO 11 D (D 10(D ( (D (D (D N (D (D - (D N 0- (D omD c Lo om oo m ma meZ s n N m r-o to on omoIo m om c o oN coN r -Z no m c o o m - ( mr mLoL om o m o om oomm mm o e mmo (Dm m oo c o ( m (oD co tlo (mN r- Lm Lm '0 r- ( Ln oo co ooo L o mmmm O mo ss mo ooo I o o oooo oZI oo (D o (D ooI ooo mo o o o oo LoooLo o oooo LOOLo o ESCE- ESCDC CD CU E-GDE-UC CD CU CE-sE-U U E-sE-C CD-EG < << < U< JU<U <L<< <<O<<LU< << O mZZ c - co C0-mD t o o no r--mI o -ZIIto m r- o o o m o n o D( Lo 0m 0 co lo m tl s(N( Om( -ZZI ooN -ZI ooL - r D ML ( OM r ZI l oooo co oo men (N an mL o coo(N r-oo m < (N m c m mco o m a ma -Zman o omo (N 'z'M N( i -i( nLnr Z LNN ooo ol (N (DNN (D oo moo oZ m N ( NoDCoD0 N ( ( N ( l -ZZ 000 0 0I -Z 0 00 0000 0 000 E0EEmmm -ZZI -ZI - L Z Z Z Z Z t mmmmmUUUU ~~- (N~- (NC rLO- '.0 '.0 rL O . L '.0CNCNI L L L LL--n L LLO LOO-HO. Ln LLLO'n '.0 L-HO Ln-H n O o i - N-ZI r or o r o ( o mN ( o m r ZImL o o tlo n oomm o oD (m o N (o (moN aoCooC C Co C Co C C ) C))C))C) C) CJ)CQC) CJ)CJ)CJ)C) WO 2006/053955 PCT/F12005/050429 60 00m

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a) - m 0 m a m- (N (D ( )0 P 4 4 4P (D (DD (D -L( D (N - - a) a) ( D a)() (Y Y 0 LO (Y LO( -ZI D 00 r- (D (Y) a) 10 ZZI Ln (O N~ O r- ( a) LO -ZI rLO -1 (N( -1 UU UC (JY)~ (Y (D r- (D a)~ C 00 -ZZI'.0 rL O a) a) r-HNCL (D- a)~ LO -1 OOOOCD 0mm m 0 01 0- 00000 N 00 0 000 (D ZZZ-Y 00 z n -ZI r- -zzz C N (DZ ( -1 0 0 10 ( ZI a)a aZZIa) a)-) r- )Ca) a) -Nr -(N D a) -) aaaaa) a)) (N~a a) -1~a WO 2006/053955 PCT/F12005/050429 61 ~~CI)~( mw~D CmH 0-x 0 0Or r 0 0~ F< F< x f~ - UD 14u . D0H 0 0 ',D~- -4 E-r4EP 0 0 UN HxU U R4 R4 U U) C D Z Z r-~o o0 oN (D oN mD (Dm (Nor (om moN( m ea ( m ( oo- mom e mmem (e m en m o mo0 (N mos oo mo - onl e m am om :Zl om0 om :Zl mo o 0 o0 om Lno m (D o m mo- o am a o o m m o m -zzl om (N om e m (N- - amm o mo e w n o0 moo (New o a m o CD e m no moo em mas aa maa s -oi o oo o (D m oN 0 zzl om oD oe m (N (D s i (D (D oo mo o o o n o0 om(N Me m(mmD -im - w a oo o o ooooo o o o Lo 0 o o r- :Zll :Zl m m (N m (o 00 o o o o Mo r (o Lo o -i :Zll r- 00 Ln m -i 10 N m 0 Zl D( D M 0 Na n m ( n( 1 1 CD( ( N N( (N (N( N )a)a 0 n 0 (N( N( N( Zl Zl:l Zl M (D (D (LD(DO i LO nL n L L LO OnLnLn-HL L LOZl '.0 L0mzlOnzl 00 1r 1 0 - N L 0 L 0r - L M ( ( (D :Zl m m i - - - (N (N -i i N :ll :Zl m (N (D D D ( M m LO - NL C LO.0'LON m-~H r- OrhO m 0 ( N 0 L 1 C D (D (D E- E- - UEC)DC (D (D (D UE-E- E-D E-(-D- E-E- 0 CD Q ) uCD u u uU uU R4 U UU LOC'.( r - :l Zl Zl0 :l -w ( - N :l :l 0 L m (N'. (N (D .m m.00 0 000 00'. '.0'.00000 00mzl a(N00 00 0 n00 (N -E-00 00 0 nr i:l (NC~C C :ZllC m mO'L~ m-H 1 N m zl )a U)U)U U ) U )U )U )U)U )U ) )U) U) U) ) C) C) U) U) ) ) U) ) WO 2006/053955 PCT/F12005/050429 62 LO r (N~ i u (N Fc z9 P R4CI H 4U 4U : 2 4D4U (D 0D (D 00 O T OL HO L N N NN(N ( I-N (D~I -ZIOr-( ( mO Hm HH 00 0c' L L LZ me (D o L0 o o (D M i O-O -om 0 m o 00 M OO N H NNO ~ I- LO C r- -H()N LOC m~ O N~ ~L o N (Do mNN om 000 00 000 0- 0N S O~H~-H T -HLO L OM Q LomL M0N (m -i -iH mo -1 mH HO G00co <U< <UU U UU< U <UU< <U <<<0 < m~~~~~ r-- -- 0 1( - (M LOO r- m CNO'mCN M M OH WN ( -r DM'zlL 0( D r 00C ZZZZ 00 (DH Wn WWWW CNN-i WW WW WWI 0-ZI o (DO- D 00 0a)m m L 0 0 00 r 000 r- -Z 00 00 00MM-zl nL N (N( ZI ZI ZI ZI (DH m l m -ZIZZ (N ZZZZZ ZZ 0 0 0r D(D( D( D 0 (D(D( NNN N a) a)mm an om aC n m O OO OOO N HOOOO OO O Cm C C CCCC CCCCCC Cm)Cmm m O O1 O O O O -1M O O O O mO O ZI ZII-ZI r -r-r -r 0 Hn - i-ZIL iL - L n0 n m m 0 0-o( (N (N (HD -1M m ( 1- D( N( O i( D - 1 -ZI( WO 2006/053955 PCT/F12005/050429 63 m ~m -i R4 R4(D(D - -4 Z4 Z ) ZCI ) 0 Z 0H Cl) Dr4 ) u uu F (D r- (D LO 0 ( i( (D (N( 1 ZI -- ZI ( LO ( HN( 0 r 0 ( 0( H0 H1 -1 ( 0 Z LO 00 Lr- LO 00 (D0 m~ H LO Hi H-m -Z 00 mLm Ln - (N LO 0 00 LO LO m H HN mN (N L 0 (D H(H 00 -ZZI (N LO-i H0 m LO r- HD 0 00 (D -ZZI (N r-( 0 0 0 0 C n H- H m~ m ~ r- L0OZI im -ZI (DmLO '.0 (N ko k.o k'.0ZI LO H0 H1 HI (N LO L-- L--H LO ZZC N (N (N 0 - m -- ZI LO (N'. m N -ZZH i - 00 (N (D00 Ln (D~ Ln -zl 0 NCDmfl (D r- H 00H-1.0~ 00 H (ZII-i -i mHi N LO 0 r-r L -i -H(D-zl ( OLO 00 -1H1 - M (DLO 00 HH(i ( r -i(N 0 m -iO' ( -HI C10 mO~ m.0 ('~N N 00'.' (N HLOO a) H DC HHHCN m m m LO '.0 L0 - (N (N -ZI ZI ZI ZI ZI I Lr- - r- Lr- r- - L LO LO LO 0 0 0 0 HHH H1 HHH -I (N (N(N (N (NN LO LO L- r-L HHH H (D D(D(ED D ( ( D( CD CD CD (DE~ -C DH - (D(D(D (D((D D (CDU( CD (U( (D (D(D U U (D (D(D( ( DD(D (DU (DUU (D (D (D CD~ (D( D( D ( D( D( D ( LO O C C C C ( ( ( ( )LO QCN C H N C (CN C C C QCN H r- J 1 000W 0 ZZ~~W WWW -ZZ)I) W0 m00m m 0000 0 ZZN (NCl m a)C)l)C) 00 0 000 m H 0 1 -1N N( 00 ~ -ZZI 00 HLO r- 0 m N m CN~ (N r- H (N-i H F<HC R40'O R4 R4 H ~ L O~H~~ ~ O L ( L O m~L- <() H CNHC L-~ H-O'H (N HN HC H L (N~ (D~c (D ) (DCJCJ (D2 4 4R4R4R R) Q)C))C)C)J (DCJ C)) C)C)C) C)) ) C)) WO 2006/053955 PCT/F12005/050429 64 0 x f C m u 4 U O LO O L SLO T LO LO LOO O O '0LOO O O O '0 '0 0LOu P NNN Nr Nr N N Nr N O~LOLO (D CL D (D ( SLl (D (D ( .zl i D mr '.0 1- (D (D (DH (D0 10 L LO LO LO 110 (N (D (MN (n Ln( D D ( a) a) L L O L N( )N O (LO . O - N'.0 N m N - 00H n -i OLO z e C N .O' o N o~ e mLO NNH O N 00Th T - 1 m m (D (D o O(N - m (D m 0 00 LD m Q OOQQ LO M T QQ n WWWWW eW W -i W W -H WW (N LN r- W 0 a 00000 000 a) 000 mU zzll 0000 000 (D(D( m ~ (D~- '0 QLOn (L zzll (ooD o ' m a m 1o 11o o O . O mN LO O - '.0 L ' 0 -i m -i zzl r- (D m LO - i on o ooo-oe m (D m ozl m m o zl o - O N L oD oD oN on o- o o mom0 m m0 D m o o o o o(DN CD oO -HzlC N LN L 11 O zzlCJ) C)0 LO N J 110 m N N (D M O i m m (D C ) zzl S (D (N (D 00 1 o m o o m a) a) o o oN (D o N m m m o zzl zzl zzl a) o o 0 (D CD M 00 (D (D oN oN oN m m (D o N 'o o m o 0 o m o 0 o o zzl m m oo N T~~~zl (N N - N0 N0 Nm L D -zl zzl N Lm Ln (N (D (D D M -i (D& M m zzl m m zzl zzl l C (DCC C C C C ( C C C ( ( ( C C M U C C C C C C LO0 M( N zzl 11 m m m (N (D M LO 0 ON m m 11 Mm10 - LO 1 0 LO LO LO (D (D (D ( O L ONN OON ziLO 0 0 0 m m (D (D T L. fQ Q x r. r. r. 4 O O4 r. r. 4 r. L. r. - - O O~ N~ 0 O WO 2006/053955 PCT/F12005/050429 65 I-o I 4-1 m m Cr u m (N LO O H ( z N W (D M- 0 4 m 1< Q R 9: Q -1 Ez. (D - (D I r | 4-1 -H 0 < u zl H H x X 4 O I uH -H 0 ( L-OZO OZI M N O HLC) (D (D 1 4-) 4-)J D (D ( I 0 -H -H r 4 -H I ( a) a) (d a) L) LC) (D (DI 0 0 r. 0 1 LC ~ I O O E C (N ( (N (DI : 0W I W- W t O H H -H - (N (N U) 4-3 4- m Q SLO |N H U H 0 LO | Z T LO | t,C -d -*Hi (N I -I 0 0 (N - H W o W H a) (N (N l> t t r H1Q, , * H -H O& O 4-) 4-) -1 0, C I Wt 0 0 (Y) LCL I w O O -H) (N (D(N LC)I ( -4P HrHI r.rI . .zl D0 LOI, 0 0 00 0 -C. P rm ( LO | ( H C H C -0 , LC (N ( I| -0 -0 r. - 0 C) O (D (N (N It >d -H -H d : H N (N I r- O01 0 U O1 t I R40 m QmQ ) 0 WW)u 0 o o o I Z -H -P -P -HH Z zzl LO (N z L) r- () ' r- N I w -P 4 4 ) -P 0a O a)H (N-( ( (N z zzlN LC | -H -H -H >i -H -H 0 00H Cf 00 zzl(D - 0 Y ( 00 00 LC) 'D ) (d (drHa ) -P ,0 N G(N N CD () LO O) LOI 0 WQ pW o ( 0 H H(Y 0HHH M 0 ' 10 ' (N (n I H Q H )H C Qa C at 0 o0 o0 0 o0 o0 ( ( H ( I m Pa) P, ) (d 0 R-1) LO L) a) ( N N N N (N (N IUH u r (d H a 2 ) a) ) a) a) a a 00 a 0o 1 0 W0 0 C) mo l -H ) U) -H -H o ) I >iLf) 4-) (d -P- (H (d > () -P ,4 a) a) ( 00 r- r oa) 1 4 -C C , - -H) -C -C 0) ( (NH f (N (N (N L CD CD (N I) | Q p ) L4 O OOO O O O OI WH0 0 M H 4- O*o o o o oo oo o o oI o a)o ol o pa0 I m (0 H H 4 ( D ( D(D(D(CD CD G CD CD (-)I r 0 W 0 (1 P - k I W -H -H 0H U a r H U( << U U < U U U 1-H 4-1H ) )H ) )O 0 H I -t 0 o -a - -P - a (d H NoN 1 O- ' p 'D - C H uH I -H OD Z ) >1 )i a 0 H S0 HHI4-1H tld, Q d ,Q Od OH (0 r I -H P r. -H M4 R4 O I -P -H (m ( t W t 4-1 -H -H I r. w rm 0 -P 0 -P 0 LO P, U)U I 0 ) CW m -P m (0 (1 -H ( a p (D~~ (D fp i f C (d fC (d j (d 4- _Q I -H o M- O - W H pa a I zzH zzHH (0 HH (N F- < Q V - O - O > ) -H o ) ( N CU W H r H r I (d p (d O HHH H H F< -H < -H P- -P 4-1 0 0 0 0 0 Z 0 0 P ZI *4 LHH H Ha) H|W p ( 0 (N (DO I Lf ) (N (Y) (Y)(NH -1 D O1 zzl a)l HI a)0 -1 w * Sa 0 z N O 0 1 H -0 - W 1 (N MCfM H-1 ' D H1 - H0 (N a) I H- r. W1 W -H 0 z(N(NLO(Y H LC H H LC) I W H H H-I H H o n r N (N I W W (H W1O (Y) aHHHH (N H H H H r- I UC) 0 1 H H > U) a a) w w w wa ) a) a) a) a) a I (o) oH H 1 O 4 M 4 4 4 4 4 M M M Z4) 0 U) F< -< P-1 R- 0 WO 2006/053955 PCT/F12005/050429 66 I H I I HI N 4 M ( ID -zzl ( T N (D M M N LO Cl. N 0 0 (D r. (D O r. -P ( fC P R4 -1 2 Q Ez. U U U R-1 M C =l U) 4g D 0 0 ) 0D IP zzl (D 1- (GON (D < zlO (N ( .- (D N- - - - - - - - - - - - (N I L ( LD I( (N O L zl O- m (N z -H I H tlo lo zllr- -i - - --------------- I L L)o -H -H LC)I t|: I(D L 0 C) -P M 4 -P (D m ~ (N 'tC' (N ) zI (N D a) *n mIm m OI LO L .o -H - - - - - - - - - - - -1 -H I '.0NT LO LO ( OL I-100(D ~-i N zzl(N (N r- (DLO Q -i -HL O R 0 () -H 0 I -H O0( M (NOM -(D(D '.0 (N(mzlCN0 I- P zl(N ~ -i t.'.o t Cflm m (N (D( D zl P - I Ln l 0 zl nzl zlzl n L zl ( D ( D ( p-i | O O LO LO -C U -, 40 r~j 0 W I~ - - - - - - - - - - - -I O-C. pm O m -a m -zl ON zzl t o o H o o 00 m zzll o S + zzl zzl N zl m M HD zzl (D (D M N O O - I LD ( ( ( LD ( ( (L LD ( ( ( ( ( ( O OHUH WH (D (D (DD- ECD C ( EE-EE-CD CD C C fC 0 - - - O - O - O - o I .H p- - - - - - - - - - - - H-HHH-HHI (D - - - - - - - - - - - -| 4 ( (D(N (D (N (N +- 10 Lr- ( ( r o - 00 -H I OO 0 O 0 0 O O 0 O O0 O 00--l H| H pI>( Hpo|O O O O O O O- O O- - O-O O I . . . .. . . . . CD l) H 0 ) I OO OOOOOOO O OOOOO U)O - - - - - - - - - - - -1 I ~ (D OP H <I W LC)(N R4 R4 R R4 R4 r ( < H H 0 U U U U U < 0 0 < 0 (d U - - - - - - - - - - - -| zzl m mm m 11 0 100 m C m 00C11C 00C( M -- 1 '00000 4 -- mmCmm(D M r- NC M ZZnm I H0O 1 1 H H z O) 2 R4 o 0- C )o H H (D LD ', (D M L (D (D zl M (D (D -- HI c o ae o H o H '.o H HHH H n -i - l - L- -- - -l - - - O IO .ANC~- N H C HH wClZ w a) HCD w w w.0 w w ) ww I 1- 14 a) a) a4a4 4 a a) WO 2006/053955 PCT/F12005/050429 67 0 0 00 00 0O '0 0 0 00 r- LO Ln () (Y (Y)(D 0 Ln a) 0 00 ( 00 -1 () .- 1 ( (N '.0 LO-1r- ZZI, 00 (N 0 D a -1 (N 00r--10n 0 a) (N - 00 a r- 0 -H'N C QCN (DOO~ CN0Q 00.'. O Cm 00L LO( -H00 NC LOLO (Y L O C O'O 000 00 0000 000 0000000 00 000 Lnr-(N - 0 N 00(Y )-10'00 (N (NLOn r- L -H1 10 -1r--ZI ( ( O00000- (Y). Ln 0000-O a (D - DM D(N ()-ZI OZI nL r- (N-H0-H O ' 00 1- -- LO( O LOLn O O1 1 a Y - -1 n (N -ZZ WO 2006/053955 PCT/F12005/050429 68 LO LO Cla) a)~H U) r. r. H H H4uuu0 r 0 H)00 R 4 P R Y P cD c2 c2 R4O c4 4 (D 00- ( D NC CD ~ ~~~ HD( D a) a D( (D HY L 0 a)N -ZZI (N 00 HD Cn -ZZI HY H1 '.N a) . Lr- H10 1- (N (N H1 a) H0 -ZZI (Y (Y) 00 -ZILCO r- (Y LO (D () (D -100(N0 00 HN Ln HN H0 a) C (N '.0 H0 -1 (Y) HY H1 -. 0 ' -O~ O H 1 (N(T CNN Hn -1 ~ -O'LO (Y H H'(DN ~~ O H, (D~ r-.0 H , CC Ln r-N( (Y) H- - DM 0 0a (Y) ~L (D OLL -. 01 (Y 0 0 C O r--LO( Y Y N (N- )- 0a n-ZI zl z D -I -1 1 C C 0 N - 0-1a) -1 - (Y (DOL '0 (Y) -1 Q'0 HHNN Q(N 00()M 0 ( -1~~ ~ ~ CNCNC 00OL a) a) Q'- LOOL'. HHC a) '0.0I()0 1 zl ()( Y 0 -ZIa ) Y 1( ZI 1 LnL L nLn( D D( L D a)-ZI n D D - HHHI (')( Ln H 1 (DC -1 M (N 00CN0Na) Cn '. (D -1 D ( ( ( ( D E- N E- Y-)C CDENC (N E-) (NC CD-1 -1C -1 DC (Y (N-1D (DU~~ CD CDD CD CDC CD CD C D ( D D ( D( ( D ( D ( (UD~ DU (D ( D( D( H L- H1 (D N a)'. (D ) - H1 M - MH MM MD() (N (Y ( ('N LO 1-1a (DO (D Q(Y) - HLOH-N (D'zzl (Y LO -H H0H-HH-H H1 -H. -H. H1 -1 HH H - a)- -H(Y)H H-' 0 00 00 00 0 00 0 C) x U) 0 00 0 0 0 0 P4P40 0 0 00 0 0 zzzzzz X D zzz Ol Ol z oz u a) L O ) 10a)a)a)(Y a- a) a) 0 N (NH-1LO (N 00(Y N r- a) N LOHO -1a) a) r- (Noa kooa o Co C) Coo-~ CoN ~ Co Co 0- - 1- WO 2006/053955 PCT/F12005/050429 69 m 1O 0~~~~0 0D 0 m m OD 0 O LO a)o 11N (D (Nm< ma (N (N m r. r. r. 0 0 00% R4 4 r. R4u > 0 0 0 2NWWW< 2O0 0 P O O O u Q (N (D a) (D (D (D (D (D (D a) OL OO M O LO HO OH (O (D -( LO HO O~ M O O LO (Dl L- a) O - ( O (N (O LO O -H LO '. O ~ M I- 00 N H 0O N Ln ~ ~~ ~~~~~ r-0 L)a 1 (Y 0- 0 N Ln ~ ~~ ~ ~ '.0 r-()0 N ). ) 0 LO LO )( Y '.0 00 -1 0n 1 ) 00 L0 O Nr- 0 O OY) LH ON r- r- r~- r- - 1 M O M 0 000 OZZ '.0O H.0 LH O'mO~ CN~Omm LN o T HoNCN T L O -H M QQo CN'0OLOO CN'.0 O NM QQ H LO% OQ Q rm (mmD ooo (D 00 oo o (N (Do e NNN NNNNN NNN N NN NN NZ N NN N N1 NY N( ON L O0 NNaH -1 (-HCN HN 00 NN (YN -ZZI ECE-CD- CDE nC E- 0 -DU CD0 (D~ -~ E-) E-~ UI U U< U<U<U <U U << U< < U <U U U < om - r-o one a) m) Lo o o H-1 -ZZI -H 0 T H Z H H H H H.H OCOCN OOOO.0L O OOO O -M OO OL -1 (D OO 0 O O N (Y) D (Y Ln D (D ,D 0 (D M (Y -ZZI , (D D -1 00 () -1-ZZI (n 00 () L 000 00000 ZII-ZII 00 0 0 a) (N(N 0 00 00 -1 L n a 00 0 0 L-Ha) L N'.0CNOa) (N (Y) -1 a) -1 (N (LO (Y (n CD-C CD~H-H-C CDH~- (D -- ZZ H( Y (NN '.0 (Y (Y) O -ZZI '.0 LH () (N aC c ~~ CD) CDCJ CDJ CD) CD) CD) (D) (D) C)) (D(C)) ( D ( D D ( D ( WO 2006/053955 PCT/F12005/050429 70 O T H O O H OH H H 00 11 m a) U0 H 00 N UO r- 1 U M0 a r ( r r MO ':l R4 ON MOO (D OON m1 mO N a)O 00 OU U ~ n 2 u oa: 0 Da -wi M (D Ex- -ZZ-i 00(N u. u >-I >- - - uu Q uuP C tDo C C ( o o (D N (D -ZI L (D (D LO (N (D (D (D N m (D (D m (D~ ( N (D -1 00 (D O( H 00 (D HD HD CD O N (N (N N (N N (N N (N m Ln t 'o 'o 0 (D (D ~00 00 (N tl 00 r m ~00 tl (D -ZI Hl H m N N m m LO O 00 (N LO H H - a H n LO 00 m m N N N N LO LO LO LO '00 O~ HO H m -Zo to coo o N 00 m H0 (DT t too 00 0 ND 00 N Hl -ZI co -ZI too m -ZI on a D H LO 00 00 m N N N N LO LO LO M M 'zl (D m m LO -ZI tl tl T H SLL 00 (D M (D tO to m m 00 000 N o 00 ( H H t, M (D M r- M (D 00 0 r H 00 tl H N m r- 00 -ZI tl 00 (N -I 00 0 00 N m m 00 C L o O m - r LO (D H O LO m m -ZI LO LO (D M - H H H N to m -ZI (N -ZZH M M ma LO a) -ZI -ZZI r- 00 00 LO m m m m Hl r- m 00 tl tl tl (N H 00 00 00 00 00 00 00 (N LO LO LO L H m L H N ( ( ( 00 (D H r- - - Q(D (D (DT (nH - N C0 Ln Ln L- LH C0 m m H HH N N (N N (N N (N (N (N - - (n (n (n (n LO LO L N (N N (N N (N N (N N (N (N N (N m m m m m m m m Mm (DE- (D ~ E- CD~ E-UCD CD~ CD -CD CD C D(CD(D ( D( DU- (D (DCD( ( ( ( UCD U C CD CD C C C C U ( (D -ZZo &n m m SD LO C ON LN NNOL '.0 M H N N N H m ON m O-~O O O LOLOCO OL O ClN'. LO O O O O~ O~ r- 00 D ( ( I R4n m m m W N on to 00 (D000 00 00 t N 0 t 0 to to m (D t m m Z 0- P- C P 0-~C 0L L LO '.40 0000 0 N0Q Ln (n1 a n H H0 H0 H a) a o (N m 00 a - ( 00 r 00 N LO -ZI -ZI LO 00 00 - a) 00 a) a) H 00 (N r- 0a - N a) LO 00 (D r- , r- 00 r- -1 -I -I (N 00 - (D (N -zl (D to to to o1 'zl 00 M (D a) a) N a) (N 00 00 (D a) a) -I a) H L (D (D a) H MY (Y -ZI r- -ZI -ZI 1 -I (N 00 00 -ZI -ZI H1 00 H1 LO N -ZI r- r H Hz Qz (D ( (N 'N .N OLO' ( O H H 00 M a U)U )U ) U) ) U U) U) U) ) C) U) ) UC) Uo) ) U) U) ) C)) C) WO 2006/053955 PCT/F12005/050429 71 LO 1O0 (N (D H 1 (N LO (N LO LO U)O (D H -ZZIH G D H W 0 0 0 0 H 0 Lz m m m D m -i LOL (D LH m N m OZI ( l m to N 00 (D N l H LO m0 L0 LO (D(00ZI l H m LO LO -ZZII co co ( co c LL L0 H tL- mO H co Lr- r- co i H H H m L LO 00O HL 0 LOL L LH 00 M N M ( L LO m CO LO (D r- (N (N M (D M O (N tO M m 00 N -ZI 00 m M M 'zl (D r- -Z M (D (D to o L m LH 00 m m r N m - L N ZZI0 m - - - o D M (D r- 00 M ( O r ( LO C0 M O LO m CO m m ' r ( m m MnOT HM ( 00 m N m Z Z t 00 ( N 0c N O 00 ( 00 HN 00 0 LO M T m H C - - - N N ( m m m 00 00 LO N m m - m LOLO O HN (N N ( ( M (0 00 00 r- - O m 0 m m m (N m m - LO m m m m (D (D (D (D (D (D (D H H 00 00 -ZI T 0 LO LO LO CO CO LO LO LOLO HHH H H H H m m m LO LO LO LO LO m ~ ~ ~ c m ' ZZI-ZI ZI LO(M m M m H LOn LO Z LO LO LO O m c m n H N CD E - E - C C ( E-CDE- C C CDE C - CDCCE- C D E-(- CDU ( ( < u u F< < U< u u < u < u u < < u u u u < < < u u < u < H (N 00 00 00 m 00 (D (N r- 00 00 (D M ' 00 LO -I m tl m LO to - -i m m LO (N m m (N H N i C m - - m - N m ( N m . cH 00 MN N C m N 00 ( LO -m LO LO LO( m LO LO(n M 0 H L ( M M LO H r- r- ( m O LO LO LO LO LO LO LO LO WWW WHHHHWW WWW (WWD 0W W (D WWW WW(D 0 0 00 00 00 0 00 0 00 0 0 00 0 00 F< 0 00 0 0 LO LO C0 O 00 ( 00NO M ( - ( M M LO M a ( N - LO - - (YM N a 00 L a N - LO O ( ( - LO LO LO ( 00 00 00 - 00 00 M O( 00 00 N - H H D a 00 H N 00 H a a a 00 00 00 HN N LOH m 0 Z m - r - N - ( M ( M -(N (D - 00 00 00 - L - 0 LO - - - - - LO - 00 (N OL QaN L O ) 0 a) L DO L - (D r L L (Dr- La (D 00 (N (D (Y (D L N ( ( ( ( WO 2006/053955 PCT/F12005/050429 72 m CDa N L O WM( W MM N(MN W H N ZUU C UU UU U U 0 C aH D m O OZI O O m Nm O N m Oi Qr OD ( 0 00 0 00 N 0 -4 0 0 CD 0 z ~ CI).JF~HU LO O LN O ND N) N1 N ND N N N N (N ND O a) 00 (o 0)o ( LO M O M M T N O O LO LOH O ) a)o LO -o om 000 a) HNNN (Y) a)O NN ZI O 00 -1 & H H Q (D 0 OO H H00 (D -1 0() 00 ooo HHH a)H HM H Y HHH -M HHH (DO - (D OOO O -1 -ZI a)a N na OO O O O O OOO. OOOON( OO~C O. OO OO OOOO O~ OOO-O OO OOO OOOO)H( OOON a O OO O- O LO -OLa) LO (D 'C -~ H- ~-H1 (Y LOLL '.'0 0'0.'. ~ <U< U UUUU U< <U< U< < C<U << UUUCD U 00 OO~ T OHQ oHoQa m o O N om m mom << oUUU WWWW W WWWW (Th NCNWN WW W NN WW WWWW W CC UUU CUD B CC a )CC C 0000 0 0000 000 ( O 00 0 00 0 00 0 -ZZ Z ZZ Z -ZI CICIZ -ZZ Zn (Y (N Z Z (N Cn~- - M' CD C)LC CD O(- (NH a) LO O rO. ~~0- (Y '.0 110-~- 00 -~H~H- r-00NLO 0 n 1 Y)(N( (Y (N (LO (D '.0 L LO ( ( 0 Ln (Y LL (D a) Q' LOLO' L O 1 0 a) 1 1 Y) 00 - 1 N ~ (D 0 -zl Qn (Y r- -1''0 -ZI n a Y 00 Y) ( n r -r 1 L -zl zl D ( 1Czo(N ( D -ZI-ZI-ZI L (D)C~ (NCJ~ Co zoz) l Con)I C)) (Y) ( ) CJ)C)) (N)CoJ (N ( ( n ZZ )( WO 2006/053955 PCT/F12005/050429 73 (()C 00 ) m N me (N O O- n H 00 Hm N Um 2 <EQ U UN m UN H U >HU u Uu f ~ 4>-I u~ x u 1<uu . u PFWOO 0 0 F< 00= 00o 0 HOn 0 0 HUM u FU RU 4 uR 4 Z Z4 < U) -1 N -ZI r N 0 (D (N (D M a) ( O O ( N (D ( r- 00 I C ( N-1 LO - (o m (Ym m (N -ZZ a o r- (N o (D o 0 (D N 0 NND 00 (D ND - LNO a) e oZZI a meno (Ye -om - om0o o (D OON Q H O HH O HH o m LOoo I O (D H NO (N NO L -1 ( -1 a) 0 ( (NLO LO (Y) m o o o m) r- e a) m m -1 - -1s s -1 e oLm - oN - OOO 0 (NHM-T O-MH N N N M-( N OO O - (N (OD OO OO OOOOO -OOOO O O- - OOOO O -O OaOOO OO~ OO- O OOOOa OOOO O~OO O~ OO O O LOO LOOLOO LOO LOO LOOOO LOOOO OOLO LO LOOOO LO LOO L O -o (Y) n o -1 (N -ZZI E-CDC 00C (D a)-~-E-C a)E~-~-C 00DC CD 00 E-n E r- DQ rO-H -(1 00 O Lr-CN D (D (N 'L'c LO (Th N 00 ~-(Y r- N~ a) OL -HL c -1 -1 -O 00()00 00 00 (D (N10 N(N-ZI 1 0000C 0n00 0n M0 0 0D L 1 0 D ( 00M(D r NL (D zz z zz Mz uuuu -ZI 1a -L -0 1 ZZ -ZZI Ln r- ZLZ (Y -1ZZ() 0 1 -( CoN o LN om- H m-m N -0OO' o0 r-HmoL (N ()-ZO oL OZ Zn Lo LOLO -1 00 (ooD - -1 (D (D - 1LOL- o me -1 o1 0 m000n m0 M M ( 0 a) -1- a)mos amm o -omen o no (N(N(N(Y 0 0 Y)(Y - - - - - a)a)a)ama OOO OO 0 0ppp0 FAAAp0p0 0E E 000 O-- HO O O 00 WO 2006/053955 PCT/F12005/050429 74 (D a) N a) (N U1 1 m zzllU UUZ -iI) on -i -i Ln a)oo oon a)O~ rQ- i (D~ -i (D (Dm IN ( a) LO a)0 00 0 LO LO 110 110 N ( (n (N~HCCC (N LOOL- (D (D r. 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'00 I- L-'O 1CO '0 11 (OO ~ H0N (nH 0000 zll00 000r n NL loL 000 00 -1 0 ZNtl ZlZlZ tZZZ r- (D(Dr ( C0D0:Zl zzl0 :Z Z( L'.0L-L L O L L n LOn LO 1 1 110 LO -10 -10 LO (-HN (n a C ( ( ( C a)C C)) C)CJ)CJ)C) C)) C)) C)) C)) -0- 0 u A00000up0 pp0pp0pF WO 2006/053955 PCT/F12005/050429 75 o~oma NC n 0 m< 00 -1 MO 44 U 4 'F A FA>A>IH 0 p u0 '.00P P R CD (D (Dm (D (D (D z &E U (D MD (NN NDr D00LD( N- ND( D00(D( a)C (D LO (N( D D( (N L( (NN NY -r 1a a)0 (N '.0 '0.n0 '.0 r-m (oD (D osoY ( o ~ ( HYN MH O HL ON H M HT n -o m -e e -1o -1e m-1 o mNN mLnO n(C om- eY) r om o - No ma) a)m mom os0 a)m a)0 -C OOOO CTTT LTT MMM 0a00 -1'.0m (N (N L-O a) n On 00 L H - 00 N HHHH HHH HHH HHH H HH CD E-E-CD E-E-E-C (DU-~- C-ZZI (N-~- (D CCD C-)N (Y) UULUnU ~ U M MMT 0QNMT OO T NH T T M 0 a)OOO CN OOOO OOO-ZZI OOO OOO 00 a OaO) -OOO QO'aOOO' OrcONO OL- O O O (D r-)~ (D aD)( ) a n a L a)Ln ) a) a)~ ) a) (D~ -1-1 -10 (N O O O00 MO Hr- H 110 (r 1 ZI nr - ) L Y - a n CD <D DC C DC C D M C (D <D (D (D( D( D D( D( 0 FA m FA 00 p p - - - - 0 00 WO 2006/053955 PCT/F12005/050429 76 m0 00o CNCN 10 0- LZ CD-Z D a) 00 0 -ZI (N r- r N N (N N O i D ( m m O 0 R0 D(0 (N ND 0 OM mO M H O m0 O1OmO NO m aNO) a) O0 m m 00 mN (D oo roo 00 mas mm m(Dno o m m MMI HHH (N NNNNNN m -TTT -I LL 0 O (o oo m (D (D L o ( TN MN H m MHN HH HOH (N H CD 00T N <UU <GGcU U U<<< UU<< << U < UU<<U . o 0 m (n o,'0 M '0OM o LOHOT OM NONLO N M M OCNO CNN O M CNLNHMM LN-oH. 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HH H HHHHH HHH (D (n o HH- 0 m (n (n z CUGU m -<< <I U) U C CCCCC (D 0000 0 ( ((( D 0000 - o r(D 00000 LL O N(D 1011 (D 000 LCO ( NLO , , '00 ( M (D( t.O~ M l D'z 0M t o )aoa a) o C))C)) )C)J mC)J mo 0 O rO-))C))C))C)) MMM mMM m MMM MM (DM (DM -i m ( Mr -r -- D( N0 N( D M (DMM WO 2006/053955 PCT/F12005/050429 77 (N OD M LO (Da (N 1 zzl m 00 C P4 a r- CDa) 00 m zTz (D 00 raU O U NU O H H D-1 m ( A N m ( G N (N (D (D (D 1 m 0 0 0 04 0 - 0 0 0 0 0 0 CD a0 LL N a ( ( ( a 00 ( n()( D ( CD LN r-Ln 1 ( Ln r 00 LO LO LL L L0 L a LOL T O LO (D (Y L n( Da L a) (N ()( - ()( (N N zzl 00 r LL CO 00 (z - 0 (D .zl (N 00 (D0) Dr 00 L N LO LO LO LO LO -1 zl L LO O O 00 r M 00 zl 00 L a) LL N a) (Y l ( ( ( - ( LO D - ( M N L z 00 N ( z N LO - - z Q L L - O - LO N D 0 LO (N '. a CD- L OO (N (Y 00 00 (D H MY r- r 00 (N a) (D r- r- ( zlN (D -l (N 00 00 (N zzl 00 r-LO L r- r- (D (D0 C O ( ( ( - D N C 0 LO LO LO LO LO 0 0 M z z z z z M ( - M M LO ( - 0 r- 00 LO (N LO L LO LO LO LO LO L LO LO LO LOn'0 'LO O LO N 00 00 00 zzll LO C0 & C & h N O QL- L& (CO C CO C C C C CO CO CO CO C a) L a N ~L OLO a) CO LO L N a) C 0 CO0 L LO r '.0 O OO OO O O O OLOLO CO~ O OO LO O~ O. 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C CDEE-E ECD- CD CD CD~ EE-CD CDE- U-CD (D~ (D ( D (CD( D D( D( D ( D (D~ (D (D < U < UUU < < U U< U < < < U 0 < < 0 < 0 < U < U 0 < H a) LO M 00 (D LO MY C0 a) MY -1 zzl - 1 a)Q LO 00 MY a) N (N zzl zzl r 00 a) zzl (D r- , M M H , (D H , (D r- H-zl - - 00 r- LO -1 N (N Y (N (N (N N 00 O OO -o o o n o 1 o zl o 1 o 1 o o- o o) on o1 o) N -o o o1 o oN oN << .0 .C O O C U U CCC H 00000 00 0 L 0 0 0 0 000 0 00 0 0 0 0 zzzz Z 4Z u u ZZZ C) I) R4Z R4Z Z CL- -1 (D0 '.0.0 Ln M C 0 z a a C (N C -H LO(Y a) -H .0 zLO (Y)( '.0 LO LO LO .0 ( D ( LO - -1 () ( (Y)M ( LO LO 00 ( 00 O(Y N (N C z (D H NHH H H H1 C (N - H -H 00 00 a) H H1 H1 H -)N WO 2006/053955 PCT/F12005/050429 78 O0 (N m mm m 0 0 4 0 0 0 0 0u0 U 0 0 (D (1 0( r Da LO r~L- LO ~ -1 (N a) D LO LO Y) 0 O O 0O O.0 N OO< I-'o 'o o- o~ o o o C Lo M o0 '.0 r- '0 -1 (D L0 0 (D (D (D (D - a) o(oD (N -ZI r- - LN (N L o- r- ( -1om o v e m o n Ln o or- r -1 o m o m e m -ZI -Z m o m a m '0 (D (o (D (D - - C0- o-oo 1- o.o ooo - - - --- -- - - -H- --- 1 e QQ (N (Ym r ao r moLoLOLO - Oco m ComNCN U< UUU UUU CD U<UU<UU< CDU<U U <UU< << oss a com ommc c os owom cv s cvo co 10 ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ . 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(DH C(N0 :l m~~L m C m D Ln~~~O m D :ll N ( 00 r- 00 0 :l n D ( (D M( Dr O( 1 rO-O.0 O L LO C L L-H CNn (D -:Zl0 -i m -m zl- m mO 11 1 10 ( D D( ( D( (DDE- (D CD CD E-nC (DE- E-E- (N (N N ( (N(N(N N ( (N N N N N ( (-( ( N -( (N QNN CN - 00 IC 00 00 C C C C C Co Con) C C Con Cn C)) ( ( C D (o o oC))C) WO 2006/053955 PCT/F12005/050429 80 00 (D m o(D - o o0 - o -l (D (D M -iLO LO LO 00 r- 11 r a) - 1 1 LO LO a LOO LO r p4 a) a)o a o 0 9 a) a) a) a)o a)o m a) a) 0 2 m o m zzl a) eeo a o l ooo /OLO m m zzl zzl H m U U U U H Z>< MU N r U R4 u u u u D U U U 0 0 0 O 2 m 4 O 0 <0 0 0 O p 4 0 0 0 u 000 4 4 0 < H H U Z 4 4 W 4 4 4 1-< LO '. LO . LO LO . . LO 'L0 '.0 LO (D LO LO LO L T L0 LO t LO O( 0 o ( e o omL m N C o o oa- O Q Q00 00 t (DLO ' HCCN LO -i -zl M O- (N m N r- m L0 -z LD N Ln tN 00 m O L m m m m o o0 (N LO (D OO to -i tO (D- M -zl LO (N m (N N LO zzl M (D r- t N m m m m L - m l zl zl 0 m LO 00 - zzll LO LO m m i t r zzl LO 00 N (N N (N m i i r- m 0 o 0 o 0 o o- o- r- - 00 0 oo0 LO LO 11 110 N N N m NH-H-HH N-H N N N CN N N LO LO m m m (nH -H L- L- LO LO LO LO LO m 00 11 m a) H 00Q N zzl 0 N to N N H (D (N M C D DC m CN N zzl N N N m zzl zzlH (CN N (D ( (~D DE D D C CD CD CD C CD- CD CD (D D D ( ( (D( ( D D( ( D D( (D~-E~- ~D D (~D C C-HO C C-C N C'0O CNCN ( ( ( ( (- - CNNC - CCN C -H CN CN (D CNO' O u 1< u u F< u u F< u F< F< < F< < u u F< u F< u < F< < u u u < u< m mWW0 mo 0 L S m r- N O (DM zz L zz LO o r 0 m M zlOl( zl O - L N O O LO m CO a o o a Cno~ o 0 OQ~ LOM O' T m- zl l(( -H(o LO e o omLO 00 LOLOOa LOL0a L o O o 0 m l Po o o 0 0 0 0 0 0 0 0 0 o ro 0 0 0 0 F< < F< 0 0 R4 R4 0 u 0 u 0 OD M (D (D -i (N (N m -i m m -i 2 % UUU M2N (N LO 00 -1 M -l - N (D M ', (N (n 11 11 r- 00 r- (n (n r- 00 11 -LO 0 m tl w -i N -i m L (N (N - 00 zzl zzl zzl LO L r- (D r- (N 00 M (D M M (N m N m m - m o0 m zzl LO 0 00 m m 0 00 0 m 00 0 00 0 11 00 m LO 00 M M (D - zzl m (N (D L (D L O (D (D (D L zzl (D (D (D r- (D (D (D O (D L (D (D r- M i zzl N zzl i -i m LO m (N 0 LO L m 00 LO LO LO (D LO L 00 LO (D L (D zzl LO LO to zzl N o - - o o N zzl ol o o O m - m a) zzl Oi zzl O O O L O L WO 2006/053955 PCT/F12005/050429 81 (D LO ) 00 R4 m m R4~ u Z Z u 0 0 H 11 0 mC C D C (D 0 a) LO (D -i ( '.0 L0 '.0 ( LO LO 1.0 1.0 00 0 (D ~ ~ NL 1.0 (n0 (D~HO N'. LO CN n CN lH tl mw w ww w m zzl (D (D w 0 UU UU 00 0 00 0 1ZZZZ 1-0 00 H O 00 0 ~ ZZZ ZZZZ Z ~ ZZ Z (D (D (D (N tO r- O '.0NtotoL - n m L n m - m zl l D Ln( zl L l Ozll'zl OOzl ( D ( m zl 10m zl 0 'O'O (N ~ ~ C L '.0l 00H 0C '.0O MLOMM L L D tozll DM ( r- ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ C 00n 00 a)a) r-~ 00MMMM(CD(oD nL nL nL - (n( n( n ( D ( WO 2006/053955 PCT/F12005/050429 82 C(D 1-im x x zl( D (D CD4 ( 00 CDD0 0 CD 0 4 U) U L 4U 4 (D a) D (N L 00 a LO LO ) Y 00 ~ ~ I a)r-(D 00LO LO Y Y 00 ~~ ~ c LO D- D( r- ( a) ~ ~ ~ 'c a) ()-0(Y Y 00 (Y)LO 00 00a) 0 zzll (Dc (D 00 a) r- ()a 00~LO zzl( - D( -1 N () 0 a) -C~ -1O '.DO-H~-- a)c a) -1 NCNNL a) NNNC UCCCD-C E- 0 1 1 a DC CD CD CD- CDEE- CD CDE- (n-CD (n zlEzl-zl 0 0E- Y ()( )a 00- 1 - 1- UL nL nL nL Ln (D (D~C U (U D U (DU (D - U(UD D( D( -o oo oo 0N 000000 ()(r r)() n00 0 000 00 00 00 00a)) (Y (D zz (D (D 1 Y) zzzlz N 1( ( D- zzll( D ( zz (Y zzl zN z D Y zz( (D ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ C (D( DC DC D CDC D( D( D(D ( D( D( D (D( D( D( D( (D D D ( ( ( (D (D (D D D (OL OD C (DI (DN (D( D( D D ( D ( ( D( D( -1 -1 -1 o Lon a) zzll L 00 00 on a) a) -1o o ~~a -1a zzl o - 1 (N -1zzl a) -1 a)- WO 2006/053955 PCT/F12005/050429 83 LO R-1 R-1 00H N m m LOi) Z ( m ( EN LO U Cl) W (D -i u Z0 -1Z Z ~ fl) U 44 u~ 0 0 o 0 00 0 R4 2oM U U4 r . p p (D 0 00 OH 0 OOLOO (D a) a) (D D a D N (D N (D N (D (D N 00 (D H '.0 LO a)~~I r- zl - L 00 r N (D 00 CO LOH L M LO CO-1 r 00 -1 r- zl (YN zzl (D MY N r- 00 (Y () a) (N zllr 00 N-- 1 00 00 () ( LO LO '. LO r- (N zzll 00 - r .zl r- O ( 00 00 .0 00( - r- zzl a 1r LO L 00 00 rO LO LO LO LO'. CO LO ' LOn 00 ( ( 00 z a T zl S z z 00 N (D ( 00 L (D C0 Q (D LO 00 (Y) a) 00 00 00 (D (D LO r- zzl zl Y T M (D M 'zl a) r- - LO zzl 1 r M Q LO LO MY 00 -1 1 r 00 00 00 00 00 00 (D -1 r- LO 1 CO CO 00 NN (N (N (Y zzl (D (D (D (Y M Y M Y M LO r- (N r- r )a )a a) a) ( (Y (N N M 00 00 HH N (N (N (N (N (Y 1 1 o o o o (D (D N (N zzl - (D 1 00 00 00 00 00 00 00 00 (D (D (D (D (D (D -H -H -H H 00 00 -H -H H LO LO LO LO LO LO LO . 0 r r r r & a) -HC (Nc LOa c-1 a)0 a) N (Y) N N N Y C (N D O H ) - H (DO D -1 E-E E- E- U E-D C C C U E- ( E- U D UU E- U D ( U U <U U U( ( ( (U<U < ( < < < < (D LO C O C LO CN O & 0 O0 Q'O T TH - LO O O HCN -H H N FA HA 0 HA HA H H 0 0 0 u p 0 p u 0 u L p 0 Ln00 00 00M 0 0 00 L OL (Y) (Y)z S0 00 0 0 00 00 0000 0 R R R R R L4 00 (LO '. zl O O- LO .zl 00- ( N (D a a '(Y DM D M 1z L O' 0 LO ( 0 zz LO LO zLO LO z a) 0 zzl z LO O LO - LO 0 O CO CO C0 S (D (D (D (D (D a) (D (N H1 - r- 00 00 (D (Y (D H (D r- Tl (Y (Y (Y WO 2006/053955 PCT/F12005/050429 84 om o om (DrQ -M i( Um C | | M UUM~bCQ<C OM O OHC OOO O (N W HD Hi -N -(Z D(G N m QH m u f I I H o uM H u D r. 0 0 0 0o 0mem0 0 4 u H4 LO '.0 0) 0 4 4 H - oo ( 00 L LO LO HHH H H- H- H HHH a" a) (D (D M H Ln -1 (N a m an ( o o 1 o0 00 -ZZ0 0 (N (D (D a)0 0 L o (D o- (D (Y)0 r- 1 o 000 (N r o c (D -1 a) (D -000 a) o0 00 r- (D c - H H r- 00 (D ( DN a) LO H1 LO 0 00 ( 'N 0 o m a Ln (D LO '.0 H LO ILn n( Y H00 '.0 0 (D - 0- L 00 o o o o H HHHH H H ooooo oo a) LO LO O ' C | | | | | | | C D C -1 (D 00 CNC C -C H C0 CC00 H LO HO~ ~ 'ON~O H OO~C 00 0 O O O -ZI H1 LO 0 '00 0 0H'.-1 0 () -ZZI (N '.oD'zl ( D D a LO L LO LO r- a ( D 00 MMM D( (D (D -1 H 0 00 ir- r- -- '. 0 a) a) a) a) (D- (D- r- HH H- r-C r-NNCC r-r 00 D( D (D( 1- 1 (~C NN (NC (NNNCC (NC (N (N(NThI H1 (N N (NH -1 ( (D -H N (N (Y)( -1 (~H N -ZI - ''( HD (N (D(E( D(D ( D D( CD CDE- ECD (D (DU(D (D (DE- (D~ (DE- (-E~ ~ D (D~ (D (D(D ( (D (D (D (D (D (D( D(D( D(D ( D(D D(D (D ~(D (D (DD(DD H0 ~0 p p 0 0~--'C 0 ~~ H HH 0OHH HA FA F F pp 0 r- 1 101 a) -1( ZI 1( 0 0 0 0 on on ono n 0 0 0 00 0 00 0 0 00 0 LO H1 C 00 C ( 00 (Y) 0 Hzzl H D - -I - LO - M -HCN H H-1 - LO'.00Q a) 00CN HH- -- Z Z L n -1O '0 -ZI LO Y Y)-1 r-(D N (N a) 0 0'0 H -ZI c() Y ir- - (Y) '0 '0 LO- 00 -ZI (0 Y (Y)h1 00 a) - - a) 'zz .0zl ( r- Lr- -ZZC N ((D00 (CN(N (N ( ( (Y) a) ' LO'.0'.00(D0( LO 00 ZI r- 00 a)-ZII LO LO 00r- r- '0 LO Lr- - (Y HH - LOH-H-Oa' a - - - H0 H (N '.0 (Y 00 0 ()( -( (N (Y LO () (Th LO ( ( -ZI ZI Y ()()0 HH HH (N (N (N -Z - (N N '.0 a) ) 1 a a HO O1'0 '0 a) a) a)O U) U) ) U)C) CJ) )) Uoo o) ) ) C) U) C) U))C)) Uo) ) ) ))) ) ) U ) ) WO 2006/053955 PCT/F12005/050429 85 m M LOm H P CDM - HR R4NH pi -ZI m i m (N R4 - i LD O4 N Ia R4 N WN W2 UN~ C) C) L) C)Z 0 (N R PC 4 ) u P -z, c c U zl c p 0 0 0E- P R4 R4r >IR4R 44 U) Z Z H uU)Ih ZD u u~ - OO O E- H0 L- H LO r- -i lo (D 00 00(D r N 00 (O m ( 00(D( (nL Ln 1 L m ( N HN 00 L H T LLO LO 00H H H) H H m L L LOL LO LO L 00 LO LH O 00 m( '.0 '.0H H H H ~LO N mC 0 LO n ' H O LO -i ( Q M- 00 N TOL H H H L O LOCO L -ZI 00 N m NL NO HC LO'.0M -H - ' H00fC M LO' .NLOT LO HO C( LO' N O TL m0 ' G Q LOLO - LO'& HCN 00 C0L (TD h m 0 00 L O' O'H L -h- - - Q O H - H O H NHM T 00 0 N'.0.0 0o 0 m HHH H H 00 0 m L LOLO N -M MMMM - (D (D CD CD O O OOO N N- N NN (N - zzl (D M 00 r- (D M (D (D MMMM L LO LO 00 N ) ( C M N N G 00 00 0 0 MLO O LO NN N H NL HrO O ( H( O N H LO M r 0 E- r ED ( D E- E- U E- E- (DU E- D U E- N N N E-1E-N (NG E-N E-1DD-1 0 E- 0 E10 U U <<U U < G ~CU G UU U U TH LO N LO m ( ( LO( (Nn ( NH ( (n (nOH H M CO- O LO LO O L 00 H r-N ( N -H ( HH H H 0 NNH LO ( 00 0 M0 0 M m ( H0 N HHH N - HHH H NDN -ZLI' HmHN mN N H ( (N (00 H H C C C C C C C C (D (D ( ( <D <D <D <D <D < C C C U U U C 0 LrL- M (D (D r- MZI L(D I- LO (N M -i O M 'zl tO n (D (N m - M- m r- tO M i -i -1OCO 00 m LnL m -i -I 00o LO N (N 00 O N i m -i -i M MN -i -Z 00 (N -i L M - M 0 -1 M r- (DQ M L toON OLOO O N O C 00 N (N (N N r- r- 00 00 00 N Lm 11 CO(n 00 CL - LO L L LO R4R 4 )( WWWW WW W WD Z Z C) U) U) U) U) ) W) W) HHH < < 00 00 0 00 00 0 0 0 0 R4EO- R H R0 H R 00 0 H O 000 LOO N - N H Z ( - H H H H H H Z H H H N ND H ) 0 H H H 0 N H N ( Z 1.0 CN (DL D'zl( (N M (D M M L- 00 00 - (N (Y .0H-1 - 0 00 CN L O.0 w L O ZZI QCC0CN H(Y) H) Q'Q 00 (D~ 00H 1 -ZI- -ZQ'ZI ) -ZQ ZI - Y -00'D( QzL-ZO ZI n n-ZI - (D() 0 ( ( Y)( D 1(D-zN D( CDO~O~ HHHH a)- HHH C NH- H HH( D(D - l( 0 L n -zHCN zl WO 2006/053955 PCT/F12005/050429 86 to o0 oD o eN eN e m 0 -1 m < 4-1 M (D C a) CD - 00 oD DZ 0 omo a <p O ':ZlZ R4 Z< z4 M< < m~ ~ i -i (D m ON ( N U)4 :l H 1< < ZZI m O m -ZI -ZZIa m - (D O O N O 0 -l R4 < Q O 0 >-A H H O O < O O O O 2 H H0 LOO OO c~ cc 00 ND LD ( ZI D( D( (1) (D (Di~ (~Da)( -D(Y In o In In c n 00 tD ( a) ( t 0 (D - (D ( In) -I (nn I0 n '.0 ~-1 (DaIan I0 to (N 00 to ( In o (N r- 00 a) a) -1 e 00 -ZZI (D '.0 (NH (Dn n ~InOt co L) a) LO 000 (~DE- CD (D~ E-n CD0- (N a)C (D c0 n In () a)n n OY (D (Y 00 -ZI 1( 00 a) a) a) eN 00 Cn N ( - N ( r- (N n ZZI ( ( HY) D a) - H N (D 00 (D L (D (D (D H LO (D LO 00 00 (N a)r M0( Y LO 0000oo n - n - 0 n L 00 r- n 00 a) aO)N a) Noo Inn n Ln nn n On 'n -ZZI Ln In (N a ()a)- )a In-1 '.0-1 (N E- E-1 N E-( -ZU U LE-n (D~c (D ) (D ( ( DD(D CD) CDC) CD) CDC)C C D) CD) u F< u F< F< 0 F< u << F< < u u < (Y (Y (Y (Y -ZI to N a) (D - M t -1 a) an ( t 0 00 -Zo co t (D (D (D M 00 Q(D (D( N Ln Ln a 00 00 00 (D -ZI r 00 00 00 (Doo r- n Ln - (D Ln cN Ln 00 a - (D to (D (N -1 00 eN( 00 M (D o t - -o 00 r- a) n Ln Ln ()00 a) o1 00 (D -z -O -O -O N N 1 N Ln r- O H 00 ON LnL c0 00 00 a) - a) - -ZI -1 -1 (Y (Y - - WO 2006/053955 PCT/F12005/050429 87 o~c o o a ' H NM HHM NO 0< W > < N HC<HUQ rGI)U UUCIU U U C 0 0 UE- HHE-UH 11 110 m N0 O HO O N OD F4 0. 0 .04 110 (D H 1 (D (0 O O. O O ~ N ON M0 HD - LH (DMD-(O r- (D0 110 0:(D)o N (D MO (D 0 HH 10 mom~~ zzl m m men a o n m e e zzll CD CD m o L o o o-i o e o m oO Lm (o m0 m m H 00 ON OOO Nm mOHNN m'. (D ( ( - m 1 100 m m~. 1'10C CN(- (n0.0 ~ O -1 O 110 1n NN- (nQ' Q( 000 N 00 m o z0 00 m moon ' 'a. 00 s -o os m o m om (oN Lso -i en s 10 .0 : ~ 1'. UG0'. C0.010 L0 -H09 C90 9 H UUU < < < <UU< <UU< <UU<UU<U (D O- NM -i o N00 ON O O OM O HON (N H mNQ CNQO mN0' m NHO'N ma (D (D -i to mo < man Ln ma UUU U (N m lO 00Oo 00z>-1 0rM(0 ( a)zzl N zl 00 0 M ~ (D tO L l 0L -- 1 0 0 00 (N 000 ( N - m H(Dn 0 zN N(Dr-MM L~-'.0H H Lm-H m r- ( n (D NM O (n~c (n) mJCJ mJC)J )C)CJ )C()()0CD(D(J()N m ( CD (N N (N(N ( WO 2006/053955 PCT/F12005/050429 88 o0 LO 0 H ( LO (N (D -I a) i m -i - (N -i o0 (D N L (D (D o0 q-|( m 0 1 3 ( Mlz'm Qp-4 R H H H H H H p- u U F- - 40 P P P .4 0 H 0 00 - O R 0 N N N N N O SLO LO . N1 CD ND ( () 00 0 0 LO LD 00 r- L L -00 (D r 00 Ln L L LO L L L 00 00 L L (N -. 0 (N () N(Da LO 00 00 L0(a LOL a)0 -1L Oa - 1r L () LO L' LO n (D a) -1 00 a)0OTQ L ONTHT& Q MC LO T0 '.0LO T O O -HLO - '.0N~ 00 HL-HC MN 0 TLO O 000N 00 (O CN ( D LON M LOCNCN LDLO . L O N (Y LO NO C 00 00 LOLOLO L OO 000'0 O L- N 0QQ 00 (QQ) (D ( N NQ & 0 '. ooo LO LO) LO L 0 LO (Y) (Y) CN- (D - 00 N0 M (D 'zz . LL LO L LO LOCNLOO LOL - NO LO 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 a) (N -I a) LO m r- a) -1 r 00 -1 N (Y (Y (Y (N tl tl (r C ) S (o ( ( -o LO o oN - - O o N ' ( ( ( o o Z o 000 0000I 0) 0000 00 oo 000 0 (D M ~ Z (D (DZZ (D( DZ 1- 1( N ZZ w zm mzzzz- ZZZZI -1mZD( Y - O O O O O OC O O O O O O O OO OO O ~L~CL~(L O 0O O O O I - L OCN -Q -- COLO- O0 00 00 L(DO&M NML 0O 0OO&QO TO O C ~-HoTh LO -LOLO o LO'.0OH O - - NC O -H LN LO~ LO CNM-H LO 00 00 00 a0 H H LO LOCNC (Y) ( ' 0 P4 4h 0 0 0 0 0 OO O O O OO O O O 0 000 O O O O O O N N OOO O O O 00 O O (D O r- LO to LO LO (Y - r a) r L - (D (D (D r- (N , (D (N (D 00 (D , 00 LO (N (Y r- (N -I (N (N -I (N N (N 00 (N r LO LO (Y LO - (N LO r- (Y LO -ZI 1 LO a) C0 a) -I - (D (Y (D (D O (D( LO (N -zl -1 00 00 (Y 00 (N LO 00 (N (D -1 M -1 - LO r -1 (Y r LO r- LO LO (N LO LO M (D a) r - (D (N (N M -(N LO (D - (N 00 -1 LO LO (N (N (D (N (D (D (D (D 00 (D (Y (D M M LO LO (Y (Y L (D r- LO - a) 00 -ZI LO LO 1 -1 1 -1 - -I -1 -1 - C0 a) 00 -1 -1 - a) - - a) -1 - -1 - LO a) -ZI - WO 2006/053955 PCT/F12005/050429 89 m m I 0 zl - a r- m 0 LO LO -i - - 4 m 0 0ZZ0 0 0 Z 2ZZ4 2 4- p Z c'-0 p H 0 0 u NE-p (D D D - (D ( (N~c (Nm N(N ( 00c -1 ( LOn I-i -ZI'r-10 LO ZI 00 a) - 0 M - 00 - '(D ( QLOC(N Lr- (D M 110 LO LO 110 ( L -o (0 N (D r- 0 110 10 (D( (D (D D (~D (D (D D D (D (D ( ( ( CD E~ (D (D (D (DU (D (D( D( D (D( D ( D( ( ( C C C (DC C -C N C C 0~~~~~CCC 0 A00F u u F< < uF< uu F<F< < u u 0N0 00 (DmC CD (D (D (D00 00 0 110 110~C m 110 L-H C 1C 0 '.0 LL m Co~( (Dn F< ooooo)C) )CC) C) C)) -ZZII -ZZIIa) U WO 2006/053955 PCT/F12005/050429 90 LO 00 (D 0 0 0r M CN(D N~ 00 WD 0 c) Lou N rcn WD I 5m 44 H OpIx 0 . ri U) R4I 0 0 0 WD (d I t H H 0D W 0 HIH0f*H *P H UNI 0 C) C)) (d U) rlL) I 0 0 0 Sm r ~ 0Cm LCN I ) - P D Cn I ( OD (nIo-Q H Q -P( 00 'H (D ( U H H 0) (D(dQ I Wt 0 0 0 0 00 0 ,Cl4 P rm Il (-H l H 0 -P l 1-O ~~~~ 0 r.-Pr.

I R40 (dQ dQ U) 0 WW(Du I -H 4-3 4- H H r. Z r- (N LO m WHW0 H >1H H 0 r m n I r) rm! Ha rm!m Jw P Izl 0 WQ (WQ (d 0 H m oa )I H (Dr- HWH Q U)U)t r I rm U)o U d 0 4( o~~~~ I UH 5r5 Hm Q- 4 Za ) I4 0 0 0 tP (d ) (d I .H,~ tH WD W CNNNC H U) 4 W -C, ) I >1L l- d - d ( 1 ( P M 0 1 4-, M H H H) a)O-Z 0) M DM I (D W2 iCI) 4-1 (D (D CD CDr.H H 14 H (D (D (D (D 0 0 0 Q~ 0 P4 )H1 CN NC ( . H WZ U) U~) r.WH (D( DI 4-P tC),C),O -P0 40 4 U) ( 0 I 4J (d W -l W I-) Wj H Co I W 0 -H r 0 4-4 r- J U) r J W) 0~ >i H I W P 0 -H( dQ ( O H t 0 M~ I m r. 0-3t- -C aH Q W u N Nm I H W0 WH ( H 2 >I ( >1 0 00000 14--H U ) oo t(dp HI - H W 4R I -P - d (D ( L L L OI H rm W W -P 0 P 0 L ) Q I~ WD0 d-P( ( H H4- I l CNNN-Il) 0 1 H H Q)) I d c 0H H 10 W 00~~ (D M U WO 2006/053955 PCT/F12005/050429 91 I LO LO WD L( LO LO LC (N I I I I I (D( H I( 0 1 - O - - -O 0 I I M -* H * 4I V v v v ( - ( t I W I ) I WI (0 O I a) |r LC CN LC DH Im HH -H , | N H N N (NHH U | ul o 1 O) I -H I O M4 1 U u u < F< u F< F< < F< < u F< u < F< < u u a) (d I W0I a) WI S H I (I S o I a) ( 1 O | U UI HI -C -H I -PII W I (D 00 0 LC) II~~~~~~~ IZI ro-r (NO( ~ r ~ n o~~- ~ - ) ~ f m o-ic '.r- LC LO() M m -im i -00 r- m r- (N (N m (N 00 -N WI a) )I Q I 0 H I H 01 01 WI 000 0 00 0 00 0 00 0 0 0 R 0 0 0 0 0Q 00R R + HI 00 m r- L- m || a I -OZL (N m O1 OH m m r- Q - 0 ( m o aL L 0 r-N W0 I Q O M L O T H H - (NCLC -LO L- O LO LOLOM M LOLO T LO LOM HH I (D O LO L M M ( O m - - m 0 N 0 ( ( N ( N M ( M N Cl) I -z,D -- i ~-i(N -i ~-i -i -i Lr- -im (N m -i -i(N -l ~-(N m-~H- m-~- O U I 4 4 I 4 4 4 - - - - - - -4 4 4 4 4 4 4 4 WO 2006/053955 PCT/F12005/050429 92 LO o rHLO LO LO (DI I I I O m o om m mI 4- C r- - - HH I C r-H0 0V V V I I t C I ci I 0Oa I O~ 0 I O~ I (D L O) rH N0 (l 04 N- I H2 N NN N IU I C C H Q) q0I IrH >1 0 4 I D R p0CD0-p 0 0 0EFA FA-pEp H0p 0HE~- p p -pCpE- pCD I w I- I- I- - - I I-C I- I I-C I (d 42 ( 100 42 rR 00 I H -| I C R H IZ -1O I -H 0 R4 I 0 -0H (d U0<0EH < < 00 &H U H&<0 <H00O <HOO G< I HQA2 OUH 4 a I U 0 C0N H N 0N N CN CN -N - Z Z 0 0 0 oH0 H0H 0 H I C -H x 4 C2 H lH-H HHr-H rHrHrHrHH rHrHH HrHH H N N N N I H a) 01 0N 0 >1 LO OuR 42 0 Q I M ,Cr H R1-H t'o I C r.aO r C I mr C Ord H I p C-H-H Q) o) t I P C P (d U) U) f I Q d -, D (d - - H 0N ICC a M A U) Hr O H 0 (D 0 4 N ( ( r C r- -1 W(W O( rH N N I 0 -H D -H 1 R H C NNN0 4 O M ( r m N NN fN mi -H r 4-1 ) 0 0 0 oDCD o NN W H L) 0 Z LOr D D -1 C DL I 2 t O -H H a p $4 P I r.WC4 (D H P ) I (CDC | H I (d 1 HO00 420>0 4 I4-1 H t rdH 0 M I - r.C -H Q) H 0 IP -H Cd HW 4 0 r.42 r- (D 0SCH(d r. rHrHrHrH I C a SoOr-H 'dC S0HHrHH I 0 O P O 4 m P01 Z ZZZIr-H WSEH r a o H H H H HH I GH p 0 ( -H I G id 04 04 04 W o o 0 0 0-0 0-R4 1 (- (- R R R R ( I R-1 r. M 00 1 H -H ) > =ZZC C=C CC CCC CC r.ZZ CCCCC I Z u ( ,CH (dC (d I R4 R4 R4 0 00 00 0 0 00 W 00 W W W W 0 0 0 0 0 1 ') 0 I) U <0> U p4 IHa H H 0 WLO m LL m m 00 0 N W 0 rLOf M LO -- i I I . 24 o o0 o N 4 N M (N (D 0 ( N mH O N0 L 0 )CD -m I a 0 01 H a) (d O OW D o m LON N LON LO LO r N LO LOW WN I $M 0 0 H 2 P (0 WWWN 0 ( LO M M (O LO 0 N -H N LO LO LO LO LO H C (d C 1 CDo0 D 0 0 r- ( f LO04 OrH04 m4-04' N 0N 4 0 40 m IN 0N IN N - H mfffO mN m mrN -H H0 (D lLO m O LOfl M CDCDN I Z m 0 00 0 -H I (d HH H N N HN 0N 4N . N a)04040 H H N 4 HrH N N N N H I U) C ) 1- a) -H > a ) ) a) a)a) a)a aa a a a a a a a a a aaa) I ,0 1 UC -H) (,C I M Mm M - M M M M M M 4 4 4 4 M4 I O U > u P4 WO 2006/053955 PCT/F12005/050429 93 Table 6. Genes associated with HT according to point wise or haplotype analyses (n = 722). Chromosome I Gene 1 DAB1 1 APG4C 1 CAPON 1 COL24A1 1 COL24A1 1 DPT 1 EAT2 1 FAAH 1 FAFl 1 FLJ20277 1 FLJ32825 1 GADD45A 1 GNG12 1 LOC116123 1 LOC128153 1 LOC200008 1 LOC284680 1 LOC388656 1 LOC391046 1 LOC391046 1 LOC399701 1 LOC400758 1 LOC440598 1 MAST2 1 MGC22960 1 MGC29875 1 MGC8902 1 MRPL37 1 MUF1 1 POGK 1 PRKCL2 1 RAD54L 1 RGS18 1 RGS7 1 SEC22L1 1 SHREWD 1 SIAT7C 1 SIPAlL2 1 SSBP3 1 SSBP3 1 TARBPi 1 TSPAN-1 1 UQCRH 1 USH2A 1 USH2A 1 XM 372814 2 COMMD1 2 CRYGEPl 2 ERBB4 2 LOC130576 2 MYO3B 2 REGL 2 SP100 2 ACVR1 2 ACVR1 WO 2006/053955 PCT/F12005/050429 94 2 AOX2 2 ASB1 2 B3GNT1 2 BARD1 2 BAZ2B 2 CRYGA 2 CRYGB 2 CRYGC 2 CRYGD 2 CTNNA2 2 FLJ13110 2 FSHR 2 GORASP2 2 GPD2 2 HMGlL3 2 IDH1 2 LANCL1 2 LHCGR 2 LOC130429 2 LOC339789 2 LOC344471 2 LOC388954 2 LOC389072 2 LOC389073 2 LOC391481 2 LOC402117 2 LOC442053 2 LOC442066 2 LOC93349 2 LPIN1 2 LRPlB 2 LTBP1 2 MGC4268 2 PAP 2 PDE11A 2 PDE11A 2 PELIl 2 POLR2D 2 PRKCE 2 PRKCN 2 QPCT 2 REG-III 2 REGlA 2 REGMB 2 TLK1 2 TRAF3IP1 2 UNQ429 2 VPS54 2 WDR33 3 LOC152225 3 AHSG 3 ATR 3 CRBN 3 CRYGS 3 DGKG 3 DNAJBll 3 FETUB 3 FLJ10560 3 FLJ12604 3 FLJ42117 3 GRM7 WO 2006/053955 PCT/F12005/050429 95 3 HRG 3 IL5RA 3 KCNAB1 3 KNG1 3 L0C152225 3 L0C389105 3 L0C389105 3 L0C442078 3 L0C442099 3 MAIL 3 MGC15606 3 MGC16471 3 MRPL47 3 PLS1 3 SCHIPi 3 TRAD 3 TRNT1 3 TRPC1 3 XRN1 4 L0C339979 4 PDGFC 4 ANK2 4 ANK2 4 FLJ20647 4 FLJ37673 4 GABRBl 4 GABRBl 4 GLRB 4 KIAA0 882 4 KIT 4 L0C152594 4 L0C255130 4 L0C339979 4 L0C345378 4 L0C389217 4 L0C391681 4 L0C391686 4 L0C441018 4 PDGFC 4 SEC24B 4 SNCA 4 STK32B 4 TDO2 4 TLL1 5 ABLIM3 5 ADAJMTS12 5 BTNL3 5 BTNL8 5 BTNL9 5 CANX 5 CCNG1 5 CDH6 5 CEI 5 CHD1 5 CMYA5 5 CMYA5 5 CTNND2 5 EBF 5 FBXL17 5 FLJ10904 5 GABRAl WO 2006/053955 PCT/F12005/050429 96 5 GABRAl 5 GABRA6 5 HMVR 5 HNRPH1 5 IL12B 5 IRX2 5 KIAA1061 5 KIAA1985 5 L0C134492 5 L0C255187 5 L0C255187 5 L0C285638 5 L0C285679 5 L0C345471 5 L0C389285 5 L0C389352 5 L0C391734 5 L0C391774 5 L0C391819 5 L0C391858 5 L0C402222 5 L0C441113 5 L0C441120 5 L0C442129 5 L0C51149 5 LTC4S 5 MML1 5 MAN2A1 5 MGAT4B 5 NDFIP1 5 PAPD4 5 PJA2 5 RGMB 5 RUFYl 5 SGCD 5 SGCD 5 SQSTM1 6 STX11 6 ACAT2 6 BA13 6 EPAGi 6 BRP44L 6 C6orfl42 6 C6orfl49 6 C6orfl57 6 C6orf2O4 6 C6orf33 6 C6orf65 6 C6orf83 6 C6orf93 6 CRISP2 6 DST 6 FARSi 6 GFOD1 6 IGF2R 6 IL17F 6 L3MBTL3 6 L0C285741 6 L0C389370 6 L0C401255 6 L0C401284 WO 2006/053955 PCT/F12005/050429 97 6 MAS1 6 MCM3 6 MGC35308 6 MRPL18 6 MRPL42P3 6 MYCT1 6 PARK2 6 PARK2 6 PHACTR1 6 PLAGL1 6 PNLDC1 6 RPS6KA2 6 SAJMD3 6 SEC5L1 6 SERPINBi 6 SERPINB9 6 SF3B5 6 SLC35F1 6 SOD2 6 SYNEl 6 T 6 TBC1D7 6 TBC1D7 6 TCBAl 6 TCPi 6 TINAG 6 UTRN 6 VIP 6 WRNIP1 6 WTAP 6 UTRN 7 DNAH11 7 ABCA13 7 AKR1B1 7 ARL4 7 ARL4A 7 BMPER 7 CNTNAP2 7 CNTNAP2 7 DGKI 7 DNAH11 7 FLJ21075 7 FLJ32786 7 HDAC9 7 HIC 7 HUSi 7 L0C136288 7 L0C340228 7 L0C340274 7 L0C389602 7 L0C401406 7 L0C402463 7 L0C402640 7 MGC16075 7 MGC33329 7 OSBPL3 7 PEEF1 7 PHF14 7 PHF14 7 PKD1L1 7 PKD1L1 WO 2006/053955 PCT/F12005/050429 98 7 SCIN 7 SDK1 7 SEMA3A 7 SEMA3E 7 SHH 7 SLC35B4 7 SP4 7 UNQ739 8 SGCZ 8 ZFPM2 8 ASAH1 8 CGI-62 8 CSMD1 8 EFA6R 8 ENTPD4 8 FGF20 8 FLJ23749 8 FLJ36980 8 HNF4G 8 L0C286097 8 L0C340357 8 L0C346887 8 L0C392262 8 LRP12 8 MATN2 8 MSCP 8 PKIA 8 PSD3 8 RBPMS 8 RIMS2 8 RRM~2B 8 ZHX2 9 NT 008470 9 ORlJ4 9 ADAJMTSL1 9 AK3L1 9 C9orflO2 9 C9orf97 9 ECM2 9 FBXW2 9 FLJ31810 9 FLJ31810 9 FRJMD3 9 GRIN3A 9 JAK2 9 L0C138882 9 L0C392282 9 L0C392371 9 L0C401492 9 L0C441384 9 MRRF 9 NR4A3 9 NTRK2 9 ORlBl 9 ORlHlP 9 ORM~ 9 ORlJ2 9 ORlJ4 9 OR1L8 9 ORM~ 9 ORlN2 WO 2006/053955 PCT/F12005/050429 99 9 OR1Q1 9 PHF19 9 PSMD5 9 PTGS1 9 RCL1 9 RCL1 9 STX17 9 TMC1 9 TRAFl 9 TRPM3 9 TXNDC4 9 XPA 10 CTNNA3 10 LOC389938 10 PCDH15 10 ACF 10 ADRA2A 10 ADRA2A 10 ANK3 10 ANK3 10 ANKRD16 10 ASCCl 10 C10orf104 10 C10orf35 10 ClOorf49 10 ClOorf59 10 Cl0orf64 10 CACNB2 10 CACNB2 10 CBARAl 10 CTNNA3 10 CXXC6 10 DDIT4 10 DNA2L 10 DNAJB12 10 EPCl 10 FBXO18 10 FBXO18 10 FLJ31958 10 GDI2 10 HNRPH3 10 IL15RA 10 IL2RA 10 LOC387630 10 LOC389970 10 LOC399707 10 LOC399708 10 LOC399713 10 LOC399773 10 LOC439946 10 LOC439953 10 LOC439978 10 LOC441550 10 LOC441551 10 LOC441561 10 MAWBP 10 MCM1o 10 MINPP1 10 MPP7 10 MYO3A 10 NEUROG3 WO 2006/053955 PCT/F12005/050429 100 10 NOPD1 10 NY-BR-1 10 OPTN 10 PARD3 10 RUFY2 11 NUMAl 11 CD44 11 DKFZP564MO82 11 FAT3 11 FAT3 11 FLJ20625 11 FN5 11 GALNTL4 11 GAS2 11 IL18BP 11 KIAA1731 11 LOC113174 11 LOC220074 11 LOC260340 11 LOC260340 11 LOC338661 11 LOC338661 11 LOC387795 11 LOC390241 11 LOC390263 11 LOC440062 11 MGC5306 11 MRGX3 11 MRGX4 11 NELL1 11 NUMAl 11 OR10Si 11 OR4D5 11 OR51R1P 11 OR52K3P 11 OR6M2P 11 OR6M3P 11 OR6T1 11 OR8D4 11 PDGFD 11 PDHX 11 PTDO12 11 RNF121 11 RRM1 11 SAA2 11 SAA3P 11 SAA4 11 SSAl 11 STIM1 11 USP47 12 EB-1 12 TPH2 12 ALG10 12 ARL1 12 DRIM 12 DRIM 12 EB-1 12 ELKS 12 FLJ36004 12 HSN2 12 LOC283392 WO 2006/053955 PCT/F12005/050429 101 12 LOC283432 12 LOC390359 12 LOC400019 12 LOC400046 12 NAV3 12 NAV3 12 PAH 12 PRKWNK1 12 RAD52 12 SLC16A7 12 TBClD15 12 TPH2 12 TRHDE 12 VPS29 12 WIFl 13 DNAJD1 13 FARP1 13 FARP1 13 FLJ10094 13 FLJ25477 13 FLJ25952 13 GPC6 13 GRK6PS 13 HSMPP8 13 IRS2 13 LOC387904 13 LOC387905 13 LOC390393 13 LOC400099 13 LOC400120 13 LOC440127 13 NUDT15 13 PABPC3 13 PSPCl 13 RFXAP 13 SCEL 13 SMAD9 13 STK24 13 SUCLA2 13 VDRIP 13 VGCNL1 13 ZNF198 13 ZNF237 14 LOC145497 14 LOC388001 14 LOC400234 14 NPAS3 14 AKAP6 14 AP4S1 14 C14orfl66B 14 C14orf68 14 COACH 14 CYCSP1 14 EEFlAlP2 14 EML1 14 KCNH5 14 KIAA0759 14 KIAA1036 14 LAMR1P3 14 LOC341965 14 LOC388001 WO 2006/053955 PCT/F12005/050429 102 14 LOC440198 14 LRFN5 14 MGC4645 14 NDUFB3P4 14 NPAS3 14 NRXN3 14 RPL22P2 14 RPLPlP1 14 RPS2P3 14 SELlL 14 SLC25A29 14 STRN3 14 STRN3 14 UBA52P3 14 WARS 15 MGC14798 15 ADAM10 15 ARNT2 15 ARRDC4 15 BG1 15 CHD2 15 CIB2 15 CYFIP1 15 DNAJA4 15 FES 15 FURIN 15 GABRA5 15 IDH3A 15 KIAA1055 15 LOC339003 15 LOC390561 15 LOC390572 15 LOC390638 15 LOC400320 15 LOC440272 15 LOC441722 15 LOC441723 15 MAN2A2 15 MEIS2 15 MGC14386 15 MGC14798 15 MGC45386 15 MTHFS 15 NANOGP8 15 NIPAl 15 NIPA2 15 NTRK3 15 pp9099 15 PRCl 15 RASGRF1 15 RGMA 15 SIAT8B 15 SLCO3A1 15 SMAP-1 15 VPS33B 16 CDYL2 16 WWOX 16 CDH13 16 CLECSFl 16 RRN3 16 WWOX WO 2006/053955 PCT/F12005/050429 103 17 CGI-125 17 FLJ12760 17 FLJ25818 17 KIAA0753 17 KRT10 17 KRT12 17 KRT20 17 KRT20 17 KRT23 17 KRT24 17 KRT25A 17 KRT25B 17 KRT25C 17 KRT25D 17 LOC342531 17 LOC342531 17 MGC21518 17 MGC21518 17 MGC45562 17 MRC2 17 PITPNM3 17 SLC13A5 17 SMARCEl 17 SOX9 17 TLK2 17 TXNL5 18 LOC388458 18 SERPINBll 18 C18orflO 18 Cl8orfll 18 C18orfl4 18 FLJ20793 18 KIAA1012 18 KIAA1328 18 LOC342705 18 LOC390848 18 LOC440491 18 MEPlB 18 NETO1 18 NPMlP1 18 PTPRM 18 RIT2 18 RIT2 18 RNF125 18 RNF138 18 SERPINB10 18 SERPINB2 18 SERPINB3 18 SERPINB4 18 SERPINB7 19 FLJO0060 19 FLJ14011 19 FLJ26175 19 GRLF1 19 ILT10 19 KIR2DL1 19 KIR2DL3 19 KIR2DS2 19 KIR3DL3 19 LILRA1 19 LILRA2 WO 2006/053955 PCT/F12005/050429 104 19 LILRB4 19 LOC284373 19 LOC388526 19 LOC388558 19 LOC400713 19 LOC440544 19 LOC441862 19 MGC9913 19 ZFP28 19 ZNF471 19 ZNF528 19 ZNF534 19 ZNF542 19 ZNF578 19 ZNF582 19 ZNF583 19 ZNF610 20 SNPH 20 C20orf149 20 C20orf46 20 EEFlA2 20 FKBPlA 20 GMEB2 20 IDH3B 20 KCNB1 20 KCNQ2 20 LOC200213 20 LOC388807 20 LOC441937 20 MGC10715 20 MGC5356 20 NOL5A 20 NSFLlC 20 PRIC285 20 PROCR 20 PSMF1 20 PTGIS 20 PTK6 20 PTPNSlL2 20 PTPNSlL3 20 SDCBP2 20 SIRPB1 20 SNPH 20 SNRPB 20 SRMS 20 TGM6 20 TMC2 20 TMC2 21 ATP50 21 C21orf94 21 ITSN1 21 ITSN1 21 LOC388819 21 MRPS6 22 DGCR2 22 FBLN1 X RPS6KA6 Gene name according to HUGO Gene Nomenclature Committee (HGNC) WO 2006/053955 PCT/F12005/050429 105 R4 ~ ( H l I (D I(VD I I (D I(D Iv I H I 0 1 I0 a) In 0\IH( (D I I W I(D WD I -Z I 44U 0(I44 - H ( .- I m( .H (D I- m H) 1 LO If) H I R (D ~ ~ ~ ~ ~ ~ ~ ~ ~ - -PH O4- )4 P 0 P Dr P> Q 4i Ii 0 Z ID -ZI ZI -14I 1 I D -ZIL-ZII -ZC - .~ H I--ZI - L 0)I 0001- 00 0( 0 I) Ol H L O (1 () (N0 (N ) WO I0CN) Y) 0 CD) CD CD) CD I- rot H 4 J~ ~ H , H I r.r . r.r WO 2006/053955 PCT/F12005/050429 106 rH rHrHrHrHrH fli oD oooo CDC DD ( o ooooo C CI vvv I o (O U) 0 O (N -- , I -WOC -HW LOW I 0 -H .O-H ,mHI H iS >1 > - -mrH No I 0 - C o1 * 4-1 r-O O I PH r-H LO ..- LO I CR 0 oNH O(d (N 'C H - * I t U~ o- - OI NO C S Z Mo m ( I Md 0 m I - 4 * 1 - t - 0) 4- H m r- r0- I r C4 -H m N M~ zzl zzl H~ ( 0 rciJU oD (D (oo r- IcN (d ' n m m m *H rH IH (d C CN 0 oLW-O O I -HH 0) N o O LO N (D D zz I a - 4-|C -Y -l - -D -D -0 N Q) om N I= - O C d CO. r-HLOCNO I -H P 4-1 A-H m ~ ~ mm~ I ,Q tCoH ON (DI Cd o4-0. 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I OI 0 O(LOD OH I100 (DI C I I O o- I I 00 I I ,C I| ' - I I (N N IC 0 00 I I I (N (N 4-a I I0 0 4- IU0 (4-14-1rHC0hHW(D - I CI I mlH -H ( | .M4W II H W I (N Wall I N f 4-G( I N -0 m SI 4 D l( I (D ' C O II HH -H -H I -4 I <HH WH WH *) -H C0 D H Q- |H 0 P H 0, Q) SOI I <OH <HG U < R I H I t I H azll m H 0 |d H WI I C P- P- P- P 42. 4 I CoWCW 00OC 4 oWC4m1 I0 O 1 1 0 ' 0 zzll I Z0 zl l r- )CI P4 H I I m m r- m Lnzl H (d o I I T H H M o e N LO HP nO H0 I D L R4 R RLO4 WC/H I 0 I 42 I H H H H It I m 1mo - H I|Z Z H > I H HH H DH I I W W W W - 11 - o H t I I C CC C N N m Q C I I 000 N z '' P4 Sr-H I C oWCW H0O OaP I ZZZ CI)CICI)CI I' Z I I' <all l I LOWO W I I W O~ W W W H LC) (f W W < - I IO m M HC NW . 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L) I- . 0o p( OH I c 0 C N~ (N C)LC oo co c o c o c oc o c 0 OD (N (D mN r- ( ( WD > U) I SN IO '.0 C C 0 1I1 4] I a). 00 a)) 0000 0 o '.o (N (N WW I L nL D m( 4-)H 0 ~ 1'.m N0 (N (N Cl) Sr- I rm tp( I r C WH 0 0 (D (D0 I> '.m0n H H( II m~ (N '.0 '.0 a I co ( oc o 0 00 0 0 o WI c ( (D H I -H m- (N (N- C 4ri I :-L- m z(mr- Dr-m zl 4-i vm mm- c U) ( I ) -. - Z Z -H ) O~~~~~U 1I Z ~ t~~ H HH ~ )- N~ U) .( I .0'0if 0 '.0 (N(I: f U) m a) a) m) a a) )I 4 F q4 H H HH H m xc ) C WO 2006/053955 PCT/F12005/050429 113 rH I i o | H I 42 -4 I (f I 42 i C | 0 N I CO * I 0 H I H I 0 S I o4 I 0 - | P CN I O 00 1 -H I (40 H I C (N I> ( 0 >. 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Claims (101)

1. A method for identification of an individual who has an altered risk of or susceptibility for 5 developing HT, the method comprising the steps of: a) providing a biological sample taken from said individual; b) collecting personal and clinical information of said individual; c) determining the nucleotides present in one or several of the polymorphic sites as set forth in tables 2 to 5 and 7 to 11 in said individual's nucleic acid; and 10 d) combining the SNP marker data with personal and clinical information to assess the risk of an individual to develop HT.

2. The method according to claim 1, wherein the altered risk is an increased risk of HT. 15

3. The method according to claim 1, wherein the altered risk is a decreased risk of HT.

4. The method according to claim 1, wherein the polymorphic sites are those present in the haplotypes presented in tables 3, 4, 5, 7 and 8. 20

5. The method according to claim 1, wherein the polymorphic sites are associated with the SNP markers set forth in tables 2 to 5 and 7 to 11.

6. The method according to claim 5, wherein the polymorphic sites are in complete linkage disequilibrium with the SNP markers set forth in tables 2 to 5 and 7 to 11. 25

7. The method according to claim 6, wherein the polymorphic sites are in complete linkage disequilibrium in the population in which the said method is used.

8. A method for identification of an individual who has an altered risk of or susceptibility for 30 developing HT, the method comprising the steps of a) providing a biological sample taken from a subject b) determining the nucleotides present in one or several of the polymorphic sites as set forth in tables 2 to 5 and 7 to 11 in said individual's nucleic acid c) combining the SNP marker data to assess the risk of an individual to develop HT 35

9. The method according to claim 8, wherein the altered risk is an increased risk of HT.

10. The method according to claim 8, wherein the altered risk is a decreased risk of HT. 40

11. The method according to claim 8, wherein the polymorphic sites are those present in the haplotypes presented in tables 3, 4, 5, 7 and 8.

12. The method according to claim 8, wherein the polymorphic sites are associated with the SNP markers set forth in tables 2 to 5 and 7 to 11. 45

13. The method according to claim 12, wherein the polymorphic sites are in complete linkage disequilibrium with the SNP markers set forth in tables 2 to 5 and 7 to 11.

14. The method according to claim 13, wherein the polymorphic sites are in complete linkage 50 disequilibrium in the population in which the said method is used. WO 2006/053955 PCT/F12005/050429 122

15. The method according to claim 1, wherein said one or several polymorphic sites reside within a HT risk gene or genes as set forth in table 6. 5

16. The method according to claim 1, wherein the HT risk genes reside in the genome regions which are defined by the haplotype pattern mining analysis, the genes set forth in tables 3, 4, 5, 7 and 8.

17. The method according to claim 1, wherein the polymorphic sites are associated with the 10 haplotype regions, haplotypes or SNP markers defining the haplotypes set forth in tables 3, 4, 5, 7 and 8.

18. The method according to claim 17, wherein the polymorphic sites are in complete linkage disequilibrium with the haplotype regions, haplotypes or SNP markers defining the 15 haplotypes set forth in tables 3, 4, 5, 7 and 8.

19. The method according to claim 18, wherein the polymorphic sites are in complete linkage disequilibrium in the population in which the said method is used.

20 20. The method according to claim 1, wherein one or several of the SNP markers are selected from the group consisting of the following haplotypes or individual SNPs: a) rs1521409 (A/G) (SEQ ID NO: 544), rs 10511365 (C/T) (SEQ ID NO: 316) and rs10511366 (C/T) (SEQ ID NO: 317) defining the haplotype ACT (or nucleotides from the complementary strand); 25 b) rs10508771 (A/T) (SEQ ID NO: 286), rs3006608 (C/T) (SEQ ID NO: 854), rs10508773 (C/T) (SEQ ID NO: 287) and rs950132 (C/T) (SEQ ID NO: 1325) defining the haplotype TCCC (or nucleotides from the complementary strand); c) rs2221511 (A/G) (SEQ ID NO: 733), rs4940595 (G/T) (SEQ ID NO: 986), rs1522723 (C/T) (SEQ ID NO: 548) and rs1395266 (C/T) (SEQ ID NO: 476) defining the haplotype 30 ATCC (or nucleotides from the complementary strand); d) rs1992906 (A/G) (SEQ ID NO: 655) defining the risk allele G; e) rs10270360 (A/G) (SEQ ID NO: 10) defining the risk allele G; f) rs1318392 (A/G) (SEQ ID NO: 438) defining the risk allele G; g) rs2209672 (A/G) (SEQ ID NO: 730) defining the risk allele A; 35 h) rs503208 (C/G) (SEQ ID NO: 989) defining the risk allele G

21. The method according to claim 1, wherein one or several of the SNP markers are selected from the group consisting of the following haplotypes or individual SNPs: a) rs1521409 (A/G) (SEQ ID NO: 544), rs10511365 (C/T) (SEQ ID NO: 316) and 40 rs10511366 (C/T) (SEQ ID NO: 317) defining the haplotype ACT (or nucleotides from the complementary strand); b) rs2221511 (A/G) (SEQ ID NO: 733), rs4940595 (G/T) (SEQ ID NO: 986), rs1522723 (C/T) (SEQ ID NO: 548) and rs1395266 (C/T) (SEQ ID NO: 476) defining the haplotype ATCC (or nucleotides from the complementary strand); 45 c) rs1997454 (A/G) (SEQ ID NO: 656) defining the risk allele G; d) rs10270360 (A/G) (SEQ ID NO: 10) defining the risk allele G; e) rs1318392 (A/G) (SEQ ID NO: 438) defining the risk allele G; f) rs2209672 (A/G) (SEQ ID NO: 730) defining the risk allele A; g) rs503208 (C/G) (SEQ ID NO: 989) defining the risk allele G 50 WO 2006/053955 PCT/F12005/050429 123

22. The method according to claim 1, wherein one or several of the SNP markers are selected from the group consisting of the following haplotypes: a) rs4845303 (A/T) (SEQ ID NO: 980), rs6428195 (C/G) (SEQ ID NO: 1030) and rs1935659 (A/G) (SEQ ID NO: 637) defining the haplotype ACG (or nucleotides from the 5 complementary strand); b) rs1997454 (A/G) (SEQ ID NO: 656), rs2139502 (A/G) (SEQ ID NO: 709) and rs1519991 (A/C) (SEQ ID NO: 542) defining the haplotype AGC (or nucleotides from the complementary strand); c) rs1521409 (A/G) (SEQ ID NO: 544), rs10511365 (C/T) (SEQ ID NO: 316) and 10 rs10511366 (C/T) (SEQ ID NO: 317) defining the haplotype ACT (or nucleotides from the complementary strand); d) rs7679959 (C/G) (SEQ ID NO: 1178), rs10517338 (C/G) (SEQ ID NO: 381) and rs959297 (A/T) (SEQ ID NO: 1338) defining the haplotype CGA (or nucleotides from the complementary strand); 15 e) rs2278677 (A/G) (SEQ ID NO: 749), rs3886091 (C/G) (SEQ ID NO: 899), rs1998167 (A/G) (SEQ ID NO: 657), rs1998168 (A/G) (SEQ ID NO: 658) and rs2235280 (A/G) (SEQ ID NO: 740) defining the haplotype GCAGG (or nucleotides from the complementary strand); f) rs10521062 (A/C) (SEQ ID NO: 404), rsl10512296 (A/G) (SEQ ID NO: 331), rs1924001 20 (C/G) (SEQ ID NO: 633) and rs2417359 (A/G) (SEQ ID NO: 784) defining the haplotype AACG (or nucleotides from the complementary strand); g) rs10508933 (C/G) (SEQ ID NO: 289), rs10509071 (A/G) (SEQ ID NO: 295) and rs10490967 (A/G) (SEQ ID NO: 94) defining the haplotype GGA (or nucleotides from the complementary strand); 25 h) rs10508771 (A/T) (SEQ ID NO: 286), rs3006608 (C/T) (SEQ ID NO: 854), rs10508773 (C/T) (SEQ ID NO: 287) and rs950132 (C/T) (SEQ ID NO: 1325) defining the haplotype TCCC (or nucleotides from the complementary strand); i) rs1386486 (C/T) (SEQ ID NO: 472), rs1386485 (A/C) (SEQ ID NO: 471), rs1386483 (A/G) (SEQ ID NO: 470) and rs7977245 (C/T) (SEQ ID NO: 1212) defining the haplotype 30 CAGT (or nucleotides from the complementary strand); j) rs276002 (A/G) (SEQ ID NO: 814) and rs274460 (A/G) (SEQ ID NO: 810) defining the haplotype AA (or nucleotides from the complementary strand); k) rs1245383 (A/G) (SEQ ID NO: 430), rs2133829 (C/T) (SEQ ID NO: 707), rs2173738 (C/T) (SEQ ID NO: 722), rs2050528 (C/T) (SEQ ID NO: 677) and rs202970 (C/T) (SEQ ID 35 NO: 671) defining the haplotype GCTTC (or nucleotides from the complementary strand); 1) rs1395266 (C/T) (SEQ ID NO: 476), rs931850 (A/G) (SEQ ID NO: 1303) and rs1522722 (C/T) (SEQ ID NO: 547) defining the haplotype TAC (or nucleotides from the complementary strand); m) rs2221511 (A/G) (SEQ ID NO: 733), rs4940595 (G/T) (SEQ ID NO: 986), rs1522723 40 (C/T) (SEQ ID NO: 548) and rs1395266 (C/T) (SEQ ID NO: 476) defining the haplotype ATCC (or nucleotides from the complementary strand); n) rs2825555 (A/G) (SEQ ID NO: 819), rs2825583 (C/T) (SEQ ID NO: 820), rs2825601 (A/G) (SEQ ID NO: 821), rs2825610 (G/T) (SEQ ID NO: 822) and rs1489734 (A/G) (SEQ ID NO: 532) defining the haplotype ATGGA (or nucleotides from the complementary strand) 45

23. A method for assessing susceptibility or predisposition to HT in an individual, the method comprising determining alteration of expression levels of one or several of the genes of table 6 in the individual, wherein a difference in expression is indicative of susceptibility to HT. WO 2006/053955 PCT/F12005/050429 124

24. The method according to claim 23, wherein alteration of expression levels is determined by assessing transcription levels of one or several of the genes of table 6 in the individual.

25. The method according to claim 23, wherein alteration of expression levels is determined 5 by assessing translation of mRNAs encoded by one or several of the genes of table 6 in the individual.

26. A method for assessing susceptibility or predisposition to HT in an individual, the method comprising determining alteration of biological activity of one or several ot the 10 polypeptides encoded by one or several of the genes of table 6 in the individual, wherein a difference in biological activity of one or several of the polypeptides is indicative of susceptibility to HT.

27. The method according to claim 26, wherein alteration of biological activity is determined 15 by assessing structure of one or several ot the polypeptides encoded by one or several of the genes of table 6 in the individual.

28. The method according to claim 26, wherein alteration of biological activity is determined by assessing amount of one or several of the metabolites of a polypeptide or polypeptides 20 encoded by one or several of the genes of table 6 in the individual.

29. The method according to claim 1, wherein the personal and clinical information, i.e. non genetic information concerns age, gender, behaviour patterns and habits, biochemical measurements, clinical measurements, obesity, the family history of HT, cerebrovascular 25 disease, other cardiovascular disease, hypercholesterolemia, obesity and diabetes, waist-to-hip circumference ratio (cm/cm), socioeconomic status, psychological traits and states, and the medical history of the subject.

30. The method according to claim 29, wherein the behaviour patterns and habits include 30 tobacco smoking, physical activity, dietary intakes of nutrients, alcohol intake and consumption patterns and coffee consumption and quality.

31. The method according to claim 29, wherein the biochemical measurements include determining blood, serum or plasma VLDL, LDL, HDL, total cholesterol, triglycerides, 35 apolipoprotein (a), fibrinogen, ferritin, transferrin receptor, C-reactive protein, glucose or insulin concentration.

32. The method according to claim 29, wherein the non-genetic measurements are those presented in table 8. 40

33. The method according to claim 29, wherein the non-genetic information contains BMI and history of obesity in the family of the subject.

34. The method according to claim 29 further comprising a step of calculating the risk of HT 45 using a logistic regression equation as follows: Risk of HT = [1 + e -(a+ r:(bi*Xi)] -1 , where e is Napier's constant, Xi are variables associated with the risk of HT, bi are coefficients of these variables in the logistic function, and a is the constant term in the logistic function. 50 WO 2006/053955 PCT/F12005/050429 125

35. The method according to claim 34, wherein a and bi are determined in the population in which the method is to be used.

36. The method according to claim 34, wherein Xi are selected among the variables that have 5 been measured in the population in which the method is to be used.

37. The method according to claim 34, wherein Xi are selected among the SNP markers of tables 2 to 5 and 7 to 11, among haplotype regions and haplotypes of tables 3, 4, 5, 7 and 8 and among non-genetic variables of the invention. 10

38. The method according to claim 34, wherein bi are between the values of -20 and 20 and/or wherein Xi can have values between -99999 and 99999 or are coded as 0 (zero) or 1 (one). 15

39. The method according to claim 34, wherein i are between the values 0 (none) and 100,000.

40. The method according to claim 1, wherein subject's short term, median term, and/or long term risk of HT is predicted. 20

41. A method for identifying compounds useful in prevention or treatment of HT comprising determining the effect of a compound on biological networks and/or metabolic pathways related to one or several polypeptides encoded by HT risk genes of table 6 in living cells; wherein a compound altering activity of one or several said biological networks and/or 25 metabolic pathways is considered useful in prevention or treatment of HT.

42. The method according to claim 41 comprising determining the effect of a compound on a biological activity of one or several polypeptides encoded by HT risk genes of table 6 in living cells; wherein a compound altering biological activity of a polypeptide is considered 30 useful in prevention and/or treatment of HT.

43. A method for prevention or treatment of HT comprising administering to a mammalian subject in need of such treatment an effective amount of a compound in a pharmaceutically acceptable carrier enhancing or reducing biological activity of one or several polypeptides 35 encoded by HT risk genes of table 6; and/or enhancing or reducing activity of one or several biological networks and/or metabolic pathways related to said polypeptides.

44. The method according to claim 43 comprising administering to a mammalian subject in need of such treatment an effective amount of a compound in a pharmaceutically acceptable 40 carrier enhancing or reducing expression of one or several HT risk genes of table 6; and/or enhancing or reducing the expression of one or several genes in biological networks and/or metabolic pathways related to polypeptides encoded by said HT risk genes.

45. The method according to claim 43 comprising administering to a mammalian subject in 45 need of such treatment an effective amount of a compound in a pharmaceutically acceptable carrier enhancing or reducing activity of one or several pathophysiological pathways involved in cardiovascular diseases and related to polypeptides encoded by HT risk genes of table 6.

46. The method according to claim 43, said method comprising the steps of: 50 a) providing a biological sample taken from a subject; WO 2006/053955 PCT/F12005/050429 126 b) determining the nucleotides present in one or several of the polymorphic sites associated with altered expression and/or biological activity and present in HT risk genes of table 6 in said individual's nucleic acid; and c) combining polymorphic site genotype data to select effective therapy for treating 5 HT in said subject.

47. The method according to claim 43, said method comprising the steps of: a) providing a biological sample taken from a subject; b) determining expression of one or several HT risk genes of table 6 and/or 10 determining biological activity of one or several polypeptides encoded by the HT risk genes of table 6 in said individual's sample; and c) combining the expression and/or biological activity data to select effective therapy for treating HT in said subject. 15

48. The method according to claim 43, wherein said treatment is gene therapy or gene transfer.

49. The method according to claim 48, wherein said treatment comprises the transfer of one or several HT risk genes of table 6 or variants, fragments or derivatives thereof. 20

50. The method according to claim 48, wherein said HT risk genes of table 6 or variants, fragments or derivatives thereof are associated with reduced risk of HT.

51. The method according to claim 48, wherein said treatment comprises treating regulatory 25 regions and/or gene containing region of one or more HT risk genes of table 6 or variants, fragments or derivatives thereof in somatic cells of said subject.

52. The method according to claim 48, wherein said treatment comprises treating regulatory regions and/or gene containing region of one or more HT risk genes of table 6 or variants, 30 fragments or derivatives thereof in stem cells.

53. The method according to claim 52, wherein said treatment comprises treating regulatory regions and/or gene containing region of one or more HT risk genes of table 6 or variants, fragments or derivatives thereof in stem cells in tissues affected by cardiovascular diseases. 35

54. The method according to claim 43, wherein said compound is a recombinant polypeptide encoded by an HT risk gene of table 6 or variant, fragment or derivative thereof.

55. The method according to claim 43, wherein said treatment is based on siRNA hybridising 40 to mRNA and/or to hnRNA of a HT risk gene of table 6.

56. The method according to claim 43, wherein said treatment is based on siRNA hybridising to mRNA and/or to hnRNA of one or several genes in biological networks and/or metabolic pathways related to polypeptides encoded by said HT risk genes of table 6. 45

57. The method according to claim 43, wherein said method of treating is a dietary treatment or a vaccination. WO 2006/053955 PCT/F12005/050429 127

58. The method according to claim 43 comprising a therapy restoring, at least partially, the observed alterations in biological activity of one or several polypeptides encoded by HT risk genes of table 6 in said subject, when compared with HT free healthy subjects. 5

59. The method according to claim 43 comprising a therapy restoring, at least partially, the observed alterations in expression of one or several HT risk genes of table 6 in said subject, when compared with HT free healthy subjects.

60. A method for monitoring the effectiveness of treatment of HT in a human subject the 10 method comprising measuring mRNA levels of HT risk genes of table 6, and/or levels of polypeptides encoded by said HT risk genes, and/or biological activity of polypeptides encoded by said HT risk genes in a biological sample taken from said subject; alteration of mRNA levels or polypeptide levels or biological activity of a polypeptide following treatment being indicative of the efficacy of the treatment. 15

61. A method for predicting the effectiveness of a given therapeutic for HT in a given individual comprising screening for the presence or absence of the HT associated SNP markers, haplotypes or haplotype regions in one or several of the HT risk genes of claim 15. 20

62. A method for predicting the effectiveness of a given therapeutic for HT in a given individual, the method comprising the steps of: a) providing a biological sample taken from a subject b) determining the nucleotides present in one or several of the polymorphic sites as set forth in tables 2 to 5 and 7 to 11 in said individual's nucleic acid; and 25 c) combining the SNP marker data to predict the effectiveness of a given therapeutic in an individual for HT.

63. A method for diagnosing of a subtype of HT in an individual having HT, the method comprising the steps of: 30 a) providing a biological sample taken from a subject; b) determining the nucleotides present in one or several of the SNP markers as set forth in tables 2 to 5 and 7 to 11 in said individual's nucleic acid; and d) combining the SNP marker data to assess the subtype of HT of an individual.. 35

64. The method according to claim 63, wherein said one or several SNP markers reside within a HT risk gene or genes as set forth in table 6.

65. The method according to claim 63, wherein the HT risk genes reside in the genome region which is defined by the haplotype pattern mining analysis, the genes and regions set 40 forth in tables 3, 4, 5, 7 and 8.

66. The method according to claim 63, wherein the polymorphic sites are associated with the haplotype regions, haplotypes or SNP markers defining the haplotypes set forth in tables 3, 4, 5, 7 and 8. 45

67. The method according to claim 63, wherein the polymorphic sites are in complete linkage disequilibrium with the haplotype regions, haplotypes or SNP markers defining the haplotypes set forth in 3, 4, 5, 7 and 8. WO 2006/053955 PCT/F12005/050429 128

68. The method according to claim 63, wherein the polymorphic sites are in complete linkage disequilibrium in the population in which the said method is used.

69. The method according to any one of claims 43 or 60 to 63 further comprising a step of 5 combining non-genetic information with the results obtained according to any one of claims 44 to 68.

70. The method according to claim 69, wherein the non-genetic information concerns age, gender, behaviour patterns and habits, biochemical measurements, clinical measurements, 10 obesity, the family history of HT, cerebrovascular disease, other cardiovascular disease, hypercholesterolemia, obesity and diabetes, waist-to-hip circumference ratio (cm/cm), socioeconomic status, psychological traits and states, and the medical history of the subject.

71. The method according to claim 69, wherein the behaviour patterns and habits include 15 tobacco smoking, physical activity, dietary intakes of nutrients, alcohol intake and consumption patterns and coffee consumption and quality.

72. The method according to claim 69, wherein the biochemical measurements include determining blood, serum or plasma VLDL, LDL, H-DL or total cholesterol or triglycerides, 20 apolipoprotein (a), fibrinogen, ferritin, transferrin receptor, C-reactive protein, glucose, serum or plasma insulin concentration.

73. The method according to claim 69, wherein the non-genetic measurements are those presented in table 8. 25

74. The method according to claim 69, wherein the non-genetic information contains the BMI and history of obesity in the family of the subject.

75. A method for measuring HT risk gene product protein expression, production or 30 concentration in a biological sample taken from a subject, wherein said HT risk gene is as defined in table 6, the method comprising the steps of: a) providing a biological sample taken from a subject to be tested; and b) detecting the expression, production or concentration of said protein in said sample, wherein altered expression, production or concentration indicates an altered risk of 35 cardiovascular disease in said subject

76. A test kit based on a method according to claim 1 for assessment of an altered risk of or susceptibility for HT in a subject. 40

77. A test kit for determining the nucleotides present in one or several of the SNP markers as set forth in tables 2 to 5 and 7 to 11 in said individual's nucleic acid for assessment of an altered risk of HT in a subject.

78. A test kit for determining the nucleotides present in one or several of the SNP markers as 45 set forth in tables 2 to 5 and 7 to 11 in said individual's nucleic acid for assessment of an altered risk of HT in a subject, containing: a) reagents and materials for assessing nucleotides present in one or several SNP markers as set forth in tables 2 to 5 and 7 to 11; and b) software to interpret the results of the determination. 50 WO 2006/053955 PCT/F12005/050429 129

79. The test kit according to claim 76 further comprising PCR primer set for amplifying nucleic acid fragments containing one or several SNP markers as set forth in tables 2 to 5 and 7 to 11 from the nucleic acids of the subject. 5

80. The test kit according to claim 76 comprising a capturing nucleic acid probe set specifically binding to one or several SNP markers present in HT associated markers and haplotype regions as set forth in tables 2 to 5 and 7 to 11.

81. The test kit according to claim 76 comprising a microarray or multiwell plate to assess 10 the genotypes.

82. The test kit according to claim 76 comprising a questionnaire for obtaining patient information concerning age, gender, height, weight, waist and hip circumference, skinfold and adipose tissue thicknesses, the proportion of adipose tissue in the body, the family history of 15 diabetes and obesity, the medical history concerning HT.

83. A test kit for detecting the presence of SNP markers in one or several of HT risk genes as set forth in table 6 in a biological sample, wherein said SNP markers are more frequently present in a biological sample of a subject susceptible to HT compared to a sample from a 20 subject not susceptible to HT, the kit comprising: a) reagents and materials for assessing nucleotides present in SNP markers in one or several of HT risk genes as set forth in table 6; and b) software to interpret the results of the determination. 25

84. The test kit of claim 83 further comprising PCR primer set for amplifying nucleic acid fragments containing said SNP markers from HT risk genes as set forth in table 6 from the nucleid acids of the subject.

85. The test kit of claim 83 comprising a capturing nucleic acid probe set specifically binding 30 to one or several SNP markers present in HT risk genes as set forth in table 6.

86. The test kit of claim 83 comprising a microarray or multiwell plate to assess the genotypes. 35

87. The test kit of claim 83 comprising a questionnaire for obtaining patient information concerning age, gender, height, weight, waist and hip circumference, skinfold and adipose tissue thicknesses, the proportion of adipose tissue in the body, the family history of diabetes and obesity, the medical history concerning HT. 40

88. A test kit based on a method according to any one of claims 46, 47, 60 to 63 or 75.

89. The test kit of claim 88 further comprising PCR primer set for amplifying nucleic acid fragments containing said SNP markers from HT risk genes as set forth in tables 2 to 5 and 7 to 11 from the nucleid acids of the subject. 45

90. The test kit of claim 88 comprising a capturing nucleic acid probe set specifically binding to one or several SNP markers present in HT risk genes as set forth in tables 2 to 5 and 7 to 11. WO 2006/053955 PCT/F12005/050429 130

91. The test kit of claim 88 comprising a microarray or multiwell plate to assess the genotypes.

92. The test kit of claim 88 comprising a questionnaire for obtaining patient information 5 concerning age, gender, height, weight, waist and hip circumference, skinfold and adipose tissue thicknesses, the proportion of adipose tissue in the body, the family history of diabetes and obesity, the medical history concerning HT.

93. The test kit of claim 76, 83 or 88, further comprising a marker set to assess the ancestry of 10 an individual.

94. The test kit of claim 93 comprising a SNP marker set to assess the ancestry of an individual. 15

95. The test kit of claim 93 comprising a microsatellite marker set to assess the ancestry of an individual.

96. The method of claim 1 further comprising a marker set to assess the ancestry of an individual. 20

97. The method of claim 1 comprising a SNP marker set to assess the ancestry of an individual.

98. The method of claim 1 comprising a microsatellite marker set to assess the ancestry of an 25 individual.

99. The method according to claim 1, wherein one or several of the SNP markers are selected from the group consisting of the following individual SNPs: a) rs1860933 (AT ) (SEQ ID NO:1366) defining the risk allele A 30 b) rs4236780 (CG) (SEQ ID NO:1367) defining the risk allele C c) rs2000112 (CT) (SEQ ID NO:660) defining the risk allele C d) rs931850 (AG) (SEQ ID NO:1303) defining the risk allele A e) rs2192947 (AG) (SEQ ID NO:728) defining the risk allele G f) rs9328292 (AG) (SEQ ID NO: 1316) defining the risk allele A 35 g) rs1409367 (CT) (SEQ ID NO:490) defining the risk allele C h) rs1893814 (CT) (SEQ ID NO:622) defining the risk allele T i) rs2263356 (CT) (SEQ ID NO:746) defining the risk allele T j) rs6826647 (CT) (SEQ ID NO:1368) defining the risk allele C k) rs1913157 (CG) (SEQ ID NO:630) defining the risk allele C 40

100. The method according to claim 99 further comprising a step of combining information from hypertension drug treatment of the subject to the genetic information of the subject.

101. The method according to claim 1, wherein one or several of the SNP markers are 45 selected from the group consisting of the following individual SNPs: a) rs6826647 (CT) (SEQ ID NO:1368) defining the risk allele C b) rs1409367 (CT) (SEQ ID NO:490) defining the risk allele C c) rs9328292 (AG) (SEQ ID NO: 1316) defining the risk allele A d) rs1395266 (CT) (SEQ ID NO:476) defining the risk allele T 50 e) rs1893814 (CT) (SEQ ID NO:622) defining the risk allele T WO 2006/053955 PCT/F12005/050429 131 f) rs931850 (AG) (SEQ ID NO:1303) defining the risk allele A g) rs1860933 (AT) (SEQ ID NO:1366) defining the risk allele A h) rs1386483 (AG) (SEQ ID NO:470) defining the risk allele A i) rs4236780 (CG) (SEQ ID NO:1367) defining the risk allele C 5 j) rs1913157 (CG) (SEQ ID NO:630) defining the risk allele C k) rs2263356 (CT) (SEQ ID NO:746) defining the risk allele T 1) rs2000112 (CT) (SEQ ID NO:660) defining the risk allele C