DE10348600A1 - Use of a gene modification in the human GNAS1 gene to predict disease risks, disease courses and to predict the response to disease therapies - Google Patents

Abstract

The invention relates to the use of a genomic gene modification in the gene for the Galphas subunit of human G protein, which is encoded by the gene GNAS1, for predicting disease risks, disease courses and for response to disease therapies by means of pharmacological and non-pharmacological measures and for predicting undesirable effects drug reactions. Furthermore, the invention relates to the provision of individual gene changes and haplotypes, with the aid of which further gene changes that can be used for the above-mentioned purposes can be detected and validated. Such gene changes can be that there is a substitution of thymine by cytosine in exon 5 at position 393, that there is a substitution of guanine by adenine at position -1211 or that thymine is replaced by guanine at position -839. The gene changes can be detected individually or in any combination by means of methods familiar to the person skilled in the art.

Description

Using a genetic modification in the human GNAS1 gene to predict disease risks, disease courses and to predict response to disease therapies

Function and meaning heterotrimeric G proteins

All cells of the human body have membrane receptors on their surface that control all cell functions. Such receptors include the so-called heptahelical receptors for e.g. B. classic hormones, neurotransmitters and chemokines. There are also a variety of receptors for growth factors and receptors with intrinsic tyrosine kinase activity, for example receptors for insulin, insulin-like growth factor, epidermal growth factor, platelet-derived growth factor and many more. There are also many receptors that are responsible for regulating blood formation, such as the receptor for erythropoietin. Such receptors control cell growth, motility, gene expression, apoptosis and chemotaxis, among others. The receptors mentioned transmit their signals into the cell interior by activating so-called heterotrimeric G proteins. These G proteins consist of a large family of different isoforms and are each composed of different α, β and γ subunits. Currently 5 β subunits, 13 γ subunits and more than 20 α subunits are known which are encoded by different genes (Farfel Z et al. The expanding spectrum of G protein diseases. N Engl J Med. 1999 Apr 1; 340 (13): 1012-20). The combination of these different α, β and γ subunits creates a large number of different heterotrimeric G proteins. The isoform composition determines which heterotrimer can be activated by a specific receptor. The βγ subunits are to be considered functionally as a monomer. At rest, the α subunit bound GDP ( 1 ). After activating a coupling receptor, the α subunit releases GDP in exchange for GTP and the βγ subunits dissociate from the α subunits. Both the free α and βγ subunits can control the activity of a variety of different effectors. These include, for example, ion channels, the adenylyl cyclase, the PI3 kinase, different MAP kinases etc. The α subunits have an intrinsic GTPase activity which hydrolyzes the GTP bound after activation to GDP. Subsequently, the released βγ subunits reassociate with the α subunit, thereby ending the activation cycle. The heterotrimer is thus available for a renewed activation cycle (Bourne HR. How receptors talk to trimeric G proteins. Curr Opin Cell Biol. 1997; 9 (2): 134-42). A scheme of the G protein cycle is in 1 shown. The activation of such G proteins is the crucial step for cell activation. Due to the paramount importance of G proteins, it is immediately clear that mutations or genetic polymorphisms in genes that code for G proteins must have a lasting influence on the activability of cells if these mutations function or express G protein - affect subunits. This also has a decisive influence on the risk of illness or the course of illness. In addition, the response to the therapy of diseases, be it through pharmaceuticals or other measures such as radiation, diets, operations, invasive interventions, etc. depends on the activability of G proteins.

importance the Gαs subunit

The Gαs subunit is expressed in all human body cells. Their stimulation leads, among other things, to activation of the adenylycyclase and thus to an increase in the intracellular cAMP concentration. For example, cAMP-dependent protein kinases can be activated. However, other cAMP-dependent signal cascades are inhibited or activated by stimulation of receptors that couple to Gαs. Furthermore, Gαs can regulate the activity of ion channels, for example potassium or calcium channels (see 2 ). A variety of receptors couple to Gαs, e.g. the receptors for adenosine, adrenaline, noradrenaline, P2-purinergic receptors, opioids, dopamine, epidermal growth factor, FSH, VIP, thyroliberine, glucagon, vasopressin, histamine and many more. After stimulation of Gαs-coupled receptors, apoptosis is induced in many cell types, so that there is a connection to tumor diseases and their course and treatability, but also a connection to inflammatory diseases and their course and treatability. In addition, diverse metabolic pathways are regulated by Gαs. Inhibition of the expression of Gαs in fibroblasts accelerates their differentiation into adipocytes (Wang Hy et al., Antisense oligodeoxynucleotides to GS protein alpha-subunit sequence accelerate differentiation of fibroblasts to adipocytes. Nature. 1992 Jul 23; 358 (6384): 334-7 ; Wang H et al. G (s) alpha repression of adipogenesis via Syk.J Biol Chem. 1999 Nov 5; 274 (45): 32159-66; overview at Neves at al, G protein pathways.Science. 2002 May 31; 296 (5573): 1636-9).

The GNAS1 gene

The human Gαs gene (GNAS1) is located on chromosome 20g13.2–13.3. A schematic representation of the gene structure can be found in 3A , The use of different promoters, P1, P2 or P3 leads to the creation of different mRNAs and thus to different gene products ( 3B ). Two long (Gαs-1 and -2) and two short (Gαs-3 and -4) isoforms of Gαs result from alternative splicing of exon 3 (T. Kozasa et al. Isolation and characterization of the human Gs alpha gene. Proc. Natl. Acad. Sci. US A 85: 2081-2085, 1988.). The use of an alternative splice acceptor site for exon 4 leads to the insertion of another serine in Gsα-2 and -4 ( 4 ; P. Bray et al. Human cDNA clones for four species of G alpha s signal transduction protein. Proc.Natl.Acad.SCi.USA 83 (23): 8893-8897, 1986). The splice variants differ in terms of their tissue distribution, their activability by G-protein coupled receptors and their activation of effectors (J. Novotny and P. Svoboda. The long (Gs (alpha) -L) and short (Gs (alpha) -S) variants of the stimulatory guanine nucleotide-binding protein. Do they behave in an identical way? J. Mol. Endocrinol. 20 (2): 163-173, 1998; R. Seifert et al. Different effects of Gsalpha splice variants on beta2- adrenoreceptor-mediated signaling. The Beta2- adrenoreceptor coupled to the long splice variant of Gsalpha has properties of a constitutively active receptor. J.Biol.Chem. 273 (18): 5109-5116, 1998).

Somatic mutations in GNAS1

There have been a number of somatic Mutations in the GNAS1 gene described that contribute to rare metabolic diseases. Most of these are activating (endocrine GH-secreting Tumors, McCune-Albright syndrome, fibrous Bone dysplasia) or inactivating (e.g. pseudohypoparathyroidism Type IA, pseudopseudohypoparathyroidism, (PPHP)) mutations in Area of the GTPase region of the protein. (Overview at: Weinstein et al., Endocrine Manifestations of Stimulatory G Protein α-Subunit Mutations and the Role of Genomic Imprinting Endocrine Reviews 22 (5): 675-705, 2001). In contrast to single nucleotide polymorphisms (SNP's), these mutations in the corresponding patients, for example, not in peripheral areas Blood cells found.

Genomic mutation in GNAS1

Jia et al. investigated in 1999 whether the GNAS1 gene contributes to the risk of essential hypertension (Jia H. et al., Association of the Gsα Gene With Essential Hypertension and Response to β-Blockade Hypertension. 1999; 34: 8–14.). They examined a common silent T393C polymorphism (SNP, single nucleotide polymorphism) in exon 5 of the GNAS1 gene (numbering according to the sequence of the cDNA; 5 ). This polymorphism is characterized by the lack of a restriction site for the endonuclease FokI (Fok- in the T allele) or the presence of this interface (Fok + in the C allele). The 393T allele was found more often in hypertensives than in normotensive controls. This finding was later confirmed by other investigators (Abe M et al., Association of GNAS1 gene variant with hypertension depending on smoking status. Hypertension. 2002; 40: 261–5). In addition, it was described that hypertensive 393C allele carriers respond better to β-blockers with regard to lowering blood pressure than 393T allele carriers Jia H. et al., Association of the Gsα Gene With Essential Hypertension and Response to β-Blockade Hypertension. 1999; 34: 8-14.). Other investigators showed a connection between the T393C status and the risk of orthostatic hypotension (Tabara, Y. et al., Polymorphisms of genes encoding components of the sympathetic nervous system but not the renin ± angiotensin system as risk factors for orthostatic hypotension Journal of Hypertension 2002, 20: 651-656). This shows that the T393C status has so far only been associated with blood pressure regulation or the response to a β-adrenoceptor blockade. A correlation to other diseases, disease courses or to response to pharmaceuticals in general has not yet been described. Since this T393C nucleotide exchange is silent and does not change the encoded amino acid (isoleucine), there must be other polymorphisms that are in coupling imbalance with the T393C polymorphism.

Object of the invention

• a. funktionsverändernde genomische Polymorphismen und Haplotypen im Gen GNAS1 bereitzustellen, die entweder zu einem Aminosäurenaustausch führen, oder
• b. die das Spleißverhalten beeinflussen, oder
• c. die zur Änderung der Proteinexpression oder zur Änderung der Expression von Spleißvarianten führen, oder
• d. die zum Auffinden und/oder Validieren weiterer Polymorphismen bzw. Haplotypen im Gen GNAS1 geeignet sind.

The invention described here aims to

• a. provide function-changing genomic polymorphisms and haplotypes in the GNAS1 gene which either lead to an amino acid exchange, or
• b. that affect the splicing behavior, or
• c. which lead to a change in the protein expression or to a change in the expression of splice variants, or
• d. those suitable for finding and / or validating further polymorphisms or haplotypes in the GNAS1 gene are not.

Because of the basic meaning by Gαs for the Signal transduction is suitable for such polymorphisms or haplotypes, to generally predict disease risks or disease courses for all diseases or therapy response / therapy failure or undesirable side effects for all pharmaceuticals or non-pharmacological To predict therapies.

Detection of new polymorphisms in the GNAS1 gene before exon 1

The main object of the present invention is to find the polymorphisms G (-1211) A and T (-839) G ( 5 ) found by systematic sequencing of DNAs from humans homozygous for the 393C allele or homozygous for the 393T allele. For this purpose, gene sequences which are in front of Exon1 of GNAS1 were amplified by means of a PCR reaction and sequenced according to the Sanger method. The person skilled in the art is familiar with the methods required for this, for example the derivation of primer pairs required for the PCR reaction and the selection of sequencing primers. New polymorphisms were found, with adenine substitution of guanine at position - 1211 (G (-1211) A polymorphism), or a guanine substitution of thymine at position -839 (T (-893) G- polymorphism). These SNPs are numbered in such a way that the nucleotide A of the start codon ATG is assigned the number +1. Since the convention does not have the number 0, the number -1 is assigned to the nucleotide in front of the A of the start codon ATG.

The detection of these SNPs in terms of their use according to the invention can with any to the person skilled in the art procedures are shown, for example, direct sequencing, PCR and subsequent restriction analysis, reverse hybridization, dot blot or slot blot method, mass spectrometry, Taqman ^® - or Light -Cycler ^® technology, Pyrosequencing ^® . Invader ^® technology, Luminex method etc. Furthermore, these gene polymorphisms can be detected simultaneously after multiplex PCR and hybridization to a DNA chip.

Distribution of T393C, G (-1211) A and T (-839) G polymorphisms and derived genotypes with different ethnicities, detection of haplotypes and use of these genotypes to find other relevant polymorphisms and haplotypes.

For this purpose, different DNA samples from Caucasians, Black Africans and Chinese were genotyped. The result is shown in the following tables

This genotype distribution is highly significant in the chi² test with a Chi 43.3 and a P <0.0001. The 393T allele frequency (% T) is highest among black Africans.

This genotype distribution is very significantly different in the chi² test with a Chi 208.30 and a P <0.0001. The (-1211) G allele frequency (% G) is highest in Black Africans, followed by Caucasians and Chinese.

This genotype distribution is included in the chi² test a Chi 72.2 and a P <0.0001 highly significant different. The (-839) T allele frequency (% T) is highest among black Africans and Chinese. From these distributions can one conclude that history of development (related to Caucasians) the 393T allele, the (-1211) G allele and the (-893) G allele represent the respective “original states” Differences in genotype distribution among different ethnic groups usually indicate that associated phenotypes for evolution were significant and the bearers brought a certain advantage. It is known to the person skilled in the art that ethnically diverse genotype distribution is an indication of this are that even today certain genotypes and haplotypes with certain diseases or physiological and pathophysiological Modes of reaction or response to therapy, e.g. with pharmaceuticals, are associated. Another analysis shows a coupling imbalance between the three polymorphisms in Caucasians. Under coupling imbalance one understands the occurrence of allele combinations (haplotypes), the statistically clearly more common or are less common than their frequency would be expected.

The following table shows the distribution of T393C genotypes stratified according to GNAS G (-1211) A genotypes for Caucasians.

A clear accumulation of the (-1211) G allele can be seen in carriers of a 393T allele (p <0.01). The following table shows the distribution of C393T genotypes stratified for GNAS1 T (-893) G genotypes for Caucasians.

One can see an accumulation of the (-893) T allele in carriers of a 393T allele. The following table shows the distribution of G (-1211) A genotypes stratified according to GNAS1 T (-893) G genotypes for Caucasians.

A clear accumulation can be seen of the (-893) T allele with carriers a -1211G allele (p <0.0001). Further analyzes show that in Caucasians preferential the haplotypes 393T + G (-1211) + T (-839) (60%) and C393 + (-1211) A + (–839) G occur to 47. In addition, all possible permutations have been found Service.

• 1. Für bestimmte Phänotypen (zelluläre Eigenschaften, Krankheitszustände, Krankheitsverläufe, Arzneimittelansprechen usw.) wird zunächst eine Assoziation mit den Polymorphismen T393C, G(-1211)A und T(-839)G hergestellt, wobei diese Assoziationen für jeden Genotyp einzeln oder unter Verwendung aller Permutationen der Haplotypen aufgestellt werden können.
• 2. Für neu detektierte Genveränderungen in GNAS1 wird untersucht, ob bereits bestehende Assoziationen unter Verwendung oben beschriebener Geno- oder Haplotypen verstärkt oder abgeschwächt werden.

It is an object of the invention that these new polymorphisms and haplotypes can now be used to detect and validate further relevant genomic gene changes in GNAS1. To do this, proceed as follows:

• 1. For certain phenotypes (cellular properties, disease states, disease courses, drug responses, etc.), an association with the polymorphisms T393C, G (-1211) A and T (-839) G is first established, these associations for each genotype individually or under Use of all permutations of the haplotypes can be established.
• 2. For newly detected gene changes in GNAS1, it is examined whether existing associations are strengthened or weakened using the genotypes or haplotypes described above.

Functional meaning of the C393T polymorphism and associated polymorphisms

It was examined which functional changes gene changes in the GNAS1 gene can be assigned. A correlation to alternative splicing, tissue-specific expression or an overexpression of the Gαs protein depending on genotypes of the C393T polymorphism or the A-1211G polymorphism are conceivable here, for example. For this purpose, mRNA was obtained from human heart and adipose tissue and transcribed into cDNA using reverse transcriptase. The process is familiar to the person skilled in the art. The expression level was subsequently determined using real-time PCR (Tagman method) and compared with the housekeeping gene β-actin. The results are in 6 shown. T allele carriers of the C393T polymorphism have significantly higher expression values than homozygous CC genotypes. Similarly, in human heart tissue, TT genotypes have relatively higher expression levels than CC and TC genotypes of the C393T polymorphism ( 6B ). The same applies to analysis according to the A (-1211) G polymorphism. With A allele carriers one finds in the adipose tissue ( 6C ), in the heart tissue ( 6D ) and in the bladder tissue ( 6E ) an increased expression of the Gαs mRNA, both absolutely and in relation to housekeeping genes (eg GAPDH, β-actin).

This proved that it in the GNAS1 gene. gene mutations that gives an expression change by Gαs effect in different tissues. This can be the C393T polymorphism, the A (-1211) G polymorphism be or polymorphisms associated with these in coupling imbalance stand. It is therefore part of the invention described here also, the expression of Gαs quantify at the mRNA level or protein level, with known ones Associate polymorphisms of GNAS1 and new, more appropriate ones Discover and validate polymorphisms.

The finding shown here of a genotype-dependent expression of Gαs is extremely important. at Transgenic animals, the overexpression of Gαs leads to increased apoptosis of cardiac myocytes (Geng Y. et al., Apoptosis of cardiac myocytes in Gsalpha transgenic mice. Circ Res. 1999, 84, 34-42) as the cause of heart failure. In such animals there is increased Ca2 + influx in myocytes (Kim SJ et al., Differential regulation of inotropy and lusitropy in overexpressed Gsalpha myocytes through cAMP and Ca2 + channel pathways. J Clin Invest. 1999 Apr; 103 (7): 1089– 97). In addition, these animals show increased heart rate and blood pressure (Uechi M et al., Depressed heart rate variability and arterial baroreflex in conscious transgenic mice with overexpression of cardiac Gsalpha. Circ Res. 1998 Mar 9; 82 (4): 416-23 ) Similar effects can therefore also be expected in people who overexpress the gas protein due to a gene change in GNAS1. Here we expect an increased cardiovascular risk, a changed response to substances whose receptors activate Gαs. The changed tendency to apoptosis can influence a variety of disease courses (e.g. tumor and immune diseases) and determine the response to pharmaceuticals.

Using a genetic modification in GNAS1 for predicting disease risks and disease courses

Because of the key function of the Gαs subunit for the Cell activation it's an integral part of the invention that using genetic modification in GNAS1 generally predicted disease risks and disease courses can be.

Human heterotrimeric G proteins are composed of the subunits α, β and γ. Again, a number of isoforms are known, which are characterized by different genes are encoded. For example, there are 13 different γ isoforms (γ1 – γ13), at least 5 different β-isoforms (β1-β5) and one Many different α isoforms (αs (short and long), αo, αi1-3, αq, α11-16, αolf etc.) Da G proteins are known to play a central role in control function of all human cells regardless of which cell receptors are activated is to be expected immediately, that the course is more diverse and very different diseases in a genetically determined, increased activatability is influenced by G proteins. Especially with the diverse functions Functional mutations of G proteins become completely different particularly outstanding importance and predictive power. This stands as opposed to mutations in other genes that are for others Proteins, e.g. Encode hormones or hormone receptors.

This means that due to gene changes in proteins that are expressed in all human body cells and bundle incoming hormone signals at a central point and thereby regulating cell functions, all physiological and pathophysiological operations be decisively influenced or at least modulated. Besides responses to pharmaceuticals are also influenced in a special way. Of these are desired but also undesirable Drug effects affected.

It was in the scientific Literature repeatedly postulates that functional changes of G proteins has a lasting impact on a wide range of diseases or more diverse on the course Have diseases. Such gene changes can change the structure Mutations in G protein subunits, for example the ability to be activated by a receptor, the enzymatic GTPase activity concern or affect the dimerization of βγ subunits. Next to it such changes change the composition of heterotrimeric G proteins. Further the level of expression of such G protein subunits may be changed or it can splice variants with changed Function occur (Farfel Z et al., The expanding spectrum of G protein diseases. N Engl J Med. 1999 Apr 1; 340 (13): 1012-20; Iiri T et al., G-protein diseases furnish a model for the turn-on switch. Nature. 1998 Jul 2; 394 (6688): 35-8; Iiri T et al., G proteins propel surprise.Nat Genet. 1998 Jan; 18 (1): 8-10. Mirror AM. Hormone resistance caused by mutations in G proteins and G protein-coupled receptors. J Pediatr Endocrinol Metab. 1999 Apr; 12 Suppl 1: 303-9. mirror AT THE. Inborn errors of signal transduction: mutations in G proteins and G protein-coupled receptors as a cause of disease. J Inherit Metab Dis. 1997 Jun; 20 (2): 113-21. Mirror AM. The molecular basis of disorders caused by defects in G proteins. Horm Res. 1997; 47 (3): 89-96. Mirror AM. Mutations in G proteins and G protein-coupled receptors in endocrine disease. J Clin Endocrinol Metab. 1996 Jul; 81 (7): 2434-42. Review. mirror AT THE. Defects in G protein-coupled signal transduction in human disease. Annu Rev Physiol. 1996; 58: 143-70).

• 1. Genveränderungen in Genen, die für ubiquitär exprimierte Proteine kodieren, beeinflussen vielfältige Erkrankungen bzw. verursachen vielfältige Erkrankungsrisiken
• 2. G-Proteine steuern nahezu alle Signaltransduktionsvorgänge im menschlichen Körper

The following can be seen from the examples mentioned:

• 1. Genetic changes in genes that code for ubiquitously expressed proteins influence a variety of diseases or cause a variety of disease risks
• 2. G proteins control almost all signal transduction processes in the human body

While it is clear from the literature cited that it can be assumed in general that G protein mutations can cause such diseases, there has been a connection with genomic mutations in the GNAS1 gene for the GαS subunit of heterotrimeric G proteins with disease risks neither described nor assumed in the literature.

• 1. Herz-Kreislauferkrankungen. Darunter fallen insbesondere Hypertonie, Schlaganfall, koronare Herzkrankheit und Myokardinfarkt, Herzinsuffizienz, Herzrhythmusstörungen, Präeklampsie bzw. Gestose. Da bereits ein Zusammenhang zwischen dem T393C-Polymorphimsmus und Hypertonie beschrieben wurde (Jia H. et al., Association of the G_sα Gene With Essential Hypertension and Response to β-Blockade Hypertension. 1999;34:8–14.), ist es ein Bestandteil der vorliegenden Erfindung; dass die hier neu beschriebenen G(-1211)A- und T(-839)G-Polymorphismen ebenfalls ein Risikofaktor für Hypertonie sind, da sie mit dem T393C-Polymorphismus im Kopplungsungleichgewicht stehen. Da Hypertonie bekanntlich ein Hauptrisikofaktor für Schlaganfall, Herzinfarkt und Herzinsuffizienz ist, ist es ein weiterer Bestandteil der vorliegenden Erfindung, dass die genannten Genveränderungen in GNAS1 auch das Risiko für solche Erkrankungen erhöhen.
• 2. Enokrinologische und Stoffwechselerkrankungen. Darunter fallen insbesondere Adipositas, metabolisches Syndrom, Typ-2 Diabetes-mellitus, Gicht, Osteoporose, Schilddrüsenerkrankungen wie Hyper- und Hypothyreose und M.Basedow, Hyper- und Hypoparathyreodismus, Morbus Cushing, Hxper- und Hypoaldosteronismus und viele mehr.
• 3. Psychiatrische Erkrankungen wie Depression, Schizophrenie, Alkoholismus und Angststörungen, Phobien, Neurosen
• 4. Neurologische Erkrankungen wie Morbus Parkinson, multiple Sklerose, Epilepsien
• 5. Dermatologische Erkrankungen wie Psoriasis, Neurodermitis
• 6. Tumorerkrankungen

Diseases which are associated with a gene change in GNAS1 and which are determined, for example, by a changed expression level of Gαs protein, can be mentioned:

• 1. Cardiovascular diseases. These include in particular hypertension, stroke, coronary heart disease and myocardial infarction, heart failure, cardiac arrhythmia, preeclampsia or gestosis. Since a connection between the T393C polymorphism and hypertension has already been described (Jia H. et al., Association of the _GSA Gene With Essential Hypertension and Response to β-Blockade Hypertension. 1999; 34: 8-14.), It is a component of the present invention; that the G (-1211) A and T (-839) G polymorphisms newly described here are also a risk factor for hypertension, since they are in coupling imbalance with the T393C polymorphism. Since hypertension is known to be a major risk factor for stroke, heart attack and heart failure, it is a further component of the present invention that the gene changes mentioned in GNAS1 also increase the risk of such diseases.
• 2. Enocrinological and metabolic diseases. These include, in particular, obesity, metabolic syndrome, type 2 diabetes mellitus, gout, osteoporosis, thyroid diseases such as hyper- and hypothyroidism and M.Basedow, hyper- and hypoparathyroidism, Cushing's disease, hypertension and hypoaldosteronism and many more.
• 3. Psychiatric disorders such as depression, schizophrenia, alcoholism and anxiety disorders, phobias, neuroses
• 4. Neurological diseases such as Parkinson's disease, multiple sclerosis, epilepsy
• 5. Dermatological diseases such as psoriasis, neurodermatitis
• 6. Tumor diseases

Basic Properties of malignant tumors

For malignant tumors, also as cancer referred to, there are characteristic changes in basic functions, that unfavorably promote the growth of such cells. cancer cells are characterized by a loss of contact inhibition and an uncontrolled cell growth. Such changes are triggered through a large number of noxae, so-called carcinogens, which are the genome damage. Such noxae belong many chemicals, tobacco smoke, but also UV light. Play next to it genetic factors play an important role in the development of cancer. Characteristic of In addition to their uninhibited growth, cancer cells also tend to metastases (Metastases) in other organs. The spread of the Metastases occur regularly through the Bloodstream or over Lymphatic vessels. cancers are with a large part of cases incurable and lead to death. The initial tumor and metastases are tried therapeutically to remove surgically. Next to it Tumors are irradiated. Using so-called cytostatics, antibodies against certain proteins or surface markers or immunomodulating substances (cytokines, interferons) tries to kill the rapidly dividing cancer cells or to be transferred to programmed cell death (apoptosis). The currently available therapeutic activities to lead in most cases only for prolonged survival, not for definitive healing.

Prognostic factors in cancer

It is of great medical relevance to define prognostic factors for the course of cancer that provide information about the response to certain forms of therapy or that are generally predictive of the occurrence of metastases, tumor progression and survival. Up to now, prognostic factors that are generally known to the person skilled in the art have been used in medicine. These include, for example, the size of the tumor, its depth of penetration into the surrounding tissue, growth beyond organs, penetration into blood or lymphatic vessels or lymph nodes, and the degree of differentiation of the tumor cells. There are also some relatively non-specific serological markers. The procedure for classifying the tumors is generally referred to as "staging" and "grading". In general, the presence of distant metastases and a low degree of differentiation are prognostically very unfavorable parameters. Nevertheless, it is the general experience in medicine that patients with the same tumor stage have drastically different disease courses. While rapid progression of the disease and the appearance of metastases and recurrences are observed in some patients, the disease comes to a standstill in other patients for unclear reasons. Metastases can occur locally, regionally and far from the mother tumor. For this it is necessary that a large number of tumor cells reach the adjacent lymphatic or blood path by simply flushing them into liquid or by clapping into neighboring tissue. Recurrence tendency is the recurrence of a tumor after incomplete or partial removal of the tumor. This is not a new ma ligne transformation, but the regrowth of a tumor tissue that has not been completely removed. Recurrence is also possible through metastases, which can remain latent for many years. Progression is the recurrence of a tumor with a higher grading (more dedifferentiation) or the reappearance of metastases.

Obviously there is one Numerous individual, unrecognized biological variables that Course of a tumor disease independent of staging and grading determine to a high degree. Such factors include genetic Host factors.

In addition, it is desirable to have genetic markers to develop the predictive for the Occurrence of tumors are. Such markers fulfill the function, the affected ones Individuals early further screening measures (serology, X-ray, ultrasound, NMR etc.) supply. So that can Early stage cancer recognized and treated therapeutically, with the healing and chances of survival in early-stage tumors are much better than with advanced tumors.

Types of tumors

Generally, all human cells body malignant and lead to cancer. The here and beyond made executions describe general mechanisms of tumor progression, metastasis and the therapeutic response. In this respect, the ones described here apply Mechanisms and claims for all Human tumors, for example for the following tumors:

Urogenital tract tumors: Here These include bladder carcinoma, renal cell carcinoma, Prostate cancer and the seminoma.

Tumors of the female genital organs: Breast cancer, carcinoma of the body, ovarian cancer, the Cervical cancer.

Tumors of the gastrointestinal tract: oral carcinoma, esophageal carcinoma, gastric carcinoma, liver carcinoma, bile duct carcinoma, Pancreatic carcinoma, colon carcinoma, rectal carcinoma.

Tumors of the respiratory tract: the Larynx carcinoma, the bronchial carcinoma.

Tumors of the skin: malignant melanoma, the basalioma, the T cell lymphoma of the hematopoietic Systems: Hodgkin and non-Hodgkin lymphomas, acute and chronic leukemia Etc.

Tumor diseases of the brain or of nerve tissue: glioblastoma, neuroblastoma, medulloblastoma, meningeal Sarcoma, astrocytoma.

Soft tissue tumors, for example sarcomas and head and neck tumors.

G proteins and malignant transformation of cells

Experimental in vitro investigations prove that mutations in G protein subunits are malignant Can transform cells. The expression of a constitutively active Gαq subunit with lack of GTPase activity leads to malignant transformation of fibroblasts (Kalinec G et al., Mutated alpha subunit of the Gq protein induces malignant transformation in NIH 3T3 cells. Mol Cell Biol. 1992 Oct; 12 (10): 4687-93). Similar observations were made for the α subunits Gα12, Gα13, Go, Gz and Gαi2 made (Xu N et al., A mutant alpha subunit of G12 potentiates the eicosanoid pathway and is highly oncogenic in NIH 3T3 cells. Proc Natl Acad Sci U S. A. 1993 Jul 15; 90 (14): 6741-5. Xu N et al Potent transforming activity of the G13 alpha subunit defines a novel family of oncogenes. Biochem Biophys Res Commun. 1994 Jun 15; 201 (2): 603-9. Pace AM et al A mutant alpha subunit of Gi2 induces neoplastic transformation of Rat-1 cells. Proc Natl Acad Sci U.S.A. 1991 Aug 15; 88 (16): 7031-5. Wong YH et al., Mutant alpha subunit of Gz transforms Swiss 3T3 cells. Oncogene. 1995 May 18; 10 (10): 1927-33). These works prove that mutations in α subunits can contribute in principle to malignant cell transformation.

G protein mutations and Cancer in humans

In humans, in some rarely adenomas detected somatic mutations in the subunits Gαs and Gαi2. These are called Gip2 (Gαi2 subunit) or. as Gsp (Gαs subunit) designated. In contrast, these are not genetic Host factors that modulate the course of the disease, but around causal factors (overview with: Farfel Z et al. The expanding spectrum of G protein diseases. N Engl J Med. 1999; 340 (13): 1012-20).

Use of gene changes in the GNAS1 gene to predict the course of tumor diseases

An essential part of the present invention is the provision of diagnostically relevant Gene changes in the GNAS1 gene as a prognostic factor for all tumor diseases in humans. Naturally, not all tumor diseases can be described here. The principle is therefore explained using selected examples that demonstrate its general applicability:

Example 1: Chronic lymphoblastic leukemia (CLL)

In chronic lymphatic leukemia is a chronic form of leukemia. Characteristic for the Illness is a big one Number of degenerate lymphocytes. A total of 30 percent of all leukaemic Diseases are chronic lymphatic leukaemias. The median age of onset is 65 years old. Only ten percent of the patients are younger than 50 years. Men are about two to three times more common affected than women. Risk factors for the development of a CLL are not known. However, the disease occurs in Japan and China rarely on. Japanese people who immigrated to the USA are also extremely rare on a CLL. This fact suggests that genetic Factors play a role. The therapy depends on the Stage of disease. A CLL can be benign for up to 20 years get lost, that is, the patient shows except enlarged lymph nodes and eventual fatigue as well as loss of appetite no symptoms. The treatment only starts then when the number of lymphocytes increases above a certain level begins, the proportion of red blood cells and the number of blood platelets decreases or other complications occur. Early treatment has none Influence on the course and outcome of the disease. The most important therapeutic measure is chemotherapy. In certain cases, the patient must also irradiate or have surgery. Patients can up to to live 20 years with the diagnosis of CLL without being serious Show symptoms. The further the disease has progressed, the more are bigger, however also the health disorders through the change of the organ system. Depending on the binet stage of the disease, the doctor can estimate the forecast. The stage of a CLL is characterized, among other things, by how many lymphocytes are in the blood and bone marrow, how large spleen and liver and whether there is anemia or not. A CLL leads to Changes in Immune system so that people who suffer from this disease are more at risk to develop other types of cancer. At the same binet stage however, patients show very different disease courses. component The invention is to show that gene changes in the GNAS1 gene are suitable are to predict the course of the CLL. For this purpose, patients with CLL regarding of the described gene changes genotyped in GNAS1 and the gene status with disease progression compared. Here we define the time interval under Progression between the initial diagnosis of CLL and the need for therapy.

7A shows a significant difference in freedom from therapy depending on the GNAS1 T393C status, whereby the CC genotype is associated with a two-fold accelerated disease progression (hazard ratio 2.07; p = 0.02). 7B shows a significant difference with regard to freedom from therapy depending on the GNAS1 G (-1211) A status, whereby the GG genotype is associated with a three-fold accelerated disease progression (hazard ratio 3.01; p = 0.01). The effect becomes particularly strong when the alleles or genotypes are combined (-1211) GG + 393 CC versus (-1211) AG / AA + 393 CT / TT ( 8A ). Similarly, analyzes of the G (-1211) A status and the T (-839) G status can be combined as shown in 8B will be shown. 9 proves that gene changes in GNAS1 can be used to predict CLL, especially in the early stages (BINET A), as is shown for the G (-1211) A status.

Example 2: Bladder cancer

Bladder cancer is a malignant tumor on the bladder mucosa. Bladder cancer most often occurs between the ages of 60 and 70. Men are affected three times as often as women. Bladder cancer is the third most common form of cancer in men after lung and prostate cancer. Bladder cancer can be caused by external influences. Risk factors include smoking, constant exposure of the organism to chemicals such as dyes, painkiller abuse. For many patients, the studies show that it is a superficial tumor. This can be surgically removed using the cystoscope. More than 70% of patients treated for superficial bladder cancer show a tumor recurrence in the course. Recurring tumors with non-muscle-invasive disease occur in more than half of them. These can be treated or controlled curatively by transurethral resection. It is therefore important to recognize these lesions early and to follow up the patient regularly and closely. The first place is cystoscopy with urine cytology. Elimination urograms are used at regular intervals to check for possible tumor manifestations in the kidney pelvis and ureters. So far there are hardly any valid markers that are predictive for the further course of the disease. Therefore, the classic factors such as depth of penetration, degree of differentiation, metastasis, lymph node involvement etc. are currently used for the prognosis. Genetic markers for tumor progression, recurrence tendency, survival probability and therapy response would be the care of Significantly improve patients with bladder cancer. Another component of the invention is that the use of gene changes in GNAS1 is suitable for predicting the further course of the disease. 10A shows the time until the occurrence of metastases depending on the GNAS1 T393C status. The risk of metastasis in patients with a TC / CC genotype is around 1.2–4.0; p = 0.015). The 2.5-fold increase (95 median time to metastasis is 29 months for the TT genotype, but 13 months for the combined TC / CC genotype. A similar relationship is found when examining the time to tumor progression ( 10B ). By this we mean the occurrence of metastases or the recurrence of the tumor with higher staging or grading. The curve shape is significantly different for the 393 TT, TC and CC genotypes (p = 0.027, logrank test, with the CC and TC genotypes being assigned the less favorable shape. Finally, the relationship between GNAS1 T393C status and Survival shown ( 10C ). Again, one recognizes that patients with the GNAS1 393 genotypes TC and CC die earlier than patients with the TT genotype. The median survival time was 44 months for patients with the TT genotype, but only 20.5 months for the combined TC / CC genotype. Because of the significant coupling imbalance to the T393C polymorphism, the G (-1211) A polymorphism can be used in a similar manner. It can be assumed that an increased tendency to apoptosis due to increased gas expression in 393T carriers or -1211A carriers has a favorable influence on the course of the disease.

Example 3: Renal Cell Carcinoma

Kidney cancer is a serious illness. The chances of recovery depend on the size and spread of the tumor. In patients without metastases, the 10-year survival rate is up to 80%, but with considerable inter-individual variability. Thanks to the widespread use of ultrasound technology today, many of the tumors are recognized at an early, non-metastatic stage and can be treated in time. If distant metastases are present, it is possible to combine surgical removal of the kidney with subsequent immunotherapy with interleukin-2 or interferon-alpha. This strengthens the body's own cancer cell defense. In metastatic renal cell carcinoma, the combination of interferon, interleukin-2 and 5-fluorouracil can be used to achieve a response rate of 36 percent in studies. There are currently no predictive markers for progression, survival, or therapeutic response in patients with renal cell carcinoma. Accelerated progression was seen in patients with the TC / CC genotypes using the GNAS1 T393C polymorphism ( 11A ). It also shows that survival in patients with renal cell carcinoma depends on the GNAS1-T393C status ( 11B ). Here the patients with the CC genotype die the earliest, followed by the TC and TT genotypes, the latter surviving the longest. In multivariate analysis, GNAS status (p = 0.012), grading (p <0.001) and stage (p <0.001) are independent predictors of survival. Because of the significant coupling imbalance to the T393C polymorphism, the G (-1211) A polymorphism can be used in a similar manner.

Example 4: Acute myeloid leukemia (AML)

The cells with the greatest importance for the AML are the granulocytes and the monocytes. The main function of this Cells is the immune defense. The AML is a degeneration of the stem cell based. This degeneration leads that the stem cell's ability for differentiation into functional blood cells. Their tendency to reproduce remains fully intact. It happens to a malicious one Proliferation of immature cells, the blasts. These proliferate in the bone marrow too fast, too uncontrolled and released into the blood too soon.

So they do not get the required Maturity and ability therefore not their tasks, such as infection control progeny. Since the space in the bone marrow is limited, the leukemia cells displace the growing red blood cells and platelets. Not enough erythrocytes and platelets are produced. consequences are anemia and an excessive tendency to bleed. Because the leukemia cells through the blood and lymphatic system circulate, they also migrate into lymph nodes, spleen and liver and to lead there to a functional impairment. This often leads to an increase in size of these organs. So acute myeloid leukemia is considered malignant Expansion and loss of the ability to differentiate hematopoietic cells in the bone marrow. The cells of the are particularly affected myeloid series, i.e. monocytes and granulocytes. basic Part of the treatment of patients with acute myeloid leukemia is chemotherapy. This will inhibit the growth of leukemia cells tried by giving medication. Such drugs will be also called cytostatics. Chemotherapy can be supplemented through two further therapy options: stem cell transplantation and bone marrow transplant.

Unfortunately, it has so far not been possible to develop drugs that selectively damage the leukemia cells ended. The side effects of chemotherapy are caused by damage to healthy, naturally rapidly dividing cells. Cytostatics, i.e. drugs that interfere with cell division, are usually administered in combinations and according to a schedule. In about 70 to 80% a complete remission, i.e. a regression of all symptoms, is achieved after 2 to 3 treatment cycles. Intensive chemotherapy destroys 99 to 99.9% of all leukemia cells. The remaining leukemic cells cannot be destroyed in the majority of patients despite intensive consolidation therapy. After 5 years, only 20 to 40% of the patients are still alive. It has now been investigated whether gene changes in GEN GNAS1 read prognostic factors can be used for survival in patients with AML. A significant correlation was found with the genotype status based on the A (-1211) G polymrophsimsu. 12 shows that patients with the AA genotype of the A (-1211) G polymorphism have the best chance of survival, while the course of the disease is accelerated in AG and GG genotypes. All patients included in the study had received intensive chemotherapy or a bone marrow transplant. This not only shows that gene changes in the GNAS1 gene are markers for the progression of AML, but that these gene changes can also be used as general pharmacogenetic markers for cancer chemotherapy drugs and bone marrow transplantation.

Possible molecular causes the GNAS dependent Tumor progression.

For this, cDNA from CLL cells was hybridized with Affymetrix gene chips as described in the literature (Dürig J et al., Expression of ribosomal and translation-associated genes is correlated with a favorable clinical course in chronic lymphocytic leukemia. Blood. 2003 Apr 1; 101 (7): 2748-55). The expression level of the genes was examined not according to the clinical course, but according to the status of the GNAS1 G (-1211) A polymorphism. The result is in 13 shown. Of course, not all of the 30,000 genes on the Affymetrix chip can be shown here. Interestingly, in (-1211) A allele carriers, compared to the GG genotype, there is a significant increase in expression of apoptosis-associated genes, e.g. from caspase 8, immediate early response 3, IL24, CDKN1a, etc. This suggests that there is an increased signal transduction via Gαs Due to the increased expression contributes to an increased propensity to apoptosis. This also shows that gene changes in the GNAS gene can be used to discover disease-relevant genes and new targets for pharmaceuticals.

Pharmacogenetics - diagnostics the effectiveness of pharmaceuticals, the potency and efficiency of pharmaceuticals and the occurrence of unwanted effects

Basics and goals of pharmacogenetics

The effectiveness of drugs and / or the occurrence of undesirable Side effects are in addition to the specific properties of the substance chemically defined products defined by a number of parameters. Two important parameters, the achievable plasma concentration and the plasma half-life largely determine the effectiveness or ineffectiveness of pharmaceuticals or the occurrence of undesirable effects. The plasma half-life is determined, among other things, by what speed certain drugs in the liver or other body organs to be effective or inactive metabolites metabolized and at what speed them out of the body are excreted, the excretion via the kidneys, via the Breathing air, over the sweat over which Sperm fluid, about the Chair or over other body secretions can be done. In addition, the effectiveness with oral administration is determined by limits the so-called "first pass effect" because after Absorption of pharmaceuticals via the intestine a certain proportion in the liver to inactive metabolites is metabolized.

Mutations or polymorphisms in Genes of metabolizing enzymes can the activity change them in the way that their amino acid composition changed becomes what the affinity is increased or decreased to the metabolizing substrate and so that the metabolism can be accelerated or slowed down. In a similar way can Mutations or polymorphisms in transport proteins affect the amino acid composition change in the way that the transport and thus the excretion from the body accelerates or is slowed down.

To select the most suitable for a patient Substance, the optimal dosage, the optimal dosage form and to avoid unwanted, z.T. health Hazards or more deadly Side effects is knowledge of genetic polymorphisms or of mutation leading to change of gene products lead of paramount importance.

The effect of hormones in the human body and the importance of polymorphisms in hormone receptors

A variety of hormones and peptide hormones of the human body but also practice receptor antagonists their effect on so-called receptors of the body cells. These are proteins different composition. After activating these receptors have to these signals are directed into the cell what about the Activation of heterotrimeric G proteins is mediated. Such G proteins are composed of different α, β and γ subunits. Depending on the activatability, these receptors can be defined by Hormones, divided into certain groups. The person skilled in the art knows that mutations or polymorphisms in certain receptors Effectiveness of certain agonists or antagonists on these receptors can determine. So one affects more often Gly16Arg polymorphism in the gene coding for the β2 adrenoceptor, the Strength responsiveness to the β2 sympathomimetic Salbutamol (Martinez FD, et al. Association between genetic polymorphisms of the beta2-adrenoceptor and response to albuterol in children with and without a history of wheezing. J Clin Invest. 1997 Dec 15; 100 (12): 3184-8). polymorphisms in the D2 receptor gene determine the frequency the occurrence of dyskinesia in the treatment of Parkinson's disease (Parkinson's disease) with Levadopa (Oliveri RL, et al .; Dopamine D2 receptor gene polymorphism and the risk of levodopa-induced dyskinesias in PD. Neurology. 1999 Oct. 22; 53 (7): 1425-30): Polymorphisms in the μ-opiate receptor gene determine the analgesic effectiveness of opiates (Uhl GR, et al. The mu opiate receptor as a candidate gene for pain: polymorphisms, variations in expression, nociception, and opiate responses. Proc Natl Acad Sci U.S.A. 1999 Jul 6; 96 (14): 7752-5).

The gene changes mentioned in specific receptors are only used to diagnose the effects of pharmaceuticals, than these drugs specific agonists or antagonists to the considered receptors. Desirable however, is an individual diagnosis of general responsiveness across from all pharmaceuticals and the individual prediction of the risk of undesirable Effects under therapy with drugs.

The diagnosis of the activatability of G proteins allows a general diagnosis of the effectiveness of pharmaceuticals, their optimal dosage and the occurrence of side effects.

We generally understand pharmaceuticals Substances to the human body from the outside be fed about certain conditions herzustelle. Such substances can Hormones, low or high molecular weight substances, peptides or proteins, antibodies and much more. his.

Most used to treat diseases, physical Malfunctions or mental disorders used pharmaceuticals are hormones, agonists on hormone receptors, antagonists on hormone receptors or other substances which the expression of receptors or directly or indirectly affect the concentration of hormones. A number of pharmaceuticals practice this influence from the fact that under therapy with such substances Physiological counter-regulations take place, which are the concentrations of hormones increase activate the G protein-coupled receptors. As a well known An example here is therapy with diuretics, especially called loop diuretics and thiazide diuretics. The loss of saline during therapy and lowering of blood pressure to lead to activate the renin-angiotensin-aldosterone system. The more educated Hormone angiotensin II stimulates an increased absorption of sodium in the kidney, stimulates salt intake, increases blood pressure through a direct vasoconstrictive effect on smooth vascular muscle cells and induced Proliferation processes. It is well known that these are caused by angiotensin II Mechanisms occur after coupling the hormone to receptors Your effect over mediate activation of heterotrimeric G proteins. The efficiency these effects are predictable if the strength of the Activability of G proteins can be diagnosed. Other drugs exert their effect by that the resumption of transmitters released from neurons, e.g. Inhibit norepinephrine, adrenaline, serotonin or dopamine. Here The pharmacon sibutramine, which is the reuptake of serotonin and noradrenaline in the central nervous system inhibits the feeling of hunger reduced and thermogenesis increased. Similarly, sibutramine can be used to treat obesity. Because norepinephrine and Diagnostics is to activate sibutramine G protein-coupled receptors the activatability of G proteins to predict effectiveness of sibutramine and the appearance of typical sibutramine-associated Side effects (e.g. increase in heart rate and blood pressure) in special measure suitable.

The invention is based on the fact that a method has been developed which is generally suitable for the diagnosis of the activatability of G proteins. For this purpose, one or more polymorphisms in the gene GNAS1 are examined, which codes for the human Gαs subunit of heterotrimeric G proteins. Polymorphisms which predict the diagnosis of the occurrence or non-occurrence of an alternative splicing process of the gene or a changed expression of Gαq are particularly suitable for this. Overexpression predictably leads to an increased activatability of heterotrimeric G proteins and to an increase All cells of the human body can be activated. A determination of the presence of polymorphisms in GNAS1 thus enables the diagnosis of the effectiveness and undesirable effects of drugs, in particular agonists and antagonists of all receptors, the effects of which are mediated by heterotrimeric G proteins. In addition, such polymorphisms in GNAS1 can be used to diagnose the effects of drugs that either indirectly or as a result of body counter-regulation mechanisms increase or decrease the concentrations of endogenous hormones whose receptors activate heterotrimeric G proteins. Thus, the invention allows a diagnosis of effects and undesirable effects of all pharmaceuticals and is not limited to pharmaceuticals that influence specific receptors in an agonistic or antagonistic manner. In addition, the diagnosis of allelic or haplotype status in GNAS1 can be used to determine the individually optimal and tolerable dosage of medicinal products.

For the diagnosis of an increased or reduced activatability of G proteins serves in particular Detection of T393T polymorphism or G (-1211) A polymorphism or of the T (-839) G polymorphism either alone or in all conceivable Combination.

In addition, all others can be used for diagnostics gene mutations used in GNAS in a coupling imbalance stand by these polymorphisms and / or the alternative splicing process or the expression additionally promote or inhibit.

These gene changes can be done with any, the expert common Procedures are detected, e.g. direct sequencing, restriction analysis, reverse hybridization, dot blot or slot blot method, mass spectrometry, Taqman or light cycler technology, Pyrosequencing etc. These can also Gene polymorphisms simultaneously after multiplex PCR and hybridization be detected on a DNA chip. In addition, an increased diagnosis can be made Activation of G proteins also uses other methods be the direct detection of the expression level of Gαs or splice variants of Enable Gαs.

• 1. Adrenerge Rezeptoren, inbesondere α- und ß-Adrenozeptoren und deren Isoformen und Untergruppen, d.h. α1- und α2-Adrenozeptoren sowie β1-, β2-, β3 und β4-Adrenozeptoren
• 2. Muskarinrezeptoren und deren Isoformen, z.B. m1-,m2-, m3-, m4 und m5-Muskarinrezeptoren und deren Subtypen. Typische Antagonisten an Muskarinrezeptoren sind beispielsweise Atropin, Scopolamin, Ipratroprium, Pirenzepin und N-Butylscopolamin. Typische Agonisten sind Carbachol, Bethanechol, Pilocarpin etc.
• 3. Dopaminrezeptoren, z.B. D1-, D2-, D3-, D4-, und D5-Rezeptoren und deren Isoformen und Spleißvarianten
• 4. Serotoninrezeptoren, z.B. 5-HT1- 5-HT2-, 5-HT3-, 5-HT4-, 5HT-5, 5HT-6 und 5-HT7-Rezeptor und deren Subtypen, Isoformen und Spleißvarianten. Typische Agonisten sind Sumatriptan und Cisaprid, Antagonsiten sind beispielsweise Ondansetron, Methysergid, Buspiron und Urapidil.
• 5. Endothelinrezeptoren und deren Subtypen, Isoformen und Spleißvarianten
• 6. Bradykininrezeptoren, z.B. B1- und B2-Rezeptoren und deren Subtypen, Isoformen und Spleißvarianten
• 7. Angiotensinrezeptoren, z.B. AT II Typ1 und Typ2 Rezeptor, typische Antagonisten am AT II-Rezeptor sind Losartan und andere Sartane.
• 8. Rezeptoren für Endorphine und Opiate, z.B. der μ-Opiatrezeptor
• 9. Chemokinrezeptoren CCR1-12 und CXCR1-8 für z.B. Interleukin-1/2/3/4/5/6/7/8/9/10/11/12, RANTES, MIP-1α, MIP-1β, stromal cell-derived factor, MCP1-5, TARC, Lymphotactin, Fractalkine, Eotaxin 1-2, NAP-2, LIX etc.
• 10. Adenosinrezeptoren und deren Subtypen, Isoformen und Spleißvarianten
• 11. Rezeptoren für Thrombin (Protease-aktivierte Rezeptoren)
• 12. Rezeptoren für Lyso-Phophatidsäure, Phosphatidsäure, Rezeptoren für Sphingosinphosphate und deren Derivate
• 13. Rezeptoren für Prostaglandine und Thromboxane, z. B. für PGE1, PGE2, PGF, PGD2, PGI2, PGF2α, Thromboxan A2, etc.
• 14. Rezeptoren für Neuropetide, z.B. NPY1-5
• 15. Histaminrezeptoren, z.B. H1-H3-Rezeptoren
• 16. Rezeptoren für Plättchen-aktivierender Faktor (PAF-Rezeptor)
• 17. Rezeptoren für Leukotriene
• 18. Rezeptoren für Insulin, Glucagon, Insulin-like growth factor (IGF-1 und IGF-2), Epidermal Growth Factor (EGF) und platelet-derived growth factor (PDGF)
• 19. Rezeptoren für Wachstumshormon (GH), Somatostatin (SSTR1-5), thyreotropes Hormon (TSH), Oxytocin, Prolactin, Gonadotropine
• 20. Rezeptoren für Zytokine, z.B. Interferone
• 21. Rezeptoren für Purine
• 22. Orphanrezeptoren, deren Wirkungen durch G-Proteine vermittelt werden.
• 23. Rezeptoren für Leptin

The method mentioned is particularly suitable for the diagnosis of the effect of agonists or antagonists on receptors, the effects of which are known to be mediated by G proteins. The following examples are given here, although the list of examples could be extended:

• 1. Adrenergic receptors, in particular α and β adrenoceptors and their isoforms and subgroups, ie α1 and α2 adrenoceptors as well as β1, β2, β3 and β4 adrenoceptors
• 2. Muscarinic receptors and their isoforms, eg m1, m2, m3, m4 and m5 muscarinic receptors and their subtypes. Typical antagonists on muscarinic receptors are, for example, atropine, scopolamine, ipratroprium, pirenzepin and N-butylscopolamine. Typical agonists are carbachol, bethanechol, pilocarpine etc.
• 3. Dopamine receptors, for example D1, D2, D3, D4 and D5 receptors and their isoforms and splice variants
• 4. Serotonin receptors, for example 5-HT1- 5-HT2, 5-HT3, 5-HT4, 5HT-5, 5HT-6 and 5-HT7 receptors and their subtypes, isoforms and splice variants. Typical agonists are sumatriptan and cisapride, antagonists are for example ondansetron, methysergide, buspirone and urapidil.
• 5. Endothelin receptors and their subtypes, isoforms and splice variants
• 6. Bradykinin receptors, for example B1 and B2 receptors and their subtypes, isoforms and splice variants
• 7. Angiotensin receptors, eg AT II type 1 and type 2 receptor, typical antagonists on the AT II receptor are losartan and other sartans.
• 8. Receptors for endorphins and opiates, for example the μ-opiate receptor
• 9. Chemokine receptors CCR1-12 and CXCR1-8 for eg interleukin-1/2/3/4/5/6/7/8/9/10/11/12, RANTES, MIP-1α, MIP-1β, stromal cell -derived factor, MCP1-5, TARC, lymphotactin, fractalkine, eotaxin 1-2, NAP-2, LIX etc.
• 10. Adenosine receptors and their subtypes, isoforms and splice variants
• 11. Receptors for thrombin (protease-activated receptors)
• 12. Receptors for lysophosphatidic acid, phosphatidic acid, receptors for sphingosine phosphates and their derivatives
• 13. Receptors for prostaglandins and thromboxanes, e.g. B. for PGE1, PGE2, PGF, PGD2, PGI2, PGF2α, Thromboxan A2, etc.
• 14. Receptors for neuropetides, eg NPY1-5
• 15. Histamine receptors, for example H1-H3 receptors
• 16. Platelet Activating Factor Receptors (PAF Receptor)
• 17. Receptors for leukotrienes
• 18. Receptors for insulin, glucagon, insulin-like growth factor (IGF-1 and IGF-2), epidermal growth factor (EGF) and platelet-derived growth factor (PDGF)
• 19. Receptors for growth hormone (GH), somatostatin (SSTR1-5), thyreotropic hormone (TSH), oxytocin, prolactin, gonadotropins
• 20. Receptors for cytokines, eg interferons
• 21. Receptors for purines
• 22. Orphan receptors, the effects of which are mediated by G proteins.
• 23. Receptors for leptin

Can also be predicted the effects of pharmaceuticals, the resumption, the breakdown or affect the resynthesis of neurotransmitters or at those undergoing therapy changes in the expression or responsiveness of the above-mentioned receptors occur (e.g. sibutramine, fluoxetine). The can also be diagnosed Effects of all pharmaceuticals, which are direct or indirect also as a result of a physiological backlash of agonists that activate the above-mentioned receptors. predicted can also be the influence of radiation therapy in cancer patients

• 1. Antihypertensiva, z.B. β-Blocker (Propanolol, Bisprolol, etc.), Diuretika (Hydrochlorothiazid und weitere Thiaziddiuretika; Furosemid, Piretanid und weitere Schleifendiuretika, Chlorthalidon), α1-Adrenozeptorblocker (z.B. Doxazosin, Prazosin), Angiotensinrezeptor-Blocker (z.B. Losartan), ACE-Hemmer (Enalapril, Captopril, Ramipril etc.), Ca²⁺-Kanalblocker (z.B. Nifedipin, Verapamil, Amlodipin, Felodipin), Clonidin, Reserpin, Renin- Inhibitoren
• 2. Pharmaka zur Behandlung der Herzinsuffizienz, z.B. β-Blocker (z.B. Propanolol, Metoprolol), ACE-Hemmer (z.B. Captopril, Enalapril, Ramipril, etc.), Angiotensin-Rezeptorblocker (z.B. Losartan), Digitalisglykoside, Katecholamine, Diuretika.
• 3. Pharmaka zur Behandlung des niedrigen Blutdrucks oder einer Herzinsuffizienz, z.B. α- und β-Smpathomimetika (Effortil, Adrenalin, Noradrenalin, Dobutamin, β-Adrenozeptorblocker, ACE-Hemmer, Angiotensin II-Rezeptorblocker.)
• 4. Pharmaka zur Behandlung der Migräne, z.B. Sumatriptan, Rizatriptan, Zolmitriptan und weitere Agonisten an Serotoninrezeptoren, β-Blocker (Propanolol, Timolol), Ergotamin und Dihydroergotamin
• 5. Analgetika vom Morphintyp (Morphin, Codein, etc.)
• 6. Pharmaka zur Behandlung der koronaren Herzkrankheit wie Adenosin, β-Blocker (z.B. Propanolol, Acebutolol), Nitrate und Ca²⁺-Kanalblocker
• 7. Pharmaka zur Behandlung psychiatrischer Erkrankungen (Schizophrenie, manisch-depressive Erkrankungen, Psychosen, Depressionen) und von Suchtkrankheiten wie Alkoholismus, (z.B. Fluoxetin, Paoxetin, Imipramin, Desipramin, Doxepin, Mianserin, Trazodon, Lofepramin), Angstsyndrome (Diazepam, etc.), welche z.B. das dopaminerge, serotonerge oder adrenerge System beeinflussen. Aber auch Pharmaka, die ihre Wirkung über Rezeptoren für GABA-, Glycin- oder Glutamat bzw. deren Derivate vermitteln.
• 8. Pharmaka zur Behandlung des Morbus Alzheimer (z.B. Tacrin) und zur Behandlung des Morbus Parkinson (z.B. Bromocriptin, L-DOPA, Carbidopa, Biperiden, Seleginil, etc.) welche Transmitterkonzentrationen an z.B. muskarinergen oder dopaminergen Substanzen, beeinflussen
• 9. Pharmaka zur Behandlung von Asthma bronchiale, welche z.B. entweder direkt bronchodilatierend oder antiinflammatorisch wirken, z.B. Salbutamol, Terbutalin, Albuterol, Theophyllin, Montelukast, Zafirlukast, Cromoglicinsäure, Ipratropiumbromid. Zu solchen Pharmaka gehören auch Antikörper die gegen bestimmte Proteine und Rezeptoren gerichtet sind.
• 10. Pharmaka zur Behandlung von Motilitätsstörungen des Magens oder Darms und Pharmaka zur Behandlung des Reizdarmsyndroms (irritable bowel syndrome) z.B. N-Butylscopolamin, Pirenzepin, Metoclopramid)
• 11. Pharmaka zur Behandlung der Adipositas, welche entweder direkt lipolytisch wirksame Rezeptoren aktivieren, z.B. β3-adrenerge Agonisten, oder zentral wirksam sind, z.B. Sibutramin, oder ähnliche Substanzen, die das Sättigungsgefühl verändern oder welche die Thermogenese beeinflussen. Dazu gehören auch Pharmaka, welche die Magenentleerung beeinflussen.
• 12. Pharmaka zur Behandlung chronischer Entzündungsvorgänge oder von Störungen des Immunsystems, z.B. Zytokine (Interferone) bei der Therapie von Virushepatitiden oder Interleukin-2 bei HIV-Infektion. Zu solchen Erkrankungen zählen auch Morbus Crohn, Colitis ulcerosa, Asthma, Psoriasis, Neurodermitits, Heuschnupfen,
• 13. Pharmaka zur Behandlung der Gestose und der Präeklampsie/Eklampsie und des HELLP-Syndroms
• 14. Pharmaka zur Behandlung von Fertilitätsstörungen oder zur Beseitigung von Zyklusstörungen bei der Frau oder zur Empfängnisverhütung
• 15. Pharmaka zur Behandlung von Herzrhythmusstörungen
• 16. Antidiabetika (Acarbose, Insulin, Troglitazone, Metformin, etc.)
• 17. Hypnotika, Antiemetika und Antiepileptika
• 18. Pharmaka zur Behandlung von Störungen des Sexuallebens, z.B. der erektilen Dysfunktion, der weiblichen sexuellen Dysfunktion, Libidomangel, Orgasmusstörungen (Phosphodiesteraseinhibitoren wie Sildenafil, Prostaglandin E1, Agonisten an Dopamin-Rezeptoren, z.B. Apomorphin, Yohimbin, Phentolamin)
• 19. Pharmaka zur Therapie von Krebserkrankungen und Chemotherapeutika, z.B. 5-Fluoruracil, Antikörper gegen Proteine (z.B. gegen HER-2), Substanzen, die Tyrosinkinasen blockieren, etc.
• 20. Pharmaka zur Behandlung von allergischen und Tumorerkrankungen, bei denen die Wirkung über die Verabreichung von CpG-Nukleotiden erzielt wird.

In particular, the effects and undesirable effects of the following drugs from the following indication areas can be diagnosed:

• 1.Antihypertensives, e.g. β-blockers (propanolol, bisprolol, etc.), diuretics (hydrochlorothiazide and other thiazide diuretics; furosemide, piretanide and other loop diuretics, chlorthalidone), α1-adrenoceptor blockers (e.g. doxazosin, prazosin receptor, angiotensin receptor, angiotensin) ), ACE inhibitors (enalapril, captopril, ramipril etc.), Ca ²⁺ channel blockers (e.g. nifedipine, verapamil, amlodipine, felodipine), clonidine, reserpine, renin inhibitors
• 2. Pharmaceuticals for the treatment of heart failure, for example β-blockers (for example propanolol, metoprolol), ACE inhibitors (for example captopril, enalapril, ramipril, etc.), angiotensin receptor blockers (for example losartan), digitalis glycosides, catecholamines, diuretics.
• 3. Pharmaceuticals for the treatment of low blood pressure or heart failure, e.g. α- and β-smpathomimetics (Effortil, adrenaline, noradrenaline, dobutamine, β-adrenoceptor blockers, ACE inhibitors, angiotensin II receptor blockers.)
• 4. Pharmaceuticals for the treatment of migraines, for example sumatriptan, rizatriptan, zolmitriptan and other agonists on serotonin receptors, β-blockers (propanolol, timolol), ergotamine and dihydroergotamine
• 5. Morphine-type analgesics (morphine, codeine, etc.)
• 6. Pharmaceuticals for the treatment of coronary heart disease such as adenosine, β-blockers (eg propanolol, acebutolol), nitrates and Ca ²⁺ channel blockers
• 7.Pharmaceuticals for the treatment of psychiatric diseases (schizophrenia, manic-depressive diseases, psychoses, depression) and addictive diseases such as alcoholism (e.g. fluoxetine, paoxetine, imipramine, desipramine, doxepin, Mianserin, trazodone, lofepramine), anxiety syndromes (diazepam, etc. ), which influence the dopaminergic, serotonergic or adrenergic system, for example. But also pharmaceuticals that mediate their effect via receptors for GABA, glycine or glutamate or their derivatives.
• 8. Pharmaceuticals for the treatment of Alzheimer's disease (eg tacrine) and for the treatment of Parkinson's disease (eg bromocriptine, L-DOPA, carbidopa, biperiden, seleginil, etc.) which influence transmitter concentrations on, for example, muscarinic or dopaminergic substances
• 9. Pharmaceuticals for the treatment of bronchial asthma which, for example, either have a direct bronchodilating or anti-inflammatory effect, for example salbutamol, terbutaline, albuterol, theophylline, montelukast, zafirlukast, cromoglicinic acid, ipratropium bromide. Such pharmaceuticals also include antibodies directed against certain proteins and receptors.
• 10. Pharmaceuticals for the treatment of gastric or intestinal motility disorders and pharmaceuticals for the treatment of irritable bowel syndrome (irritable bowel syndrome), for example N-butylscopolamine, pirenzepin, metoclopramide)
• 11. Pharmaceuticals for the treatment of obesity, which either directly activate lipolytically active receptors, for example β3-adrenergic agonists, or are centrally active, for example sibutramine, or similar substances which change the feeling of satiety or which influence thermogenesis. This includes pharmaceuticals that affect gastric emptying.
• 12. Pharmaceuticals for the treatment of chronic inflammatory processes or disorders of the immune system, for example cytokines (interferons) in the therapy of viral hepatitis or interleukin-2 for HIV infection. Such diseases also include Crohn's disease, ulcerative colitis, asthma, psoriasis, neurodermatitis, hay fever,
• 13. Pharmaceuticals for the treatment of gestosis and preeclampsia / eclampsia and HELLP syndrome
• 14. Pharmaceuticals for the treatment of fertility disorders or for the elimination of menstrual disorders or for contraception
• 15. Pharmaceuticals for the treatment of cardiac arrhythmias
• 16.Antidiabetics (acarbose, insulin, troglitazone, metformin, etc.)
• 17. Hypnotics, antiemetics and antiepileptics
• 18. Pharmaceuticals for the treatment of disorders in sexual life, for example erectile dysfunction, female sexual dysfunction, libido deficiency, orgasm disorders (phosphodiesterase inhibitors such as Sil denafil, prostaglandin E1, agonists on dopamine receptors, e.g. apomorphine, yohimbine, phentolamine)
• 19. Pharmaceuticals for the treatment of cancer and chemotherapeutic agents, e.g. 5-fluorouracil, antibodies against proteins (e.g. against HER-2), substances that block tyrosine kinases, etc.
• 20. Pharmaceuticals for the treatment of allergic and tumor diseases, in which the effect is achieved by the administration of CpG nucleotides.

Of course it is not possible in the Within the scope of the invention described here to provide evidence that all pharmacological effects are determined by the GNAS1 gene status. In contrast, examples of Pharmaceuticals with different mechanisms of action are shown, so that these findings can be generalized.

Example 1: Efficacy of sibutramine

Obese patients were treated with 15 mg sibutramine / die as part of a placebo-controlled, double-blind study over a period of 1 year. Sibutramine is a centrally effective reuptake inhibitor of norepinephrine and serotonin, both of which exert their effect via G protein-coupled receptors. Sibutramine (trade names: Reductil, Meridia) increases the feeling of satiety and facilitates weight loss as part of structured weight reduction measures (increase in physical activity, reduction in calorie intake). The clinical end goal of the study was weight loss after 1 year. The genotype-dependent comparison of placebo versus sibutramine is in 14A shown. In the placebo group, patients with the GNAS1 393 CC or CT genotype lost significantly more weight (7.0 and 4.7%) than those with the TT genotype (1.5%). The administration of sibutramine leads to a significant (p = 0.027) increase in weight loss only in carriers of the TC and TT genotype, whereas this beneficial medicinal effect was not observed in patients with the CC genotype. Therapy with sibutramine is therefore indicated primarily in patients with a TT / TC genotype, while patients with a CC genotype can also lose weight well under placebo.

In addition, the study population was genotyped for the GNAS1 G (-1211) A polymorphism ( 14B ). In the placebo group, patients with the GNAS1 (-1211) GG genotype lost significantly more weight (7.3%) than those with the AA and AG genotype (1.8 and 3.1%). The administration of sibutramine leads to a significant (p = 0.027) increase in weight loss only in carriers of the AG and AA genotype, whereas this beneficial drug effect was not observed in patients with the GG genotype. Therapy with sibutramine is indicated primarily in patients with AG / AA genotype, while patients with CC genotype can also lose weight well under placebo. The weight loss curves for each genotype of the C393T polymorphism (sibutramine versus placebo) are shown in the 15A . 15B and 15C shown.). It can be seen that only TC and TT genotypes benefit from therapy with 15 mg sibutramine per day, while CC genotypes can lose weight without medication. This also shows that it is also possible to predict the success of non-pharmacological measures for weight loss using gene changes in the GNAS gene. This includes structured weight loss programs (e.g. Optifast, Weight Watchers, other training programs), the intake of satiating source substances (CM3, BMI23) or low-calorie foods.

Example 2: Efficacy oral anti-diabetic to lose weight and improve insulin sensitivity

Polycystic Ovarian Syndrome (PCOS) is a common one endocrine disease caused by chronic anovulation and hyperandrogenism is characterized.

About 5% of all premenopausal women are affected. Most women with PCOS have an insulin resistance and thus an increased risk of a metabolic syndrome (obesity, type 2 diabetes, fat metabolism disorder and hypertension). Treatment of PCOS patients with metformin improves fertility, androgen levels, reduces insulin resistance and facilitates weight loss. Here women with PCOS syndrome were treated with metformin for 12 months. The change in body mass index (BMI) and insulin resistance was examined using the HOMA-IR method (homeostasis model assessment for insulin resistance) using GNAS genotypes of the C393T polymorphism. 16A clearly shows the greatest improvement in the HOMA-IR index in individuals with a CC genotype (2.9) in contrast to 2.2 in the TC genotype and only 0.9 in the TT genotype. CC and TC genotypes show the most significant increase in insulin sensitivity under metformin therapy, while TT genotypes benefit only slightly. The changes in the BMI (measure of body fat) behave in the same way. There is a decrease of 15.8% for the CC genotype, while the BMI for TC and TT genotypes was reduced by only 4.8 and 1.5%, respectively ( 16B ). This shows that gene changes in the GNAS gene can be used to predict the effectiveness of oral antidiabetic agents. In addition to metformin, this includes the so-called

Glitazone, tolbutamide, sulfonylureas and its derivatives and aracabose.

Example 3: Prediction the effects of catecholamines

For this purpose, human fat cells were stimulated ex vivo with isoprenaline and lipolysis was quantified based on the release of glycerol (Hauner H et al. Effects of the G-protein beta3 subunit 825T allele on adipogenesis and lipolysis in cultured human preadipocytes and adipocytes. Horm Metab Res. 2002 ; 34 (9): 475-80). This shows the strongest stimulation of lipolysis in fat cells of individuals with the GNAS 393 TT genotype ( 17 ). This is consistent with the observation of increased expression of Gαs protein in this genotype. This shows that gene changes in the GNAS gene can be used to predict the effects of catecholamines. Accordingly, this concerns substances which directly stimulate α- and / or β-adrenoceptors and also substances which induce an increased release of catecholamines. In addition, side effects can be predicted, whereby the administration of any substance causes sympathetic activation (sildenafil and similar PDE inhibitors).

Example 4: Prediction of cardiovascular Side effects with sibutramine therapy

Due to the mechanism of action (central Inhibition of the reuptake of norepinephrine and serotonin) comes it taking typical side effects while taking sibutramine such as dry mouth, trouble sleeping and constipation. dangerous however, are the cardiovascular Side effects, e.g. Increase in heart rate and blood pressure, which can lead to tachycardia and myocardial infarction. So far there is no prediction possible to with which people this occurs.

The relationship between gene changes in the GNAS gene and changes in heart rate when taking sibutramine versus placebo was examined ( 18 ). You can see at 393TC- ( 18B ) and TT genotypes ( 18C ) a heart rate significantly increased by sibutramine (versus placebo) by 10 (TC) up to 20 beats per minute. This effect does not occur with CC genotypes ( 18A ). Furthermore, a significant increase in blood pressure was observed in 393 TT genotypes during therapy with sibutramine ( 19C ) at CC- ( 19A ) and TC genotypes ( 19B ) does not occur.

The use of gene changes is therefore suitable for identifying patients in whom Cardiovascular side effects occur during therapy with medication. These effects can directly caused by such drugs stimulating receptors, mediates the gas cause a vasoconstriction or an increase in heart rate. This includes for example sibutramine, triptans and noradrenaline / serotonin reuptake inhibitors. Furthermore belong medicines that inhibit the breakdown of catecholamines (MAO inhibitors) and tricyclic antidepressants. It also includes medication that a Cause drop in blood pressure and reflex activation of Induce sympathetic nervous system (e.g. sildenafil and other inhibitors of Phosphodiesterases, nitrates).

figure description

1 : The G protein cycle NA, norepinephrine; β1, β1 adrenergic receptor; Gαs, stimulatory α subunit, AC, adenylyl cyclase; PKA, protein kinase A; Details on G protein activation in the text.

2 : The Gαs signal path. The diagram shows how the demonstrated cAMP pathway is linked to a variety of signal transduction components, including ion channels, transcription factors and metabolic enzymes. AC, adenylyl cyclase; PKA, protein kinase A; PDE, phosphodiesterase; L-Ca ⁺⁺ channel, L-type Ca ²⁺ channel; CNGC, cyclic nucleotide-gated channel; PhosK, phosphorylase kinase; GlyPhos, Glycogen Phosphorylase; CREB, cAMP response element-binding protein; EPAC, the cAMP and AMP regulated exchange factor for Rap1; Rap1, a small GTPase; MAPK, mitogen-activated protein kinase; Raf1 and B-Raf, MAP kinase kinase kinases; MEK, MAPK / ERK kinase; MEKK, MAPK / ERK kinase kinase; GRK, G protein receptor kinase; RGS, "regulators of G protein signaling"; βAR, β-adrenergic receptor (from: Neves at al, G protein pathways. Science. 2002 May 31; 296 (5573): 1636-9).

3A : Intron / exon structure of human GNAS1

3B : Using different promoters P1, P2 or P3 in GNAS1 leads to the creation of different gene products (pre-mRNAS).

4 : Splice variants from Gαs. The use of different splice sites leads to the inclusion of a CAG triplet (serine) from exon 4. The exclusion of exon 3 leads to the formation of Gαs-short.

5 : Structure of GNAS1 and location of new and known polymorphisms. The T393C polymorphism in exon 5 and the polymorphisms G (-1211) A and T (-839G) before exon 1 are shown. ATG denotes the start codon in exon 1. The numbering of the promoter / enhancer region polymorphisms is carried out in this way that the nucleotide A of the start codon ATG is assigned the value +1.

6A : Relative expression of Gαs mRNA (ratio Gαs / β-actin) in human adipose tissue depending on genotypes of the C393T polymorphism. An increased expression is found in T allele carriers by Gαs.

6B : Relative expression of Gαs mRNA (ratio Gαs / β-actin) in human adipose tissue depending on genotypes of the C393T polymorphism. In TT genotypes there is an increased expression of Gαs.

6C : Relative expression of Gαs mRNA (ratio GαsGAPDH) in human adipose tissue depending on genotypes of the G (-1211) A polymorphism. A allele carriers show an increased expression of Gαs.

6D : Relative expression of Gαs mRNA (ratio Gαs / GAPDH) in human heart tissue depending on genotypes of the G (-1211) A polymorphism. A allele carriers show an increased expression of Gαs.

6E : Relative expression of gas mRNA (ratio Gαs / GAPDH) in human urinary bladder tissue depending on genotypes of the G (-1211) A polymorphism. A allele carriers show an increased expression of Gαs.

7A : GNAS1 T393C status and disease progression in patients with CLL; ED = initial diagnosis.

7B : GNAS1 G (-1211) A status and disease progression in patients with CLL; ED = initial diagnosis.

8A : Combined analysis of GNAS1 T393C status and GNAS1 G (-1211) A status for disease progression in patients with CLL; ED = initial diagnosis.

8B : Combined analysis of GNAS1 (-839) TT status and GNAS1 G (-1211) A status for disease progression in patients with CLL; ED = initial diagnosis.

9 : GNAS1 G (-1211) A status and disease progression in patients with CLL Binet stage A; ED = initial diagnosis.

10A : GNAS1 T393C status and time to metastasis in patients with bladder cancer.

10B : GNAS1 T393C status and time to tumor progression in patients with bladder cancer.

10C : GNAS1 T393C status and survival in patients with bladder cancer.

11A : GNAS1 T393C status and time to tumor progression in patients with renal cell carcinoma.

11B : GNAS1 T393C status and survival in patients with renal cell carcinoma.

12 : GNAS A (-1211) G status and survival of patients with acute myeloid leukemia. All patients had received chemotherapy and / or bone marrow transplantation.

The data suggest that besides survival also the response to chemotherapy drugs or bone marrow transplantation can be predicted.

13 : Gene expression in lymphocytes from patients with CLL depending on the GNAS G (-1211) A genotype CDNA was hybridized on Affymetrix gene chips and the strength of the hybridization signal was quantified. The method for quantitative gene analysis is known to the person skilled in the art. A statistically significant, stronger expression of apoptosis-relevant genes in AA / AG genotypes compared to the GG genotype can be seen.

14A : Dependence of the effect of sibutramine on placebo in the context of therapy for obesity. Placebo and sibutramine (15 mg / day) were administered over 1 year. The weight loss after 1 year (%) is shown depending on the GNAS1 C393T genotype. Only patients with TT and TC genotypes benefit significantly (p = 0.027) from sibutramine therapy compared to placebo. In the CC genotype, there was no significant effect of the drug compared to placebo.

14B : Dependence of the effect of sibutramine on placebo in the context of therapy for obesity. Placebo and sibutramine (15 mg / day) were administered over 1 year. The weight loss after 1 year (%) is shown depending on the GNAS1 G (1211) A genotype. Only patients with AA and GA genotypes benefit significantly (p = 0.027) from sibutramine therapy compared to placebo. In the GG genotype, there was no significant effect of the drug compared to placebo.

15 : Time course of weight changes under placebo or sibutramine for the GNAS 393 CC genotype (A), the TC genotype (B) and the TT genotype (C). It can be seen that only TC and TT genotypes benefit from therapy with 15 mg sibutramine per day, while CC genotypes can lose weight without medication.

16 : Influence of therapy with an oral anti-diabetic (insulin sensitizer; metformin) on insulin resistance (A) and body mass index (BMI; B) in women with polycystic ovaries. Therapy with metformin continued for 12 months. A .: The change in the HOMA-IR index is shown as a measure of insulin resistance. The strongest decrease is found in GNAS 393 CC and TC genotypes. B. Changing the Body Mass Index (BMI)

17 : Isoprenaline-stimulated lipolysis in human fat cells depending on GNAS C393T status. Increased lipolysis, measured as free, is found in fat cells of people with a TT genotype setting of glycerol.

18 : Use of a gene modification in the human GNAS1 gene to predict an increase in heart rate during therapy with sibutramine. The effects are shown depending on genotypes of the C393T polymorphism.

19 : Use of a gene modification in the human GNAS1 gene to predict an increase in diastolic blood pressure during therapy with sibutramine.

The time-dependent changes are shown of diastolic blood pressure (DBP) over time of genotypes of the C393T polymorphism.

Claims (7)

Use of a gene modification in the human GNAS1 gene to predict disease risks Use of a gene modification in the human GNAS1 gene to predict disease courses Use of a gene modification in the human GNAS1 gene to predict drug effects Use according to claims 1-3 provided that the T393C polymorphism is examined. Use according to claims 1-3 provided that the G (-1211) A polymorphism is examined. Use according to claims 1-3 provided that the G (-839) T polymorphism is examined. Use according to claim that a combination of these Polymorphisms is examined.