EP1392854A2

EP1392854A2 - Dna chip for performing causal diagnosis of hypertension

Info

Publication number: EP1392854A2
Application number: EP01985807A
Authority: EP
Inventors: Rudolf Wiesner
Original assignee: Individual
Current assignee: Individual
Priority date: 2000-12-29
Filing date: 2001-12-28
Publication date: 2004-03-03
Also published as: US20040053239A1; DE10065666A1; DE10065666B4; WO2002053769A3; WO2002053769A2

Abstract

The invention relates to a DNA chip for performing causal diagnosis of hypertension (hypertonia), which enables an early diagnosis and a highly effective therapy that is tailored to each patient and which has the fewest possible side effects. The invention also relates to a DNA chip for developing novel antihypertensive pharmaceutical agents and for stratifying patients in early phases of the clinical testing of novel pharmaceutical agents of this type.

Description

DNA chip for the causal diagnosis of high blood pressure

The present invention relates to a DNA chip for the causal diagnosis of high blood pressure (hypertension), which enables an early diagnosis and a highly effective therapy which is tailored to the respective patient and as free of side effects as possible. The present invention further relates to a DNA chip for the development of new blood pressure-lowering pharmaceuticals and for the stratification of patients in the early phases of clinical testing of such new pharmaceuticals.

Around 15 million people in Germany suffer from high blood pressure, of which about 5 million are undetected (German Hypertension League, 2000). Hypertension, along with its complications (arteriosclerosis, heart attack, stroke, cardiac hypertrophy and heart failure) is by far the most common cause of illness and death in all western industrialized nations, even before malignancies (cancer). Hypertension is based on excessive constriction of the blood vessels or inadequate fluid excretion by the kidney. Both in the regulation of muscle tone of the smooth muscle in blood vessels and the vessel size as well as to the liquid excretion by the kidney, a plurality of central nervous ^mechanisms' and hormone systems are involved. These control and regulate blood pressure, which physiologically increases during physical work, fear, stress, arousal etc. Derailment of one or more of these systems ultimately leads to high blood pressure. In nine out of ten cases, the actual causes of high blood pressure are not known (essential hypertension). However, a genetic predisposition (predisposition) through mutation of genes coding for proteins that are involved in blood pressure regulating systems, sometimes only in combination with external factors (stress, smoking, obesity, lack of physical activity, malnutrition) is highly probable.

Hypertension can currently only be diagnosed symptomatically, routinely by simple measurement according to Riva-Rocci (arm cuff, stethoscope) or in intensive care blood-invasive via arterial catheter. This means that when diagnosed, the disease and possibly negative side effects have already been manifested. The diagnosis of limit hypertension is also not unproblematic, since physiologically large fluctuations in blood pressure can already be observed (e.g. daily rhythm) and many other factors can vary blood pressure (e.g. fear of the examination; before the diagnosis; "white coat effect") There is no early diagnosis, that is to say the possibility of diagnosing the onset of the disease before a significant increase in blood pressure, and it is completely unsatisfactory to have only one physical method of diagnosis, ie pathological, in the case of a nolks disease with such a large number of people affected Essential clinical hypertension can only be treated symptomatically by trying out different antihypertensive agents as monotherapy or very often in combination therapy ("discontinue") because the primary causal cause is n is not known. However, some of the patients remain resistant to therapy, and even with successfully treated patients, the findings often deteriorate again with increasing age. It is therefore urgently necessary to establish an early detection method that already recognizes the development of hypertension even before the physically measurable blood pressure has increased significantly in order to prevent anatomical changes in the blood vessels, which are then irreversible, and further consequential damage. Second, it would be desirable to know the primary cause of hypertension in each patient.

DΝA chip technology is a very efficient method, among other things for the detection of DNA polymorphisms. This detection with the aid of DNA chip technology is used by an already existing method for the causal diagnosis of high blood pressure, ie differences in the DNA sequence of genes between different individuals are detected, which code for proteins that are responsible for mechanisms regulating blood pressure. However, this procedure has the disadvantage that only in a few cases is essential hypertension monogenic, most cases will be due to a combination of mutations, each of which has little predictive value. Another disadvantage is that, for ethical reasons, DNA analysis with regard to the predisposition to a serious illness such as high blood pressure will not be possible. Nevertheless, it is urgently desirable to clarify with each patient which of the many possible blood pressure regulating mechanisms is disturbed and is therefore responsible for their individual high pressure. Only the causal diagnosis of the causes that lead to the high blood pressure of each individual patient, i.e. the hormonal system or central nervous system Mechanism or a corresponding combination that has been derailed in the individual case will enable a tailor-made therapy that is oriented towards the respective patient. Even undesirable side effects of unsuitable medication can be avoided as far as possible. This would significantly improve the healing success of this major widespread disease and thus significantly reduce morbidity and mortality, but also treatment costs.

The present invention is therefore based on the object of providing means for causally diagnosing high blood pressure. Another object of the present invention is to provide a method for the early detection of hypertension. Another object of the present invention is to provide a method for dividing hypertension patients into subclasses, e.g. also for the stratification of patients in the early phases of the clinical trial of new pharmaceuticals.

The tasks are solved by the subject defined in the patent claims.

The term "subclass" used here means a group of patients with high blood pressure, the causal cause of the high blood pressure being due to a disruption of the same blood pressure regulating system. The patients in a subclass therefore react immediately to a blood pressure lowering drug.

The term "blood pressure regulating systems" used here denotes central nervous mechanisms or hormonal systems which are involved in the regulation of the contraction state of the smooth muscles of the blood vessels (vessel width) or in the absorption capacity of the renal epithelium. Disrupted blood pressure regulating systems are those that are causally responsible for high blood pressure.

The term "hypertension" as used herein means any type of stage I, II, III and IN hypertension. Depending on age, there is usually already hypertension with a systolic value above 140 mm Hg and a diastolic value from 90 to 94 mm Hg. Hypertension is usually also present when there is only an increase in the diastolic value, whereby the following classifications can be made: mild hypertension at 95 to 104 mm Hg, moderate hypertension at 105 to 114 mm Hg, severe hypertension at over 115 mm Hg. Switching genes on and off is the basis of all biological processes and also an extremely sensitive response to changes in external conditions. DNA chips will therefore revolutionize biomedicine and molecular biology in the next few years. With the extraction of the RNA from a biological sample, the action of labeled cDNA or the RNA itself on a DNA chip (hybridization) and its analysis, a large amount of information about the gene expression profile and thus about the state of the cells in the cell is available within a very short time biological sample possible under changed conditions. The technology based on the hybridization of nucleic acids has the advantage of extremely high specificity, sensitivity and relatively easy feasibility, in contrast to, for example, the analysis of protein patterns.

If the switching on and off of genes in blood vessels or kidney tubule cells occurs in a non-physiological manner, it can be the cause of high blood pressure. The background to this is that all intracellular signaling cascades involved in the regulation of vascular tone or renal epithelial transport either directly or indirectly influence transcription factors and thus the activity of genes. If the expression of one or more genes coding for proteins which are involved in these blood pressure regulating systems is disturbed, or if these genes carry mutations which impair the function of the corresponding proteins, this affects the blood pressure. The switching on and off of genes in blood vessels or kidney tubule cells in a non-physiological manner can also be used as a measurable sign of the disrupted blood pressure regulating mechanisms (surrogate marker).

One aspect of the present invention therefore consists in the provision of a DNA chip for the causal diagnosis of hypertension, comprising at least two different nucleic acids, each coding for a polypeptide, the polypeptides being involved in different blood pressure regulating systems, and the nucleic acids as an indicator for at least two disturbed blood pressure regulating systems are used, and optionally also include one or more nucleic acids coding for polypeptides that are expressed in each cell and are part of the basic equipment of a cell (so-called housekeeping genes) for normalizing the signals obtained. A DNA chip is preferred, wherein one, two, three, four, five, six or more nucleic acids of at least two blood pressure regulating systems are contained. Also preferred is a DNA chip with at least one each Nucleic acid, coding for one polypeptide each, of three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen or seventeen blood pressure regulating systems.

The nucleic acids used on the DNA chip code for peptides that are involved in biochemical processes that are particularly involved in one or more of the following blood pressure regulating systems or also in other blood pressure regulating systems to be discovered in the future:

The sympathetic-adrenergic nervous system and the adrenergic system of the adrenal medulla • The parasympathetic nervous system The renin-angiotensin system The aldosterone system

The pituitary adiuretin (vasopressin) system The atrial natriuretic peptide system • The renal kallikrein-kinin system The NO (nitric oxide) system

The EDHF system (endothelial derived hyperpolarization factor) The prostaglandin system The endothelin system • The PTHrP system (parathormone-related peptide) The histamine system The serotonin system «The purinergic system • Local growth factors and / or • The calcium Smooth vascular muscle cell or heart household

The nucleic acid used on the DNA chip can be any nucleic acid that is known to encode a peptide that is involved in blood pressure regulating systems or that is expressed or expressed at an increased rate in hypertension. In particular, nucleic acids selected from the following nucleic acids are used for the DNA chip according to the invention (the GenBank accession number is given in each case in the brackets):

smooth muscle α-actin (J05192); cardiac α-actin (AF053356); skeletal α-actin (AF053356); adrenergic receptor β 1 (J03019); adrenergic receptor ß 2 (M15169), ß-adrenergic Receptor kinase II (M80776); Adrenomedullin (D14874); Angiotensin II receptor type I (AT 1) (D13814); Angiotensin converting enzyme (ACE) (J04144); Arginine vasopressin (AVP) (M25647); Arginine vasopressin receptor (VI) (L25615); Atrial natriuretic factor (M30262); Brain natriuretic peptides (M25296); Ca ²⁺ sarcoplasmic reticulum ATPase (SERCA) (M63603); Calcium Release Channel (Ryanodine Receptor II) (J05200); Collagen α-1 (I) (AF017178); Collagen α-1 (III) (Ml 1134); Collagen α-1 (IV) (NM_001845); Collagen α-2 (IV) (M33653); Type V collagen (BC008760); Connexin 43 (Cx 43) (M65188); Cytochrome P 450 CYP11-B2 (D13752); Cytocl rom P 450 CYP11-B1 (aldosterone synthase) (NM_000497); Desmin (U59167); Elastin (tropoelastin) (M36860); Endothelin I (ET-I) (NM J01955); Endothelin m (ET DT) (J05081); Endothelin receptor (ETA) (L06622); Endothelin receptor (ETB) (L06623); soluble E-selectin (D38257); Fibroblast growth factor (bFGF) (M27968); Fibronectin (X02761); G protein, α subunit of inhibitory (Gl-α) (BC014627); G protein coupled receptor kinase II (L16862); Guanylate cyclase, αl subunit of soluble (X66534); Guanylate cyclase, β1 subunit of soluble (X66533); Heat shock protein 70 (hsp 70) (L12723); Insulin Like Growth Factor 1 (IGFl) (M29644); hisulin like growth factor 1 receptor (NM_000875); Interleukin 6 (M54894); Intercellular Adhesion Molecule (ICAM-I) (J03132); Cardiotrophin 1 (CT 1) (BC012939); Laminin (M20206); Medium chain acyl coenzyme A dehydrogenase (MCAD) (M16827); Mitochondrial DNA (J01415); Monocyte chemoattractant protein I (MCP I) (D29984); Natriuretic peptide receptor A (XI 5357); Myosin, heavy α chain (D00943); Myosin, heavy ß chain (NM_000257); Myosin light chain 2 (AB046614); Sodium potassium (Na + / K +) ATPase alpha-1 subunit (U16798); Sodium proton exchanger (NHE-I) (S68616); Nerve growth factor (NGF) (M57399); NO synthase endothelial (ENOS) (M95296); NO synthase inducible (LNOS) (AF051164); Ornithine decarboxylase (ODC) (MI 6650); Osteopontin (J04765); Plasminogen activator inhibitor (PAI-I) (MI 6006); Parathyroid hormone-like protein (PTH / parathyroid hormone related protein) (M17183); Phosphodiesterase (L20965); Phospholamban (M63603); Platelet derived growth factor (PDGF-alpha) (L20965); Platelet derived growth factor (PDGF-beta) (X98706); Platelet derived growth factor PDGF-alpha receptor (NM_006206); Platelet derived growth factor PDGF-beta receptor (L20965); Prostacyclin synthetase (PGI-JI synthetase) (D38145); S-100 beta protein (P04271); Superoxide dismutase (K00065); Thrombospondin (NM_003246); Tissue plasminogen activator (tPA) (M15518); Transforming growth factor β (M38449); Tropomyosin, α-skeletal (M19713); Troponin I, cardiac (M38449); Troponin I, skeletal (J04760); Vascular Cell Adhesion Molecule (VCAM-I) (X53051); Vascular endothelial growth factor / permeability factor (VEGF) (AY047581); Voltage-gated-K + channel (KV1.2) (XM_010737); Voltage-gated-K + channel (KV1.5) (M83254); Voltage-Gated-K + Channel ß subunit (X83127).

A DNA chip according to the invention with the nucleic acids listed below is particularly preferred, the nucleic acids encoding polypeptides which are involved in the respectively mentioned blood pressure regulating systems or which are differentially regulated in the event of disorders of these systems: cytochrome P 450 CYP11-B1 (aldosterone synthase) and cytochrome P 450 CYP11-B2, involved in the aldosterone system, angiotensin LT receptor type I (AT 1), angiotensin converting enzyme (ACE), intercellular adhesion molecule (ICAM-I), vascular cell adhesion molecule (VCAM-I), vascular endothelial growth factor / permeability factor (VEGF), monocyte chemoattractant protein I (MCP I), plasminogen activator inhibitor (PAI - I), transforming growth factor ß and fibroblast growth factor (FGF-II), involved in the renin-angiotensin system, adrenergic receptor ßl , adrenergic receptor ß2, ß-adrenergic receptor kinase II and G-protein coupled receptor kinase II (GRK 2), involved in the sympathetic / a drenergen system, and inducible NO synthase (INOS), interleukin 6 and monocyte chemoattractant protein I (MCP-I), involved in the Ca ²⁺ household.

For the purposes of the present invention, “proteins involved in blood pressure regulating systems” on the one hand means that the polypeptides are involved in biochemical processes in blood vessels, in the heart muscle or in the renal tubular epithelium, which lead to hypertension in the event of a malfunction. On the other hand, it also means those proteins whose expression is down-regulated or up-regulated due to high blood pressure (surrogate marker. Therefore, in tissue samples from hypertensive patients (e.g. white blood cells, blood-rich tissue, heart muscle tissue, kidney tissue), the expression of genes can also be changed in their products do not causally cause high blood pressure, such genes can also be indicative of the blood pressure regulating systems that are disturbed in the respective patient.

The nucleic acids which may be used to normalize the signals on the DNA chip according to the invention can be those which are known to be expressed in every cell and are part of the basic equipment of the cells. The genes that code for these nucleic acids are usually also referred to as housekeeping genes. An exemplary selection for such nucleic acids is given below: cellular β-actin, cyclophilin, glyceraldehyde phosphate dehydrogenase (GAPDH), glycero-3-phosphate dehydrogenase (GPDH), histones, phospholipase, ribosomal proteins, tubulin, Ubiquitin. If a tissue rich in blood vessels is used for diagnosis, smooth muscular housekeeping genes such as the caldesmon or the calponin can be used to standardize the signals.

The nucleic acid sequences of the nucleic acids used on the DNA chip can be taken from the generally accessible databases (e.g. GeneBank, EMBL) and selected according to the usual criteria. Preference is given to sequences which are unique, have no GC-rich regions, do not allow intramolecular hybridization and do not form loops. A single-stranded nucleic acid is preferably applied to the DNA chip according to the invention. Both the sense and the antisense strand can be used. However, a double-stranded nucleic acid can also be applied to the DNA chip according to the invention. There is no upper or lower limit on the length of the nucleic acids, as long as the length is sufficient for a specific hybridization. The nucleic acids can also be DNA, RNA or hybrid molecules. The production of such nucleic acids is state of the art and it is possible to apply and immobilize oligonucleotides, PCR products, cloned DNA or cDNA fragments or cRNAs after in vitro transcription (natural or modified) using standard techniques. In the case of single-stranded nucleic acids, the sequence complementary to the cDNA (when using cDNA from the biological sample) or the sequence complementary to the RNA (when using RNA from the biological sample) is preferably applied.

The carrier on which the nucleic acids are applied can be any carrier which is usually used for DNA arrays or DNA chips. The methods for applying and immobilizing the nucleic acids are also prior art and known to the person skilled in the art.

The DNA chip according to the invention can be used, for example, for the sensitive early detection of developing hypertension, even before hypertension manifests itself clinically. The DNA chip can also be used to divide the patients into sub-nipples. The genes on the DNA chip can be used to detect the blood pressure regulatory systems that are disrupted in the respective hypertension patients. The patient subgroups can then be treated with individually suitable active ingredients or combinations of active ingredients, which are not only the symptom of high blood pressure eliminated, but also counteracts the causal cause of high blood pressure. Furthermore, the DNA chip according to the invention can be used for the development of new blood pressure-lowering pharmaceuticals or for the stratification of patients in the early phases of clinical testing of such new pharmaceuticals. One aspect of the present invention therefore relates to the use of the DNA chip according to the invention for the diagnosis of high blood pressure, also for early diagnosis, for dividing high blood pressure patients into subnibs with different causes that trigger high blood pressure and for monitoring the therapy of high blood pressure.

For this purpose, blood or tissue samples are taken from the patients to be examined. RNA can be isolated from these patient samples using standard techniques and used either as total RNA or after fractionation as PolyA ⁺ RNA. The RNA can be rewritten into cDNA with the aid of reverse transcriptase and thereby provided with a label, for example with a fluorescent group or an isotope etc. Starting from the cDNA, PCR products can also first be produced and labeled, which are then hybridized with the DNA chip. The RNA can also be used directly or unlabeled for hybridization on the DNA chip. After hybridization of the sample nucleic acids with the target nucleic acids on the support, subsequent washing steps etc., the binding of sample nucleic acids with the target nucleic acids on the support is analyzed. This can be done by optical methods in the case of fluorescence labeling, by autoradiography or by any other method that is able to recognize a successful hybridization of sample nucleic acids with target nucleic acids.

The tissue removed from the patient can be used as a biopsy, e.g. from the blood vessel-rich mouth-nose area or from any other blood vessel-rich but relatively low-risk tissue, or from the heart muscle, e.g. as part of cardiac catheterization to determine contractility and cardiac output. Lymphocytes (white blood cells) can also be taken from the patient's blood.

According to the invention, the patients can be divided into subgroups based on their gene expression profile (gene activity: ON-OFF; high-low), that is, in sub-nipples in which one or more of the hormonal or central nervous pathways described above (blood pressure regulating systems) are disturbed. For example, in hypertension patients with hyperaldosteronism (aldosterone system), the genes for cytochrome P 450 CYP11-B1 (aldosterone synthase) and for cytochrome P 450 CYP11-B2 are activated in white blood cells. These patients should be treated with an aldosterone antagonist, such as spirolactone.

For example, in hypertension patients with increased circulating angiotensin JJ (increased plasma renin activity; renin-angiotensin system), on the one hand the expression of the angiotensin II receptor (type 1, ATI) in white blood cells, on the other hand the expression of the angiotensin converting enzyme, of the intercellular adhesion molecule (ICAM-I), the nascular adhesion molecule (NCAM-I), the nascular endothelial growth factor / permeability factor (VEGF), the monocyte chemoattractant protein (MCP I), the PAI-1, the transforming growth factor ß and of fibroblast growth factor (FGF-IJ) in blood vessels or the expression of brain natriuretic peptide (BNP) in the heart muscle is increased. These patients should be treated with an angiotensin II receptor antagonist, e.g. with irbesartan, or an angiotensin converting enzyme (ACE) inhibitor, e.g. with enalapril or captopril.

For example, in hypertension patients with a stimulated sympathetic-adrenergic system (sympathetic-adrenergic nervous system and the adrenergic system of the adrenal medulla), on the one hand the expression of the G protein-coupled receptor kinase II in white blood cells, on the other hand the expression of the adrenergic receptors (ßl and ß2) and the ß-adrenergic receptor kinase in blood vessels or the expression of omithine decarboxylase (ODC) in the heart muscle is increased. These patients should be treated with a beta-adrenergic receptor blocker, e.g. with atenolol or bisoprolol.

For example, in hypertension patients with disturbed Ca ^ balance of the vascular muscle cells (calcium balance of the smooth vascular muscle cell or the heart), the expression of interleukin 6 in blood vessels is reduced. These patients should be treated with a Ca ⁺⁺ channel blocker, eg with verapamil, nifedipine or amlodipine.

For example, in hypertension patients with a disturbed NO system, the expression in blood vessels on the one hand of the endothelial NO synthase (eNOS) is reduced, and on the other hand the angiotensin converting enzyme (ACE) or type I collagen are increased. These patients should be treated with NO donors, such as nitrates. The following examples are illustrative only and in no way limit the scope of the invention.

A submucosal biopsy is taken from the mouth or the lower turbinate (tissue rich in blood vessels), from the heart or from white blood cells

Both the lower lip and the lower turbinate are suitable for biopsy removal, as they are easily accessible, spraying with a local anesthetic is sufficient for anesthesia, the tissue is very rich in arterioles and nenoles, but does not have any large vessels or nerves and the removal defects are not visible and heal quickly.

1st lower lip

In 8 patients, after application of 1% xylocaine spray with epinephrine additive (Xylocaine 1% ®, Astra Chemie), the lower lip was raised and a 2 - 4 mm long superficial mucosal incision was made at the gingiva with a size 15 scalpel. With the cutting forceps according to Grünwald (Storz), a tissue fragment of approx. 250 mg weight was taken through the cut submucously and snap-frozen in liquid nitrogen. As a rule, a further treatment of the removal defect was not necessary, the re-epithelization was completed after 2 to 3 days. Occasionally there was increased bleeding perioperatively, so that the defect was closed with a single button seam with resorbable suture material (Vicryl 3/0, Ethicon).

2. Lower turbinate In 10 patients, the nasal mucosa was first sprayed with Pantocam Privin (1/1, Pantoprivin 2% ®, Astra Chemie). After surface anesthesia, the cutting forceps according to Grünwald (Storz) were used to remove a tissue fragment measuring approx. 250 mg submucously from the lateral part of the head of the inferior concha. The tissue was immediately snap frozen in liquid nitrogen. Bipolar coagulation occurred in cases of heavy bleeding. A tissue suture was not necessary. The defect healed in 3-4 days. 3. Heart muscle biopsy

A biopsy of approximately 500 mg was taken from the free chamber wall with a punch in three patients as part of a left-heart catheterization to diagnose left ventricular function. The tissue was immediately snap frozen in liquid nitrogen.

4. White blood cells:

100 ml of blood were drawn from 15 patients and subfractionated in a conventional manner into erythrocytes, thrombocytes and lymphocytes (Lymphoprep, Nycomed, Pharma AS or by density gradient centrifugation on a Ficoll / Metrizoat gradient). The lymphocyte fraction was immediately processed further.

5. RNA extraction:

The total RNA was obtained from the tissue samples or the white blood cells using various extraction methods with the aid of commercially available “kits”. The yield was about 0.5 mg RNA per g heart muscle, 0.1 mg RNA per g submucosal tissue and about 0.1 mg RNA per lymphocyte preparation. The RNA was examined with conventional methods (blots, hybridization with radioactive or non-radioactive labeled probes) with regard to the expression of the genes involved.

6. Gene Expression Analysis Using a DNA Array:

RNA from tissue samples from 5 hypertensives or a reference RNA sample, pooled from the individual RNA samples from tissue from 5 healthy volunteers, was analyzed with the help of oligo-dT primers in the presence of Cy5 or Cy3 dUTP ( Amersham) reverse transcribed in cDNA and thereby fluorescence-labeled. The cDNA samples (subject or reference) were each mixed in the same ratio and hybridized with oligonucleotide arrays or cDNA arrays, these were washed after hybridization and the fluorescence signals were evaluated with a DNA array reader.

Claims

claims

1. Device comprising a support to which at least two different nucleic acids coding for one polypeptide each have been applied, the polypeptides being involved in different blood pressure regulating systems, and the nucleic acids serving as an indicator for at least two disturbed blood pressure regulating systems, and optionally further comprising one or more nucleic acids, coding for polypeptides, which are expressed in each cell and belong to the basic equipment of a cell.

2. Device according spoke 1, wherein the blood pressure regulating systems are selected from the sympathetic-adrenergic nervous system and adrenergic system of the adrenal medulla, parasympathetic nervous system, renin-angiotensin system, aldosterone system, pituitary adiuretin (vasopressin) system, atrial natriuretic - peptide system, renal kallikrein-kinin system, NO (nitrogen oxide) system, EDHF system (endothelial derived hyperpolarization factor), prostaglandin system, endothelin system,

PTHrP system (parathormone related peptide), histamine system, serotonin system, purinergen system, local growth factors, and calcium balance of the smooth vascular muscle cell or the heart.

3. Device according spoke 1 or 2, wherein the nucleic acids are selected from smooth muscle α-actin (J05192); cardiac α-actin (AF053356); skeletal α-actin (AF053356); adrenergic receptor β 1 (J03019); adrenergic receptor ß 2 (M15169), ß-adrenergic receptor kinase II (M80776); Adrenomedullin (D14874); Angiotensin II receptor type I (AT 1) (D13814); Angiotensin converting enzyme (ACE) (J04144); Arginine vasopressin (AVP) (M25647); Arginine vasopressin receptor (VI) (L25615); atrial

Natriuretic factor (M30262); Brain natriuretic peptides (M25296); Ca ²⁺ sarcoplasmic reticulum ATPase (SERCA) (M63603); Calcium Release Channel (Ryanodine Receptor II) (J05200); Collagen α-1 (I) (AF017178); Collagen α-1 (DI) (MI 1134); Collagen α-1 (IV) (NM XH845); Collagen α-2 (IN) (M33653); Collagen type N (BC008760); Connexin 43 (Cx 43) (M65188); Cytochrome P 450 CYP11-B2 (D13752);

Cytochrome P 450 CYP11-B1 (aldosterone synthase) (ΝMJ300497); Desmin (U59167); Elastin (tropoelastin) (M36860); Endothelin I (ET-I) (NM_001955); Endothelin DI (ET III) (J05081); Endothelin receptor (ETA) (L06622); Endothelin receptor (ETB) (L06623); soluble E-selectin (D38257); Fibroblast growth factor (bFGF) (M27968); fibronectin (X02761); G protein, α subunit of inhibitory (Gl-α) (BC014627); G protein coupled receptor kinase II (L16862); Guanylate cyclase, αl subunit of soluble (X66534); Guanylate cyclase, β1 subunit of soluble (X66533); Heat shock protein 70 (hsp 70) (L12723); Insulin Like Growth Factor 1 (IGFl) (M29644); Insulin Like Growth Factor 1 Receptor (NM_000875); Interleukin 6 (M54894); Intercellular adhesion molecule

(ICAM-I) (J03132); Cardiotrophin 1 (CT 1) (BC012939); Laminin (M20206); Medium Chain Acyl Coenzyme A Dehydrogenase (MCAD) (MI 6827); Mitochondrial DNA (J01415); Monocyte chemoattractant protein I (MCP I) (D29984); Natriuretic peptide receptor A (XI 5357); Myosin, heavy α chain (D00943); Myosin, heavy ß chain (NM_000257); Myosin light chain 2 (AB046614); Sodium potassium (Na + / K +) ATPase alpha-1 subunit (U16798); Sodium proton exchanger (NHE-I) (S68616); Nerve growth factor (NGF) (M57399); NO synthase endothelial (ENOS) (M95296); NO synthase inducible (TNOS) (AF051164); Omithine decarboxylase (ODC) (M16650) osteopontin (J04765); Plasminogen activator inhibitor (PAI - I) (M16006) Parathyroid hormone-like protein (PTH / parathyroid hormone related protein) (Ml 7183)

Phosphodiesterase (L20965); Phospholamban (M63603); Platelet derived growth factor (PDGF-alpha) (L20965); Platelet derived growth factor (PDGF-beta) (X98706); Platelet derived growth factor PDGF-alpha receptor (NM_006206); Platelet derived growth factor PDGF-beta receptor (L20965); Prostacyclin synthetase (PGI-H synthetase) (D38145); S-100 beta protein (P04271); Superoxide dismutase (K00065); Thrombospondin (NM_003246);

Tissue plasminogen activator (tPA) (M15518); Transforming growth factor β (M38449); Tropomyosin, α-skeletal (M19713); Troponin I, cardiac (M38449); Troponin I, skeletal (J04760); Nasular Cell Adhesion Molecule (NCAM-I) (X53051); Nascular endothelial growth factor / permeability factor (VEGF) (AY047581); Noltage-Gated-K + Channel (KV1.2) (XM_010737); Voltage-gated-K + channel (KV1.5) (M83254); Voltage-

Gated-K + Channel ß subunit (X83127).

4. Use of the device according to one of claims 1 to 3 for the development of new blood pressure-lowering pharmaceuticals, for stratifying patients in the early phases of clinical testing of such new pharmaceuticals, for the causal diagnosis of high blood pressure, for

Early diagnosis of high blood pressure, for dividing high blood pressure patients into subgroups with different causes that trigger high blood pressure, or for monitoring the therapy for high blood pressure.