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WO1999053052A2 - Lignees cellulaires srebp-2-deficientes - Google Patents

Lignees cellulaires srebp-2-deficientes Download PDF

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Publication number
WO1999053052A2
WO1999053052A2 PCT/DE1999/001143 DE9901143W WO9953052A2 WO 1999053052 A2 WO1999053052 A2 WO 1999053052A2 DE 9901143 W DE9901143 W DE 9901143W WO 9953052 A2 WO9953052 A2 WO 9953052A2
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WO
WIPO (PCT)
Prior art keywords
nucleic acid
acid sequence
seq
srebp
listed
Prior art date
Application number
PCT/DE1999/001143
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German (de)
English (en)
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WO1999053052A3 (fr
Inventor
Wilhelm Krone
Dirk Müller-Wieland
Original Assignee
Wilhelm Krone
Mueller Wieland Dirk
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Wilhelm Krone, Mueller Wieland Dirk filed Critical Wilhelm Krone
Priority to EP99927674A priority Critical patent/EP1071778A2/fr
Priority to AU44969/99A priority patent/AU4496999A/en
Publication of WO1999053052A2 publication Critical patent/WO1999053052A2/fr
Publication of WO1999053052A3 publication Critical patent/WO1999053052A3/fr

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
    • C07K14/4702—Regulators; Modulating activity
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/06—Antihyperlipidemics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy

Definitions

  • the invention relates to SREBP (stero / regulatory element binding grotein) -2-negative eukaryotic cell lines, the use of these cell lines for screening and testing substances and medicaments for the treatment or prevention of e.g. Hypercholesterolemia, arteriosclerosis, or coronary heart disease.
  • SREBP stero / regulatory element binding grotein
  • Cholesterol is an essential component of the cell membrane and the starting compound for the synthesis of steroid hormones and bile acids. It is vital, although cholesterol deposition in arteries with its complications (e.g. heart attack, stroke) is the most common cause of death in humans.
  • the cells meet their cholesterol requirements preferably by ingesting LDL cholesterol from the blood plasma. Since cholesterol is insoluble in water, it must be brought into a water-soluble form of transport for transport in the blood plasma.
  • various lipid protein complexes also called lipoproteins
  • VLDL Very Low Density Lipoproteins
  • IDL Intermediate Density Lipoproteins
  • LDL Low Density Lipoproteins
  • HDL High Density Lipoproteins
  • the LDL cholesterol is introduced by the cells mainly via the LDL receptor, which sits on the surface of many cells, into the cell interior by means of endocytosis.
  • the cholesterol bound in the LDL is broken down into free cholesterol in the cell.
  • the free cholesterol then reduces cholesterol biosynthesis by inhibiting activity Hydroxymethylglutaryl-CoA reductase (HMG-CoA reductase), the key enzyme in cholesterol biosynthesis.
  • HMG-CoA reductase Hydroxymethylglutaryl-CoA reductase
  • free cholesterol inhibits the expression of the HMG-CoA reductase gene as well as the expression of the LDL receptor gene. If the cell is supplied with enough LDL, less cholesterol is synthesized and less LDL receptor is expressed. The concentration of the LDL and thus the cholesterol in the blood plasma is therefore primarily regulated by the activity of the LDL receptor and the HMG-CoA reductase.
  • sre-1 sterol-regulated cis element
  • sterol regulatory element-1 sterol regulatory element-1
  • SREBP-1 and -2 cholesterol-sensitive transcription factors which are localized as precursor protein of approx.
  • LDL cholesterol was found to have the highest predictive value, although it can be seen that, given the LDL level, there is considerable variability in the risk depending on the other risk factor load. If the LDL cholesterol concentration in the blood plasma is increased, this leads to an increased uptake of LDL cholesterol in the cells of the blood vessels, which causes vasoconstrictions and unstable plaques (deposits), which ultimately lead to an increased risk of coronary heart disease.
  • hyperlipidemia In addition to the increased LDL concentration caused by high-fat food, other causes of an abnormally caused increased LDL concentration (hyperlipidemia, Hypercholesterolaemia) in the blood plasma. A distinction is made in the latter between primary and secondary hyperlipidemia. Secondary hypercholesterolemia occurs in hypothyroidism and diseases of the kidney, pancreas or liver and is only treated in exceptional cases with drugs that affect the metabolism, since they can be improved by therapy for the underlying disease. The primary hyperlipidemias, however, are based on mono- and polygenetically inherited defects. The most common monogenetically inherited hyperlipidemia is familial hypercholesterolaemia (FH), which is the cause of an LDL receptor dysfunction.
  • FH familial hypercholesterolaemia
  • Hypercholesterolemia is currently predominantly caused by the administration of HMG-CoA reductase inhibitors, ion exchangers or the like. a. also treated nicotinic acid derivatives.
  • the therapy of first choice is currently HMG-CoA reductase or cholesterol synthesis inhibitors.
  • a disadvantage of therapy with HMG-CoA reductase inhibitors and also ion exchangers is that it is dependent on the expression of the LDL receptor. Accordingly, the effect in patients with defects in the expression of the LDL receptor or its structural change is also absent.
  • the invention has for its object to provide cell lines that are used to find drugs for the prevention or treatment of e.g. Hypercholesterolemia, arteriosclerosis or coronary heart disease can be used.
  • the invention is also based on the object of medicaments for the prevention or treatment of
  • Arteriosclerosis with its complications, e.g. Stroke, coronary artery disease, peripheral arterial occlusive disease
  • cardiovascular risk factors e.g. B. glucose intolerance or diabetes mellitus, arterial hypertension, obesity, fat metabolism disorders
  • a first aspect of the invention relates to a eukaryotic cell line, characterized in that a polypeptide is expressed in reduced form, which is encoded by the nucleic acid sequence listed in SEQ ID NO: 1 (SREBP-2).
  • SREBP-2 the nucleic acid sequence listed in SEQ ID NO: 1
  • the term "a polypeptide is” expressed reduced means that the expression is reduced by more than 50%, preferably by about 70% and particularly preferably by about 90-100% compared to the normal expression of the polypeptide in the cell.
  • the eukaryotic cell line is preferably characterized in that a nucleic acid sequence stably integrated into the genome of the cell line, comprising the nucleic acid sequence as listed in SEQ ID NO: 1 or a fragment thereof and a promoter at which the transcription of the nucleic acid sequence as listed in SEQ ID NO : l or a fragment thereof is initiated, is transcribed so that an antisense RNA to the endogenous nucleic acid sequence listed in SEQ ID NO: 1 is transcribed and no polypeptide encoded by the endogenous nucleic acid sequence is expressed.
  • nucleic acid sequence as used herein means that expression is reduced by more than 50%, preferably about 70%, and most preferably about 90-100% compared to that normal expression of the polypeptide in the cell.
  • the skilled worker is aware that both the sense and the complementary antisense strand must be used to integrate a nucleic acid. Therefore, the nucleic acid sequence listed in SEQ ID NO: 1 encompasses both strands.
  • a human cell line is particularly preferred.
  • a liver cell line is particularly preferred.
  • the eukaryotic cell line is provided for screening and testing substances used to treat or prevent arteriosclerosis with its complications, e.g. Stroke, coronary artery disease, peripheral arterial occlusive disease, cardiovascular risk factors, e.g. Glucose intolerance or diabetes mellitus, arterial hypertension, obesity, lipid metabolism disorders, genetic liver diseases, which can be modulated by the activation of SREBP-sensitive genes, e.g. B. genetic hypertriglyceridemia with ApoC2 deficiency, Alzheimer's disease or neurodegenrative disorders can be used.
  • arteriosclerosis with its complications, e.g. Stroke, coronary artery disease, peripheral arterial occlusive disease, cardiovascular risk factors, e.g. Glucose intolerance or diabetes mellitus, arterial hypertension, obesity, lipid metabolism disorders, genetic liver diseases, which can be modulated by the activation of SREBP-sensitive genes, e.g. B. genetic hypertriglyceridemia with Apo
  • the following steps are used to test substances, which should contribute, for example, to new cholesterol-regulated genes or those that work via SREBPs or through the latter are modified in their effect to identify.
  • the cells are incubated in the presence or absence of cholesterol and then incubated over a period of time with a concentration of the substances listed above.
  • the expression of different genes for example “differential display” / PCR-Select TM / subtraction banks or relative promoter activity of defined genes by means of reporter gene analyzes
  • the effect on previously defined end points such as expression or promoter activity of the LDL receptor
  • nucleic acid sequence as listed in SEQ ID NO: 1 or a fragment thereof is used to produce a cell line which expresses reduced expressed a polypeptide which is encoded by the nucleic acid sequence as listed in SEQ ID NO: 1.
  • Another aspect of the invention relates to an antisense nucleic acid of the nucleic acid sequence as listed in SEQ ID NO: 1 or a fragment thereof as
  • Manufacture of a medicament for the prevention or treatment of arteriosclerosis with its complications e.g. Stroke, coronary artery disease, peripheral arterial occlusive disease, cardiovascular risk factors, e.g. Glucose intolerance or diabetes mellitus, arterial hypertension, obesity, fat metabolism disorders, genetic
  • Liver diseases that can be modulated by the activation of SREBP-sensitive genes e.g. B. Genetic hypertriglyceridemia with ApoC2 deficiency, Alzheimer's
  • antisense nucleic acid in combination with cholesterol synthesis inhibitors is particularly preferred.
  • Another embodiment relates to the use of the nucleic acid sequence as listed in SEQ ID NO: 1 or a fragment thereof as a tool for somatic gene therapy.
  • Preference is given to using the nucleic acid sequence in somatic gene therapy for the treatment of arteriosclerosis with its complications, for example stroke, coronary heart disease, peripheral arterial occlusive disease, cardiovascular risk factors, for example glucose intolerance or diabetes mellitus, arterial hypertension, obesity due to the genetic disorders of the liver, fat disorders, the Activation of SREBP-sensitive genes can be modulated, e.g. B. genetic hypertriglyceridemia with ApoC2 deficiency, Alzheimer's disease or neurodegenrative disorders.
  • FIG 1 shows schematically promoter-reporter gene constructs of the LDLR and their relative promoter activity.
  • HepG2 cells were transiently transfected by lipofection with various 5 'deleted or mutated constructs. Before harvesting, the cells were cultured in serum with 0.5% LPDS for 16 h and then either uninduced (SF) or stimulated for 4 h with 10 '7 M insulin or 3.3 x 10 "10 M PDGF.
  • the promoter activity is determined by the Luciferase activity measured in the cell extract was represented. The luciferase activity was corrected for the transfection efficiency by means of cotransfection.
  • the mean values ( ⁇ SD, n 4) of four different determinations are shown, which were each carried out in triplicate determinations.
  • the cis elements for the Transcription factors SP1 (spl), SREBP-1 and -2 (sre-l) as well as for the RNA polymerase (tata) and the transcription start point (TS) are shown.
  • FIG. 2 shows schematically the LDL receptor promoter activity (phLDL4) in SREBP 1 (-) and SREBP2 (-) cells.
  • HepG2 and SREBP 1 (-) and SREBP2 (-) cells were transiently transfected with the promoter construct phLDL4.
  • Prior to harvesting the cells for 16 h in serum were 0.5% LPDS cultured, and then either uninduced (SF) or 4 h with 10 -7 M insulin, 10 -8 M IGF-1, l, 15 M EGF 5xl0- or 3,3xl0- 10 M PDGF stimulates.
  • the promoter activity is represented by the luciferase activity, which was measured in the cell extract. Luciferase activity was corrected for transfection efficiency by means of cotransfection.
  • FIG. 3 schematically shows the influence of specific inhibitors on the action of insulin or PDGF on the LDLR.
  • HepG2 cells were transiently transfected with the phLDL4 promoter construct. Before harvesting, the cells were exposed to 0.5% serum for 16 h
  • the nucleotide sequence coding for the N-terminus (amino acid 1 to 460) of SREBP-la was amplified by PCR after reverse transcription of 1 ⁇ g RNA with the primers SREBP-la (NTl) and SREBP-la (NT2) and then cloned into the BamHI / EcoRI site of the cloning vector pUC19.
  • the N-terminus (amino acid 1 to 468) of SREBP-2 was cloned after amplification with the primers SREBP-2 (NT1) and SREBP-2 (NT2).
  • the N-terminal end of SREBP-lc was reached by modifying the SREBP-la clone.
  • the vector containing SREBP-la was cut with BamHI and Hgal (nucleotide 230). Then a PCR was carried out with the primers SREBP-lc (NTl) and SREBP-la (NT2) to obtain SREBP-lc.
  • the nucleotide sequences were verified using DNA sequencing.
  • Oligonucleotides were made according to the solid phase phosphoamidite method with the Pharmacia
  • oligonucleotides were treated with NFLOH conc. decoupled from the column material, the column material by centrifugation (15000xg; 2 min; room temperature (RT)) removed and the supernatant concentrated in vacuo (1 h).
  • the oligonucleotides were washed with 1/10 vol. NaAc pH 4.0 and 3 vol. EtOH. Precipitated for 30 min at -20 ° C, centrifuged (15000xg; 15 min; 4 ° C;) and in 100 ⁇ l H 2 O bidistilled . resuspended.
  • the following oligonucleotides were constructed to generate the specific cDNA fragments.
  • NT2 5'-GGAATTCGTCAGGCTCGGAGTCACTGCC-3 'SREBP-2 (NT1) 5'-GTAGGATCGCGATGGACGACAGCGGCGAGCTGGG-3' SREBP-2 (NT2) 5'-GGAATTCATCTTGGAC3
  • RNA 1 ⁇ g was analyzed using H 2 Obidest. Heat denatured for 5 min at 70 ° C. Then dNTP mix (0.5 mM), pd (N) 6 primer (0.2 ⁇ g), Mu-MLVRT (200 U) and lx reverse transcriptase buffer (50 mM Tris / HCl; 40 mM KCl; 1 mM DTT; 6 mM MgCl 2 ; 0.1 mg / ml BSA; pH 8.3) was added and the synthesis was carried out at 37 ° C. for 1 h.
  • dNTP mix 0.5 mM
  • pd (N) 6 primer 0.2 ⁇ g
  • Mu-MLVRT 200 U
  • lx reverse transcriptase buffer 50 mM Tris / HCl; 40 mM KCl; 1 mM DTT; 6 mM MgCl 2 ; 0.1 mg / ml BSA; pH 8.3 was added and the synthesis was
  • PCR buffer 10 mM Tris / HC1; 50 mM KCl; 0.1 mg / ml BSA pH 8.3), MgCl 2 (1.5 mM), dNTP- was added to 10 ⁇ l of the first-strand synthesis approach.
  • Mix (0.2 mM), 3 'primer (0.1 uM), 5' primer (0.1 uM) and 1.25 U Taq polymerase added.
  • the PCR was carried out in a sample volume of 100 ⁇ l; Mineral oil was used to prevent evaporation.
  • amplification was carried out in 32 cycles (94 ° C. for 1 min; 60 ° C. for 2 min; 72 ° C. for 3 min) and an elongation step (72 ° C., 7 min) was connected.
  • SREBP-1 deficient cells The production of SREBP-1 deficient cells is described; see. Streicher et al, 1996, J. Biol. Chem. 271, 7128-7133. HepG2 cells were deficiently produced for SREBP-2.
  • SREBP-2-deficient cells a cDNA fragment with a length of 421 bp was generated with the primers SREBP-2 (antil) and SREBP-2 (anti2) by means of PCR; the cDNA of the N-terminal domain of SREBP-2 served as template (see above). This fragment was cloned into an expression vector (pcDNA3) such that SREBP- y
  • 2-antisense RNA is expressed (described for SREBP-1). This antisense construct is complementary to the SREBP-2 specific nucleotide sequence 257 to 678.
  • oligonucleotides were constructed to create the specific cDNA fragment.
  • SREBP-2 (antil) 5'-GTAGGATCCAGAACAGCTGTGTAGCTCC-3 * SREBP-2 (anti2) 5'-ATTGAAGCTTGCTTTGGACTTGAGGCTGAAC-3 '
  • the HepG 2 cells were transfected either as with the method already described or by electroporation.
  • the cells were cultured as a monolayer in RPMI-1640 (Sigma, Germany) complemented with 10% (v / v) FCS (Gibco, Germany) and antibiotics (Sigma, Germany, A9909, 1% solution).
  • the cells were trypsinized and washed and suspended in Opti-MEM (Gibco, Germany).
  • the cell suspensions (2 x 10 5 cells per well) were mixed with the corresponding vectors below.
  • the ⁇ -galactosidase activity was determined by the Galaktolight assay (Tropix) according to the manufacturer's instructions. The data show the relative luciferase activity as x-fold induction due to either endogenous or exogenous stimulation compared to unstimulated cells.
  • the expression vectors (Stratagene), the regulatory domains of transcription factors, fused to heterologous DNA binding domains of Gal-4 (amino acids 1-147) under Control of an MLV promoter were used to produce constructs with the corresponding N-terminal domains (see above) of the SREBPs in the open reading frame (for this purpose, the vectors with BamHI / EcoRI subcloned fragments were cut with BamHI, filled in and blunt-ended in frame "religiert). To determine the transactivation of the reporter plasmid, the luciferase gene was used, which is under the control of 5 Gal-4-binding elements (Stratagene, Germany).
  • HepG2 cells were transiently transfected by lipofection with various 5 'deleted or mutated constructs. Before harvesting, the cells were cultured in serum with 0.5% LPDS for 16 h and then either uninduced (SF) or stimulated for 4 h with 10 "7 M insulin or 3.3x10 " 10 M PDGF.
  • the promoter activity is represented by the luciferase activity, which was measured in the cell extract. Luciferase activity was corrected for transfection efficiency by means of cotransfection.
  • the mean values ( ⁇ SD, n 4) of four different determinations are shown, each of which was carried out in triplicate determinations.
  • the cis elements for the transcription factors SP1 (spl), SREBP-1 and -2 (sre-l) as well as for the RNA polymerase (tata) and the transcription start point (TS) are shown.
  • Example 5 In order to further characterize the need for SREBP-1 in the hormone-induced transcription activation of the LDLR, hepG2 cells were generated by means of stable transfection of pcSREBP-1 (-), the content of SREBP-1 of which was significantly reduced (SREBP-1 (-) - cells). A transient transfection of these cells with the promoter construct phLDL4 showed that the transcription-activating effect of insulin or IGF-1 was eliminated in comparison to mock-transfected HepG2 cells. Interestingly, the stimulating effect of PDGF and EGF is also significantly reduced in SREBP-1 (-) cells (FIG. 2). The Regulation of sre-1 mediated transcription activation appears to be mediated specifically by SREBP-1.
  • the cells were cultured in serum with 0.5% LPDS for 16 h and then either uninduced (SF) or 4 h with 10 "7 M insulin, 10 '8 M IGF-1, 1.5 x 10 " 15 M EGF or 3.3xl0 '10 M PDGF stimulated.
  • the promoter activity is represented by the luciferase activity, which was measured in the cell extract. Luciferase activity was corrected for transfection efficiency by means of cotransfection.
  • the mean values ( ⁇ SD, n 4) of four different determinations are shown, each of which was carried out in triplicate determinations.
  • the hormonal activity on the LDLR promoter could be restored by transient expression of SREBP-la as well as of SREBP lc in SREBP-1 (-) cells or of SREBP-2 in SREBP-2 (-).
  • HepG2 cells were transiently transfected with the phLDL4 promoter construct. Before harvesting, the cells were cultured for 16 h in serum with 0.5% LPDS and then either uninduced (SF) or 4 h with 10 '7 M insulin or 3.3xl0 "10 M PDGF with and without inhibitors (75 ⁇ M PD98056 resp 50nM Wortmannin).
  • the promoter activity is represented by the luciferase activity measured in the cell extract.
  • the luciferase activity was corrected for the transfection efficiency by means of cotransfection.
  • the mean values ( ⁇ SD, n 4) of four different determinations are shown, each were carried out in triplicate determinations.
  • GTTCCCACCA CACCCAGGGC AACTCCTATT CTTCAGCCCC GCCCCCAGCC CCAGCCTCAA 420 CCTCAAACTC AGCTGCAACA ACAGACGGTA ATGATCACGC CAACATTCAG CACCACTCCG 480 CAGACGAGGA TCATCCAGCA GCCTTTGATA TACCAGAATG CAGCTACTAG CTTTCAAGTC 540
  • CTTCAGCCTC AAGTCCAAAG CCTGGTGACA TCCTCCCAGG TACAGCCGGT CACCATTCAG 600
  • ATCGAGGACT TTAATCAGAA TGTCCTTCTG ATGTCCCCCC CAGCCTCTGA CTCAGGGTCC 1320 CAGGCTGGCT TCTCTCCCTA CTCCATTGAC TCTGAGCCAG GAAGCCCTCT ATTGGATGAT 1380
  • GTCCTGTCAT TCGAGTCAGG TTCTGGGGGC TGGTTTGACT GGATGATGCC
  • ACTCTTCTC 1620 TTATGGCTGG TAAATGGTGT GATTGTCCTG AGCGTCTTTG TGAAGCTGCT GGTTCATGGG 1680
  • GCCAAGGAGA GTCTATACTG TGCCCAGAGG AACCCAGCTG ACCCCATTGC GCAGGTCCAC 2400 CAGGCCTTCT GCAAGAACCT GCTGGAGCGA GCTATAGAGT CCTTGGTGAA ACCTCAGGCC 2460
  • MOLECULE TYPE synthetic DNA
  • HYPOTHETICAL NO
  • MOLECULE TYPE synthetic DNA
  • HYPOTHETICAL NO
  • SEQUENCE DESCRIPTION SEQ ID NO: 4: GTAGGATCGC GATGGACGAC AGCGGCGAGC TGGG 34
  • MOLECULE TYPE synthetic DNA
  • HYPOTHETICAL NO
  • SEQUENCE DESCRIPTION SEQ ID NO: 5:

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Abstract

L'invention concerne des lignées cellulaires eucaryotes SREBP(Sterol Regulatory Element Binding Protein)-2-négatives, leur utilisation par échantillonnage et test des substances, ainsi que des médicaments utilisés pour traiter ou assurer la prophylaxie par ex. de l'hypercholestérolémie, de l'artériosclérose ou de maladies cardiaques coronariennes.
PCT/DE1999/001143 1998-04-16 1999-04-15 Lignees cellulaires srebp-2-deficientes WO1999053052A2 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
EP99927674A EP1071778A2 (fr) 1998-04-16 1999-04-15 Lignees cellulaires srebp-2-deficientes
AU44969/99A AU4496999A (en) 1998-04-16 1999-04-15 Srebp-2-deficient cell lines

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE19816894.2 1998-04-16
DE19816894A DE19816894C1 (de) 1998-04-16 1998-04-16 SREBP-2-defiziente Zellinien

Publications (2)

Publication Number Publication Date
WO1999053052A2 true WO1999053052A2 (fr) 1999-10-21
WO1999053052A3 WO1999053052A3 (fr) 2000-01-20

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PCT/DE1999/001143 WO1999053052A2 (fr) 1998-04-16 1999-04-15 Lignees cellulaires srebp-2-deficientes

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EP (1) EP1071778A2 (fr)
AU (1) AU4496999A (fr)
DE (1) DE19816894C1 (fr)
WO (1) WO1999053052A2 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1743523A4 (fr) * 2004-04-09 2008-05-14 Univ Tsukuba Animal transgénique non humain en tant que modèle pour le diabète de type 2

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5527690A (en) * 1987-03-30 1996-06-18 Board Of Regents, The University Of Texas System Methods and compositions relating to sterol regulatory element binding proteins

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1743523A4 (fr) * 2004-04-09 2008-05-14 Univ Tsukuba Animal transgénique non humain en tant que modèle pour le diabète de type 2

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WO1999053052A3 (fr) 2000-01-20
EP1071778A2 (fr) 2001-01-31
AU4496999A (en) 1999-11-01
DE19816894C1 (de) 2000-03-02

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