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WO1987002059A1 - POLYMORPHISMES LIES AU METABOLISME DES LIPIDES ET PERMETTANT DE DECELER UNE PROPENSION A L'ARTERIOSCLEROSE: ApoB, ApoCII, ApoE, ApoAIV - Google Patents

POLYMORPHISMES LIES AU METABOLISME DES LIPIDES ET PERMETTANT DE DECELER UNE PROPENSION A L'ARTERIOSCLEROSE: ApoB, ApoCII, ApoE, ApoAIV Download PDF

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Publication number
WO1987002059A1
WO1987002059A1 PCT/US1986/002048 US8602048W WO8702059A1 WO 1987002059 A1 WO1987002059 A1 WO 1987002059A1 US 8602048 W US8602048 W US 8602048W WO 8702059 A1 WO8702059 A1 WO 8702059A1
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probe
restriction endonuclease
substantial equivalent
apob
dna
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Philippe M. Frossard
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Biotechnology Research Partners Ltd
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Biotechnology Research Partners Ltd
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/156—Polymorphic or mutational markers

Definitions

  • the invention relates to the use of genetic polymorphisms to determine disease states. More particularly, the invention concerns the use of polymorphisms of the apolipoprotein 6, CII, E, and AIV genes to diagnose susceptibilities to atherosclerosis.
  • the degree of morbidity and mortality associated with atherosclerosis in developed countries is higher than that associated with any other particular disorder, even cancer.
  • the disorder manifests itself in the form of cholesterol deposition in arterial cell walls.
  • the deposition is slow and irreversible and starts at an early age.
  • Clinical symptoms may take years to manifest themselves and are extremely serious; they include coronary heart disease and stroke. Generally, the disease process will have begun long before these clinical manifestations appear.
  • a diagnostic technique which provides an early warning of the onset of the deposition.
  • the present technique depends on measuring cholesterol or triglyceride levels in serum, and while these levels can be measured quite accurately, they do not offer the desirable high correlation to true susceptibility.
  • More reliable predictive methods which rely on detection of atheromatous lesions, use highly invasive procedures, which are sufficiently painful and expensive that they cannet be employed on a screening basis, or even applied to specific groups selected on the basis of family histories. These techniques also offer too little, too late; by the time the atheromatous lesions have appeared, the most effective time for treatment has been passed.
  • the present invention provides polymorphisms located in genes related to lipid metabolism, those encoding apolipoproteins B, CII, E, and AIV, which are useful in predicting susceptibility to atherosclerosis.
  • Other polymorphisms in the apoAI/CIII gene complex also useful in atherosclerosis prediction are disclosed in U.S. Serial No. 724,192. filed 17 April 1985, and its continuation-in-part application United States Serial No. 782.666. filed 30 Sept. 1985.
  • the invention provides identification of polymorphisms which are useful as predictors of the subsequent development of atherosclerosis. Since most of these polymorphisms are located in the genomic sequences which regulate lipid metabolism, their pattern of presence or absence correlates to propensity to develop this disease. In addition, the presence or absence of these polymorphisms is useful as a form of genetic identification or fingerprinting of an individual and in ascertaining familial relationships.
  • the invention is directed to a method of predicting the likelihood of development of atherosclerosis in an individual, or of genetically identifying said individual, which method comprises detecting one or more of: the presence or absence of PvuII, Stul. EcoRVa, EcoRVb, EcoRVc, Hpal, or Dral polymorphisms in the apoB gene; the presence or absence of "BamHI”, “BanI”, “Bgll”, or “Ncol” polymorphisms in the apoCII gene; the presence or absence of the "Hpal” polymorphism in the apoE gene; the presence or absence of four Xbal polymorphisms in the apoAI/CIII/AIV gene complex detected with the apoAIV probe; the presence or absence of a "TaqI” polymorphism in the apoAI/CIII/AIV gene complex detected with the apoAIV probe; the presence or absence of a "Dral" polymorphism in the apoAI/CIII/A
  • the invention is directed to a method for predicting the susceptibility of an individual to atherosclerosis or of providing genetic identification of said individual by digesting human genomic DNA of an individual subject and detecting one or more of: the presence or absence of a 5.5 kb PvuII digestion fragment which hybridizes to a 970 bp apoB probe; the presence or absence of a 5.2 kb StuI fragment which hybridizes to a 2 kb apoB probe; the presence or absence of a 4.8 kb EcoRV fragment which hybridizes to a 3 kb apoB probe; the presence or absence of 11.0 and 7.0 kb EcoRV fragments which hybridize to a 3 kb apoB probe; the presence or absence of 3.6 and 2.7 kb EcoRV fragments which hybridize to a 3 kb apoB probe; the presence or absence of 8.3 and 6.2 kb Hpal fragments which hybridize to a 3 kb apoB probe; the presence or absence of 8.3
  • the invention relates to a method to predict the susceptibility of an individual to atherosclerosis by digesting genomic DNA from this individual and detecting the presence or absence of a 600-1600 bp insert or a 1600-2200 bp insert 5' of the insulin gene.
  • the invention thus relates to determination of a genetic fingerprint of a subject, which fingerprint may relate to disorders of lipid metabolism and transport, using polymorphisms of the genes associated with proteins involved in these functions.
  • the genetic fingerprint is also useful in identification of particular individuals and in assessing familial relationships.
  • the invention is also directed to kits suitable for performing the method of the invention.
  • Figure 2 is the DNA sequence of the apoCII probe
  • Figure 3 is the DNA sequence of the apoE probe
  • Figure 4a is the DNA sequence of the two-part apoAIV probe
  • Figure 4b shows the apoAI/CIII/AIV gene complex.
  • A. Techniques for Detection of Polymorphisms Application of the method of the invention to predict potential atherosclerotic individuals or to obtain a genetic "fingerprint" based on some or all of the polymorphisms associated with the designated genomic regions, employs standard techniques of DNA extraction, purification, restriction enzyme digestion, and size separation. Techniques for hybridization with probe and detecting successfully hybridized substrate arranged according to molecular weight are also well known to those in the art. The general approach to finding and detecting the significant polymorphisms is the following: DNA is extracted from the somatic cells of the individual to be tested, for example from leukocytes, placental cells, cultured fibroblasts, or, in the case of fetal individuals, from cells of the amniotic fluid.
  • the high molecular weight DNA fraction is separated, and subjected to treatment with a particular, selected restriction enzyme, such as, for example, EcoRl, BamHI, MstI, XmnI, and the like.
  • a particular, selected restriction enzyme such as, for example, EcoRl, BamHI, MstI, XmnI, and the like.
  • the digest is applied to a polyacrylamide or agarose gel and subjected to electrophoresis to obtain separation of the DNA fragments resulting from restriction enzyme digestion into positions on the gel determined by the size (length) of the fragment.
  • the contents of the gel are then replicated by transferring to a nitrocellulose filter or other suitable matrix for use as a probe hybridization support.
  • the DNA fragments, either before or after transfer to the nitrocellulose filter replica are treated with a denaturant such as sodium hydroxide/salt.
  • the denatured, single-stranded DNA, replicated electrophoresis patterns are probed with labeled (usually by
  • fragments will be detected which derive from a particular region on the genome.
  • a cDNA sequence from the apolipoprotein B (apoB) or apolipoprotein CII (apoCII) or apolipoprotein E (apoE) gene sequences is used as a probe. Therefore, the only fragments which will appear on the hybridized filters are those which contain sequences complementary to the designated probe—i.e., only those which have not been severed either in the genome itself or by the restriction enzyme cleavage from the complementary apoB or apoCII or apoE fragment. Stated in another way, by using a particular probe, alterations in the genome which are proximal to sequences corresponding to that probe are detected.
  • probes useful in the present invention are selected from the apoB, apoCII, and apoE genes.
  • the fragment pattern obtained is diagnostic for a particular polymorphism if the probe selected is complementary to a DNA sequence sufficiently proximal to the polymorphism on the genome that it is not severed from the polymorphism by the restriction cleavage, and has a low probability of being segregated from the polymorphism by crossing over. Acceptable distance limits between the region of probe complementarity and the polymorphism are therefore arbitrary. Generally, probes which hybridize to DNA sequences within 10 kb upstream or downstream of the polymorphism give acceptable results. Occasionally, the pattern of restriction enzyme cleavage may place a distal probe hybridization site on a fragment irrelevant to the polymorphism. The closer the probe to the polymorphism, the greater the range of usable restriction enzymes.
  • a probe which is a "substantial equivalent" to a specified probe is one which gives the same fragment length in digests of DNA from individuals for a particular polymorphism when the same restriction enzyme is used.
  • slightly shorter or longer probes could be used which hybridize to the same region as the designated probe without altering the results; a probe which hybridizes closer to the site of the polymorphism could also be substituted.
  • genes encoding other proteins related to lipid metabolism are also useful.
  • the gene regions which are of interest with respect to the polymorphisms herein are those of the apoB, apoCII, apoE, and apoAIV genes.
  • Apolipoprotein B is the major protein component of very low density lipoproteins (VLDL) and of chylomicrons. It is the sole protein in low density lipoproteins (LDL). and is essential for the assembly and secretion of chylomicrons and VLDL. It also functions as the ligand for removal of LDL from circulation by receptor-mediated uptake into a variety of cells. (Lusis, A.J., et al. Proc Natl Acad Sci (USA) (1985) 82: 4597-4601.) Four major plasma species of apoB have been described (Kane, J.P., et al. Proc Natl Acad Sci (USA) (1980) 77: 2465-2469).
  • apoB-48 is synthesized by the intestine and is a component of chylomicrons; the other primary form, which is apparently attacked by the plasma protease, apoB-100, is the protein ligand on LDL that binds to the LDL receptor and results in uptake and catabolism of LDL by the liver (Deeb, S.S., et al. Proc Natl Acad Sci (USA) (1985) 82: 4983-4986). In any event, the apolipoproteins encoded by the apoB gene are integral to cholesterol and fat metabolism.
  • apoCII apolipoprotein
  • apoCII Another apolipoprotein. apoCII, also plays an important role in the relevant metabolic pathways. It is a 79 amino acid peptide associated with the circulating triglyceride-rich lipoproteins, chylomicrons, and VLDL (Myklebost, O., et al, J Biol Chem (1984) 7 : 4401-4404). It is known to activate lipoprotein lipase (LaRosa. J.C., et al, Biochem Biophys Commun (1970) 41: 57-62; Breckenridge, W.C., et al. New Enq J Med (1978) 298: 1265-1272).
  • a third relevant gene sequence is that encoding human apolipoprotein E.
  • ApoE is also a component of chylomicrons and chylomicron remnants, and is found in VLDL and HDL.
  • the sequence of this 299 amino acid protein is known, and a cDNA clone has been prepared (McLean. J.W., et al. J Biol Chem (1984) 25: 6498-6504).
  • ApoE appears to mediate the uptake of lipoproteins through specific receptors (Mahley, et al, Biochem Biophys Acta (1983) 737: 197-222; Mahley, R.W., Klin Klin Klischr (1983) 61: 225-232) and to bind to LDL receptors (Innerarity, T.L., Biochemistry (1978) 17: 1440-1447).
  • Variable forms of apoE protein have been found, and certain abnormal forms of apoE2 seem to be associated with a genetically determined hyperlipoproteinemia (Mahley, R., et al, Adv Intern Med (1983) 29: 385-411).
  • a fourth protein whose coding sequence serves as the basis for a useful probe is human apolipoprotein AIV. Genetic mapping has shown that the apoAIV and apoAI/CIII gene regions are, in fact, closely linked (Schamaun, O., et al. Hum Genet (1984) 68:181-184; Karathanssis, S.K., Proc Natl Acad Sci USA (1985) 82:6374-6378; Elshourbagy. N.A., et al, J Biol Chem (1986) 261:1998-2002), and a number of structural and organizational similarities have been noted between the apoAI and apoAIV genes.
  • ApoAIV is a 376 amino acid protein whose complete amino acid sequence is known (Elshourbagy, et al, (supra)). ApoAIV is believed to mediate various metabolic steps associated with cholesterol and other lipid metabolism in a manner similar to apoAI/CIII. It should be noted that due to the proximity of the apoAI/CIII gene to the apoAIV gene, polymorphisms detected with the apoAIV probe may in fact detect changes in DNA sequence which reside in the apoAI/CIII complex. Therefore, the polymorphisms detected with this probe will be referred to as polymorphisms of the "apoAI/CIII/AIV gene complex". The relative positions and reading directions of these protein encoding regions are shown in Figure 4b.
  • each of the foregoing gene sequences encoding apoB, apoCII, apoE, and apoAIV appear to be intimately involved with the metabolic steps that determine cholesterol and other lipid metabolism, and are thus relevant to prognosis of atherosclerosis. Accordingly, probes designed to hybridize to regions of these genes are useful in the method of the invention. A description of appropriate probes and restriction enzymes for detection of insertion polymorphisms 5' of the insulin gene is found in Bell, C.I., et al. Nature (1980) 284:26-32; Proc Natl Acad Sci USA (1981) 78.: 5759-5763; Diabetes (1984) 33:176-183.
  • Probes are labeled by nick translation using ⁇ [ 32 P] dCTP and ⁇ [ 32 P] dGTP, which results in fragmentation of the probe.
  • cDNA probes which are complementary only to the exon regions of the gene and which span over intron regions are workable in the method of the invention.
  • the reagents suitable for applying the method of the invention to detect the appropriate polymorphisms may be packaged into convenient kits providing the necessary materials, packaged into suitable containers, and, optionally, suitable containers or supports useful in performing the assay.
  • the essential components of the assay include the restriction enzyme associated with the polymorphism, and a suitable probe.
  • packages containing concentrated forms of reagents used for hybridization, prehybridization, DNA extraction, etc. may be included if desired.
  • labeled probe, or reagents suitable to form conveniently labeled probe are useful in facilitating the conduct of the method of the invention.
  • kits Instructions regarding the conduct of the method are also included in the kit. Said instructions describe the operations which constitute the assay—i.e., the manner of detecting the relevant genomic fragments and indicating the correlation of results to atherosclerosis prediction.
  • Polymorphisms in the apoB, apoCII, apoE, and apoAIV regions may be correlated with a propensity to exhibit the symptoms of atherosclerosis. Such correlations are discerned by screening samples of patient and control populations. One useful criterion for separating patients from controls is the presence or absence of atheromatous plaques, as detected by angiography. Thus, a sample population may be divided into those showing atheromatous plaques using this technique and those lacking, them. DNA samples are then obtained from the leukocytes or other convenient source of both patient and control groups and subjected to the methods of detecting the relevant polymorphisms, as described herein. Correlations can then be made using any convenient statistical method. One particularly convenient method which results in the calculation of a relative incidence of atherosclerosis is illustrated below-. However, any other convenient correlation method may also be used.
  • the essential features of the invention as it relates to detection of a particular polymorphism are selection of enzyme and probe.
  • PvuII/B embodiment for atherosclerosis prediction one may use PvuII digestion of the genomic DNA and probe with a sequence complementary to the genomic sequence (in the nonrepeating regions) proximal (i.e., in this case within ⁇ ⁇ 5.5 kb) to the site of the polymorphism.
  • other restriction enzymes may be used in conjunction with a probe which hybridizes in particularly close proximity to the polymorphism.
  • the fingerprinting polymorphisms may employ other specific restriction enzymes.
  • a variety of substantially equivalent probes could be designed with respect to this region, and the particular restriction enzyme and cDNA probe chosen are arbitrary.
  • the efficacy of the probe is enhanced as it moves closer to the site of the polymorphism. Otherwise, additional cleavage points may be encountered between the polymorphism and the probe, and also the probing site may be separated from the site of the polymorphism during replication by crossing-over events.
  • Leukocytes were obtained from freshly drawn blood collected from each of the human subjects, and high molecular weight genomic DNA was isolated by the procedure of Law, D. J., et al. Gene (1984) 28:153-158.
  • DNA fragments were denatured in situ in 0.5 M NaOH/1.5 M NaCl for 2 x 10 min, neutralized in 1 M ammonium acetate pH 7.2 for 2 x 10 min, and transferred overnight onto nitrocellulose paper (Schleicher and
  • apoB (0.97), apoB (2 kb) and apoB (3 kb); apoCII probe; the apoE probe and a two-part, mixed apoAIV probe.
  • EcoRI/EcoRI insert fragment which contains 70 bp of 5' untranslated region and 900 bp of sequence encoding the
  • the filters are prewashed for 2 hr in 3 x NaCl/Cit (1 x NaCl/Cit is 150 mM NaCl/15 mM sodium citrate, pH 7.0), 0.1% SDS at 55°C, and then prehybridized in 6 x NaCl/Cit, 200 ⁇ g/ml denatured salmon sperm DNA, 5 x Denhardt's, 0.05% sodium pyrophosphate for 1 hr at 50°C.
  • a 192-fold degenerate 23 base oligonucleotide probe which encodes, taking account of codon redundancy, the first 8 amino acids of the previously determined sequence of apoB-26 (Asp-Glu-Pro-Pro-Gln-Ser-Pro-Trp) was used as a probe.
  • the probe was 5' end labelled with
  • T4 polynucleotide kinase (PL Biochemicals) and ⁇ -[ 32 P]-ATP. added to the filters and incubated for
  • LB25-1 One positive plaque, designated LB25-1, was purified and the cDNA insert was subcloned in both orientations into M13/mp8 for sequencing.
  • the EcoRI insert was subcloned into pBR322 to obtain pB25-1 for amplification.
  • pB25-l thus contains some 5' untranslated region, the signal sequence, and the first 266 amino acids of the mature protein, i.e., apoB (0.97kb) probe.
  • apoB probes For the remaining two apoB probes, additional portions of the apoB encoding sequence were obtained using linearized denatured pB25-l insert as initial probe.
  • a 2 x 10 5 member human adult intestine cDNA library in ⁇ gt10 was screened using this insert and a cDNA designated IB7, containing an approximately 1.3 kb insert, about 800 bp of which extended beyond the 3' end of clone pLB25 was obtained.
  • Isolated, denatured IB7 insert was subcloned into pBR322 for amplification, creating pIB7.
  • the purified pIB7 insert was denatured and used to screen the intestine library.
  • One positive cDNA fragment designated 110 contained an approximately 3 kb insert, about 2.5 kb of which extended beyond the 3' end of IB7. This cDNA insert was subcloned into the EcoRI site of pBR322. creating pB10. This insert provided the second apoB probe and was designated apoB (3 kb).
  • Linearized, denatured pB10 insert was used as a probe to obtain still another cDNA fragment designated IB-(2)1, containing an approximately 2 kb insert, about 1 kb of which extends in the 3' direction beyond the IB-10 sequence.
  • the EcoRI cDNA insert was also subcloned into the EcoRI site of pBR322. creating pB(2)1. This insert represents the third apoB probe and is designated apoB (2kb).
  • the apoCII probe is a 1.03 kb EcoRI/EcoRL insert fragment which corresponds to a portion of the Myklebost cDNA (supra).
  • This fragment was obtained from a human fetal liver cDNA library constructed in ⁇ gt-10 (by providing EcoRI linkers and inserting into the EcoRI site of the phage) and screened with a 51 base synthetic oligonucleotide containing the coding sequence of nucleotides 73-122 as published by Myklebost. Two positive clones were obtained from 500,000 screened, and one was sequenced; it spans nucleotides 10-432 encompassing amino acid -14 of the signal sequence through 38 bases of the 3' untranslated region. The complete sequence of this probe (without the linkers) is shown in Figure 2.
  • the apoE probe is a 1 kb EcoRI/EcoRI fragment which covers the entire mature protein-encoding sequence and corresponds to the sequence published by McLean et al, supra. It was obtained from a human fetal liver cDNA library prepared in ⁇ gt-10 (by providing EcoRI linkers and inserting into the EcoRI site of the phage) and screened with a synthetic 46 base oligonucleotide containing the coding sequence of nucleotides 469-514 of the published DNA sequence. Of 10 positives obtained from 450,000 phage, one was sequenced and encodes the protein spanning nucleotides -14 to +1020, which encompasses amino acid -4 of the signal sequence through 120 bases of the 3' untranslated region.
  • the complete sequence of this probe (without the linkers) is shown in Figure 3.
  • the apoAIV probe is a mixture of two DNA segments which together encode most of the apoAIV protein.
  • the complete sequence of these "apoAIV-5'" and "apoAIV-3'” probes is shown in Figure 4a.
  • the apoAIV-5' probe extends past the N-terminus of the protiin encoding sequence with the additional sequence there shown. It slightly overlaps the 5' end of the apoAIV-3' probe which extends from the codon for amino acid 187 through part of the codon for amino acid 358, only 18 amino acids short of the C-terminus of the protein.
  • apoAIV probes were prepared starting with the ⁇ A1.9 genomic clone described by Protter, A.A., et al. DNA (1984) 3:449-456. ⁇ A1.9 was digested with
  • 1.2 kb fragment containing a portion of the apoAIV gene was isolated by electrophoresis. The identity of the 1.2 kb fragment was confirmed to correspond to the coding sequence for the apoAIV protein.
  • the 1.2 kb fragment was labeled by nick translation and used to screen a human intestinal cDNA library (human jejunum) in ⁇ gt-10 containing about 3 x 10 5 recombinant and stored in CsCl.
  • the 661 bp apoAIV-5' probe and the 513 bp apoAIV-3' probe hybridized to the labeled 1.2 kb fragment.
  • control and patient groups were set up using as a criterion positive or negative results relating to atheromatous plaque formation as determined by angiography. Persons were classified as “patients” who showed plaques in this assay, whether or not they had suffered heart attacks. They were designated “controls” if the results of this test were negative; none of these persons had had heart attacks.
  • a standard x-squared analysis was used to determine a significance level.
  • the significance level represents the probability that an association is due to chance alone. Therefore, the results obtained would not hold up if high numbers of subjects were used or a large number of independent trials were made.
  • a significance level of less than 0.05 means that there is a greater than 95% probability that the observed results are true - i.e., that the tested hypothesis is different from the null hypothesis. Therefore it is likely that testing additional or larger numbers of subjects would yield the same results.
  • a significant level of 0.10 means that there is one chance in 10 that the results would be different if a larger or different sample were tested.
  • PP is the number of patients having the polymorphism
  • PN is the number of patients not having the polymorphism
  • CP is the number of controls having the polymorphism
  • CN is the number of controls not having the polymorphism.
  • the value calculated by this ratio if greater than 1, indicates that the persons having the polymorphism are at a greater risk of having the disease; a value less than 1 shows protection against the disease.
  • the Taql/CII polymorphism reported by others appears to have no correlation.
  • the Banl/CII polymorphism appears to exert a protective effect.
  • On the other hand, for the Banl/CII polymorphism 19 of 35 patients, or 54%, had the polymorphism, whereas 3 of 5, or 60%, of controls showed this "abnormality". This leads to a calculated relative incidence value of 0.8 for a slightly protective effect. While the significance level (0.6) is unfavorable, there is still an appreciable probability that this ratio will be maintained upon further testing.

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Abstract

Des polymorphismes présents dans des gènes liés au métabolisme des lipides, spécifiquement des apolipoprotéines B, CII, E et apoAIV ont été identifiés. La présence ou l'absence de ces polymorphismes chez certains sujets peut être corrélée avec la tendance à montrer des symptômes d'artériosclérose. Une corrélation est également établie de cette manière entre deux polymorphismes d'insertion (5') du gène d'insuline.
PCT/US1986/002048 1985-09-30 1986-09-29 POLYMORPHISMES LIES AU METABOLISME DES LIPIDES ET PERMETTANT DE DECELER UNE PROPENSION A L'ARTERIOSCLEROSE: ApoB, ApoCII, ApoE, ApoAIV Ceased WO1987002059A1 (fr)

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DK277087A DK277087A (da) 1985-09-30 1987-05-29 Fremgangsmaade til forudsigelse af aterosclerose i et individ baseret paa detektering af forekomst eller fravaer af polymorfier i relevante gener

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US78266385A 1985-09-30 1985-09-30
US782,663 1985-09-30
US86917786A 1986-05-30 1986-05-30
US869,177 1986-05-30
US06/900,593 US4772549A (en) 1986-05-30 1986-08-26 Polymorphisms related to lipid metabolism: ApoB, ApoCII, ApoE, ApoAIV
US900,593 1986-08-26

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1988003175A1 (fr) * 1986-10-29 1988-05-05 Biotechnology Research Partners, Ltd. POLYMORPHISMES DE RECEPTEURS D'apoAI-CIII-AIV, apoAII, apoB, apoCI ET LDL POUR L'IDENTIFICATION GENETIQUE ET LE DIAGNOSTIC PREDICTIF DE L'ATHEROSCLE
ES2142248A1 (es) * 1997-11-20 2000-04-01 Univ Madrid Autonoma Polimorfismos en el promotor del gen humano de la apolipoproteina e, y sus usos para aplicaciones diagnosticas y terapeuticas.
US7972802B2 (en) 2005-10-31 2011-07-05 University Of Washington Lipoprotein-associated markers for cardiovascular disease
US8460889B2 (en) 2008-07-08 2013-06-11 University Of Washington Methods and compositions for diagnosis or prognosis of cardiovascular disease

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA1186991A (fr) * 1982-03-03 1985-05-14 David Owerbach Methode de depistage de la propention au diabete non insulino-dependant et (ou) a l'atherosclerose

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA1186991A (fr) * 1982-03-03 1985-05-14 David Owerbach Methode de depistage de la propention au diabete non insulino-dependant et (ou) a l'atherosclerose

Non-Patent Citations (9)

* Cited by examiner, † Cited by third party
Title
Molecular Biology and Medicine, Vol. 1, issued June 1984 (London NW1 7DX England) S.E. HUMPHRIES et al., "A DNA Polymorphism Adjacent...", pages 463-471, see Summary; page 465, Lines 32-37. *
Nucleic Acids Research, Vol 13, No. 18, issued September 25, 1985 (IRL Press Ltd., Eynsham, Oxford, England) L. PRIESTLEY et al., "RFLP for the Human Apolipoprotein B Gene: II: Ecori", page 6790, see all of page 6790. *
Nucleic Acids Research, Vol. 13, No. 18, issued September 25, 1985 (IRL Press Ltd., Eynsham, Oxford, England) L. PRIESTLEY et al., "RFLP for the Human Apolipoportein B Gene: V; XbaI", page 6793, see all of page 6793. *
Nucleic Acids Research, Vol. 13, No. 18, issued September 25, 1985 (IRL Press Ltd., Eynsham, Oxford, England) L. PRIESTLEY et al., "RFLP for the Human Apolipoprotein B Gene: IV; MspI", page 6792, see all of page 6792. *
Nucleic Acids Research, Vol. 13, No. 18, issued September 25, 1985 (IRL Press Ltd., Eynsham, Oxford, England) L.PRIESTLEY et al., "RFLP for the Human Apolipoprotein B Gene: III; Eco RV", page 6791, see all of page 6791. *
Nucleic Acids Research, Vol. 14, No. 4, issued February 25, 1986 (IRL Press Ltd., Eynsham, Oxford, England) T. COHEN et al., "DNA Polymorphic Site in the Human Apoai-CIII-AIV Cluster; Taq I and Ava I", page 1924, see all of page 1924. *
Nucliec Acids Research, Vol. 13, No. 18, issued September 25, 1985 (IRL Press Ltd., Eynsham, Oxford, England) L. PRIESTLEY et al., "RFLP for the Human Apolipoprotein B Gene: I; BamHI", page 6789, see all of page 6789. *
See also references of EP0239629A4 *
The Lancet, Vol. 1, issued February 4, 1984 (London WC2N GAD England) T. MANDRUP-POULSON et al., "DNA Sequences Flanking the Insulin Gene...", pages 250-252, see page 250, column 1, Lines 12-17; page 251, column 2, Lines 45-57; page 252, column 1, Lines 7-24 *

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1988003175A1 (fr) * 1986-10-29 1988-05-05 Biotechnology Research Partners, Ltd. POLYMORPHISMES DE RECEPTEURS D'apoAI-CIII-AIV, apoAII, apoB, apoCI ET LDL POUR L'IDENTIFICATION GENETIQUE ET LE DIAGNOSTIC PREDICTIF DE L'ATHEROSCLE
EP0269260A3 (fr) * 1986-10-29 1988-06-22 Biotechnology Research Partners, Ltd. Polymorphisme des apoAI-C-III-AIV, apoAII, apoB, apoCI et récepteur LDL pour fingerprinting génétique et annonciateur de l'arthérosclérose
ES2142248A1 (es) * 1997-11-20 2000-04-01 Univ Madrid Autonoma Polimorfismos en el promotor del gen humano de la apolipoproteina e, y sus usos para aplicaciones diagnosticas y terapeuticas.
US7972802B2 (en) 2005-10-31 2011-07-05 University Of Washington Lipoprotein-associated markers for cardiovascular disease
US8420337B2 (en) 2005-10-31 2013-04-16 University Of Washington Lipoprotein-associated markers for cardiovascular disease
US8460889B2 (en) 2008-07-08 2013-06-11 University Of Washington Methods and compositions for diagnosis or prognosis of cardiovascular disease

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ES2003359A6 (es) 1988-11-01
DK277087A (da) 1987-07-30
AU6546886A (en) 1987-04-24
EP0239629A4 (fr) 1988-05-10
DK277087D0 (da) 1987-05-29
EP0239629A1 (fr) 1987-10-07

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