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WO1997038133A1 - Variants du gene preseniline 2 - Google Patents

Variants du gene preseniline 2 Download PDF

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Publication number
WO1997038133A1
WO1997038133A1 PCT/US1997/004683 US9704683W WO9738133A1 WO 1997038133 A1 WO1997038133 A1 WO 1997038133A1 US 9704683 W US9704683 W US 9704683W WO 9738133 A1 WO9738133 A1 WO 9738133A1
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Prior art keywords
seq
gene
exon
sequence
nucleic acid
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Application number
PCT/US1997/004683
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English (en)
Inventor
John Hardy
Alison M. Goate
Rebecca A. Fuldner
Original Assignee
University Of South Florida
Washington University
The Institute Of Genomic Research
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Application filed by University Of South Florida, Washington University, The Institute Of Genomic Research filed Critical University Of South Florida
Priority to AU25414/97A priority Critical patent/AU2541497A/en
Priority to JP53622997A priority patent/JP2002515740A/ja
Priority to EP97916930A priority patent/EP0833947A4/fr
Publication of WO1997038133A1 publication Critical patent/WO1997038133A1/fr

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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
    • 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
    • C07K14/4711—Alzheimer's disease; Amyloid plaque core protein
    • 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

  • AD Alzheimer's Disease
  • the major pathological feature of AD is the presence of numerous neurofibrillary tangles and senile plaques composed prir ⁇ arily of the amyloid protein. These plaques contain beta-amyloid, a peptide varying from 39 to 43 amino acids in length, derived from a larger amyloid precursor protein (APP) (Goate et al. Nature 1991, 349, 704-6; Masters et al. PNAS 1985, 82, 4245-4249; Kang et al. Nature 1987, 325, 733-736). Studies have shown that increases in the generation of the 42 arriino acid peptide lead to AD in APP encoded disease. AD is typically a disease of the elderly, afflicting up to 6% of those aged 65 and up to
  • APP amyloid precursor protein
  • AD This type of AD is termed late-onset.
  • pedigrees a small number of pedigrees have been described wherein the disease is inherited as an autosomal dominant with age dependent penetrance. Most commonly, the age of onset of the disease is below 60 years in these families (presenile). Thus, this type of AD is termed early-onset. Genetic factors have been implicated in both early and late onset AD.
  • presenile AD segregates as fully penetrant autosomal dominant trait.
  • Linkage analysis studies in presenile AD families have identified four genes, on chromosomes 1, 14, 19, and 21, that when mutated, cause presenile AD.
  • the first presenile AD gene identified maps to chromosome 21 and codes for the beta A4-amyloid protein precursor (APP) (Goate et al. Nature 1991, 349, 704-706; Murrell et al. Science 1991, 254, 97-99; Chartier-Harlin et al. Nature 1991, 353, 844-846). Mutations in this gene account for approximately 5% of the families.
  • APP beta A4-amyloid protein precursor
  • F ⁇ esenilin-1 Q°S-1 located on chromosome 14 harbors an estimated 70% of the disease causing mutations, making it the major gene for familial presenile AD (estimated 10% of all AD cases) (Sherrington et al. Nature 1995, 375, 754-760; van Broeckhoven et al. Nature Genet. 1992. 2, 335-339; St. George-Hyslop et al. Nature Genet. 1992, 2, 330-334; Schellenberg et al. Science 1992, 258, 668-670).
  • This gene, designated S182 or PS-1 is made up of 10 coding exons (numbered 3 to 12). PS-1 is predicted to be an integral membrane protein with at least 7 transmembrane domains.
  • a third gene for presenile AD Q?S-2 maps to chromosome 1 in the Volga-German AD families, a group of farnUies in which AD is the result of a founder effect fl-evy-Lahad et al. Science 1995, 269, 970-973).
  • This gene (STM-2 or E5-1) was identified as a direct result of its high homology to PS-1.
  • the same missense mutation was found in 7 Volga-German AD families and more recently, a second missense mutation has been found in an Italian AD family O ⁇ evy- Lahad et al. Science 1995, 269,970-973; Rogaev et al.
  • PS-1 and PS-2 have approximately 450 amino acids and share an overall homology of 67% with the highest similarity observed in the TM domains fl-evy-Lahad et al. Science 1995, 269,973-977; Rogaev et al. Nature 1995, 376,775-778. This degree of homology is indicative of a similar biological function.
  • the putative seven transmembrane domain structure of the presenilins is compatible with a function as a receptor molecule, an ion channel or a membrane structural protein. It has been determined that mutations in PS-1 and PS-2 cause an increase in the generation of Ab 2 thus indicating biological interaction with amyloid.
  • An object of the present invention is to provide novel, variant PS-2 sequences. Another object of the present invention is to provide a method of diagnosing Alzheimer's disease using these novel PS-2 sequences or the exonic or intronic sequences of the PS-2 gene. Yet another object of this invention is to provide a model system for Alzheimer's disease comprising variant PS-2 genes.
  • FIG. 1 provides a schematic comparison of the organization of the PS-1 and PS-2 gene. Labeled arrows indicate sites of known mutations. Unlabeled arrows indicate intron/exon boundaries. Hatched areas in PS-2 indicate sites of alternate splicing. USF#15 contains exons 3, 4 and 8. W.U.#2 lacks exons 3, 4 and 8. W.U.#15 lacks only exon 8.
  • Figure 2 provides the gene sequence of the PS-2 gene (SEQ ID NO: 31 ).
  • the Volga-German families are a culturally distinct subpopulation in Russia, whose members did not marry into the Russian population.
  • the relative onset of AD in this group is exceptionally early, ranging from 50 to 70 years of age.
  • clinically and pathologically AD in these families is indistinguishable from typical AD.
  • the autosomal dominant locus, responsible for AD in the Volga-German kindreds has been localized to chromosome lq31-42 (Levy-Lahad et al. Science 1995, 269, 970-973).
  • Candidate genes which map to this locus have also been identified.
  • Levy-Lahad et al. isolated STM2 whose predicted amino acid sequence is homologous to that of S 182 (PS-1).
  • PS-2 is encoded by 10 coding exons.
  • the PS-2 gene sequence was determined by both sequencing the EST sequence T03796 and isolating PS-2 cDNAs using the GeneTrapper kit (Gibco BRL, Gaithersburg, MD).
  • the intron/exon structure of the PS-2 gene is shown in Table 1. Positions of introns that interrupt the PS-2 cDNA are shown. Exonic sequence is presented in upper case and intronic sequence in lower case letter. Exons are numbered from the 5' end of the cDNA sequence. TABLE 1
  • EXON4 cacgatgtggtttcccacagAGAAGCCAGGAGAACGAGGA EXON4 (141 to 355) ACAGAGAAGAATGGACAGCTgtgagttggggggctggggg
  • EXON5 acagagaagaatggacagctCATCTACACGACATTCACTG
  • EXON5 (356 to 497) ACAAGTACCGCTGCTACAAGgtgaggccctggccctgccc
  • EXON8 aggatgtctctgtcttcctaGATCTCGTGGCTGTGCTGTG EXON8 (787 to 885) CCCTGCCCTGATATACTCATgtgagtgagcccccgtgcc
  • CTTCGTGGCC ATCCTCATTGtgagtggctggggatgcgtc
  • Splicing events in Table 2 are listed with respective cDNAs in which they were identified as well as with corresponding amino acid residues. Splicing events detected by RT-PCR are denoted by a "+" in the PCR column. All events are relative to the full length cDNA clone, referred to as USF# 15. Simplified structures of the four cDN A clones are shown in Figure 1.
  • the identification of the intronic sequences is also useful in the early detection of variant forms of the PS-2 gene.
  • the genomic analysis of the PS-2 gene (see Figure 1) has led to the development of a method for identification of intronic polymorphisms which are predictive of disease. Elucidation, detection, and diagnosis of mutations in both intronic sequences associated with splice variation and in the open reading frames proximal to these intron-exon boundaries of the PS-2 gene can be performed through use of intronic sequences. Identification and analysis of mutants or variants arising from mutations in splice donor or acceptor sites are enabled by knowledge of these intronic sequences. Furthermore, a complete analysis of the intron-exon boundaries makes possible sequencing primers that would allow accurate sequence determination of the first or last 10 to 20 nucleotides of coding exons especially near cDNA termini.
  • AD Alzheimer's disease
  • LCR ligase chain reaction
  • PCR polymerase chain reaction
  • oligonucleotide probes which may be prepared synthetically or by nick translation.
  • the probes are complementary to at least a portion of the variant PS-2 genes identified herein.
  • the DNA probes may be suitably labelled using, for example, a radiolabel, enzyme label, fluorescent label, or biotin-avidin label, for subsequent visualization in for example a Southern blot hybridization procedure.
  • the labelled probe is reacted with a sample of DNA from a patients suspected of having AD bound to nitrocellulose or Nylon 66 substrate.
  • the areas that carry DNA sequences complementary to the labeled DNA probe become labelled themselves as a consequence of the reannealing reaction.
  • the areas of the filter that exhibit such labeling may then be visualized, for example, by autoradiography.
  • ligase chain reaction involves the use of a mismatch probe, i.e., probes which have full complementarity with the target except at the point of the mutation or variation.
  • the target sequence is then allowed to hybridize both with the oligonucleotides having full complementarity, i.e., oUgonucleotides complementary to the PS-2 variants of the present invention, and oligonucleotides containing a mismatch under conditions which will distinguish between the two.
  • LCR ligase chain reaction
  • PCR polymerase chain reaction
  • oligonucleotides complementary to the sequences which flank the DNA of interest are complementary to one of the two strands.
  • the DNA is then denatured at high temperatures (e.g., 95°C) and then reannealed in the presence of a large molar excess of oUgonucleotides.
  • the end product is then denatured again for another cycle. After this three-step cycle has been repeated several times, amplification of a DNA segment by more than one milhon fold can be achieved.
  • the resulting DNA may then be directly sequenced in order to locate any genetic alterations.
  • the identified PS-2 variants of the present invention make it possible to prepare oUgonucleotides that will only bind to altered DNA, so that PCR will only result in the multipUcation of the DNA if the mutation is present.
  • FoUowing PCR, aUele-specific oUgonucleotide hybridization may be used to detect the AD point mutation.
  • PAS A amplification of specific alleles
  • PAS A involves amplification with two oUgonucleotide primers such that one is allele specific.
  • the desired aUele is efficiently amplified, while the other aUele(s) is poorly ampUfied because it mismatches with a base at or near the 3' end of the aUele specific primer.
  • PASA or the related method PAMSA can be used to specifically amplify one or more mutant PS-2 aUeles. Where such amplification is performed on genetic material obtained from a patient, it can serve as a method of detecting the presence of one or more mutant PS-2 aUeles in a patient.
  • PCR-induced mutation restriction analysis often referred to as EMRA, can also be used in the detection of mutants.
  • model systems can be used to screen for agents that alter the degenerative course of AD. Having identified specific mutations in the PS-2 gene as a cause of early onset famiUal Alzheimer's disease, it is possible using genetic manipulation, to develop transgenic model systems and/or whole ceU systems containing a mutated PS-2 gene or a portion thereof.
  • the model systems can be used for screening drugs and evaluating the efficacy of drugs in treating Alzheimer's disease.
  • these model systems provide a tool for defining the underlying biochemistry of PS-2 and its relationship to AD thereby providing a basis for rational drug design.
  • ceU system which can be used in the present invention can be naturaUy derived.
  • blood samples from an affected individual are obtained and permanently transformed into a lymphoblastoid ceU line using, for example, Epstein-Barr virus.
  • Epstein-Barr virus Once established, such cell lines can be grown continuously in suspension cultures and can be used in a variety of in vitro experiments to study PS-2 expression and processing.
  • Another ceU line used in these studies comprises skin fibroblasts derived from patients. Since the FAD mutation is dominant, an alternative method for constructing a ceU line is to genetically engineer a PS-2 mutated gene, or portion thereof, as described herein, into an estabUshed ceU line of choice.
  • Baculovirus expression systems have also been found to be useful for high level expression of heterologous genes in eukaryotic ceUs.
  • the mutated gene can also be excised for use in the creation of transgenic animals containing the mutated gene.
  • a PS-2 gene of the present invention can be cloned and placed in a cloning vector.
  • cloning vectors which can be used include, but are not limited to, lCharon35, cosmid, or yeast artificial chromosome.
  • the variant PS-2 gene can then be transferred to a host nonhuman animal such as a mouse. As a result of the transfer, the resultant transgenic nonhuman animal will preferably express one or more of the variant PS-2 polypeptides.
  • minigenes encoding variant PS-2 polypeptides can be designed.
  • Such minigenes may contain a cDNA sequence encoding a variant PS-2 polypeptide, preferably fuU- length, a combination of PS-2 exons, or a combination thereof, linked to a downstream polyadenylation signal sequence and an upstream promoter (and preferably enhancer).
  • Such a minigene construct will, when introduced into an appropriate transgenic host, such as a mouse or rat, express a variant PS-2 polypeptide.
  • One approach to creating transgenic animals is to target a mutation to the desired gene by homologous recombination in an embryonic stem (ES) ceU in vitro foUowed by microinjection of the modified ES ceU line into a host blastocyst and subsequent incubation in a foster mother.
  • ES embryonic stem
  • the technique of microinjection of the mutated gene, or portion thereof, into a one-ceU embryo followed by incubation in a foster mother can be used. Additional methods for producing transgenic animals are weU known in the art.
  • Transgenic animals are used in the assessment of new therapeutic compositions and in carcinogenicity testing, as exemplified by U.S. Patent 5,223,610. These animals are also used in the development of predictive animal models for human disease states, as exemplified in U.S. Patent 5,221,778. Transgenic animals have now been developed for assessing Alzheimer's disease (U.S. Patent 7,769,626), multi-drug resistance to anticancer agents (U.S. Patent 7,260,827), and carcinogenic substances (U.S. Patent 4,736,866).
  • the PS-2 genes of the present invention which are beUeved to cause early onset Alzheimer's disease in Chromosome 14-Unked pedigrees provide a useful means for developing transgenic animals to assess this disease.
  • Site directed mutagenesis and or gene conversion can also be used to a mutate a non human PS-2 gene aUele, either endogenously or via transfection, such that the mutated gene encodes a polypeptide with an altered amino acid as described in the present invention.
  • antibodies to the PS-2 gene and variants thereof can be raised for use in the examination of the function of the truncated transcripts of the PS-2 gene.
  • These antibodies can be, for example, polyclonal or monoclonal antibodies.
  • the present invention also includes chimeric, single chain, and humanized antibodies, as weU as Fab fragments, or the product of an Fab expression Ubrary. Various procedures known in the art may be used for the production of such antibodies and fragments.
  • Antibodies generated against the PS-2 genes of the present invention can be obtained by direct injection into an animal or by administering the gene to an animal, preferably a nonhuman. The antibody so obtained will then bind the PS-2 gene or itself. In this manner, even a fragment of the gene can be used to generate thee antibodies.
  • any technique which provides antibodies produced by continuous cell line cultures can be used. Examples include the hybridoma technique (Kohler and Milstein, Nature 1975, 256, 495-497), the trioma technique, the human B-ceU hybridoma technique (Kozbor et al., Immunology Today 1983, 4, 72), and the EBV-hybridoma technique to produce human monoclonal antibodies (Cole et al, in Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, Inc., 1985, pp. 77-96). Techniques described for the production of single chain antibodies (U.S. Patent
  • 4,946,778 can be adapted to produce single chain antibodies to the PS-2 genes of this invention. Also, transgenic mice may be used to express humanized antibodies to the PS-2 genes of this invention.
  • PS-2 cDNA's were isolated using the Gene Trapper kit (Gibco BRL) according to the manufacturer's directions.
  • a human superscript brain library (Gibco BRL, Gaithersburg, MD) in pCMV.SPORT was probed with the primer 5 -CATTCACTGAGGACACACCC-3' (SEQ ID NO: 1) (derived from the EST sequence T03796).
  • RNA from brain, heart, hver, lung, placenta, and skeletal muscle was obtained from Clontech 0°alo Alto, CA)
  • First strand cDNA was synthesized foUowing the Superscript Preamplification System for First Strand cDNA Synthesis (Gibco BRL)
  • Two mg of total RNA was combined with 1 mg of oUgonucleotide (dT)i 2 is primer, 100 ng of random hexamer p ⁇ mer and DEPC treated water in a 05 ml tube
  • Samples were incubated at 70°C for 10 minutes and placed on ice Kit components were added in accordance with the manufacturer's protocol, and samples were incubated at 37°C for 2 minutes
  • Supersc ⁇ pt II Reverse Transc ⁇ ptase (RT, 200 U) was added and samples were incubated at 37°C for 1 hour Samples were then incubated at 70°C for 15 minutes, chilled on ice, and 2 U of RNAse H was added
  • 3PS2X3 35 GCT GGT GTG GAG CTG CAG GTA CAG 405 TG
  • 3PS2X5 39 TGG GGA AGA CTG GAG CTC GAT G 263
  • 3PS2X6 41 GTG CAG CAC TGG GGA CGA TTT 360
  • 3PS2X8 45 AGC TGG TCA GAG TGT TAC 510
  • AUquots of tissue cDNAs were ampUfied in a Perkin Elmer DNA Thermal Cycler 480 (Perkin Elmer, Norwalk, CT). Each PCR reaction contained 1 ml of final cDNA product, 25 pmol of each primer (forward and reverse), 12.5 nmol of dNTP (Pharmacia, Columbus, OH), 1.25 U of Taq polymerase (Promega, Madison, WI) for a total reaction volume of 50 ml overlaid with 60 ml mineral oil O ⁇ isher, Pittsburgh, PA). Primers used were designed to span at least two putative intron/exon boundaries of the PS-2 gene.
  • LP313F forward, exon 2
  • LP676R reverse, exon 4
  • samples were denatured at 94°C for 5 minutes. Samples then underwent 35 cycles of 0.5 minutes at 94°C, 0.5 minutes at the relevant anneaUng temperature for the given primer pair, and 0.75 minutes at 72°C. This was foUowed by a final extension for 10 minutes at 72°C. Products were visualized on a 2% agarose gel 0?romega) using ethidium bromide staining. Product bands were excised and purified using the Wizard PCR Preps DNA Purification System 07 ⁇ omega). Five microUters of the final 50 ml purified product was used for sequencing.
  • PCR product was treated with Exonuclease 1 and Shrimp Alkaline Phosphatase 0?CR sequencing kit-USB, Cleveland, OH). Five microUters of PCR product was incubated with 1 U of Exonuclease 1 for 15 minutes at 37°C. The sample was then held at 80°C for 15 minutes after which 2 U of Shrirnp Alkaline Phosphatase was added. As before, the sample was held at 37°C for 15 minutes and then at 80°C for 15 minutes. This final 7 ml product was used in the sequencing protocol described in the Sequenase PCR product Sequencing Kit. The forward primer used in the original PCR reaction was used for manual sequencing.
  • Example 5 PAC isolation PI derived artificial chromosomes 0?ACs were isolated by screening a gridded Ubrary (Genome Systems, Inc.) with PCR products ampUfied with primers R05822F and R05822R. Three PACs containing the PS-2 gene were digested with NotI and sized using pulse field gel electrophoresis (PFGE). PAC DNA was run at 200 V for 21 hours at 14°C with switch times varying from 5-20 seconds. Sizes ranged from 90 kb to 1 10 kb.
  • PFGE pulse field gel electrophoresis
  • Primers used in the detection of the 5' untranslated sequence was 5'-GCTTCTGTCTCAGGTTTCTTC-3'(SEQ ID NO: 3) and 5'- CC ⁇ JTGTTTGGCTGTTTTATCA-3' (SEQ ID NO: 4). Int ronic primers were used to detect presence of exons 4 and 7. Primers R05822F and R05822R were used to detect exon 12. PCR reaction conditions were as foUows: 5 minutes denaturation foUowed by 35 cycles of 94°C for 0.5 minutes, the respective anneaUng temperature for 0.5 minutes, and 72°C for 0.5 minutes. A 10 minute extension at 72°C concluded each reaction. AnneaUng temperature for each primer pair are provided in Table 2.
  • Example 6 Identification of exon intron boundaries Exon/intron boundaries were obtained by PCR/Ugation techniques. Purified PAC DNA was digested with a variety of blunt-cutting enzymes and ligated to a specificaUy designed linker. Sequences were then specificaUy ampUfied by PCR using a linker-derived primer and a PS-2 derived primer for boundary sequencing. Sequencing of products was performed directly in low- melting point agarose by using a modified dideoxynucleotide sequencing method with a 32 P end labeUed primer and Taq DNA polymerase temperature cycled reactions.
  • the Pharmacia ALF fragment manager was used to detect smaU sequence changes not detectable by standard agarose gel/ethidium bromide visualization. Relevant cDNA sequences were ampUfied by PCR using a 5'-fluorescein tagged primer.
  • CTACAGTGTG CTGGTGGGCA AGGCGGCTGC CACGGGCAGC GGGGACTGGA 1550

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Abstract

L'invention concerne des variants du gène préséniline 2, ainsi que des méthodes permettant d'utiliser lesdits variants pour le diagnostic de la maladie d'Alzheimer.
PCT/US1997/004683 1996-04-04 1997-03-20 Variants du gene preseniline 2 WO1997038133A1 (fr)

Priority Applications (3)

Application Number Priority Date Filing Date Title
AU25414/97A AU2541497A (en) 1996-04-04 1997-03-20 Variant presenilin-2 genes
JP53622997A JP2002515740A (ja) 1996-04-04 1997-03-20 プレセニリン−２遺伝子の変種
EP97916930A EP0833947A4 (fr) 1996-04-04 1997-03-20 Variants du gene preseniline 2

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US1486096P 1996-04-04 1996-04-04
US60/014,860 1996-04-04

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1078987A4 (fr) * 1998-05-21 2003-10-01 Tanabe Seiyaku Co Methode d'examen des maladies du snc et de criblage des remedes associes
EP1724588A3 (fr) * 2001-03-12 2007-09-05 Isis Innovation Limited Méthodes de diagnostic de la maladie d'Alzheimer

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5525714A (en) * 1992-03-20 1996-06-11 N. V. Innogenetics S.A. Mutated form of the β-amyloid precursor protein gene

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5986054A (en) * 1995-04-28 1999-11-16 The Hospital For Sick Children, Hsc Research And Development Limited Partnership Genetic sequences and proteins related to alzheimer's disease

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5525714A (en) * 1992-03-20 1996-06-11 N. V. Innogenetics S.A. Mutated form of the β-amyloid precursor protein gene

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
AMERICAN J. PATHOLOGY, December 1996, Vol. 149, No. 6, MURPHY G.M. et al., "Antibodies to Presenilin Proteins Detect Neurofibrillary Tangles in Alzheimer's Disease", pages 1839-1845. *
NATURE MEDICINE, February 1996, Vol. 2, No. 2, KOVACS et al., "Alzheimer-Associated Presenilins 1 and 2: Neuronal Expression in Brain and Localization to Intracellular Membranes in Mammalian Cells", pages 224-229. *
See also references of EP0833947A4 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1078987A4 (fr) * 1998-05-21 2003-10-01 Tanabe Seiyaku Co Methode d'examen des maladies du snc et de criblage des remedes associes
EP1724588A3 (fr) * 2001-03-12 2007-09-05 Isis Innovation Limited Méthodes de diagnostic de la maladie d'Alzheimer
JP2008245654A (ja) * 2001-03-12 2008-10-16 Isis Innovation Ltd アルツハイマー病の診断スクリーニング
JP2010166922A (ja) * 2001-03-12 2010-08-05 Isis Innovation Ltd アルツハイマー病の診断スクリーニング

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CA2222813A1 (fr) 1997-10-16
JP2002515740A (ja) 2002-05-28
EP0833947A4 (fr) 2003-07-02
EP0833947A1 (fr) 1998-04-08

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