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WO2015001113A1 - Procédés pour le diagnostic de maladies neurodégénératives cérébrales - Google Patents

Procédés pour le diagnostic de maladies neurodégénératives cérébrales Download PDF

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WO2015001113A1
WO2015001113A1 PCT/EP2014/064382 EP2014064382W WO2015001113A1 WO 2015001113 A1 WO2015001113 A1 WO 2015001113A1 EP 2014064382 W EP2014064382 W EP 2014064382W WO 2015001113 A1 WO2015001113 A1 WO 2015001113A1
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dpp6
mutations
protein
disease
mutation
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Christine Van Broeckhoven
Kristel SLEEGERS
Rita CACACE
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Universiteit Antwerpen
Vlaams Instituut voor Biotechnologie VIB
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Universiteit Antwerpen
Vlaams Instituut voor Biotechnologie VIB
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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 present application relates to the field of human genetics, particularly in relation to the field of neurodegenerative brain diseases.
  • the present invention relates to methods and materials to detect human neurodegenerative brain diseases, more particularly dementia (Alzheimer's disease (AD), Frontotemporal lobar degeneration (FTLD), dementia with Lewy bodies dementia (DLB)), amyotrophic lateral sclerosis (ALS) and Parkinson's disease (PD).
  • human neurodegenerative brain diseases more particularly dementia (Alzheimer's disease (AD), Frontotemporal lobar degeneration (FTLD), dementia with Lewy bodies dementia (DLB)), amyotrophic lateral sclerosis (ALS) and Parkinson's disease (PD).
  • AD Alzheimer's disease
  • FTLD Frontotemporal lobar degeneration
  • ALS amyotrophic lateral sclerosis
  • PD Parkinson's disease
  • diseases include AD, FTLD and diseases of the FTLD/ALS spectrum.
  • diagnostic assays for the detection of neurodegenerative brain diseases kits for performing
  • Neurodegenerative brain diseases are a group of disorders characterized by a progressive loss of structure or function of neurons, including death of neurons.
  • Many neurodegenerative diseases including Alzheimer's disease (AD), Frontotemporal lobar degeneration (FTLD), amyotrophic lateral sclerosis (ALS) and Parkinson's disease (PD) occur as a result of neurodegenerative processes.
  • AD Alzheimer's disease
  • FTLD Frontotemporal lobar degeneration
  • ALS amyotrophic lateral sclerosis
  • PD Parkinson's disease
  • Many of these diseases share similarities, which relate these diseases to one another on a sub-cellular level.
  • different neurodegenerative disorders feature atypical protein assemblies as well as induced cell death. More specifically, it is known that the clinical presentations of AD, FTLD and dementia with Lewy bodies (DLB) overlap considerably, despite the different pathologic processes (Piquet et al., 2009).
  • NBD neurodegenerative brain disease
  • the NBD is selected from the group of Alzheimer disease (AD), frontotemporal lobar degeneration (FTLD), Amyotrophic Lateral Sclerosis (ALS) and FTLD/ALS. More particularly, the neurodegenerative brain disease is a neurodegenerative dementia. Most particularly, the NBD is selected from the group of AD and FTLD.
  • DPP6 altered expression is indicative of the presence of a NBD.
  • Expression levels can be assessed using methods well known in the art. Without being limited to a particular technology, this can be using quantitative RT-PCR (e.g. for determining mRNA levels of DPP6) or using ELISA or Western Blot (e.g. for determining protein levels of DPP6).
  • quantitative RT-PCR e.g. for determining mRNA levels of DPP6
  • ELISA e.g. for determining protein levels of DPP6
  • Western Blot e.g. for determining protein levels of DPP6
  • the presence of one or more mutations in the DPP6 gene may be determined. Most particularly, the mutations will have a deleterious effect on DPP6 gene function, either by altering (e.g. decreasing) expression levels of the gene, or by decreasing or altering its function.
  • the one or more mutations are selected from mutations affecting the coding regions of DPP6, particularly exonic mutations, and complex genomic mutations affecting gene expression of DPP6.
  • Mutations affecting the coding regions of DPP6 typically are selected from missense mutations, nonsense mutations and frame-shift mutations.
  • Genomic mutations affecting gene expression are typically selected from inversions, deletions or duplications (the latter two can be indicated as CNVs).
  • mutations affecting translation into DPP6 protein or transcript stability that are not in the coding regions of DPP6 are also envisaged. Typically, these are mutations in the translation initiation codon or in regulatory sequences such as those regulating splicing, transcript stability or turn-over.
  • the DPP6 mutations are selected from mutations affecting transcript stability, translation or protein function and/or conformation.
  • At least one of the mutations of which the presence is determined is a mutation in the extracellular domain of DPP6.
  • At least one of the mutations is selected from those listed in Table 6 (particularly 6A and 6C, more particularly 6A), Table 8, and an inversion or CNV affecting DPP6. More particularly, it is envisaged that at least one of the mutations whose presence is determined is a mutation selected from those listed in Table 6 and in Table 8, and an inversion affecting DPP6. More particularly, it is envisaged that at least one of the mutations whose presence is determined is a mutation selected from those listed in Table 6 and in Table 8. According to even further embodiments, at least one of the mutations is selected from the mutations listed in Table 6.
  • At least one mutation is a null mutation and is causative of the neurodegenerative brain disease (i.e., is a high penetrant causal mutation).
  • the functional expression levels of DPP6 are decreased by at least 20%, and this is causative of the neurodegenerative brain disease.
  • the decrease in expression levels is at least 30% or at least 40%.
  • the one or more mutations when diagnosis is based on mutations, are not null mutations and are indicative of an increased risk of developing or presence of the neurodegenerative brain disease. I.e., they are mutations with incomplete penetrance.
  • diagnosis when diagnosis is based on expression levels, the functional expression levels of DPP6 are decreased by less than 20%, and this is indicative of an increased risk of developing or presence of the neurodegenerative brain disease.
  • a decrease of less than 30%, or less than 40% is indicative of an increased risk of developing or presence of the neurodegenerative brain disease.
  • the methods can be performed using direct sequencing methods or others.
  • detection of mutations is done using (e.g.
  • kits comprising at least one primer or probe suitable to determine the presence of one or more mutations in DPP6.
  • kits are provided comprising suitable means for determining the expression levels of DPP6, e.g. T-qPC primers and Taqman probes.
  • the DPP6 protein, or nucleic acid encoding said protein is provided for use as a medicament.
  • the DPP6 protein, or nucleic acid encoding said protein is provided for use as a diagnostic.
  • the DPP6 protein, or nucleic acid encoding said protein is provided for use in treating NBD as described herein. Treating can also mean delay the onset of, or prevent the onset of, the NBD.
  • the NBD is selected from AD and FTLD.
  • methods of treating NBD (particularly AD or FTLD) in a subject in need thereof comprising restoring the levels of DPP6 in said subject (e.g. increasing DPP6 levels in case of a loss-of-function).
  • the levels of DPP6 are increased by administering the DPP6 protein, or nucleic acid encoding said protein, to said subject. This can for instance be achieved using a gene therapy method.
  • levels of DPP6 can also be restored to normal levels by administering a compound that restores DPP6 expression. For instance, in case of reduced DPP6 levels, the levels can be increased. In case of excessive DPP6 expression (e.g. because of a gain-of-function), the levels can be decreased.
  • Figure 1 shows the pedigree of family 1270.
  • a black filled symbol characterizes a patient and a white unfilled symbol an unaffected individual or an at-risk individual with unknown phenotype.
  • the Roman numbers on the left of the pedigree denote generations. Arabic numbers above the symbols denote individuals. The Arabic numbers below the symbols denote age-at-onset for patients or either age at last examination or age at death for unaffected individuals and at-risk individuals with unknown phenotype.
  • An asterisk (*) indicates an individual who was included in the linkage analysis.
  • the arrow identifies the proband with early age at onset AD (47 years). Patients used in WGS are circled.
  • FIG. 1 Disguised linkage pedigree of family 1270 (fig. 1). Haplotypes are based on 17 informative polymorphic genetic microsatellite markers at chromosome band 7q36. Haplotypes for individuals from the first and second generations were inferred from genotype data of siblings and offspring. The risk haplotype was arbitrarily set for individual 1-2. For confidentiality reasons, haplotypes are shown only for patients; the number of at-risk individuals included in the genotyping is indicated by numbers within diamonds.
  • Figure 3 Linkage pedigrees of Dutch families with early-onset AD who share haplotypes at 7q36.
  • a black filled symbol represents a patient and a white symbol represents an unaffected individual or an at-risk individual with unknown phenotype.
  • the Roman numbers on the left of the pedigree denote generations; Arabic numbers above the symbols denote individuals.
  • the age (in years) at onset is shown below the symbol.
  • An asterisk (*) indicates that DNA was available for genetic analysis.
  • the arrow identifies the proband.
  • Haplotypes for individuals 111-20 in family 1242 and individual 111-4 in family 1034 were reconstructed from genotype data obtained from their siblings and offspring. For family 1125, only genotype data are given, since segregation data were not informative for deduction of haplotype.
  • FIG. 4 Schematic presentation of alternate transcriptional splice variants of DPP6.
  • DPP6 has 3 transcripts resulting in 3 different isoforms. Both isoforms 2 and 3 have a distinct N-terminus, compared to isoform 1. Note: The gene spans an assembly gap in intron 5 of 100 kb.
  • Figure 5 Schematic presentation of the location of the inversion breakpoint relatively to DPP6 isoforms and promoters.
  • the blue bar denotes part of the linked region of family 1270, the dark blue bar part of the shared haplotype between the 4 families and the green bar part of the inversion.
  • the 3 DPP6 isoforms have each a different core promoter predicted by FirstEF (First-Exon and Promoter Prediction) based on high GC content, and a distinct translation initiation codon within their own exon 1.
  • FirstEF First-Exon and Promoter Prediction
  • Figure 6 Graphical presentation of the haplotype sharing between families 1270, 1034, 1125 and 1242.
  • the differently colored horizontal bars represent (part of) he shared haplotype region in each family based on allele sharing (Table 2).
  • the red horizontal bar is the minimal shared haplotype derived for allele and haplotype sharing in all 4 families ( Figure 3).
  • FIG. 7 Structure of DPP6 (Strop et al., 2004).
  • Figure 8 Schematic presentation of DPP6 isoforms and location of the rare non-synonymous variants.
  • Figure 9 protein structure details of DPP6 showing how mutation may affect interaction with functional residues.
  • A interaction of p.322 (wt) with p.N319;
  • B interaction of p.D659 (wt) with p.N566.
  • Figure 10 Scatter plot of pooled relative expression levels of DPP6 in patients vs control individuals.
  • Figure 11 Relative expression levels of total DPP6.
  • the two blue bars on the left represent the average levels in controls vs DPP6 carriers.
  • the middle three red bars are the expression levels in the separate carrier individuals, the green bar on the right represents the expression levels in the VCP mutation carrier only.
  • Panel A and B reports the relative expression levels obtain from two independent experiments.
  • Figure 12 DPP6 antibody optimization. Two concentrations tested (1/500 and 1/1000). Right panel shows reactivity of the antibody vs human and mouse DPP6.
  • Figure 13 Western Blot of patient and control DPP6 levels. Normalization vs beta-actin (lower pa confirms lowered expression in patients.
  • NBD neurodegenerative brain disease
  • FTLD frontotemporal lobar degeneration
  • FDD frontotemporal dementia
  • Pick disease G31.0
  • dementia with Lewy bodies DLB
  • Parkinson's disease PD
  • ALS amyotrophic lateral sclerosis
  • DPP6 refers to the dipeptidyl-peptidase 6 gene (Gene ID: 1804 in humans), and its encoded protein, as well as the m NA that is transcribed from the gene.
  • the DPP6 gene encodes a single-pass type II membrane protein that is a member of the S9B family in clan SC of the serine proteases. This protein has no detectable protease activity, most likely due to the absence of the conserved serine residue normally present in the catalytic domain of serine proteases. However, it does bind specific voltage-gated potassium channels and alters their expression and biophysical properties. Alternate transcriptional splice variants, encoding (at least) three different isoforms, have been characterized.
  • DPP6 encompasses the different isoforms.
  • mutation in the DPP6 gene or “mutation in DPP6” as used herein refers to mutations in the coding sequence of the gene as well as mutations in the non-coding regions (e.g. introns, promoter region, UTR). Examples of mutations include, but are not limited to, substitutions, insertions, deletions, indels, amplifications, inversions, copy-number-variations (CNV). Mutations can have different effects, e.g. loss-of-function (up to complete loss-of-function, i.e. amorphic mutations), gain-of-function, dominant negative mutation, and so on.
  • loss-of-function up to complete loss-of-function, i.e. amorphic mutations
  • gain-of-function dominant negative mutation, and so on.
  • mutations resulting in decreased transcription or translation are also regarded as loss-of-function mutations.
  • mutations as used herein covers both mutations that are causative of the disease (high penetrant, pathogenic mutations) as those that only confer an increased risk of developing the disease (i.e. low to median penetrant mutations).
  • the mutations are null mutations.
  • a mutation in the DPP6 gene will typically be a small scale mutation, i.e. only comprise one or a few nucleotides (note that, in case of frameshift or nonsense mutations, a change of even one nucleotide in the genomic sequence may lead to a much larger difference in the resulting gene product). Larger mutations are envisaged as well within the definition.
  • a particular class of mutations are complex genomic mutations.
  • the term "complex genomic mutation” or “genomic mutation” is used herein to indicate structural variations that affect more than one gene (as opposed to single nucleotide changes, which are limited to one gene).
  • genomic mutations may also be indicated as large-scale mutations in chromosomal structure, and typically affect at least one kilobase (1000 nucleotides) but may be much larger (up to several megabases).
  • Typical examples include, but are not limited to: amplifications (or gene duplications) leading to more than one copy of a chromosomal region, deletions of large chromosomal regions (leading to loss of the genes within those regions), chromosomal translocations (interchange of genetic parts from nonhomologous chromosomes), interstitial deletions (an intra-chromosomal deletion that removes a segment of DNA from a single chromosome), chromosomal inversions (reversing the orientation of a chromosomal segment).
  • Copy-number variations is the term used to indicate the group of structural variations formed by amplifications and deletions.
  • DPP6 When a complex genomic mutation affects part of the (coding or non-coding) sequence of the DPP6 gene, it is envisaged within the definition of "mutation in DPP6".
  • DPP6 is located in a low copy repeat (LC ) region in the human genome, and it is known that such regions, due to their high sequence identity as a result of segmental duplication, are prone to non-allelic homologous recombination (NAHR), leading to genomic mutations (deletions, duplications and inversions).
  • NAHR non-allelic homologous recombination
  • mutations in the DPP6 genomic region may arise that do not affect the DPP6 gene sequence, but cause instability of DPP6 (e.g. because transcription of DPP6 is affected).
  • mutations are not a mutation in DPP6, they do affect functional expression of DPP6 and are envisaged under that terminology.
  • “functional expression” of DPP6 it is meant the transcription and/or translation of functional gene product.
  • “Functional expression” can be deregulated on at least three levels. First, at the DNA level, e.g. by absence or disruption of the gene, or lack of transcription taking place (in both instances preventing synthesis of the relevant gene product). The lack of transcription can e.g. be caused by epigenetic changes (e.g. DNA methylation) or by loss of function mutations.
  • a “loss-of-function” or “LOF” mutation as used herein is a mutation that prevents, reduces or abolishes the function of a gene product as opposed to a gain-of-function mutation that confers enhanced or new activity on a protein.
  • LOF can be caused by a wide range of mutation types, including, but not limited to, a deletion of the entire gene or part of the gene, splice site mutations, frame-shift mutations caused by small insertions and deletions, nonsense mutations, missense mutations replacing an essential amino acid and mutations preventing correct cellular localization of the product. Also included within this definition are mutations in promoters or regulatory regions of the DPP6 gene if these interfere with gene function.
  • a null mutation is an LOF mutation that completely abolishes the function of the gene product.
  • a null mutation in one allele will typically reduce expression levels by 50%, but may have severe effects on the function of the gene product. Note that functional expression can also be deregulated because of a gain of function mutation: by conferring a new activity on the protein, the normal function of the protein is deregulated, and less functionally active protein is expressed.
  • RNA level e.g. by lack of efficient translation taking place - e.g. because of destabilization of the mRNA (e.g. by UTR variants) so that it is degraded before translation occurs from the transcript.
  • lack of efficient transcription e.g. because a mutation introduces a new splicing variant.
  • proteins with reduced functionality or activity e.g. enzymatic activity, or binding activity, such as the binding to voltage-gated potassium channels
  • truncated proteins e.g. as a result of a gain of function mutation
  • proteins with altered function e.g. as a result of a gain of function mutation
  • a truncated protein may be equally expressed as the wild type counterpart, this will typically result in a decrease in functional expression (or functional expression levels), since the truncated protein will be less active (or less functional).
  • DPP6 may be decreased by affecting post-translational modifications, such as glycosylation.
  • post-translational modifications such as glycosylation.
  • glycosylation is important for protein function, and we have identified several mutations in NBD patients that affect glycosylation.
  • mutations affecting glycosylation of DPP6 and thereby its function are also envisaged as mutations affecting functional levels of DPP6.
  • the essence is that a decrease in DPP6 functionality and/or its levels will increase risk of, or even cause, a neurodegenerative brain disease.
  • the present application is the first to show that mutations in DPP6 (which have an effect on function or expression) or affecting DPP6 expression are found in patients with different NBD, most particularly AD and FTLD (including FTLD/ALS), and are causative of the disease, or predispose to an increased risk of having or developing the NBD. Accordingly, in a first aspect, methods are provided of diagnosing the presence of and/or risk of developing a NBD in a subject, comprising determining the presence of one or more mutations in the DPP6 gene and/or determining the functional expression levels of DPP6 in a sample of said subject.
  • the NBD is Alzheimer disease, FTLD, FTLD/ALS, or ALS.
  • mutations in DPP6 were primarily identified in AD or FTLD, it is well established that carriers of such mutations are often also at a higher risk of other NBD, cf. e.g. the progranulin missense mutations found in AD. Further characterization of the DPP6 gene is ongoing in patients with ALS, dementia with Lewy bodies, Parkinson's disease and others.
  • the neurodegenerative brain disease does not encompass multiple sclerosis (MS).
  • the one or more mutations whose presence is determined are typically mutations affecting the function of DPP6, or the amount of functional DPP6 protein. Most typically, the mutations will decrease the function of DPP6 or the amount of functional DPP6 protein. Note that mutations affecting DNA or NA are included within the mutations that affect function or amount of protein: absence of protein is also a decrease in function or amount. Thus, in these embodiments, the one or more mutations will be deleterious mutations, or even null mutations or loss-of-function mutations. The presence of these deleterious mutations is indicative of an increased risk of developing, or of the presence of neurodegenerative disease (depending i.a. of the age of the subject when the presence of mutations is determined).
  • DPP6 mutations or expression levels on NBD is already noticeable in the heterozygous state, and is dosage sensitive.
  • any decrease in the amount of functional DPP6 will have an effect: the mutation does not need to lead to a complete loss of function.
  • the dosage sensitivity implies that the more severe the effect of the mutation on the gene product, the stronger the effect on neurodegenerative brain disease.
  • a mutation that decreases expression of functional DPP6 protein by 10% will likely confer just an increased risk to develop neurodegenerative disease, while a null mutation (or null allele) decreasing functional protein by 50% or more is likely high-penetrant and can cause early-onset neurodegenerative disease.
  • Penetrance of DPP6 mutations refers to the proportion of individuals carrying a particular variant of DPP6 (allele or genotype) that also exhibits (or will develop) clinical symptoms of a NBD. For example, if a mutation in DPP6 has 95% penetrance, then 95% of those with the mutation will develop a neurodegenerative brain disease, while 5% will not.
  • the frequencies indicated to characterize penetrance are typically age-related cumulative frequencies: while an individual carrying a highly penetrant mutation will normally develop disease, it is very well possible that the individual does not have the disease yet at time of diagnosis.
  • High penetrance or “high-penetrant mutations” as used herein refers to mutations with an age-related cumulative frequency of at least 80%.
  • Median penetrance or “incomplete penetrance” as used herein refers to mutations with an age-related cumulative frequency of between 20 and 80%, or particularly between 30 and 80% or between 40 and 80%.
  • the one or more mutations whose presence is determined do not encompass rsl0260404.
  • this SNP in the DPP6 genomic region has been identified in a GWAS study as being associated with susceptibility to ALS (Van Es et al., 2008).
  • the difference between the present findings and GWAS is that genome-wide association studies identify SNPs and other variants in DNA which are associated with a disease, but cannot on their own specify which genes are causal.
  • the SNP identified in an intronic region has no effect on protein function and expression. This is distinct from the present invention, where it could be shown that altered levels or function of DPP6 (e.g.
  • the neurodegenerative disease is ALS
  • the one or more mutations whose presence is determined are not CNVs.
  • the one or more mutation whose presence is determined are not in intron 3.
  • the one or more mutation whose presence is determined is not in the 5' region of DPP6.
  • the one or more mutation whose presence is determined does not encompass a CNV in intron 3.
  • the one or more mutation whose presence is determined does not encompass a CNV in the 5' region of DPP6.
  • the one or more mutations whose presence is determined are exonic mutations.
  • the neurodegenerative disease that is diagnosed is not ALS, with the exception of FTLD/ALS cases.
  • the neurodegenerative disease that is diagnosed is not ALS.
  • a subject typically is a vertebrate subject, more typically a mammalian subject, most typically a human subject.
  • a sample of the subject will typically contain cells (or at least cellular material) of the subject, to evaluate the expression or presence of mutations in the genetic material.
  • a sample can be obtained from any tissue to establish the presence of germline mutations, and can be a tissue sample, or a fluid sample (e.g. blood, saliva).
  • a tissue sample e.g. blood, saliva
  • a tissue which can be linked to the presence of neurodegenerative disease such as a brain sample or a CSF sample.
  • samples that are particularly envisaged include neuronal cells, neuronal cell lines (particularly primary cell cultures, most particularly primary cell cultures derived from the subject to be diagnosed), iPS cells and fibroblasts.
  • the samples can be used as such or can be pre-processed (e.g. lysed) using methods routinely used in the art.
  • these methods will further include a step involving correlating the levels of DPP6 gene product to the risk of presence or development of neurodegenerative brain disease, particularly correlating decreased levels (or even absence) of DPP6 gene product to increased risk of neurodegenerative disease.
  • the reverse can also be true: concluding from an observation that the DPP6 gene product levels are not decreased, or are even increased, in the sample, that there is no increased risk of neurodegenerative disease, or in some instances even a decreased risk of neurodegenerative disease.
  • Decreased levels of DPP6 gene product are typically decreased versus a control. The skilled person is capable of picking the most relevant control.
  • Suitable controls include, but are not limited to, similar samples from subjects not having neurodegenerative disease, the average levels in a control group, or a set of clinical data on average DPP6 gene product levels in the tissue from which the sample is taken.
  • the control may be from the same subject, or from one or more different subjects or derived from clinical data.
  • the control is matched for e.g. sex, age etc.
  • DPP6 gene product With 'decreased' levels of DPP6 gene product as mentioned herein, it is meant levels that are lower than are normally present. Typically, this can be assessed by comparing to control. According to particular embodiments, decreased levels of DPP6 are levels that are 10%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 90%, 100%, 150%, 200% or even more low than those of the control. According to further particular embodiments, it means that DPP6 gene product is absent, whereas it normally (or in control) is expressed or present. In other words, in these embodiments determining the presence of DPP6 gene product is equivalent to detecting the absence of DPP6 gene product. Typically, in such cases, a control will be included to make sure the detection reaction worked properly.
  • DPP6 gene product needs to be lower in order to allow a reliable and reproducible diagnosis may depend on the type of sample tested and of which product (mRNA, protein) the levels are assessed. However, assessing the correlation itself is fairly straightforward.
  • DPP6 gene product levels measured in the sample are similar to those of a sample with neurodegenerative disease, (or are e.g. comparable to DPP6 gene product levels found in a clinical data set of neurodegenerative disease), this may be considered equivalent to decreased DPP6 gene product levels compared to a healthy control, and be correlated to an increased risk of neurodegenerative disease.
  • DPP6 gene product levels are higher than those of a control with neurodegenerative disease, this can be said not to correlate with an increased risk of neurodegenerative disease, or even to be correlated with a decreased risk of disease.
  • DPP6 gene product levels may be compared to both a negative and a positive control in order to increase accuracy of the diagnosis.
  • the DPP6 gene product whose levels are determined will typically be DPP6 mRNA and/or DPP6 protein.
  • DPP6 mRNA is chosen as the (or one of the) DPP6 gene product whose levels are determined, this can be the total of all DPP6 mRNA isoforms, or one or more specific mRNAs (e.g. isoform 1 of DPP6).
  • the DPP6 gene product of which the levels are determined may be DPP6 protein.
  • the total DPP6 levels may be determined, or those of specific isoforms only (e.g. using an antibody against the different N-termini).
  • DPP6 protein isoforms may be detected (e.g. using an antibody against a common epitope).
  • the isoforms to be detected can be all isoforms for both mRNA and protein, identical isoforms (wholly overlapping), or different isoforms (partly or not overlapping), depending on the setup of the experiment.
  • identical isoforms it is meant that the mRNA isoform encodes for the corresponding protein isoform.
  • at least one mutation is a null mutation and is causative of the neurodegenerative brain disease.
  • a 'null allele' or 'null mutation' as used herein is a mutant copy of a gene that completely lacks that gene's (i.e. typically DPP6) normal function. This can be the result of the complete absence of the gene at the genomic level, gene product (protein, RNA) at the molecular level, or the expression of a nonfunctional gene product. In the latter case, expression of gene product may be quite high, but this still corresponds to a significant decrease of functional expression, since no functional product is formed.
  • a null allele may be a protein null or a RNA null (either at RNA or DNA level).
  • the neurodegenerative brain disease is early-onset brain disease.
  • Early-onset brain disease is typically disease with an age at onset of 65 years or younger.
  • early-onset brain disease is brain disease with an age at onset (AAO) of 60 years or younger.
  • AAO age at onset
  • the effect of the DPP6 gene is dosage sensitive, and the more severe the mutation (i.e., the more severe its effect on function of DPP6 or on functional levels of DPP6 gene product), the stronger the effect on disease.
  • the one or more mutations are not null mutations and are indicative of an increased risk of developing or presence of the neurodegenerative brain disease. I.e., they are mutations with incomplete penetrance.
  • the functional expression levels of DPP6 are decreased by less than 20%, 30% or even 40%, and this is indicative of an increased risk of developing or presence of the neurodegenerative brain disease.
  • the neurodegenerative brain disease is late-onset brain disease. This typically will be the case when monitoring DPP6 risk alleles, or when functional expression levels of DPP6 are still relatively high (e.g. 90% of control, 80% of control, 75% of control, 70% of control, 60% of control) as compared to null alleles.
  • Risk alleles are alleles that themselves are not causative of the disease, but confer an increased risk of developing the disease, e.g. because they produce less functional gene product.
  • the risk of presence or of developing a neurodegenerative disease is even larger if a mutation is present in more than one allele of DPP6, or if there is also a mutation present in another neurodegenerative brain disease-linked gene.
  • the methods may further comprise determining the presence of mutations in other genes.
  • deleterious mutations include mutations that affect the coding regions of DPP6, particularly exonic mutations and splice site mutations. Most particularly, such exonic mutations are missense mutations, nonsense mutations and frame-shift mutations.
  • exonic mutations are missense mutations, nonsense mutations and frame-shift mutations.
  • the extracellular domain of DPP6 is highly structured, and non-synonymous mutations in this domain have a high chance of affecting DPP6 function, by changing the DPP6 conformational structure. Accordingly, such mutations are explicitly envisaged.
  • Alternative deleterious mutations are mutations affecting functional expression levels of DPP6, e.g.
  • CNVs copy number variants
  • the copy number variants that are evaluated do not include the CNVs described by Blauw et al. (Blauw et al., 2010).
  • inversions and duplications adjacent to, but not including, the DPP6 gene may also have effects on expression levels
  • the inversions and/or duplications involve the DPP6 genomic region. According to particular embodiments, they involve the region of isoform 1 of DPP6. According to most particular embodiments, they involve the 5' region of isoform 1 of DPP6.
  • DPP6 different deleterious mutations of DPP6 are listed that are found in patients with neurodegenerative disease. It is particularly envisaged that at least one of the mutations whose presence is determined is a mutation selected from those listed in Table 6, Table 8, and an inversion or CNV affecting DPP6. More particularly, it is envisaged that at least one of the mutations whose presence is determined is a mutation selected from those listed in Table 6 and in Table 8, and an inversion affecting DPP6. More particularly, it is envisaged that at least one of the mutations whose presence is determined is a mutation selected from those listed in Table 6 and in Table 8. According to even further embodiments, at least one of the mutations is selected from the mutations listed in Table 6.
  • detection of mutations is done using sequencing, a hybridisation assay or PCR-based assay, such as the MastRTM assay (Multiplex Amplification of Specific Targets for Resequencing, Multiplicom).
  • these assays can be used for sequencing or determining the presence of mutations in both exons and regulatory regions.
  • Other methods include e.g. cDNA sequencing to detect e.g. exon(s) deletions other than caused by splice site mutations.
  • genomic rearrangements including inversions, deletions or duplications (CNVs) determining the presence will typically be done in other ways.
  • cytogenetic methods may be used, such as fluorescence in situ hybridization (FISH) on stretched-chromosomes, which is suitable to detect inversions, deletions or duplications (as e.g. described by Salomon-Nguyen et al., 1998; Gijselinck et al., 2008).
  • FISH fluorescence in situ hybridization
  • MAQ Multiplex Amplicon Quantification
  • PFGE pulsed field gel electrophoresis
  • the skilled person is capable of selecting the most appropriate assay, and the listed methodologies should not be interpreted as limiting.
  • kits are provided suitable for practicing the methods presented herein, i.e. kits for determining the expression levels of DPP6 and/or determining the presence of one or more mutations in the DPP6 gene in a sample of said subject.
  • these kits will typically contain at least one agent specifically binding to DPP6 protein (e.g. a DPP6 antibody), or to DPP6 mRNA (e.g. primers).
  • the kits will typically comprise at least one primer or probe suitable to determine the presence of one or more mutations in the DPP6 gene.
  • larger mutations e.g.
  • kits may provide the material to perform FISH, MAQ or PFGE, as described above. Obviously, kits may contain further material, such as pharmaceutically acceptable excipients or buffers, as is established in the art.
  • the DPP6 protein, or nucleic acid encoding said protein is provided for use as a medicament.
  • the DPP6 protein, or nucleic acid encoding said protein is provided as a diagnostic. Indeed, as described in the methods above, the levels and/or presence of mutations in DPP6 may be used as diagnostic tool to establish the presence of neurodegenerative disease.
  • the DPP6 protein, or nucleic acid encoding said protein is provided as a diagnostic for diagnosing AD or FTLD (including FTLD/ALS).
  • the DPP6 protein, or nucleic acid encoding said protein is provided for use as a medicament to treat neurodegenerative disease. Indeed, restoring correct levels of DPP6 levels will prevent the onset of, delay the onset of, or treat, neurodegenerative disease.
  • the DPP6 protein, or nucleic acid encoding said protein is provided for use in treating AD or FTLD (including FTLD/ALS).
  • the levels of DPP6 are increased by administering the DPP6 protein, or nucleic acid encoding said protein, to said subject.
  • the levels of DPP6 are increased using gene therapy, particularly gene therapy wherein the nucleic acid encoding said protein is administered to a subject in need thereof.
  • DPP6 levels can also be increased by administering a compound to a subject.
  • compounds can increase DPP6 expression e.g. by increasing DPP6 transcriptional activity, by stabilizing the DPP6 m NA or protein, or by influencing other genes or gene networks (e.g. by blocking DPP6 inhibitors).
  • methods to screen for compounds that increase DPP6 expression comprising:
  • the current sample comprises 101 patients from the original group and 22 additional patients from the extended study. Median age at onset in this sample was 58.0 years (range 33-65) and 77% were women. In this extended sample, mutations were excluded in the prion gene (PRNP) (Dermaut et al., 2003). Conclusion, family 1270 was negative for the 3 causal AD genes APP, PSEN1 and PSEN2, plus for the PRNP and MAPT genes in which occasionally mutations have been observed in clinical diagnosed AD patients.
  • PRNP prion gene
  • a clinical follow-up of family 1270 was performed by neurological examination of incident patients, interviews of first-degree relatives and review of medical records (Rademakers et al., 2005). For five patients who were deceased or could not be examined, diagnosis was complemented by information obtained through a family informant. Since the first linkage report (van Duijn et al., 1994b), four family members (111-19, 111-21, 111-41, and 111-43) became symptomatic (Rademakers et al., 2005). The updated pedigree of family 1270 is shown in figure 1; clinical data is summarized in table l(Rademakers et al., 2005).
  • the finemapping of the linked region on chromosome 7 with additional STR markers identified a disease haplotype present in all patients.
  • Obligate meiotic recombinants defined a candidate region of 19.7 cM between STR markers pRR8 and D7S559 with an estimated genomic size of 5.44 Mb (Rademakers et al., 2005). Age (in years) at
  • Table 1 Clinical characteristics of patients of family 1270.
  • Patient 111-45 is the index patient of Dutch family 1270 (Rademakers et al. 2005) and was part of the Rotterdam early-onset Alzheimer disease cohort (Hofman et al. 1989).
  • a APOE genotypes of available patients in generation III (figure 1)
  • b Diagnosis obtained through family informant c Newly diagnosed patients. Patients in red have been included in WGS.
  • Table 2 Allele-Sharing Analysis of Microsatellite Markers at 7q36.
  • the linked alleles of family 1270 are in bold italics. In red the STR markers that showed allele sharing in all 4 families.
  • PAXIP1, p.Ala626 PAXIP1, p.Ala626
  • aCGH array-based comparative genomic hybridization
  • a dotplot was made using the NCBI hgl9 reference sequence, for comparative sequence analysis of the genomic region of chromosome 7 ranging from 149169800 (where ZNF746 is located) to 154794690 (where PAXIP1 is located) (data not shown).
  • the dotblot confirmed the presence of Low-Copy-Repeat (LC s) in the 5' region of the DPP6 locus and provided evidence for the presence of inverted repeats in the LCRs (Table 8).
  • LC s Low-Copy-Repeat
  • IP- LCR inverted paralogous LCRs
  • NAHR non-allelic homologous recombination
  • Structural variations were called on the WGS data of the 4 sequenced members of family 1270. Investigation of the SV data for the DPP6 locus demonstrated the presence of a heterozygous inversion in two of the WGS patients (111-38 and 111-12, Pedigree figures 1 and 2). The inversion has an estimated size of ⁇ 4 Mb (Table 3) and the breakpoints are located in IP-LCRs. The 3 SNVs (vl, v2, v3) in intron 1 in family 1270 are located outside the inversion.
  • the distance between the two ends of a mate pair is expected to be approximately 350 bp (distance inferred from documentation information available for the WGS).
  • the first end of the mate pair is located within the region spanning chr7:149331000-149332000 and the second end of the mate pair is located at a distance of ⁇ 4 Mb with opposite strand orientation, which is indicative of an inversion.
  • a similar pattern of distribution was observed in all four affected relatives (not shown).
  • DPP6 has 3 major isoforms (Figure 4).
  • DPP6 isoform 1 encodes the longest protein isoform (865 amino acids, also referred to as L).
  • Isoform 2 and 3 include an alternate in-frame exon 1, compared to variant 1, resulting in shorter protein products that have a shorter and distinct N-terminus, compared to isoform 1 (isoform 2 has 803 amino acids while isoform 3 has 801 amino acids).
  • the inversion breakpoint is positioned within intron 1 of isoforms 1 and 3 ( Figure 5).
  • the inversion is predicted to dislocate the exon 1 and promoter of these 2 isoforms and as such prevent their expression. It is also possible that the inversion breakpoint by its position affects the expression of isoform 2. We can also not exclude that the inversion affects an upstream enhancer regulating expression of some of all isoforms.
  • the distal breakpoint of the inversion is part of the priority haplotype 201-80-232 delineated by the STR markers D7S2439 and D7S2546 shared between family 1270 and the 3 families 1034, 1125 and 1242 ( Figure 6).
  • DPP6 is a single-pass type II transmembrane protein. It is a member of the S9B family that belongs to the group of serine proteases. Differently from the other family members, this protein has no detectable protease activity, due to the absence of the conserved serine residue normally present in the catalytic domain of serine proteases (NCBI, Gene database, http://www.ncbi.nlm.nih.gov/gene). The X-ray crystal structure of the extracellular domain of human DPP6 was determined at 3.0 A resolution (Strop et al., 2004). Two monomers associate to form a homodimer. Each monomer consists of eight-bladed ⁇ -propeller domain and a ⁇ / ⁇ hydrolase domain.
  • the protein has seven predicted N- glycosylation sites and 4 disulfide bridges in each monomer ( Figure 7) (Strop et al., 2004)
  • the protein is expressed predominantly in brain, with high levels in amygdala, cingulate cortex, cerebellum and parietal lobe (http://biogps.org) where it can be expected to be involved in physiological processes of brain function. It modulates the function and expression of potassium channels and excitability at the glutamatergic synapse (Wong et al., 2002). It may be involved in proteolysis and cell death (information inferred from http://www.uniprot.org/uniprot/P42658).
  • the in house available dataset contains genomes of individuals with different neurological phenotypes: 23 independent AD patients, 40 FTLD genomes, 8 independent PD/DLB genomes and 11 independent non-CNS neurodegenerative diseases (e.g. CMT, epilepsy).
  • the DPP6 inversion, with breakpoint in the intron 1 of the gene was observed in 3 unrelated FTLD patients (d5713, dlll88, dl2170), in 5 FTLD patients and 1 healthy at risk individual (d2029 - DR2.8), from 5 different branches of the GRN founder family (frequency in FTLD patients 20%) and in 1 young CMT patient (cmtl54.06), who, due to the young age at the last examination, might still develop dementia at older age (Table 5).
  • the aim of this experiment was to examine whether the inversion carriers are sharing a common haplotype for the genomic inversion.
  • all the individuals who showed the genomic inversion in the WGS data have been genotyped using the same marker panel selected previously (Table 2), that in this particular case is including the DPP6 breakpoint (downstream or 3' breakpoint).
  • DPP6 breakpoint downstream or 3' breakpoint
  • the Belgian AD subgroup consisted of 288 patients with an early onset age, mean age at onset (AAO) of 63.99 ⁇ standard deviation (SD) of 6.58 yea rs (57.3% women) . Additionally we screened 82 EOAD patients from the Rotterdam cohort (RotlOO), AAO ( ⁇ SD) of 56.99 ⁇ 5.60 years (82.3% women).
  • the Belgian frontotemporal lobar degeneration (FTLD) cohort consisted of 351 patients with a mean ( ⁇ SD) age at onset of 62.44 ⁇ 11.42 years (45% women).
  • the Belgian ALS cohort consisted of 124 patients with mean AAO ( ⁇ SD) 58.97 ⁇ 11.91 years (39% women).
  • the control cohort consisted of 408 neurologically healthy individuals with a mean ( ⁇ SD) age at inclusion (AAI) of 72.56 ⁇ 10.21 years (60.5% women). For exon 1 of isoform 1 an extra group of 423 control persons were screened, mean AAI ( ⁇ SD) of 62.70 ⁇ 13.38 years (53% women). Mutation spectrum and frequencies
  • FTLD patient IB5850 carries a VCP mutation p.Argl59His and received a pathological diagnosis of FTLD TDP-D (van der Zee et al. 2009).
  • Patient IB5941 with a clinical diagnosis of FTLD received a pathological diagnosis of AD but TDP-43 immunohistochemistry was not yet performed.
  • the mutation spectrum includes missense mutations, nonsense and frameshift mutations predicting premature stop codons, and amino acid deletions/duplications (indels). In control subjects only missense mutations have been identified. These are generally considered less deleterious than nonsense or frameshift mutations.
  • the spectrum is consistent with a loss-of function either by affecting the protein structure and function (missense mutations and indels) or loss of transcript by non-sense mediated mRNA decay (nonsense and frameshift mutations).
  • missense mutations and indels loss of transcript by non-sense mediated mRNA decay
  • Table 6 Clinical characteristics of carriers of the rare non-synonymous variants.
  • FH family history: "F” indicates positive family history; "U” indicates unknown family history and "S” indicates a sporadic patient.
  • N total number of carriers.
  • Panel A shows the variations exclusively identified in patients, panel B in control individuals only, and pa nel C in both patients and control individuals.
  • Table 7 Overall frequency of rare non-synonymous coding variants in DPP6 (MAF ⁇ 1%) in FTLD, AD, ALS and control individuals. Data are presented as counts and frequencies on the total number of individuals screened, Relative risk (RR) with 95% confidence intervals (CI) and p-values are calculated after collapsing rare variant allele.
  • RR Relative risk
  • CI 95% confidence intervals
  • p-values are calculated after collapsing rare variant allele.
  • a Mantel - Haenszel approach was used to compute RR and 95% CI, to take into account the different number of controls screened for exon 1 of isoform 1 versus the remainder of the gene.
  • Table 8 Analysis of the mutation located in the cytoplasmic domain.
  • ConSurf negative score indicates conserved residue, high score indicates non-conserved residue. Residue found back in alignment are indicated between brackets.
  • FoldX Positive DDG indicates that mutation destabilizes structure, negative DDG indicates that mutation stabilizes structure. Mutations in TM domain (96-116)
  • Table 9 Analysis of the mutation located in the TM domain.
  • the extracellular domain is the largest domain of the protein.
  • the majorities of the patient specific mutations destabilizes the protein structure (negative value of DDG) and have a high risk to result in non-functional protein.
  • DDG negative value of DDG
  • DPP6 has 7 glycosylated residues (Figure 8), several amino acids interact with these residues.
  • Figure 8 two mutated residues can have an effect on this interaction.
  • p.R322 interacts with the glycosylated p.N319 ( Figure 9A).
  • Figure 9B A detrimental effect can be determined by the mutation p.D569N.
  • the mutated residue p.D569N can compete for the glycosylation site ( Figure 9B).
  • mutations p.P229T, p.R274H, p.R322H, p.P509R and p.D569N result in a nonfunctional protein through destabilization of the native structure. Moreover, mutations p.R322H and p.D569N can affect the glycosylation.
  • GAP glyceraldehyde 3-phosphate dehydrogenase
  • GPDH glyceraldehyde 3-phosphate dehydrogenase
  • YWHAZ tyrosine 3- monooxygenase/tryptophan 5-monooxygenase activation protein
  • YWHAZ zeta polypeptide
  • HPRT1 hypoxanthine phosphoribosyltransferase 1
  • TATA box binding protein TATA box binding protein
  • Normalization of the reference genes was done by geometric averaging of the expression levels, as described by Vandesompele and colleagues (Vandesompele et al., 2002). Each sample was measured in triplicate and at least two independent experiments were performed. To increase statistical power, the results were pooled and a non parametric Mann Whitney U test was used to compare the expression of DPP6 between carriers patients and control individuals.
  • RNA quality was not optimal, on average in both patients and controls the RNA integrity number (RIN) was 5 (Intact RNA scores >9). For these reasons, no conclusions could yet be drawn on the altered levels of separate isoforms.
  • DPP6 protein expression analysis in protein extracts of human brain is ongoing.
  • TEVC voltage clamp
  • DPP6 is causally associated with FTLD and AD. Frequency of mutations in DPP6 is high and ranging between 3.5% in AD to 5.7% in FTLD, much higher than the frequency of very rare non-synonymous variants in DPP6 in control individuals.
  • missense mutations were identified of which many are located in the large extracellular domain of the protein that is extremely structured as shown by the crystal structure (figure 7) and which is present in all three DPP6 isoforms.
  • modelling data show that they can have a detrimental effect, resulting in a non-functional protein through destabilization of the native structure.
  • mutations p.R322H and p.D569N can affect the glycosylation.
  • DPP6 protein expression is basically restricted to brain. Transcript studies show a ⁇ 20% reduced levels of DPP6 in brain of mutation carriers compared to control individuals, but this decrease can be even higher in individual mutation carriers ( Figure 11). Currently, this is further being evaluated.
  • the haploinsufficiency is supported genetically (inversion, non-sense, frameshift mutations), in silico modelling support the detrimental effect of many missense mutations located in the extracellular domain of the protein. Also on a transcription level we observe ⁇ 20% reduction of the total DPP6. Experiments are ongoing to further support the role of DPP6 in NBD.
  • the CD26-related dipeptidyl aminopeptidase-like protein DPPX is a critical component of neuronal A-type K+ channels. Neuron 37:449-461.
  • Amyloid precursor protein gene mutation in early-onset Alzheimer's disease Lancet 337:978. van Duijn CM, Hendriks L, Farrer LA, Backhovens H, Cruts M, Wehnert A, Hofman A, Van Broeckhoven C. 1994b. A population-based study of familial Alzheimer disease: linkage to chromosomes 14, 19, and 21. Am J Hum Genet 55:714-727.

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Abstract

La présente invention concerne le domaine de la génétique humaine, en particulier le domaine des maladies neurodégénératives cérébrales. Plus particulièrement, la présente invention concerne des procédés et des matériaux pour détecter des maladies neurodégénératives cérébrales humaines, plus particulièrement la démence (maladie d'Alzheimer (MA)), la dégénérescence lobaire frontotemporale (DLFT), la démence à corps de Lewy (DCL), la sclérose latérale amyotrophique (SLA) et la maladie de Parkinson (MP). Des maladies plus particulièrement envisagées comprennent la MA, la DLFT et les maladies du spectre DLFT/SLA. L'invention concerne des essais diagnostiques pour la détection de maladies neurodégénératives cérébrales, des kits pour réaliser ces essais et des procédés pour traiter ces maladies.
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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2017106363A1 (fr) * 2015-12-14 2017-06-22 Parkinson's Institute Affinement du diagnostic et du traitement de troubles neurologiques complexes à plusieurs symptômes
WO2017182603A1 (fr) * 2016-04-22 2017-10-26 Université Libre de Bruxelles Nouveau biomarqueur exprimé dans les cellules bêta pancréatiques utilisé pour l'imagerie ou le ciblage des cellules bêta

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005108997A1 (fr) * 2004-05-11 2005-11-17 Bayer Healthcare Ag Diagnostic et therapeutique de maladies associees a la dipeptidyl-peptidase 6 (dpp6)
WO2011012672A1 (fr) * 2009-07-29 2011-02-03 Pharnext Nouveaux outils diagnostiques pour la maladie d’alzheimer
WO2013041577A1 (fr) * 2011-09-20 2013-03-28 Vib Vzw Procédés de diagnostic de la sclérose latérale amyotrophique et de la dégénérescence lobaire frontotemporale
WO2013067451A2 (fr) * 2011-11-04 2013-05-10 Population Diagnostics Inc. Méthodes et compositions pour le diagnostic, le pronostic et le traitement d'états neurologiques

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005108997A1 (fr) * 2004-05-11 2005-11-17 Bayer Healthcare Ag Diagnostic et therapeutique de maladies associees a la dipeptidyl-peptidase 6 (dpp6)
WO2011012672A1 (fr) * 2009-07-29 2011-02-03 Pharnext Nouveaux outils diagnostiques pour la maladie d’alzheimer
WO2013041577A1 (fr) * 2011-09-20 2013-03-28 Vib Vzw Procédés de diagnostic de la sclérose latérale amyotrophique et de la dégénérescence lobaire frontotemporale
WO2013067451A2 (fr) * 2011-11-04 2013-05-10 Population Diagnostics Inc. Méthodes et compositions pour le diagnostic, le pronostic et le traitement d'états neurologiques

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
ANONYMOUS: "Ensembl genome browser 71: Homo sapiens - Explore this variation - rs2533730 (SNP)", 1 April 2013 (2013-04-01), XP055142346, Retrieved from the Internet <URL:http://apr2013.archive.ensembl.org/Homo_sapiens/Variation/Explore?db=core;g=ENSG00000130226;r=7:153887097-154894285;source=dbSNP;t=ENST00000377770;v=rs2533730;vdb=variation;vf=2135129> [retrieved on 20140924] *
ANONYMOUS: "Ensembl genome browser 71: Homo sapiens - Explore this variation - rs2533731 (SNP)", 1 April 2013 (2013-04-01), XP055142344, Retrieved from the Internet <URL:http://apr2013.archive.ensembl.org/Homo_sapiens/Variation/Explore?db=core;g=ENSG00000130226;r=7:153887097-154894285;source=dbSNP;t=ENST00000377770;v=rs2533731;vdb=variation;vf=2135130> [retrieved on 20140924] *
DEL BO R ET AL: "DPP6 gene variability confers increased risk of developing sporadic amyotrophic lateral sclerosis in Italian patients", JOURNAL OF NEUROLOGY, NEUROSURGERY, AND PSYCHIATRY,, vol. 79, no. 9, 1 September 2008 (2008-09-01), pages 1085, XP009180266 *
MICHAEL A VAN ES ET AL: "Genetic variation in DPP6 is associated with susceptibility to amyotrophic lateral sclerosis", NATURE GENETICS, vol. 40, no. 1, 16 December 2007 (2007-12-16), pages 29 - 31, XP055141405, ISSN: 1061-4036, DOI: 10.1038/ng.2007.52 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2017106363A1 (fr) * 2015-12-14 2017-06-22 Parkinson's Institute Affinement du diagnostic et du traitement de troubles neurologiques complexes à plusieurs symptômes
WO2017182603A1 (fr) * 2016-04-22 2017-10-26 Université Libre de Bruxelles Nouveau biomarqueur exprimé dans les cellules bêta pancréatiques utilisé pour l'imagerie ou le ciblage des cellules bêta
US11243214B2 (en) 2016-04-22 2022-02-08 Université Libre de Bruxelles Biomarker expressed in pancreatic beta cells useful in imaging or targeting beta cells

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