KR20150015226A - A Marker for Detecting a Neuro-Degenerative Disease and a Method for Detecting the Neuro-Degenerative Disease by Using the Same - Google Patents

Abstract

In the present invention, provided are a diagnosis marker of neurodegenerative disease including at least one among TAR-binding protein of 43kDa (TDP-43) missense mutations whose sequence number is between 1 and 3, and a diagnosis method of neurodegenerative diseases, comprising the steps of i) obtaining samples from materials to be tested; and ii) checking whether the TDP-43 missense mutations are expressed or not wherein the sequence number of the TDP-43 missense mutations is between 1 and 3.

Description

[0001] The present invention relates to a marker for diagnosing degenerative neurological diseases and a diagnostic method using the same. More particularly, the present invention relates to a marker for diagnosing a neurodegenerative disease,

The present invention relates to a marker for diagnosing a degenerative neurological disease and a method for diagnosing a neurodegenerative disease using the same. More specifically, the present invention relates to a TDP-43 mismatch mutation of TDP-43 (TDP-43) and a TDP-43 mismatch mutation of a specific sequence invented on the basis of a pathological mechanism of a degenerative nervous system disease A marker for diagnosing a neurological disease including a mutation, and a method for diagnosing a neurodegenerative disease using the same.

TDP-43 is an RNA / DNA binding protein that is involved in transcription, RNA splicing, mRNA stability, and nucleocytoplasmic shuttling of mRNA. TDP-43 has recently been found to be a major component of ubiquitinated inclusion in amyotrophic lateral sclerosis or frontotemporal lobar degeneration.

Amyotrophic Lateral Sclerosis is a degenerative nervous system disease that progresses to degeneration of motor neurons in the spinal cord and cerebral cortex, leading to paralysis and death. Frontal temporal lobe degeneration induces degeneration of the frontal and temporal lobes, which is associated with executive function, social function and lack of cognitive function. Clinically, it is possible to distinguish between amyotrophic axillary sclerosis and frontal lobe lobe degeneration, but there is overlap in the molecular state of the patient. A number of patients with amyotrophic axillary lesions and frontal lobe laceration show positive TDP-43 implants in the central nervous system.

TDP-43 has features that indicate toxicity when it is diseased, such as abnormal hyperphosphorylation, ubiquitination, and the production of abnormal C-terminal fragments.

TDP-43 is located predominantly in the nucleus of healthy neurons, which can be abnormally located in the cytoplasm and neurites when it is diseased and has been reported to cause aggregation, suggesting that TDP-43 loss and / Which may contribute to the onset of the disease.

The inventors of the present application studied the effect of the TDP-43 mismatch mutation on differentiated cultured neurons and found that the presence of the TDP-43 mismatch mutation having a specific sequence would be a marker for diagnosing degenerative neurological diseases And completed the present invention.

The present invention provides a marker for diagnosing degenerative neurological diseases comprising any one or more of the TSP-43 (TAR-binding protein of 43 kDa) mismatch mutations of SEQ ID NOS: 1 to 3.

In addition,

i) obtaining a sample from a specimen;

ii) confirming whether one or more of the TSP-43 (TAR-binding protein of 43 kDa) mismatch mutations of SEQ ID NOS: 1 to 3 is expressed.

The marker for diagnosing degenerative neurological diseases according to the present invention and the method for diagnosing degenerative neurological diseases using the same can be accurately diagnosed.

Figure 1 shows an overview of human TDP-43, wherein TDP-43 contains a nuclear locus signal (amino acids 78-84), two RNA recognition motifs, a nuclear export signal, and a glycine-rich domain.
Figure 2 shows the results of analysis of TDP-43 wild-type and mismatch mutant expression levels in HEK 293T cells through Western blot.
3 is a photograph showing the result of transfection of GFP into nerve cells.
4 is a graph showing the ratio of the total neurite length.
Figure 5 is an image of TDP-43 expressing neuron showing the location of flag-TDP-43 immunostained with anti-flag antibody.
FIG. 6 is an image showing TDP-43 with mNLS using GFP for morphological observation of neurons. FIG.
7 is a graph showing the ratio of the total length of the nerve processes.
Figure 8 shows immunostaining of flagged TDP-43 or endogenous TDP-43 with anti-FLAG antibody or anti-TDP43 antibody, showing TDP-43 reduction in the nucleus by cytoplasmic expression of mNLS with TDP-43 Image (the circle indicates the nucleus of the neuron).

Despite recent studies on TDP-43, there is very little understanding of how TDP-43 is involved in degenerative neurological diseases. In addition, many studies of TDP-43 have used yeast, Drosophila, and mammalian cell lines. In many cases, TDP-43 in these cells often fails to properly reflect the pathological characteristics in human brain cells. Furthermore, there have been no studies on the effect of TDP-43 mutant proteins on the structure of mammalian neurons and pathological mechanisms of action.

The inventors of the present application analyzed the effect of mutant proteins of TDP-43 found in degenerative neurological diseases on the structure formation and maintenance of neurons, and revealed the pathological mechanisms of them. Thus, TDP-43 mismatch mutations were found in degenerative nervous system diseases And it can be used as a biomarker for diagnosis.

The biomarker for diagnosing a neurodegenerative disease according to the present invention comprises any one or more of the TDP-43 mismatch mutations of SEQ ID NOS: 1 to 3.

SEQ ID NOS: 1 to 3 are DNA sequences of a TDP-43 missense mutation named A315T, Q331K, and M337V, respectively.

To investigate the effect of each mutant protein on the structure and function of neurons, we compared the effects of these mutants on neuronal expression. As a result, each mutant protein formed an abnormal branch compared to the wild type, thereby increasing the overall length of the nerve cell.

In addition, by examining them over time, abnormal dendritic structures were observed as a result of the segmentation of neurites in the late period and dying. The neurons undergo various signal transduction in the dendritic region. The formation of abnormal dendrites and the increase in length are likely to disturb the signals of neuronal cells, and these neurons, which are seen in brain or animal models of patients with Alzheimer's dementia, Abnormal changes in cell structure are associated with early pathological mechanisms of degenerative neurological diseases, such as degenerative brain diseases.

Thus, abnormal structural changes of these neurons may be an important predictor of the initial pathologic process of mutation-associated frontotemporal dementia or Lou Gehrig's disease. Most degenerative nervous system diseases are found in the late stage of the disease, and they observe the death of nerve cells. However, the treatment of degenerative nervous system disorders can provide clues to the treatment of degenerative neurological diseases associated with the TDP-43 mutation in that it is very important to understand the initial pathological process.

In order to investigate the pathological mechanism of how TDP-43 mutant proteins alter the structure of neurons, the mutant proteins of TDP-43 were found to be located outside the nucleus of the cell, And the tendency to relocate to the branches of neurons increases. Since many proteins must be in a specific location to perform an intracellular role, their normal intracellular location is essential to function. The mutant proteins of TDP-43 can migrate within the cell, resulting in the loss of the inherent function involved in RNA transcription or processing of genes involved in cell structure in the nucleus.

The mutant protein of TDP-43 induced loss of TDP-43 function and induced abnormal structure of neurons. As a result, abnormal dendrites were formed and length was increased due to lack of function of TDP-43. It was confirmed that the mutant protein has a similar structure to that of the mutant protein.

Based on the above results, the inventors of the present application confirmed that the TDP-43 mismatch mutation can be a marker for diagnosing a neurodegenerative disease.

In the present invention, "diagnosis" means identifying the presence or characteristic of a pathological condition. For the purpose of the present invention, the diagnosis is to ascertain the likelihood or incidence of a degenerative neurological disease, for example, anterior temporal dementia or Lou Gehrig's disease.

In one embodiment, the present invention provides a method comprising: i) obtaining a sample from a sample; ii) determining whether one or more of the TDP-43 mismatch mutations of SEQ ID NOS: 1 to 3 is expressed, and a method for diagnosing a neurodegenerative disease.

After taking a biological sample, for example blood, from the sample through step i), it is determined whether or not any one or more of SEQ ID NOS: 1 to 3 representing the TDP-43 mismatch mutant DNA sequence of step ii) And can be confirmed by a known method such as Polymerase Chain Reaction (PCR).

Example

EXAMPLES The present invention will be further illustrated by the following examples and experimental examples, but the present invention is not limited by these examples and experimental examples.

[Experimental Method and Condition]

1. Plasmid

Human or mouse coding cDNAs were amplified by RT-PCR using the primers in Table 1 throughout the HEK293T or mouse thymus RNA extracts.

[Table 1]

¹ S, Sense; A, Antisense; hTDP-43, human TDP-43; mTDP-43, mouse TDP-43

Recombinant PCR was performed using the primers of Table 2 below, each containing a mismatch mutation, to make a TDP-43 missense mutation (A315T, Q331K, M337V).

[Table 2]

The nuclear localization signal was modified by PCR with the specific primers described in Table 1 above to produce the TDP-43 structure with a mutated nuclear localization signal (mNLS). For expression in animal cells, each cDNA was prepared by PCR using specific primers and the cDNA was then subcloned into 3xflag-CMV7.1 vector (Sigma) using the HindIII-BamHI site.

2. TDP-43 siRNA production

Five sections of siRNA against mouse TDP-43 were selected using siRNA Target Finder program to generate TDP-43 siRNA expression structure. Then, pSUPER-GFP vector (Oligoengine) was prepared using HingIII-BglII. Lt; / RTI > The TDP-43 synthetic siRNA sequence for mouse TDP-43 is sense: 5'-GAGAGGAUUUGAUCAUUAA-3 '(SEQ ID NO: 20), antisense; 5'-UUAAUGAU-CAAAUCCUCUC-3 '(SEQ ID NO: 21) (Bioneer Co., Korea).

3. Cell culture and transfection

Neuronal cells were cultured in E17 ICR mouse embryos and each DNA plasmid was incubated with HYP293T cells or manufacturer's protocol together with Lipofectamine 2000 (Invitrogen, USA) or calcium phosphate reagent (Clontech USA) And transfected into neurons.

To down-regulate TDP-43, TDP-43 siRNA was transfected with or without siRNA-resistant human TDP-43 at day 5 after culture. With respect to the rescue experiment, flag-human TDP-43 (or flag-TDP-43 [mNLS]) was transfected with scrambled siRNA or TDP-43 siRNA on day 5 after incubation.

Each siRNA (75-90 pmol / μl / well) aliquot was incubated in cell culture medium using RNAi MAX preparation (Invitrogen, Carlsbad, CA, USA) with TDP-43 siRNA transfection, Were added to trypsinized neurons.

4. Immunocytochemistry

Two days after transfection, transfected neurons were fixed with 4% paraformaldehyde (PFA) for 10 minutes. With respect to immunostaining, transfected neurons were permeabilized with 0.1% Triton X 100 for 5 minutes and then blocked with 3% bovine serum albumin (BSA) at room temperature for 1 hour. Cells were then incubated for 1 hour in anti-flag (Sigma, 1: 100) or anti-mouse TDP-43 antibody (Abnova, 1: 100) for 1 hour followed by anti-cy3 or anti-DyLight 488 secondary antibody ) At room temperature for 1 hour.

5. Image analysis and quantification of total neurite length and cell viability

(Millipore, Billerica, USA, 1: 200) positive and anti-MAP2 (Millipore, Billerica, MAUSA, 1: 200) positive neurites were used to measure the length of axons and dendrites. Neuronal images for morphometric analysis were obtained using an LSM 510 confocal laser scanning microscope (LSM700, Carl Zeiss, Oberkochen, Germany) at 24 hours, 48 hours and 72 hours after transfection. Confocal images of transfected neurons were obtained through sequential acquisition, and each image was 5-10 images of the Z series. Images from the stack were flattened into a single image using a maximal projection. To measure total neurite length, images were obtained using a 20X objective. All neurite development processes, including the aqueducts and axons of individual neurons, were traced. Image J analysis program, Adobe Photoshop CS3, and metamorphic image analysis were used to quantify the total neurite length. The length of more than 20 neurons in each experimental group was measured and quantified from three independent experiments. Statistical analysis (one way AVOVA, Student's t-test) was performed using the GraphPad Prism 5 program.

Concerning cell survival assays, neurons were transfected with either mNLS TDP-43 or siRNA and then stained with propidium iodide after 72 hours of transfection. Surviving cells were then counted on the basis of GFP-positive and PI-negative neurons. Statistical analysis (one way AVOVA, Tukey's post-hoc test or Student's t-test) was performed using the GraphPad Prism 5 program. In each experiment, a total of 100-200 cells were counted for each condition and then measured and quantified from three independent experiments.

6. Western blot

(Sigma, 1: 2000), anti-GFP (anti-GFP) antibody to detect the expression of TDP-43 (wild type or a mismatch mutant protein) or to check whether flagged mouse TDP-43 was knocked down by TDP- Western blotting was performed using antibodies (SantaCruz, 1: 500) and HRP-conjugated anti-mouse or anti-rabbit secondary antibody (1: 10,000-20,000).

[Example 1] Influence of TDP-43 mismatch mutations on neurite outgrowth morphology

Although there have been many studies on the effects of TDP-43 on cytotoxicity and neurodegeneration, little is known about the early onset of the disease by disease-associated mismatch mutant proteins.

Thus, in order to further confirm early cell defects due to TDP-43 mismatch mutations, specific TDP-43 mismatch mutations (A315T (SEQ ID NO: 1)) in the glycine-rich domain (FIG. 1) via recombinant PCR using specific primers containing mutations , Q331K, M337V).

The expression levels of the respective mismatch mutant proteins in HEK 293T cells were measured and the results are shown in Fig. Referring to Figure 2, the expression levels of each mutant TDP-43 protein were similar to wild-type.

To confirm the effect of TDP-43 mismatch mutations in neurite morphology, mutant or wild-type proteins were expressed in differentiated neurons. As shown in FIGS. 3 and 4, abnormal neurites were observed in the neurons expressing the missense mutation after 24 hours of transfection, and the total neurite was measured and quantified.

This suggests that the mismatch mutant protein increases abnormal neuronal morphology compared to wild-type TDP-43.

[Example 2] Location abnormality caused by TDP-43 mismatch mutant protein

Wild-type TDP-43 was basically located in the nucleus and marked by DAPI (4 ', 6'-diamidine-2'-phenylindole dihydrochloride) in neurons. However, the neurons expressing the mutant protein showed that TDP-43 was located in some cytoplasm and neurite along with the aggregate (FIG. 5). The number of cells showing abnormal TDP-43 position from nucleus to cytoplasm was increased in mutant TDP-43-expressing neurons compared with wild-type TDP-43-expressing neurons.

Thus, it can be confirmed that the TDP-43 missense mutant protein exhibits defects in the nuclear-cytoplasmic round trip.

[Example 3] Confirmation of the relation between position and defect of cell morphology

To determine whether the intracellular location of wild-type or mismatch mutant TDP-43 is associated with early cell morphological defects in differentiated neurons and to compare the effects of cytoplasmic expression of wild-type or mutant proteins in neurite morphology, The mutant structure was transfected into differentiated neurons. The cytoplasmic expression of wild-type TDP-43 as well as the missense mutation was increased in abnormal neurites (Figs. 6 and 7), indicating that cytoplasmic expression of TDP-43 is responsible for abnormal neurite structure.

In fact, it was confirmed that the cell viability after transfection 72 hours was decreased by the cytoplasmic expression of TDP-43 (FIG. 8). In addition, it was suggested that TDP-43 may be decreased by nuclear cytoplasmic expression of TDP-43, which may affect nuclear function.

<110> Hannam University Institute for Industry-Academia Cooperation <120> A Marker for Detecting a Neuro-Degenerative Disease and a Method          Detecting the Neuro-Degenerative Disease by Using the Same <130> p130739 <160> 21 <170> Kopatentin 2.0 <210> 1 <211> 1245 <212> DNA <213> Artificial Sequence <220> <223> Artificial sequence of A315T <400> 1 atgtctgaat atattcgggt aaccgaagat gagaacgatg agcccattga aataccatcg 60 gaagacgatg ggacggtgct gctctccacg gttacagccc agtttccagg ggcgtgtggg 120 cttcgctaca ggaatccagt gtctcagtgt atgagaggtg tccggctggt agaaggaatt 180 ctgcatgccc cagatgctgg ctggggaaat ctggtgtatg ttgtcaacta tccaaaagat 240 aacaaaagaa aaatggatga gacagatgct tcatcagcag tgaaagtgaa aagagcagtc 300 cagaaaacat ccgatttaat agtgttgggt ctcccatgga aaacaaccga acaggacctg 360 aaagagtatt ttagtacctt tggagaagtt cttatggtgc aggtcaagaa agatcttaag 420 actggtcatt caaaggggtt tggctttgtt cgttttacgg aatatgaaac acaagtgaaa 480 gtaatgtcac agcgacatat gatagatgga cgatggtgtg actgcaaact tcctaattct 540 aagcaaagcc aagatgagcc tttgagaagc agaaaagtgt ttgtggggcg ctgtacagag 600 gacatgactg aggatgagct gcgggagttc ttctctcagt acggggatgt gatggatgtc 660 ttcatcccca agccattcag ggcctttgcc tttgttacat ttgcagatga tcagattgcg 720 cagtctcttt gtggagagga cttgatcatt aaaggaatca gcgttcatat atccaatgcc 780 gaacctaagc acaatagcaa tagacagtta gaaagaagtg gaagatttgg tggtaatcca 840 ggtggctttg ggaatcaggg tggatttggt aatagcagag ggggtggagc tggtttggga 900 aacaatcaag gtagtaatat gggtggtggg atgaactttg gtacgttcag cattaatcca 960 gccatgatgg ctgccgccca ggcagcacta cagagcagtt ggggtatgat gggcatgtta 1020 gccagccagc agaaccagtc aggcccatcg ggtaataacc aaaaccaagg caacatgcag 1080 agggagccaa accaggcctt cggttctgga aataactctt atagtggctc taattctggt 1140 gcagcaattg gttggggatc agcatccaat gcagggtcgg gcagtggttt taatggaggc 1200 tttggctcaa gcatggattc taagtcttct ggctggggaa tgtag 1245 <210> 2 <211> 1245 <212> DNA <213> Artificial Sequence <220> <223> Artificial sequence of Q331K <400> 2 atgtctgaat atattcgggt aaccgaagat gagaacgatg agcccattga aataccatcg 60 gaagacgatg ggacggtgct gctctccacg gttacagccc agtttccagg ggcgtgtggg 120 cttcgctaca ggaatccagt gtctcagtgt atgagaggtg tccggctggt agaaggaatt 180 ctgcatgccc cagatgctgg ctggggaaat ctggtgtatg ttgtcaacta tccaaaagat 240 aacaaaagaa aaatggatga gacagatgct tcatcagcag tgaaagtgaa aagagcagtc 300 cagaaaacat ccgatttaat agtgttgggt ctcccatgga aaacaaccga acaggacctg 360 aaagagtatt ttagtacctt tggagaagtt cttatggtgc aggtcaagaa agatcttaag 420 actggtcatt caaaggggtt tggctttgtt cgttttacgg aatatgaaac acaagtgaaa 480 gtaatgtcac agcgacatat gatagatgga cgatggtgtg actgcaaact tcctaattct 540 aagcaaagcc aagatgagcc tttgagaagc agaaaagtgt ttgtggggcg ctgtacagag 600 gacatgactg aggatgagct gcgggagttc ttctctcagt acggggatgt gatggatgtc 660 ttcatcccca agccattcag ggcctttgcc tttgttacat ttgcagatga tcagattgcg 720 cagtctcttt gtggagagga cttgatcatt aaaggaatca gcgttcatat atccaatgcc 780 gaacctaagc acaatagcaa tagacagtta gaaagaagtg gaagatttgg tggtaatcca 840 ggtggctttg ggaatcaggg tggatttggt aatagcagag ggggtggagc tggtttggga 900 aacaatcaag gtagtaatat gggtggtggg atgaactttg gtgcgttcag cattaatcca 960 gccatgatgg ctgccgccca ggcagcacta aagagcagtt ggggtatgat gggcatgtta 1020 gccagccagc agaaccagtc aggcccatcg ggtaataacc aaaaccaagg caacatgcag 1080 agggagccaa accaggcctt cggttctgga aataactctt atagtggctc taattctggt 1140 gcagcaattg gttggggatc agcatccaat gcagggtcgg gcagtggttt taatggaggc 1200 tttggctcaa gcatggattc taagtcttct ggctggggaa tgtag 1245 <210> 3 <211> 1245 <212> DNA <213> Artificial Sequence <220> <223> Artificial sequence of M337V <400> 3 atgtctgaat atattcgggt aaccgaagat gagaacgatg agcccattga aataccatcg 60 gaagacgatg ggacggtgct gctctccacg gttacagccc agtttccagg ggcgtgtggg 120 cttcgctaca ggaatccagt gtctcagtgt atgagaggtg tccggctggt agaaggaatt 180 ctgcatgccc cagatgctgg ctggggaaat ctggtgtatg ttgtcaacta tccaaaagat 240 aacaaaagaa aaatggatga gacagatgct tcatcagcag tgaaagtgaa aagagcagtc 300 cagaaaacat ccgatttaat agtgttgggt ctcccatgga aaacaaccga acaggacctg 360 aaagagtatt ttagtacctt tggagaagtt cttatggtgc aggtcaagaa agatcttaag 420 actggtcatt caaaggggtt tggctttgtt cgttttacgg aatatgaaac acaagtgaaa 480 gtaatgtcac agcgacatat gatagatgga cgatggtgtg actgcaaact tcctaattct 540 aagcaaagcc aagatgagcc tttgagaagc agaaaagtgt ttgtggggcg ctgtacagag 600 gacatgactg aggatgagct gcgggagttc ttctctcagt acggggatgt gatggatgtc 660 ttcatcccca agccattcag ggcctttgcc tttgttacat ttgcagatga tcagattgcg 720 cagtctcttt gtggagagga cttgatcatt aaaggaatca gcgttcatat atccaatgcc 780 gaacctaagc acaatagcaa tagacagtta gaaagaagtg gaagatttgg tggtaatcca 840 ggtggctttg ggaatcaggg tggatttggt aatagcagag ggggtggagc tggtttggga 900 aacaatcaag gtagtaatat gggtggtggg atgaactttg gtgcgttcag cattaatcca 960 gccatgatgg ctgccgccca ggcagcacta cagagcagtt ggggtatggt gggcatgtta 1020 gccagccagc agaaccagtc aggcccatcg ggtaataacc aaaaccaagg caacatgcag 1080 agggagccaa accaggcctt cggttctgga aataactctt atagtggctc taattctggt 1140 gcagcaattg gttggggatc agcatccaat gcagggtcgg gcagtggttt taatggaggc 1200 tttggctcaa gcatggattc taagtcttct ggctggggaa tgtag 1245 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Artificial primer sequence <400> 4 ggcctagccg gaaaagtaaa 20 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Artificial primer sequence <400> 5 caaccaccac cccactgtct 20 <210> 6 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> Artificial primer sequence <400> 6 tccacactga acaaaccaat tt 22 <210> 7 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Artificial primer sequence <400> 7 ggcctagcgg agatttaagc 20 <210> 8 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> Artificial primer sequence <400> 8 cccccattct aaatctacct aacc 24 <210> 9 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> Artificial primer sequence <400> 9 ccacactgaa caaaccaatc tg 22 <210> 10 <211> 35 <212> DNA <213> Artificial Sequence <220> <223> Artificial primer sequence <400> 10 cgcccaagct tggcaccatg rcrgaatata ttcgg 35 <210> 11 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> Artificial primer sequence <400> 11 cgcggatccc attccccagc cagaaga 27 <210> 12 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> Artificial primer sequence <400> 12 aactttggta cgttcagca 19 <210> 13 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> Artificial primer sequence <400> 13 tgctgaacgt accaaagtt 19 <210> 14 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> Artificial primer sequence <400> 14 gcagcactaa agagcagt 18 <210> 15 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> Artificial primer sequence <400> 15 actgctcttt agtgctgc 18 <210> 16 <211> 16 <212> DNA <213> Artificial Sequence <220> <223> Artificial primer sequence <400> 16 tatggtgggc atgtta 16 <210> 17 <211> 16 <212> DNA <213> Artificial Sequence <220> <223> Artificial primer sequence <400> 17 taacatgccc accata 16 <210> 18 <211> 39 <212> DNA <213> Artificial Sequence <220> <223> Artificial primer sequence <400> 18 tatccaaaag ataacgctgc tgctatggat gagacagat 39 <210> 19 <211> 39 <212> DNA <213> Artificial Sequence <220> <223> Artificial primer sequence <400> 19 atctgtctca tccatagcag cagcgttatc ttttggata 39 <210> 20 <211> 19 <212> RNA <213> Artificial Sequence <220> <223> Artificial siRNA sequence <400> 20 gagaggauuu gaucauuaa 19 <210> 21 <211> 19 <212> RNA <213> Artificial Sequence <220> <223> Artificial siRNA sequence <400> 21 uuaaugauca aauccucuc 19

Claims (4)

A marker for diagnosing a degenerative nervous system disease comprising any one or more of TSP-43 (TAR-binding protein of 43 kDa) mismatch mutations consisting of SEQ ID NOS: 1 to 3; The method according to claim 1,
Wherein said TDP-43 mismatch mutation is abnormally located in cytoplasmic and nerve cell branches, or induces dysfunction of normal TDP-43 to cause abnormal structural change of nerve cells. The method according to claim 1,
Wherein the degenerative neurological disease is anterior temporal dementia or Lou Gehrig's disease. i) obtaining a sample from a specimen;
ii) confirming whether one or more of the TSP-43 (TAR-binding protein of 43 kDa) mismatch mutations of SEQ ID NOS: 1 to 3 is expressed.

Images