Method of diagnosis, prognostic or treatment of neurodegenerative diseases

Abstract

The present invention relates to a method of diagnosis, prognostic or treatment of neurodegenerative diseases, in particular Alzheimer's disease.

1 . Composition comprising at least one agent which directly or indirectly affects an activity and/or a level of:
 a) at least one gene selected from the group consisting of the nucleotidic sequences set forth by SEQ ID NO: 1 and SEQ ID NO: 3 and coding for an heparin-glucosamine 3-O-sulfotransferase 2 and 4 respectively, and/or   b) at least one transcription product of a gene selected from the group consisting of the nucleotidic sequences set forth by SEQ ID NO: 1 and SEQ ID NO: 3 and coding for an heparin-glucosamine 3-O-sulfotransferase 2 and 4 respectively, and/or   c) at least one translation product of said genes, said products being set forth respectively by SEQ ID NO: 2 and 4, and/or   d) a fragment or derivative or variant of said gene or said transcription or translation product, and/or   e) heparan sulfate, in particular 3-O-sulfated heparan sulfate, notably 3-O-sulfated heparan sulfate disaccharides,   f) abnormal phosphorylation of the Tau protein and/or total Tau protein,   
       for its use in the treatment or the prevention of a neurodegenerative disease, advantageously a Tauopathy, in particular Alzheimer's disease, provided that said neurodegenerative disease is different from a prion disease, and provided that when said agent directly or indirectly affects the activity and/or the level:
 of heparan sulfate, or 
 of abnormal phosphorylation of the Tau protein and/or total Tau protein, 
 
       therefore, both activities and/or levels of heparan sulfate and of abnormal phosphorylation of the Tau protein and/or total Tau protein are affected by said agent, 
       or the activity and/or the level of at least one other element chosen among said gene, said transcription product of said genes, said translation product of said genes, or said fragment or derivative or variant of said gene or said transcription or translation product is also affected. 
     
     
         2 . The composition for its use in the treatment or the prevention of a neurodegenerative disease according to  claim 1 , wherein said at least one agent is selected from the group consisting of:
 a) a polysaccharide or an oligosaccharide having a molecular weight from about 2000 Daltons to about 20 000 Daltons, preferably of about 20 000 Daltons, in particular an heparan sulfate mimetic, such as F6 molecule, CR36, HM 100 or HM 2602, or in particular a pentasaccharide, such as Arixtra® (fondaparinux), fucoidan, or pentosan polysulfate, and/or low molecular weight heparins as enoxaparin (Lovenox®), and/or ultralow molecular weight heparins and/or   b) an oligonucleotide, in particular a siRNA such as a siRNA selected from the list consisting of: sense siRNA set forth by SEQ ID NO: 71 and antisense siRNA set forth by SEQ ID NO: 72, sense siRNA set forth by SEQ ID NO: 73 and antisense siRNA set forth by SEQ ID NO: 74, sense siRNA set forth by SEQ ID NO: 75 and antisense siRNA set forth by SEQ ID NO: 76, sense siRNA set forth by SEQ ID NO: 77 and antisense siRNA set forth by SEQ ID NO: 78, or a morpholino antisense oligonucleotide, in particular selected from the group consisting of SEQ ID NO: 5 and SEQ ID NO: 6, and/or   c) a small molecule, inhibitor of heparin and heparan sulfate actions, such as protamine or protamine sulfate.   
     
     
         3 . An in vitro method of diagnosis or prognostic or follow-up of a neurodegenerative disease in a subject, or determining whether a subject is at increased risk of developing said neurodegenerative disease, comprising determining a level and/or an activity of:
 a) at least one gene selected from the group consisting of the nucleotidic sequences set forth by SEQ ID NO: 1 and SEQ ID NO: 3 and coding for an heparan sulfate glucosamine 3-O-sulfotransferase 2 and 4 respectively, and/or   b) at least one transcription product of a gene selected from the group consisting of the nucleotidic sequences set forth by SEQ ID NO: 1 and SEQ ID NO: 3 and coding for an heparin-glucosamine 3-O-sulfotransferase 2 and 4 respectively, and/or   c) at least one translation product of said genes, said products being set forth respectively by SEQ ID NO: 2 and 4, and/or   d) a fragment or derivative or variant of said gene or said transcription or translation product, and/or   e) heparan sulfate, in particular 3-O-sulfated heparan sulfate, notably 3-O-sulfated heparan sulfate disaccharide, and/or   f) abnormal phosphorylation of the Tau protein and/or total Tau protein,   
       in a sample of a biological fluid previously obtained from said subject and comparing said level and/or said activity to a respective reference value representing a known disease or health status,
 an increased level and/or activity of at least one of said gene and/or said transcription product and/or said translation product and/or said heparan sulfate, in particular said 3-O-sulfated heparan sulfate and/or said abnormal phosphorylation of the Tau protein and/or total Tau protein in the subject sample being indicative of a neurodegenerative disease or a risk of developing said neurodegenerative disease, advantageously a Tauopathy, in particular Alzheimer's disease, provided that said neurodegenerative disease is different from a prion disease, and provided that when said activity and/or the level determined is the one of:
 heparan sulfate, or 
 of abnormal phosphorylation of the Tau protein and/or total Tau protein, 
 
 therefore, the activities and/or levels heparan sulfate and abnormal phosphorylation of the Tau protein and/or total Tau protein are both determined, or 
 the activity and/or the level of at least one other element chosen among said gene, said transcription product of said genes, said translation product of said genes, or said fragment or derivative or variant of said gene or said transcription or translation product is also determined. 
 
     
     
         4 . The in vitro method according to  claim 3 , wherein said neurodegenerative disease is a Tauopathy. 
     
     
         5 . The in vitro method according to  claim 4 , wherein the level of the abnormal phosphorylation of Tau protein and/or total Tau protein is increased by at least 10% compared to a respective reference value. 
     
     
         6 . The in vitro method according to  claim 4 , wherein said biological fluid is the CSF and said activity and/or level of:
 a) said gene selected from the group consisting of the nucleotidic sequences set forth by SEQ ID NO: 1 and SEQ ID NO: 3 and coding for an heparin-glucosamine 3-O-sulfotransferase 2 and 4 respectively, and   b) said transcription product of a gene selected from the group consisting of the nucleotidic sequences set forth by SEQ ID NO: 1 and SEQ ID NO: 3 and coding for an heparin-glucosamine 3-O-sulfotransferase, and   c) said translation product of said genes, said products being set forth respectively by SEQ ID NO: 2 and 4, and   d) a fragment or derivative or variant of said gene or said transcription or translation product, and   e) heparan sulfate, in particular 3-O-sulfated heparan sulfate, notably 3-O-sulfated heparan sulfate disaccharides,   
       in a subject are substantially the same as the respective reference value and, 
       said activity and/or level of abnormal phosphorylation of the Tau protein and/or total Tau protein is increased in said subject compared to said respective reference value, in particular by at least 10%. 
     
     
         7 . The in vitro method according to  claim 3 , wherein said neurodegenerative disease is an Alzheimer's disease. 
     
     
         8 . The in vitro method according to  claim 7 , wherein said level of the transcription product of the gene set forth by SEQ ID NO: 1 is increased by at least 100% as determined by real time PCR. 
     
     
         9 . The in vitro method according to  claim 7 , wherein said level of the transcription product of the gene set forth by SEQ ID NO: 3 is increased by at least 100% as determined by real time PCR. 
     
     
         10 . The in vitro method according to  claim 7 , wherein said level of the transcription product of the gene set forth by SEQ ID NO: 1 is increased by at least 100% and said level of the transcription product of the gene set forth by SEQ ID NO: 3 is increased by at least 100% as determined by real time PCR. 
     
     
         11 . The in vitro method according to  claim 7 , wherein said level of the translation product of the gene and set forth by SEQ ID NO: 2 is increased by at least 50% as determined by Western Blot, ELISA, mass spectrometry or immunohistochemistry methods. 
     
     
         12 . The in vitro method according to  claim 7 , wherein said level of the translation product of the gene and set forth by SEQ ID NO: 4 is increased by at least 50% as determined by Western Blot, ELISA, mass spectrometry or immunohistochemistry methods. 
     
     
         13 . The in vitro method according to  claim 7 , wherein said level of the translation product of the gene and set forth by SEQ ID NO: 2 is increased by at least 50% as determined by ELISA or Western Blot and wherein said level of the translation product of the gene and set forth by SEQ ID NO: 4 is increased by at least 50% as determined by Western Blot, ELISA, mass spectrometry or immunohistochemistry methods. 
     
     
         14 . The in vitro method according to  claim 8 , comprising further determining the level and/or activity of heparan sulfate, an increase of at least 50% of said level and/or activity compared to a reference value being indicative of Alzheimer's disease. 
     
     
         15 . The in vitro method according to  claim 8 , comprising further determining the level and/or activity of abnormal phosphorylation of Tau protein and/or total Tau protein, an increase of at least 10% compared to a respective reference value being indicative of Alzheimer's disease. 
     
     
         16 . The in vitro method according to  claim 7 , wherein said biological fluid is the CSF and said activity and/or level of:
 a) at least one gene selected from the group consisting of the nucleotidic sequences set forth by SEQ ID NO: 1 and SEQ ID NO: 3 and coding for an heparin-glucosamine 3-O-sulfotransferase 2 and 4 respectively, and   b) at least one transcription product of a gene selected from the group consisting of the nucleotidic sequences set forth by SEQ ID NO: 1 and SEQ ID NO: 3 and coding for an heparin-glucosamine 3-O-sulfotransferase, and   c) translation product of said genes, said products being set forth respectively by SEQ ID NO: 2 and 4, and   d) heparan sulfate, in particular 3-O-sulfated heparan sulfate, notably 3-O-sulfated heparan sulfate disaccharides, and   
       said activity and/or level of abnormal phosphorylation of the Tau protein and/or total Tau protein is increased in said subject compared to said respective reference value. 
     
     
         17 . A kit of diagnosing or prognosticating or following-up of a neurodegenerative disease, advantageously a Tauopathy, in particular Alzheimer's disease, in a subject, or determining the propensity or predisposition of a subject to develop such a disease, provided that said neurodegenerative disease is different from a prion disease,
 consisting of means for detecting in a sample of a biological fluid obtained from said subject, a level and/or an activity of:
 at least one transcription product of a gene selected from the group consisting of the nucleotidic sequences set forth by SEQ ID NO: 1 and SEQ ID NO: 3 and coding for an heparin-glucosamine 3-O-sulfotransferase, and/or
 at least one translation product of said genes, said products being set forth respectively by SEQ ID NO: 2 and 4, and comparing said level and/or activity obtained in said sample with the one obtained with a control representing a known disease or health status. 
 
   
     
     
         18 . The kit according to  claim 17 , consisting further of means for determining the level of phosphorylation of Tau protein in a sample obtained from said subject, and
 comparing said level obtained with said sample with the one obtained with a control representing a known disease or health status,   and comparing the abnormal phosphorylation Tau level and/or total Tau levels, and the levels of the SEQ ID NO: 2 and SEQ ID NO: 4 translation products with a control representing a known disease or health status.   
     
     
         19 . A method for an in vitro screening for a modulator of neurodegenerative diseases, advantageously a Tauopathy, in particular Alzheimer's disease, provided that said neurodegenerative disease is different from a prion disease, comprising:
 a) contacting a sample of a biological fluid previously collected from an animal, in particular a mammal, being subject to a neurodegenerative disease, with a compound to test,   b) determining the activity and/or a level of:
 i. at least one gene selected from the group consisting of the nucleotidic sequences set forth by SEQ ID NO: 1 and SEQ ID NO: 3 and coding for an heparin-glucosamine 3-O-sulfotransferase, and/or 
 ii. at least one transcription product of a gene selected from the group consisting of the nucleotidic sequences set forth by SEQ ID NO: 1 and SEQ ID NO: 3 and coding for an heparin-glucosamine 3-O-sulfotransferase, and/or 
 iii. at least one translation product of said genes, said products being set forth respectively by SEQ ID NO: 2 and 4, and/or 
 iv. a fragment or derivative or variant of said gene or transcription or translation product, and/or 
 v. heparan sulfate, in particular the level of 3-O-sulfated heparan sulfate, notably the level of 3-O-sulfated heparan sulfate disaccharide 
 vi. abnormal phosphorylation of the Tau protein and/or total Tau protein, 
   c) determining said activity and/or a level in a control sample of a biological fluid previously collected from a mammal with a neurodegenerative disease not contacted with said compound,   d) comparing the difference of said activity and/or a level in the contacted sample of a biological fluid with the one in the non contacted sample of a biological fluid, wherein an alteration in said activity and/or level of the contacted cell indicates that the test compound is a modulator of said disease,   
       provided that when said activity and/or the level determined in step b) is the one of:
 heparan sulfate, or 
 of abnormal phosphorylation of the Tau protein and/or total Tau protein, 
 
       therefore, the activities and/or levels heparan sulfate and abnormal phosphorylation of the Tau protein and/or total Tau protein are both determined, or 
       the activity and/or the level of at least one other element chosen among said gene, said transcription product of said genes, said translation product of said genes, or said fragment or derivative or variant of said gene or said transcription or translation product is also determined. 
     
     
         20 . A method according to  claim 19 , wherein said mammal is a human or a mouse such as a SAMP8 mouse and/or the 3×Tg-AD mice model of AD or said animal is Zebra fish.