IDENTIFICATION OF A JAK2 MUTATION INVOLVED IN VAQUEZ POLYGLOBULIA

• US 20130189683A1
• Filed: 02/15/2013
• Published: 07/25/2013
• Est. Priority Date: 10/27/2004
• Status: Active Grant

• Current Assignee: Institut National De La Sante Et De La Recherche Medicale (Inserm) (Allianz Global Investors GmbH), Assistance Publique - Hopitaux De Paris, IGR&D SA, Institut Gustave Roussy, Universite De Versailles - St. Quentin En Yvelines, Universite Paris - Sud
• Original Assignee: Institut National De La Sante Et De La Recherche Medicale (Inserm) (Allianz Global Investors GmbH), Assistance Publique - Hopitaux De Paris, IGR&D SA, Institut Gustave Roussy, Universite De Versailles - St. Quentin En Yvelines, Universite Paris - Sud
• Inventors: Vainchenker, William, Ugo, Valrie, Chloe, James, Le Couedic, Jean-Pierre, Casadevall, Nicole
• Granted Patent: US 8,852,931 B2
• Time in Patent Office: Days
• US Class Current: 435/6.11
• CPC Class Codes: A61P 19/08 for bone diseases, e.g. rachitism, Paget's disease
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 9/1205 Phosphotransferases with an alcohol group as acceptor (2.7.1), e.g. protein kinases
• CPC Class Codes: C12Q 1/6813 Hybridisation assays
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 2600/156 Polymorphic or mutational markers
• CPC Class Codes: G01N 2333/91215 with a definite EC number (2.7.1.-)
• CPC Class Codes: G01N 2500/04 Screening involving studying the effect of compounds C directly on molecule A (e.g. C are potential ligands for a receptor A, or potential substrates for an enzyme A)
• CPC Class Codes: G01N 33/57426 leukemia
• CPC Class Codes: G01N 33/57496 involving intracellular compounds

Abstract

The present invention concerns the V617F variant of the protein-tyrosine kinase JAK2, said variant being responsible for Vaquez Polyglobulia. The invention also relates to a first intention diagnostic method for erythrocytosis and thrombocytosis allowing their association with myeloproliferative disorders, or to the detection of the JAK2 V617F variant in myeloproliferative disorders allowing their reclassification in a new nosological group.

Claims

1-20. -20. (canceled)

21. A method, comprising:

a) obtaining and analyzing a nucleic acid sample from a human patient, wherein the analyzing comprises;

a1) sequencing a region of the nucleic acid sample comprising position 2343 of SEQ ID NO;

2, or

a2) hybridizing under stringent conditions the nucleic acid sample with at least one probe or primer comprising at least 10 consecutive nucleotides of SEQ ID NO;

2 comprising position 2343 of SEQ ID NO;

2 or a probe or primer having at least approximately 90% homology to said at least 10 consecutive nucleotides;

b) detecting the presence or absence of a thymine (T) in the JAK2 gene at position 2343 of SEQ ID NO;

2 in the nucleic acid sample;

c) if T is present at position 2343, treating the patient for Vaquez polyglobulia, thrombocytaemia, or myelofibrosis without quantifying red cell mass, quantifying erythroid endogenous colonies (EEC), or testing bone marrow of the patient; and

d) if T is absent at position 2343, quantifying red cell mass, quantifying erythroid endogenous colonies (EEC), or testing bone marrow of the patient.

22. The method of claim 21, wherein the at least one probe is labeled with at least one marker and detecting comprises detection of the signal produced by the at least one probe.

23. The method of claim 21, wherein the hybridizing comprises hybridizing with at least two probes.

24. The method of claim 21, wherein the hybridizing occurs with a probe or primer having at least approximately 95% homology to said at least 10 consecutive nucleotides.

25. The method of claim 21, wherein the hybridizing occurs with a probe or primer having at least approximately 98% homology to said at least 10 consecutive nucleotides.

26. The method of claim 21, wherein the human patient has a hematocrit level of more than 51% or is presenting with polyglobulias.

27. The method of claim 26, wherein the human patient has a hematocrit level of more than 51%.

28. The method of claim 26, wherein the human patient is presenting with polyglobulia.

29. The method of claim 21, wherein the human patient is suspected of having symptoms of Vaquez polyglobulia, thrombocytaemia, and/or myelofibrosis.

30. An isolated nucleic acid comprising at least 12 consecutive nucleotides of sequence SEQ ID NO 3 or 4, or having at least approximately 90% homology to said at least 12 consecutive nucleotides, wherein the isolated nucleic acid comprises the nucleotide t²⁶¹ in SEQ ID NO 3 or t⁵⁰ in SEQ ID NO 4, wherein the isolated nucleic has the functional properties of a probe or primer.

31. The isolated nucleic acid according to claim 30, having at least approximately 95% homology to said at least 12 consecutive nucleotides.

32. The isolated nucleic acid according to claim 30, having at least approximately 98% homology to said at least 12 consecutive nucleotides.

33. The isolated nucleic acid according to claim 30, wherein said isolated nucleic acid has at least approximately 90% homology to SEQ ID NO:

11 with a g²¹t mutation.

34. The isolated nucleic acid according to claim 30, consisting essentially of 15 to 30 consecutive nucleotides.

35. The isolated nucleic acid according to claim 30, consisting essentially of at least 15 consecutive nucleotides.

36. The isolated nucleic acid according to claim 30, consisting essentially of at least 12 consecutive nucleotides.

37. A kit for detecting a G¹⁸⁴⁹T mutation in the human JAK2 gene in a human tumor, wherein the kit comprises one or more primers or probes, as defined in claim 30, for the specific detection of the presence or absence of the G ^I849T mutation in the human JAK2 gene, wherein the G¹⁸⁴⁹T mutation is nucleotide t²⁶¹ in SEQ ID NO 3 or t⁵⁰ in SEQ ID NO 4.

38. A kit for determining whether a patient is suffering from a myeloproliferative disorder involving a G¹⁸⁴⁹T mutation of the human JAK2 gene, wherein the kit comprise one or more probes or primers, as defined in claim 30, for the specific detection of the presence or absence of the G¹⁸⁴⁹T mutation in the human JAK2 gene.

39. The kit according to claim 38, further comprising at least one element chosen from among a heat resistant polymerase for PCR amplification, one or more solutions for amplification and/or hybridisation and any reagent allowing said specific detection.