Claims for Patent: 6,150,091

Summary for Patent: 6,150,091

Title:	Direct molecular diagnosis of Friedreich ataxia
Abstract:	This invention relates generally to methods for the diagnosis and therapeutic treatment of Friedreich Ataxia. Friedreich ataxia (FRDA) is an autosomal recessive, degenerative disease that involves the central and peripheral nervous system and the heart. A gene, X25, was identified in the critical region for the FRDA locus on chromosome 9q13. The gene encodes a 210 amino acid protein, frataxin, that has homologues in distant species such as C. elegans and yeast. A few FRDA patients have been found to have point mutations in X25, but the vast majority are homozygous for a variable, unstable GAA trinucleotide expansion in the first X25 intron. Mature X25 mRNA was severely reduced in abundance in individuals with FRDA. Carriers and individuals at risk for developing FRDA can be ascertained by the methods of the present invention. Further, the methods of the present invention provide treatment to those individuals having FRDA.
Inventor(s):	Pandolfo; Massimo (Graglia, IT), Montermini; Laura (Milan, IT), Molto; Maria D. (Valencia, ES), Koenig; Michael (Plobesheim, FR), Campuzano; Victoria (Strasbourg, FR), Cossee; Mireille (Strasbourg, FR)
Assignee:	Baylor College of Medicine (Houston, TX) INSERM (Paris, FR)
Application Number:	08/611,587
Patent Claims:	1. A method of screening individuals for a mutation that leads to Friedreich's ataxia, comprising the steps of: digesting DNA from an individual to be tested with a restriction endonuclease; and measuring the length of a restriction fragment length polymorphisn (RFLP) by hybridzation to probes that recognize a region encompassing a GAA repeat in the first intron of an X25 gene and performing Southern Blot analysis, wherein an RFLP having said GAA expansion of more than about 120 is an indication of said mutation that leads to Friedreich's ataxia. 2. The method of claim 1, wherein the restriction endonuclease is EcoRI. 3. The method of claim 1, wherein the probe used for performing said Southern Blot is SEQ ID NO 2. 4. The method of claim 1, wherein the probe used for performing said Southern Blot is an amplification product obtained by performing PCR on said DNA with SEQ ID NO 16 and SEQ ID NO 17. 5. A method of screening individuals for a mutation that leads to Friedreich's ataxia, comprising the steps of measuring expression of an X25 gene by determining an amount of mRNA expressed from said X25 gene and from known controls, and comparing the amount of mRNA from said X25 gene to the amount of mRNA from the known controls wherein reduced levels of said mRNA identify individuals with said mutation. 6. The method of claim 5, wherein the amount of mRNA is determined by the steps of: extracting mRNA from individuals to be tested; preparing cDNA from mRNA; amplifying said cDNA to produce amplification products; and comparing relative amounts of X25 and control amplification products present, wherein a reduced amount of mRNA from the X25 gene indicates individuals having said mutation that leads to Friedreich's ataxia. 7. The method of claim 6, wherein the comparing step includes electrophoresis of said amplification products; transfering said amplification products to a solid support; hybridizing said amplification products to a probe; and quantifying of X25 amplification products versus control gene amplification products. 8. The method of claim 7, wherein said probe is SEQ ID NO 14. 9. The method of claim 7, wherein said control gene is serine hydroxymethyltransferase (SHMT). 10. A method of screening individuals for a mutation that leads to Friedreich's ataxia, comprising the step of detecting a variation in a size of a (GAA).sub.n repeat in a first intron of a X25 gene by measuring a length of said repeat, wherein n for normal individuals ranges from 1-22 and n for affected individuals is 120 and wherein a value of n of at least 120 identifies individuals with said mutation. 11. The method of claim 10, wherein said size of said repeat is measured by restriction endonuclease digestion of sample DNA and Southern Blot analysis. 12. The method of claim 10, wherein said size of said repeat is determined by pulsed field gel electrophoresis. 13. The method of claim 10, wherein SEQ ID NO 29 and SEQ ID NO 30 are used in said detecting step. 14. The method of claim 10, wherein SEQ ID NO 31 and SEQ ID NO 32 are used in said detecting steps. 15. A method for detecting a GAA polymorphism in a first intron of an X25 gene comprising the steps of performing a PCR assay to produce amplified products of said first intron of said X25 gene and measuring the length of said amplified products wherein an increase in length of said amplified products as compared to a control amplified product indicates the presence of said GAA polymorphisms. 16. The method of claim 15, wherein SEQ ID NO 29 and SEQ ID NO 30 are used in said PCR assay. 17. The method of claim 15, wherein SEQ ID NO 31 and SEQ ID NO 32 are used in said PCR assay. 18. A method of screening individuals for a mutation that leads to Friedreich's ataxia, comprising the steps of sequencing DNA from an individual, and comparing said sequence from said individual to SEQ ID NOS 1-12 to determine what differences there are between said sequence from said individual and SEQ ID NOS 1-12 wherein said differences are indicative of said mutation. 19. As a composition of matter, the molecule having SEQ ID NO 1. 20. As a composition of matter, the molecule having SEQ ID NO 2. 21. As a composition of matter, the molecule having SEQ ID NO 3. 22. As a composition of matter, the molecule having SEQ ID NO 6. 23. As a composition of matter, the molecule having SEQ ID NO 7. 24. As a composition of matter, the molecule having SEQ ID NO 8. 25. As a composition of matter, the molecule having SEQ ID NO 9. 26. As a composition of matter, the molecule having SEQ ID NO 10. 27. As a composition of matter, the molecule having SEQ ID NO 11. 28. As a composition of matter, the molecule having SEQ ID NO 12. 29. As a composition of matter, the molecule having SEQ ID NO 13. 30. As a composition of matter, the molecule having SEQ ID NO 14. 31. As a composition of matter, the molecule having SEQ ID NO 15. 32. As a composition of matter, the molecule having SEQ ID NO 16. 33. As a composition of matter, the molecule having SEQ ID NO 17. 34. As a composition of matter, the molecule having SEQ ID NO 18. 35. As a composition of matter, the molecule having SEQ ID NO 19. 36. As a composition of matter, the molecule having SEQ ID NO 20. 37. As a composition of matter, the molecule having SEQ ID NO 21. 38. As a composition of matter, the molecule having SEQ ID NO 22. 39. As a composition of matter, the molecule having SEQ ID NO 23. 40. As a composition of matter, the molecule having SEQ ID NO 24. 41. As a composition of matter, the molecule having SEQ ID NO 25. 42. As a composition of matter, the molecule having SEQ ID NO 26. 43. As a composition of matter, the molecule having SEQ ID NO 27. 44. As a composition of matter, the molecule having SEQ ID NO 28. 45. As a composition of matter, the molecule having SEQ ID NO 29. 46. As a composition of matter, the molecule having SEQ ID NO 30. 47. As a composition of matter, the molecule having SEQ ID NO 31. 48. As a composition of matter, the molecule having SEQ ID NO 32.

Details for Patent 6,150,091

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132	06/04/1986		2039-02-26
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132			2039-02-26
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	Injection	103132			2039-02-26