Methods and Reagents for Treatment and Diagnosis of Vascular Disorders and Age-Related Macular Degeneration

Abstract

Disclosed are screening methods for determining a human subject's propensity to develop a vascular disorder and/or age-related macular degeneration (AMD), therapeutic or prophylactic compounds for treating disease or inhibiting its development, and methods of treating patients to alleviate symptoms of the disease, prevent or delay its onset, or inhibit its progression. The inventions are based on the discovery that persons with a genome having a deletion of the CFHR-1 and/or CFHR-3 gene, which normally lie on human chromosome 1 between DNA encoding CFH and CFHR-4, are at reduced risk of developing AMD, and elevated risk of developing vascular disease such as aneurysm.

1 . A screening method for determining a human subject's propensity to develop a vascular disorder and/or age-related macular degeneration (AMD) comprising:
 analyzing a biological sample from the subject to detect the presence or absence of a deletion in the region of chromosome 1 between the 3′ end of exon 22 of the complement factor H (CFH) gene and the 5′ end of exon 1 of complement Factor H-related 4 (CFHR4) gene   wherein the presence of a deletion indicates the subject is at increased risk of developing a vascular disorder and is at decreased risk of developing AMD.   
     
     
         2 . The method of  claim 1  wherein the presence or absence of the deletion is detected by assaying for a gene product encoded in chromosome 1 between the 3′ end of exon 22 of the complement factor H (CFH) gene and the 5′ end of exon 1 of complement Factor H-related 4 (CFHR4) gene, where the absence of the gene product, or a reduced level of expression of the gene product, indicates the presence of deletion. 
     
     
         3 . The method of  claim 2  wherein the presence or absence of a CFHR1 gene product and/or a CFHR3 gene product is detected, where the absence of a gene product is indicative of a deletion. 
     
     
         4 . The method of  claim 2  wherein the gene product is a protein. 
     
     
         5 . The method of  claim 3  wherein entire protein coding region of the CFHR3 gene is deleted. 
     
     
         6 . The method of  claim 3  wherein entire protein coding region of the CFHR1 gene is deleted. 
     
     
         7 . The method of  claim 1  comprising detecting a deletion of an intragenic sequence selected from a sequence between the CFHR3 gene and the CFHR1 gene and a sequence between the CFHR1 gene and the CFHR4 gene. 
     
     
         8 . The method of  claim 1  wherein the subject is homozygous for the deletion. 
     
     
         9 . The method of  claim 1  wherein the biological sample is blood, serum, urine or a tissue sample. 
     
     
         10 . The method of  claim 4  wherein the detection step comprises detecting a gene product using an immunoassay or mass spectroscopy. 
     
     
         11 . The method of  claim 1  wherein the presence or absence of the deletion is detected by assaying for a truncated CFHR1 or CFHR3 gene product, where detection of a truncated gene product is indicative of a deletion. 
     
     
         12 . The method of  claim 1  wherein the step comprising detecting the presence or absence of a deletion is performed by analyzing a chromosome or nucleic acid from the subject. 
     
     
         13 . The method of  claim 12  wherein the nucleic acid is DNA or RNA. 
     
     
         14 . The method of  claim 1 , wherein the vascular disorder is aneurysm. 
     
     
         15 . The method of  claim 1  wherein the subject has a genotype of T at position 1277 of the coding region of the CFH gene of the chromosome comprising the deletion. 
     
     
         16 . The method of  claim 1  further comprising detecting genetic variants of complement factor H (CFH) gene comprising detecting one or a plurality of polymorphic sites selected from the group consisting of:
 a) any one or more of rs529825; rs800292; rs3766404; rs1061147; rs1061170; and rs203674; 
 b) any one of more of intron 2 (IVS2 or insTT); rs2274700; exon 10A; and rs375046; 
 c) one or both of rs529825 and rs800292; 
 d) one or more of rs1061147, rs1061170 and rs203674; 
 e) at least one of rs529825 and rs800292; and rs3766404; and at least one of rs1061147, rs1061170 and rs203674; 
 f) at least rs529825, rs800292, rs3766404, rs1061170, and rs203674; 
 g) exon 22 (R1210C); and 
 h) exon 22 (R1210C) and any of (a)-(g). 
 
     
     
         17 . (canceled) 
     
     
         18 . The method of  claim 1  further comprising detecting in a sample from the subject genetic variants of the HTRA1 gene comprising detecting a polymorphic site selected from the group consisting of: at least one of rs10490924, rs11200638, rs760336, and rs763720. 
     
     
         19 . The method of  claim 1  further comprising detecting in a sample from the subject genetic variants of the complement factor B (BF) gene and/or the complement component 2 (C2) gene comprising detecting a polymorphic site selected from the group consisting of:
 a) A or G at rs641153 of the BF gene, or R or Q at position 32 of the BF protein; 
 b) A or T at rs4151667 of the BF gene, or L or H at position 9 of the BF protein; 
 c) G or T at rs547154 of the C2 gene; and 
 d) C or G at rs9332379 of the C2 gene, or E of D at position 318 of the C2 protein. 
 
     
     
         20 . A method of treating a subject having or at risk for developing a vascular disorder, comprising administering a CFHR1 polypeptide, a CFHR3 polypeptide, at least one portion of a CFHR1 and/or a CFHR3 polypeptide, or mixtures thereof, to the subject. 
     
     
         21 .- 37 . (canceled) 
     
     
         38 . A kit for conducting the screening method of  claim 1  comprising reagents for detecting the presence or absence of a deletion in the DNA sequence between the 3′ end of exon 22 of the CFH gene and the 5′ end of exon 1 of the CFHR4 gene on human chromosome 1 and/or the presence or absence of a CFHR1 and/or CFHR3 gene product. 
     
     
         39 .- 44 . (canceled)