Estrogen receptor alleles that are predictive of increased susceptibility to bone fracture

Abstract

In one aspect, the present invention provides methods of determining susceptibility to bone fracture in a mammalian subject, wherein the methods comprise analyzing nucleic acid molecules obtained from the mammalian subject to determine which of the P, p X, and x alleles of the estrogen receptor α gene are present, wherein the presence of a haplotype comprising the p and x alleles is indicative of an increased susceptibility to bone fracture. The present invention also provides kits for determining susceptibility to bone fracture in a mammalian subject.

1 - 30 . (canceled) 
     
     
         31 . A method of detecting p and x alleles of a PvuII site and a XbaI site in an estrogen receptor α gene in a mammalian subject comprising at least one estrogen receptor α gene comprising a PvuII site and a XbaI site, wherein the PvuII site can exist as a P or p allelic form, and the XbaI site can exist as an X or x allelic form, said method comprising analyzing nucleic acid molecules obtained from the mammalian subject to determine which of the P, p X, and x alleles of the estrogen receptor α gene are present. 
     
     
         32 . The method of detecting p and x alleles of a PvuII site and a XbaI site in an estrogen receptor α gene according to  claim 31 , said method comprising analyzing nucleic acid molecules of the mammalian subject to determine whether the px haplotype of the estrogen receptor α gene is present. 
     
     
         33 . The method of detecting p and x alleles of a PvuII site and a XbaI site in an estrogen receptor α gene according to  claim 31 , said method further comprising analyzing nucleic acid molecules of the mammalian subject to determine whether an allele of a vitamin D receptor gene is present. 
     
     
         34 . The method of detecting p and x alleles of a PvuII site and a XbaI site in an estrogen receptor α gene according to  claim 33 , wherein the vitamin D receptor gene comprises a BsmI site, an ApaI site and a TaqI site, wherein the BsmI site can exist as a B or b allelic form, the ApaI site can exist as an A or a allelic form, and the TaqI site can exist as a T or t allelic form, the method comprising analyzing nucleic acid molecules of the mammalian subject to determine which of the B, b, A, a, T, and t alleles of the BsmI, ApaI and TaqI sites of the vitamin D receptor gene are present. 
     
     
         35 . The method of detecting p and x alleles of a PvuII site and a XbaI site in an estrogen receptor α gene according to  claim 34 , said method comprising analyzing nucleic acid molecules of the mammalian subject to determine whether a baT haplotype of the vitamin D receptor gene is present. 
     
     
         36 . The method of detecting p and x alleles of a PvuII site and a XbaI site in an estrogen receptor α gene according to  claim 34 , said method further comprising determining the copy number of a member of the group consisting of the P, p, X and x alleles of the estrogen receptor α. gene and the B, b, A, a, T and t alleles of the vitamin D receptor gene. 
     
     
         37 . The method of detecting p and x alleles of a PvuII site and a XbaI site in an estrogen receptor α gene according to  claim 31 , wherein the subject is a human female. 
     
     
         38 . The method of detecting p and x alleles of a PvuII site and a XbaI site in an estrogen receptor α gene according to  claim 31 , wherein said method is performed in vitro. 
     
     
         39 . The method of detecting p and x alleles of a PvuII site and a XbaI site in an estrogen receptor α gene according to  claim 38 , wherein said method is performed on a blood or tissue sample of a subject. 
     
     
         40 . The method of detecting p and x alleles of a PvuII site and a XbaI site in an estrogen receptor α gene according to  claim 31  wherein the mammalian subject is suffering from low bone mineral density. 
     
     
         41 . The method of detecting p and x alleles of a PvuII site and a XbaI site in an estrogen receptor α gene according to  claim 31  wherein the mammalian subject has a normal level of bone mineral density. 
     
     
         42 . The method of detecting p and x alleles of a PvuII site and a XbaI site in an estrogen receptor α gene according to  claim 31 , wherein the presence of the haplotype is determined by amplification of a portion of the first intron of the estrogen receptor α gene to yield an amplified fragment, followed by restriction enzyme digestion of the amplified fragment. 
     
     
         43 . The method of detecting p and x alleles of a PvuII site and a XbaI site in an estrogen receptor α gene according to  claim 42 , further comprising determining the haplotype of a vitamin D receptor gene by amplification of a portion of the vitamin D receptor gene between exon 7 and the 3′ untranslated region to yield an amplified fragment, followed by restriction enzyme digestion of the amplified fragment. 
     
     
         44 . A method of determining the susceptibility of bone fracture and treating the same in a mammalian subject to prevent or reduce the risk of bone fracture, wherein the mammalian subject comprises at least one estrogen receptor α gene comprising a PvuII site and a XbaI site, wherein the PvuII site can exist as a P or pallelic form, and the XbaI site can exist as an X or x allelic form, the method comprising analyzing nucleic acid molecules obtained from the mammalian subject to determine which of the P, p, X and x alleles of the PvuII and XbaI sites of the estrogen receptor α gene are present, wherein the presence of a haplotype comprising the p and x alleles is indicative of an increased susceptibility to bone fracture, and treating the mammalian subject to reduce the risk of bone fracture if the subject has a haplotype comprising the p and x alleles. 
     
     
         45 . The method of  claim 44  further comprising analyzing nucleic acid molecules of the mammalian subject to determine whether the px haplotype of the estrogen receptor α gene is present, wherein the presence of the px haplotype is indicative of an increased susceptibility to bone fracture. 
     
     
         46 . The method of  claim 44  further comprising analyzing nucleic acid molecules of the mammalian subject to determine which of the B, b, A, a, T and t alleles of the BsmI, ApaI and TaqI sites of the vitamin D receptor gene are present, wherein the presence of a haplotype comprising at least one of the b, a, and T alleles is indicative of an increased susceptibility to bone fracture. 
     
     
         47 . The method of  claim 44  further comprising analyzing nucleic acid molecules of the mammalian subject to determine whether the baT haplotype of the vitamin D receptor gene is present, wherein the presence of the baT haplotype is indicative of an increased susceptibility to bone fracture. 
     
     
         48 . The method of  claim 44 , said method further comprising determining the copy number of a member of the group consisting of the P, p, X and x alleles of the estrogen receptor α gene and the B, b, A, a, T and t alleles of the vitamin D receptor gene, before treating the subject to reduce the risk of bone fracture if the subject has a haplotype comprising the p and x alleles of the estrogen receptor α gene. 
     
     
         49 . The method of  claim 44 , wherein the treatment comprises at least one of modifications to lifestyle, regular exercise, changes in diet and administration of a pharmaceutical preparation effective to prevent or reduce the risk of bone fracture. 
     
     
         50 . The method of  claim 44 , wherein the subject is a human female.