Lmna gene and its involvement in hutchinson-gilford progeria syndrome (hgps) and arteriosclerosis

Abstract

Disclosed herein are point mutations in the LMNA gene that cause HGPS. These mutations activate a cryptic splice site within the LMNA gene, which leads to deletion of part of exon 11 and generation of a mutant Lamin A protein product that is 50 amino acids shorter than the normal protein. In addition to the novel Lamin A variant protein and nucleic acids encoding this variant, methods of using these molecules in detecting biological conditions associated with a LMNA mutation in a subject (e.g., HGPS, arteriosclerosis, and other age-related diseases), methods of treating such conditions, methods of selecting treatments, methods of screening for compounds that influence Lamin A activity, and methods of influencing the expression of LMNA or LMNA variants are also described. Oligonucleotides and other compounds for use in examples of the described methods are also provided, as are protein-specific binding agents, such as antibodies, that bind specifically to at least one epitope of a Lamin A variant protein preferentially compared to wildtype Lamin A, and methods of using such antibodies in diagnosis, treatment, and screening. Also provided are kits for carrying out the methods described herein.

1 . A substantially purified mutant human Lamin A protein, comprising an amino acid sequence as set forth in
 (a) SEQ ID NO: 7;   (b) sequences having at least 80% sequence identity to SEQ ID NO: 7 and containing Mutation 1 (G608G(GGC>GGT)) or Mutation 2 (G608S(GGC>AGC));   (c) a conservative variant of (a) or (b) and containing Mutation 1 (G608G(GGC>GGT)) or Mutation 2 (G608S(GGC>AGC));   (d) sequences having at least 95% sequence identity to SEQ ID NO: 2 and comprising the variant E145K;   (e) sequences having at least 95% sequence identity to SEQ ID NO: 2 and comprising the variant R471C;   (f) sequences having at least 95% sequence identity to SEQ ID NO: 2 and comprising the variant R527C; or   (g) sequences having at least 95% sequence identity to SEQ ID NO: 2 and comprising the variant A269V.   
     
     
         2 . The substantially purified mutant human Lamin A protein of  claim 1 , further comprising a heterologous peptide fused thereto. 
     
     
         3 . The substantially purified mutant human Lamin A protein of  claim 1 , further comprising a label. 
     
     
         4 . A monoclonal or polyclonal antibody or fragment thereof that binds to a mutant human Lamin A protein of  claim 1  but not to wildtype Lamin A (SEQ ID NO: 2). 
     
     
         5 . A mutant Lamin A protein-specific monoclonal or polyclonal antibody or fragment thereof that binds to an epitope within the sequence SGSGAQSPQNC (positions 601 to 611 of SEQ ID NO: 7) and not to an epitope in wildtype Lamin A (SEQ ID NO: 2). 
     
     
         6 . A kit for determining whether or not a subject has Hutchinson-Gilford Progeria Syndrome (HGPS), arteriosclerosis, atherosclerosis, predisposition to arteriosclerosis, or predisposition to atherosclerosis by detecting a mutant Lamin a protein in the subject, wherein determining whether the subject has a mutant Lamin a protein comprises reacting at least one Lamin A protein contained in a sample from the subject with a reagent comprising a mutant Lamin A protein-specific binding agent to form a Lamin A:binding agent complex and wherein the amino acid sequence of the mutant Lamin A protein comprises:
 (a) SEQ ID NO: 7;   (b) a sequence having at least 95% sequence identity to SEQ ID NO: 7 and containing Mutation 1 (G608G(GGC>GGT)) or Mutation 2 (G608S(GGC>AGC));   (c) a conservative variant of (a) containing Mutation 1 (G608G(GGC>GGT)) or Mutation 2 (G608S(GGC>AGC));   (d) sequences having at least 95% sequence identity to SEQ ID NO: 2 and comprising the variant E145K;   (e) sequences having at least 95% sequence identity to SEQ ID NO: 2 and comprising the variant R471C;   (f) sequences having at least 95% sequence identity to SEQ ID NO: 2 and comprising the variant R527C; or   (g) sequences having at least 95% sequence identity to SEQ ID NO: 2 and comprising the variant A269V,   
       the kit comprising:
 a container; and 
 contained therein, the mutant Lamin A-specific antibody of  claim 4 . 
 
     
     
         7 . A kit for determining whether or not a subject has Hutchinson-Gilford Progeria Syndrome (HGPS), arteriosclerosis, atherosclerosis, predisposition to arteriosclerosis, or predisposition to atherosclerosis by detecting a mutant Lamin a protein in the subject, wherein determining whether the subject has a mutant Lamin a protein comprises reacting at least one Lamin A protein contained in a sample from the subject with a reagent comprising a mutant Lamin A protein-specific binding agent to form a Lamin A:binding agent complex the kit comprising:
 a container; and   contained therein, the mutant Lamin A-specific antibody of  claim 5 .   
     
     
         8 . A kit for determining whether or not a subject has Hutchinson-Gilford Progeria Syndrome (HGPS), arteriosclerosis, atherosclerosis, predisposition to arteriosclerosis, or predisposition to atherosclerosis by detecting a mutant Lamin A protein in the subject, the kit comprising a container, and contained therein, a mutant Lamin A protein-specific binding agent, wherein the mutant Lamin A protein-specific binding agent comprises an antibody that binds to an epitope within the sequence SGSGAQSPQNC (positions 601 to 611 of SEQ ID NO: 7) but not to an epitope in wildtype Lamin A (SEQ ID NO: 2).