Macular degeneration diagnostics and therapeutics

Abstract

Therapeutics and diagnostics based on the identification of genetic mutations, which cause Macular Degeneration (MD) is disclosed.

1 . A method for identifying a compound that modulates a FBNL bioactivity, comprising the steps of: 
 (a) contacting an appropriate amount of the compound with a cell or cellular extract, which expresses an FBNL gene; and    (b) determining the resulting FBNL bioactivity, wherein an increase or decrease in the FBNL bioactivity in the presence of the compound as compared to the bioactivity in the absence of the compound indicates that the compound is a modulator of an FBNL bioactivity.    
     
     
         2 . A method of  claim 1 , wherein the FBNL gene is a human FBNL gene.  
     
     
         3 . A method of  claim 1 , wherein the FBNL gene is a wildtype gene.  
     
     
         4 . A method of  claim 1 , wherein the FBNL gene is a mutant gene.  
     
     
         5 . A method of  claim 1 , wherein the modulator is an agonist of an FBNL bioactivity.  
     
     
         6 . A method of  claim 1 , wherein the modulator is an antagonist of an FBNL bioactivity.  
     
     
         7 . A method of  claim 1 , wherein in step (b), the FBNL bioactivity is determined by determining the expression level of an FBNL gene.  
     
     
         8 . A method of  claim 7 , wherein the expression level is determined by detecting the amount of mRNA transcribed from an FBNL gene.  
     
     
         9 . A method of  claim 7 , wherein the expression level is determined by detecting the amount of FBNL gene product produced.  
     
     
         10 . A method of  claim 9 , wherein the expression level is determined using an anti-FBNL antibody in an immunodetection assay.  
     
     
         11 . A method of  claim 1 , which additionally comprises the step of preparing a pharmaceutical composition from the compound.  
     
     
         12 . A method of  claim 1 , wherein said cell is contained in an animal.  
     
     
         13 . A method of  claim 12 , wherein the animal is transgenic.  
     
     
         14 . A method of  claim 13 , wherein the transgenic animal contains a human FBNL gene.  
     
     
         15 . A compound identified by the method of  claim 1 .  
     
     
         16 . A compound of  claim 15 , which is selected from the group consisting of: a small molecule, a polypeptide, a nucleic acid and a peptidomimetic.  
     
     
         17 . A compound of  claim 16 , wherein the nucleic acid is selected from the group consisting of: an antisense molecule, a ribozyme and a triplex nucleic acid.  
     
     
         18 . A compound of  claim 16 , wherein the polypeptide is a FBNL polypeptide.  
     
     
         19 . A method for identifying whether a test molecule is an FBNL binding partner or measuring the strength of an interaction between an FBNL polypeptide and said FBNL binding partner comprising: 
 (a) allowing (i) a first molecule comprising a FBNL polypeptide operably linked to a heterologous DNA binding domain to interact with (ii) a second molecule comprising a test molecule operably linked to a polypeptide transcriptional activation domain and (iii) a hybrid reporter gene comprising a nucleic acid encoding a reporter operably linked to a DNA sequence comprising a binding site for said heterologous DNA binding domain; and    (b) detecting or measuring the expression of the hybrid reporter gene as an indication of the existence or strength of an interaction between the first molecule and the second molecule wherein high levels of hybrid reporter expression indicate a strong interaction between FBNL and said test molecule thereby identifying a test molecule which is an FBNL binding partner.    
     
     
         20 . A method of  claim 19 , wherein said second molecule is encoded by a nucleic acid and comprises a test polypeptide operably linked to a polypeptide transcriptional activation domain, and which further comprises the step of isolating the nucleic acid encoding said second molecule from a cell expressing the hybrid reporter gene.  
     
     
         21 . A method for identifying a molecule which is a downstream or an upstream component of an FBNL biochemical pathway or for measuring the strength of the interaction between a FBNL biochemical pathway component and a FBNL binding partner comprising: 
 (a) allowing (i) a first molecule comprising a FBNL binding partner polypeptide operably linked to a heterologous DNA binding domain to interact with (ii) a second molecule comprising a test molecule operably linked to a polypeptide transcriptional activation domain and (iii) a hybrid reporter gene comprising a nucleic acid encoding a reporter operably linked to a DNA sequence comprising a binding site for said heterologous DNA binding domain; and    (b) detecting or measuring the expression of the hybrid reporter gene as an indication of the existence or strength of an interaction between the first molecule and the second molecule wherein high levels of hybrid reporter expression indicate a strong interaction between a FBNL binding partner and said test molecule thereby identifying a test molecule which is a downstream or an upstream component of the FBNL biochemical pathway.    
     
     
         22 . A method of  claim 21 , wherein said second molecule is encoded by a nucleic acid and comprises a test polypeptide operably linked to a polypeptide transcriptional activation domain, and which further comprises the step of isolating the nucleic acid encoding said second molecule from a cell expressing the hybrid reporter gene.  
     
     
         23 . A method for identifying a compound, which interacts with a FBNL polypeptide or FBNL binding partner, comprising the steps of: 
 (a) contacting an appropriate amount of the compound with a FBNL polypeptide and a FBNL binding partner under conditions wherein, but for the test compound, the FBNL polypeptide and FBNL binding partner are able to interact; and    (b) detecting the extent to which a FBNL polypeptide/FBNL binding partner complex is formed in the presence of the compound, wherein an increase or decrease in the amount of complex formed in the presence of the compound relative to in the absence of the compound indicates that the compound interacts with a FBNL polypeptide or FBNL binding partner.    
     
     
         24 . A method of  claim 23 , wherein the FBNL polypeptide is a human FBNL polypeptide.  
     
     
         25 . A method of  claim 23 , wherein the FBNL polypeptide is a wildtype polypeptide.  
     
     
         26 . A method of  claim 23 , wherein the FBNL polypeptide is a mutant polypeptide.  
     
     
         27 . A method of  claim 23 , wherein the compound, which interacts with a FBNL polypeptide or FBNL binding partner is a FBNL agonist.  
     
     
         28 . A method of  claim 23 , wherein the compound, which interacts with a FBNL polypeptide or FBNL binding partner is a FBNL antagonist.  
     
     
         29 . A method of  claim 23 , which additionally comprises the step of preparing a pharmaceutical composition from the compound.  
     
     
         30 . A compound identified by the method of  claim 29 .  
     
     
         31 . A compound of  claim 30 , which is selected from the group consisting of: a small molecule, a polypeptide, a nucleic acid and a peptidomimetic.  
     
     
         32 . An isolated FBNL nucleic acid which is operably linked to a FBNL transcriptional regulatory sequence.  
     
     
         33 . A nucleic acid of  claim 32 , wherein the FBNL transcriptional regulatory sequence is selected from the group consisting of. a FBNL enhancer, a FBNL promoter, and a FBNL initiator element.  
     
     
         34 . An isolated nucleic acid of  claim 32 , wherein the FBNL nucleic acid is functionally fused to a heterologous gene.  
     
     
         35 . An isolated nucleic acid of  claim 34 , wherein said heterologous gene encodes a protein selected from the group consisting of: a positive selectable marker, a negative selectable marker and a reporter gene.  
     
     
         36 . An isolated nucleic acid of  claim 35 , wherein the coding sequence of FBNL is disrupted by a positive selectable marker.  
     
     
         37 . An isolated nucleic acid of  claim 36 , wherein said nucleic acid is further flanked by a negative selectable marker or markers.  
     
     
         38 . An isolated nucleic acid of  claim 37 , wherein the reporter gene is selected from the group consisting of: beta-galactosidase and luciferase.  
     
     
         39 . A cell line comprising an isolated nucleic acid of  claim 39 .  
     
     
         40 . An animal comprising an isolated nucleic acid of  claim 39 .  
     
     
         41 . An animal of  claim 40 , which is transgenic.  
     
     
         42 . An animal of  claim 41 , which contains a human FBNL gene.  
     
     
         43 . An isolated nucleic acid comprising a FBNL responsive regulatory sequence operably linked to a reporter gene.  
     
     
         44 . An isolated nucleic acid of  claim 43 , wherein the reporter gene is selected from the group consisting of: beta-galactosidase and luciferase.  
     
     
         45 . A cell line comprising an isolated nucleic acid of  claim 43 .  
     
     
         46 . An animal comprising an isolated nucleic acid of  claim 43 .  
     
     
         47 . An animal of  claim 46 , which contains a human FBNL gene.  
     
     
         48 . A cell in which the biological activity of one or more FBNL proteins is altered by a chromosomally incorporated transgene.  
     
     
         49 . A cell of  claim 48 , wherein said transgene disrupts at least a portion of a genomic FBNL gene.  
     
     
         50 . A cell of  claim 48 , wherein said transgene deletes all or a portion of the genomic FBNL gene by replacement recombination.  
     
     
         51 . A cell of  claim 48 , wherein said transgene comprises: (i) at least a portion of the genomic FBNL gene, and (ii) a marker sequence which provides a detectable signal for identifying the presence of the transgene in a cell.  
     
     
         52 . A transgenic animal comprised of a cell of  claim 48.