Methods of Novel Therapeutic Candidate Identification Through Gene Expression Analysis in Vascular-Related Disease

Abstract

Multiple treatment regimens for vascular-related diseases and disorders. Methods of treating vascular-related disorders based on gene expression studies from samples collected from individuals having symptoms of vascular-related disorders. Additionally, methods for diagnostic techniques to focus treatment regimens. Finally, methods of treating vascular-related disorders involving targeting microRNAs.

We claim: 
     
         1 . A method of diagnosing a vascular-related disease in an individual comprising the steps of:
 a) identifying at least one gene that is upregulated or downregulated in the vascular-related disease comprising the steps of:
 1) obtaining a biopsy sample from the individual's artery during progression of the vascular-related disease; 
 2) obtaining an artery sample from a non-diseased control; 
 3) extracting RNA from the samples in steps 1) and 2); 
 4) obtaining gene products from the RNA extracted in steps 3); and 
 5) comparing gene expression levels from the biopsy sample with the non-diseased control; and 
   b) associating the genes upregulated in the biopsy sample with an inhibitor of the gene products for administration to the individual and genes downregulated in the biopsy sample with a promoter of the gene products for administration to the individual.   
     
     
         2 . The method of  claim 1 , wherein the vascular-related disease is pulmonary arterial hypertension. 
     
     
         3 . The method of  claim 1 , wherein the biopsy sample is extracted using an endoarterial catheter. 
     
     
         4 . A method of identifying microRNA dysregulated in an individual having a vascular-related disease comprising the steps of:
 a) obtaining a biopsy sample from the individual's artery during progression of the vascular-related disease;   b) obtaining an artery sample from a non-diseased control;   c) extracting RNA from the samples in steps a) and b);   d) converting the RNA to cDNA;   e) comparing levels of microRNA expression from the biopsy sample with the non-diseased control; and   f) identifying the microRNA dysregulated in the vascular-related disease relative to baseline.   
     
     
         5 . The method of  claim 4 , wherein the vascular-related disease is pulmonary arterial hypertension. 
     
     
         6 . The method of  claim 4 , wherein the microRNA is measured according to stages of progression of the vascular-related disease. 
     
     
         7 . A use of targeting microRNAs in the preparation of a medicament for the treatment of a vascular-related disease comprising the following steps:
 a) assessing a stage of the vascular-related disease in the individual;   b) identifying whether microRNAs are upregulated or downregulated;   c) selecting the microRNAs to target based on the stage of the vascular-related disease and whether the microRNAs are upregulated or downregulated; and   d) administering an agent known to inhibit an upregulated microRNA or an agent known to promote downregulated microRNA to the individual,   
       wherein the stage of the vascular-related disease is based on flow rates and blood pressure within an artery of the individual. 
     
     
         8 . The use of  claim 7 , wherein the vascular-related disease is pulmonary arterial hypertension. 
     
     
         9 . The use of  claim 7 , wherein the microRNAs to target are selected from the group consisting of Tables 12-17. 
     
     
         10 . The use of  claim 7 , wherein the stage of the vascular-related disease of one of high flow and high pressure within the artery of the individual. 
     
     
         11 . The use of  claim 10 , wherein the upregulated microRNAs to inhibit under the stage of high flow and high pressure comprise at least one member selected from the group consisting of hsa-miR-520g, hsa-miR-331-5p, hsa-miR-410, has-let-7d, hsa-miR-187, hsa-miR-16-2, hsa-miR-130a, hsa-miR-548n, hsa-miR-127-5p, solexa-2580-353, HS_206, hsa-miR-663, solexa-7534-111, hsa-let-7g, HS_157, hsa-miR-610, HS_251.1, hsa-miR-519e, hsa-miR-1237, hsa-miR-1185, HS_151.1, HS_135, solexa-3126-285, hsa-miR-30a, hsa-miR-192, hsa-miR-568, hsa-miR-1203, hsa-miR-28-5p, HS_262.1, HS_170, hsa-miR-1273, hsa-miR-602, hsa-miR-374a and hsa-miR-29a. 
     
     
         12 . The use of  claim 7 , wherein the stage of the vascular-related disease is one of high flow and low pressure within the artery of the individual. 
     
     
         13 . The use of  claim 12 , wherein the upregulated microRNAs to inhibit under the stage of high flow and low pressure comprise at least one member selected from the group consisting of HS_70, hsa-let-7d, hsa-miR-187, solexa-2580-353, HS_157, solexa-7534-111, hsa-miR-130a, HS_135, hsa-miR-192, hsa-miR-154, hsa-miR-151:9.1, hsa-miR-1203, hsa-miR-525-3p, and HS_199. 
     
     
         14 . The use of  claim 10 , wherein the downregulated microRNAs to promote under the stage of high flow and high pressure comprise at least one member selected from the group consisting of solexa-603-1846, hsa-miR-586, hsa-miR-1201, hsa-miR-33a, HS_56, hsa-miR-520d:9.1, hsa-miR-521, hsa-miR-519a, HS_134, HS_169, HS_221, hsa-miR-496, hsa-miR-935, hsa-miR-542-5p, solexa-5620-151, hsa-miR-99a, hsa-miR-212, hsa-let-7f-2, hsa-miR-494, hsa-miR-1321, hsa-miR-219-2-3p, hsa-miR-95, hsa-miR-128b:9.1, hsa-miR-33a, hsa-miR-495, hsa-miR-1229, hsa-miR-1205, hsa-miR-524-3p, HS_150, hsa-miR-135a, hsa-miR-483-5p, hsa-miR-124a:9.1, hsa-miR-29b-1, hsa-miR548c-5p, hsa-miR-551a, hsa-miR-377 and hsa-mir-1304. 
     
     
         15 . The use of  claim 12 , wherein the downregulated microRNAs to promote under the stage of high flow and low pressure comprise at least one member selected from the group consisting of HS_104, hsa-miR-548a-5p, hsa-miR-556-5p, HS_169, HS_140, hsa-miR-542-5p, HS_134, hsa-miR-212, hsa-miR-363, HS_59, hsa-miR-586, hsa-miR-494, hsa-miR-1205, hsa-let-7f-2, hsa-miR-483-5p, HS_150, hsa-miR-1229, HS_156, hsa-miR-619, hsa-miR-10b and hsa-miR-371-5p. 
     
     
         16 . A method of diagnosing an individual having a vascular-related disease according to  claim 1 , further wherein the individual is categorized based on progression of the vascular-related disease. 
     
     
         17 . The method of  claim 16 , wherein the progression of the vascular-related disease is selected from the group consisting of early stage, mid stage and late stage. 
     
     
         18 . The method of  claim 1 , wherein the diagnosing of the individual may be modified over the course of disease progression.