Methods and compositions for assessing patients with reproductive failure using immune cell-derived microrna

Abstract

The invention is directed to methods and compositions for collecting immune cells, preferably peripheral blood mononuclear cells (PBMCs), before or after an intervention, extracting microRNA-comprising RNA from said cells, quantifying microRNAs within the extracted RNA, determining one or more microRNAs that display a bimodal response amongst a statistically sufficient number of patient samples. Patients are then preferably segregated into groups according to their response.

1 . A method for identifying at least two characteristic groups in a patient population on the basis of microRNA expression comprising the steps of:
 a) collecting immune cells;   b) extracting microRNA-comprising RNA from the immune cells;   c) quantifying at least one microRNA within the extracted RNA; and   d) segregating the patient population into the groups on the basis of expression of the at least one microRNA.   
     
     
         2 . The method of  claim 1 , wherein the step of collecting immune cells comprises collecting peripheral blood mononuclear cells. 
     
     
         3 . The method of  claim 1 , wherein the step of segregating the patient population comprises assigning patients expressing a relatively high level of the at least one microRNA to a first group and assigning patients expressing a relatively low level of the at least one microRNA to a second group. 
     
     
         4 . The method of  claim 1 , wherein the step of collecting immune cells comprises collecting the cells before an immunotherapy treatment. 
     
     
         5 . The method of  claim 1 , wherein the step of collecting immune cells comprises collecting the cells after an immunotherapy treatment. 
     
     
         6 . The method of  claim 1 , wherein the step of collecting immune cells comprises collecting the cells before and after an immunotherapy treatment and wherein the step of segregating the patient population includes determining the change in expression level of the at least one microRNA after the immunotherapy treatment. 
     
     
         7 . The method of  claim 6 , wherein the step of segregating the patient population comprises assigning patients exhibiting a first change in the expression level to a first group and assigning patients exhibiting a second change in the expression level to a second group. 
     
     
         8 . The method of  claim 7 , wherein the first change is a relatively large change in expression level and the second change is a relatively small change in expression level. 
     
     
         9 . The method of  claim 7 , wherein the first change is a positive change in expression level and the second change is a negative change in expression level. 
     
     
         10 . The method of  claim 7 , wherein the absolute value of the mean of the change in the expression level of the at least one microRNA in the first group divided by the standard deviation is greater than or equal to one. 
     
     
         11 . The method of  claim 7 , further comprising the step identifying a subset of microRNAs within the group of known microRNAs that exhibit a change in expression level in the first group such that the absolute value of the mean of the change in expression level divided by the standard deviation is greater than or equal to one. 
     
     
         12 . The method of  claim 11 , further comprising the step of identifying a microRNA within the group of known microRNAs that exhibits the greatest change in expression level in the first group. 
     
     
         13 . The method of  claim 1 , further comprising the steps of:
 a) collecting immune cells from an additional patient;   b) extracting microRNA-comprising RNA from the immune cells of the additional patient;   c) quantifying at least one microRNA within the extracted RNA from the additional patient; and   d) identifying the additional patient as belonging to one of the segregated groups on the basis of expression of the at least one microRNA.   
     
     
         14 . The method of  claim 13 , further comprising the step of administering a treatment to the additional patient based on the identification. 
     
     
         15 . The method of  claim 14 , wherein the step of administering a treatment comprises administering IVIG. 
     
     
         16 . The method of  claim 1 , further comprising the step of diagnosing a patient as having a condition based on membership in a segregated group. 
     
     
         17 . The method of  claim 16 , wherein the condition is a reproductive disorder. 
     
     
         18 . The method of  claim 1 , further comprising the step of monitoring treatment of a patient belonging to one of the segregated groups by collecting immune cells, extracting at least one microRNA and quantifying the at least one microRNA at a subsequent time. 
     
     
         19 . The method of  claim 1 , wherein the at least one microRNA is selected from the group consisting of hsa-let-7e, hsa-mir-1181, hsa-miR-1183, hsa-miR-1224-5p, hsa-miR-127-3p, hsa-mir-1296, hsa-mir-132, hsa-mir-136, hsa-miR-139-3p, hsa-mir-141, hsa-miR-142-3p, hsa-mir-142-5p, hsa-mir-144, hsa-mir-153, hsa-mir-1537, hsa-miR-154, hsa-miR-191, hsa-mir-193a-3p, hsa-miR-19a, hsa-mir-219-5p, hsa-mir-29b, hsa-mir-301a, hsa-miR-301b, hsa-miR-30e, hsa-mir-32, hsa-mir-33a, hsa-miR-340, hsa-miR-362-3p, hsa-miR-371-5p, has-377, hsa-miR-423-3p, hsa-miR-432, hsa-mir-513a-5p, hsa-mir-545, hsa-miR-548a-5p, hsa-miR-574-5p, hsa-mir-582-3p, hsa-mir-590-5p, hsa-mir-15a, hsa-mir-548c-5p, hsa-mir-1225-3p, hsa-mir-29b, hsa-mir-21, hsa-mir-1237, hsa-mir-101, hsa-mir-1539, hsa-mir-557, hsa-mir-125a-3p and hsa-mir-423-5p. 
     
     
         20 . The method of  claim 19 , wherein the at least one microRNA is selected from the group consisting of hsa-mir-136, hsa-mir-141, hsa-mir-142-5p, hsa-mir-144, hsa-mir-153, hsa-mir-1537, hsa-mir-193a-3p, hsa-mir-219-5p, hsa-mir-29b, hsa-mir-301a, hsa-mir-32, hsa-mir-33a, hsa-mir-545, hsa-mir-582-3p, hsa-mir-590-5p, hsa-mir-1181, hsa-mir-513a-5p, hsa-mir-132 and hsa-mir-1296.