Targeting capn9/capns2 activity as a therapeutic strategy for the treatment of myofibroblast differentiation and associated pathologies

Abstract

Provided herein are, inter alia, compositions and methods for identifying and using agents capable of inhibiting myofibroblast transition as well as methods for treating diseases associated with the same in a subject in need thereof.

1 - 15 . (canceled) 
     
     
         16 . A method for screening potential compounds or agents to identify a compound or agent that decreases the level and/or activity of CAPN9 and/or CAPNS2, the method comprising:
 contacting a population of cells or an extract thereof with at least one potential compound or agent, wherein the population of cells or an extract thereof express CAPN9 and/or CAPNS2; and   assessing the ability of the at least one potential compound or agent to decrease level and/or activity of CAPN9 and/or CAPNS2 in the cells or extract thereof of the population, wherein if contact with the at least one potential compound or agent decreases the level and/or activity of CAPN9 and/or CAPNS2 relative to contact with a control compound or agent, the at least one potential compound or agent is identified as a compound or an agent that decreases the level and/or activity of CAPN9 and/or CAPNS2.   
     
     
         17 . The method of  claim 16 , wherein the population of cells is isolated from a subject with a fibrotic disease. 
     
     
         18 . A method of inhibiting myofibroblast differentiation comprising contacting a cell with at least one agent that decreases the expression level and/or activity of a calpain. 
     
     
         19 . The method of  claim 18 , wherein the calpain is CAPN9 and/or CAPNS2. 
     
     
         20 . The method of  claim 18 , wherein the at least one agent specifically inhibits CAPN9 and/or CAPNS2. 
     
     
         21 . The method of  claim 18 , wherein the at least one agent decreases TRPM7-mediated calcium influx. 
     
     
         22 . The method of  claim 18 , wherein the at least one agent specifically inhibits TRPM7. 
     
     
         23 . The method of  claim 18 , wherein the at least one agent inhibits Fibroblast-to-Myofibroblast Transition (FMT). 
     
     
         24 . The method of  claim 18 , wherein the at least one agent inhibits Epithelial to Mesenchymal Transition or Endothelial to Mesenchymal Transition. 
     
     
         25 . The method of  claim 18 , wherein the myofibroblast differentiation is a TGFβ-mediated myofibroblast differentiation. 
     
     
         26 . The method of  claim 18 , wherein the cell is in a fibrotic tissue. 
     
     
         27 . The method of  claim 18 , wherein the cell is in a cancerous tissue. 
     
     
         28 . The method of  claim 18 , wherein the cell is in a tissue with high TGFβ signaling. 
     
     
         29 . The method of  claim 18 , wherein the at least one agent is a small molecule chemical compound. 
     
     
         30 - 50 . (canceled) 
     
     
         51 . A method of using a three-dimensional structure of a CAPN9/CAPNS2 heterodimer in a drug screening assay comprising: (a) selecting a potential drug by performing rational drug design with the three-dimensional structure of the CAPN9/CAPNS2 heterodimer determined from one or more sets of atomic coordinates, wherein the selection is performed in conjunction with computer modeling; (b) contacting the potential drug with the CAPN9/CAPNS2 heterodimer; (c) detecting the binding of the potential drug with the CAPN9/CAPNS2 heterodimer; and (d) detecting the inhibition of CAPN9/CAPNS2 heterodimer activity by the potential drug; wherein a potential drug is selected as a drug if the potential drug binds to and inhibits the CAPN9/CAPNS2 heterodimer;
 or
 A method for identifying an agent capable of modulating the activity of CAPN9 and/or CAPNS2 in a cell that has yet to undergo, is undergoing, or has undergone myofibroblast differentiation, the method comprising: 
 a. contacting the cell with the agent, wherein the cell expresses CAPN9 and/or CAPNS2; and
 identifying whether the agent modulates the activity of CAPN9 and/or CAPNS2 
 
   
     
     
         52 - 98 . (canceled) 
     
     
         99 . A method of treating a subject diagnosed with or suspected of having one or more diseases associated with myofibroblast differentiation, the method comprising:
 a. assaying for the presence of a CAPN9 and/or CAPNS2 mRNA or protein in a biological sample obtained from the subject; and   b. administering a CAPN9 and/or CAPNS2 inhibitor to the subject;   or   A method for identifying an agent capable of inhibiting of a TRP calcium channel, wherein inhibition of the TRP calcium channel prevents myofibroblast differentiation, the method comprising:   a. contacting a cell with the agent, wherein the cell expresses (i) CAPN9 and/or CAPNS2 and (ii) a TRP calcium channel; and   b. identifying whether the agent prevents myofibroblast differentiation.   
     
     
         102 - 111 . (canceled)