Epicardial-derived paracrine factors for repairing cardiac tissue

Abstract

Provided herein, inter alia, are compositions and kits comprising epicardial-derived paracrine factors (such as, hypoglycosylated follistatin-like 1 (FSTL1)) for treating and repairing damage to cardiac tissue caused by cardiovascular disease, myocardial infarction (MI), other ischemic events, or cardiac-growth deficiency, as well as methods for using the same.

1 - 72 . (canceled) 
     
     
         73 . A pharmaceutical composition, comprising a hypoglycosylated follistatin-like 1 (FSTL1) polypeptide and one or more pharmaceutically acceptable excipients, wherein the pharmaceutical composition is formulated for subcutaneous administration. 
     
     
         74 . The pharmaceutical composition of  claim 73 , wherein the hypoglycosylated FSTL1 is
 (a) an FSTL1 polypeptide lacking N-linked glycosylation at one or more glycosylation-competent residues; and/or   (b) an FSTL1 polypeptide comprising substitution of the one or more glycosylation-competent residues with one or more glycosylation-incompetent residues.   
     
     
         75 . The pharmaceutical composition of  claim 74 , wherein the one or more glycosylation-competent residues comprises an arginine (R) residue and/or an asparagine (N) residue. 
     
     
         76 . The pharmaceutical composition of  claim 75 , wherein the asparagine (N) residue is located at positions 144, 175, 180, and/or 223 in an FSTL1 amino acid sequence comprising a polypeptide sequence of SEQ ID NO: 1. 
     
     
         77 . The pharmaceutical composition of  claim 76 , wherein the asparagine (N) residue is located at position 180 in an FSTL1 amino acid sequence comprising a polypeptide sequence of SEQ ID NO: 1. 
     
     
         78 . The pharmaceutical composition of  claim 73 , formulated for repairing cardiac tissue following an injury. 
     
     
         79 . The pharmaceutical composition of  claim 78 , wherein the repairing comprises one or more of
 (a) increasing the number of cardiomyocytes in the cardiac tissue;   (b) recovery of damaged cardiac tissue, including cardiomyocytes and/or cardiac vasculature;   (c) improved percent fractional shortening of cardiac tissue compared to the amount of percent fractional shortening in cardiac tissue that is not contacted by the hypoglycosylated FSTL1 polypeptide following an injury;   (d) a reduction in scar area (fibrosis) of at least a 2% compared to the amount of fibrosis in the same heart prior to the treatment by contacting, delivering or genomic editing of the hypoglycosylated FSTL1 polypeptide following an injury;   (e) an increase in vascular perfused area of at least a 2% compared to the amount of perfused area in the same heart prior to the treatment by contacting, delivering or genomic editing of the hypoglycosylated FSTL1 polypeptide following an injury;   (f) an increase in the amount of cardiomyocyte cytokinesis in the cardiac tissue compared to the amount of cardiomyocyte cytokinesis in cardiac tissue that is not contacted by the hypoglycosylated FSTL1 polypeptide following an injury, wherein the amount of cardiomyocyte cytokinesis is optionally determined by expression of Aurora B kinase; and/or   (g) decreased cardiomyocyte apoptosis.   
     
     
         80 . A method of producing a hypoglycosylated follistatin-like 1 (FSTL1) polypeptide, comprising culturing a host cell, wherein the host cell comprises a nucleic acid encoding an FSTL1 polypeptide, wherein the FSTL1 polypeptide is a hypoglycosylated FSTL1 polypeptide, wherein the hypoglycosylated FSTL1 polypeptide (a) lacks N-linked glycosylation at one or more glycosylation-competent residues; and/or (b) comprises substitution of the one or more glycosylation-competent residues with one or more glycosylation incompetent residues. 
     
     
         81 . The method of  claim 80 , wherein the one or more glycosylation-competent residues comprises an arginine (R) residue and/or an asparagine (N) residue. 
     
     
         82 . The method of  claim 81 , wherein the asparagine (N) residue is located at positions 144, 175, 180, and/or 223 in an FSTL1 amino acid sequence comprising a polypeptide sequence of SEQ ID NO: 1. 
     
     
         83 . The method of  claim 82 , wherein the asparagine (N) residue is located at position 180 in an FSTL1 amino acid sequence comprising a polypeptide sequence of SEQ ID NO: 1. 
     
     
         84 . The method of  claim 80 , wherein the host cell is treated with an inhibitor of glycosylation, optionally the inhibitor is tunicamycin. 
     
     
         85 . The nucleic acid of  claim 80 , encoding the FSTL1 polypeptide comprising the substitution of the one or more glycosylation competent residues, wherein the one or more glycosylation competent residue comprises one or more asparagine (N) residues at positions 144, 175, 180, and/or 223 of a polypeptide sequence of SEQ ID NO: 1. 
     
     
         86 . A vector encoding the nucleic acid of  claim 85 . 
     
     
         87 . A host cell comprising the vector of  claim 86 . 
     
     
         88 . The host cell of  claim 87 , wherein the host cell is a prokaryotic cell. 
     
     
         89 . The host cell of  claim 88 , wherein the prokaryotic cell is a bacterial cell. 
     
     
         90 . The host cell of  claim 87 , wherein the host cell is a eukaryotic cell, optionally treated with an inhibitor of glycosylation. 
     
     
         91 . The host cell of  claim 90 , wherein the inhibitor is tunicamycin. 
     
     
         92 . A hypoglycosylated follistatin-like 1 (FSTL1) polypeptide, produced by a process comprising the steps of
 (a) providing a nucleic acid encoding an FSTL1 polypeptide, wherein the FSTL1 polypeptide comprises substitution of one or more glycosylation-competent residues with one or more glycosylation incompetent residues;   (b) contacting the nucleic acid with a host cell, optionally treating the host cell with an inhibitor of glycosylation; and   (c) optionally isolating the hypoglycosylated FSTL1 polypeptide.