Brown Adipocyte Progenitors in Human Skeletal Muscle

Abstract

Brown adipose tissue (“BAT”) progenitor cells and methods for identifying BAT progenitor cells in a population of cells are provided. Methods are also provided for inducing differentiation of BAT progenitor cells into differentiated brown adipocytes, inducing expression or increased activity levels of BAT uncoupling protein-1 (“UCP1”), and for identifying agents capable of inducing differentiation of BAT progenitor cells into brown adipocytes and/or inducing expression or increased activity levels of UCP1. Differentiated brown adipocytes and agents and methods for inducing differentiation of BAT progenitor cells can be used for treatment of or the making of medicaments for the treatment of metabolic diseases or conditions in a patient such as obesity, overweight, impaired glucose tolerance, insulin-resistance, type 2 diabetes, dyslipidemia, hypertension, cardiovascular diseases, metabolic syndrome, and the like. Differentiated brown adipocytes and agents and methods for inducing differentiation of BAT progenitor cells can be used for prevention of hypothermia.

1 . A method for inducing differentiation of brown adipose tissue (BAT) progenitor cells into brown adipocytes comprising:
 providing a BAT progenitor cell isolated from skeletal muscle; and   exposing the BAT progenitor cell to a differentiation agent in vitro;   wherein the differentiation agent induces the BAT progenitor cell to differentiate into a brown adipocyte.   
     
     
         2 . The method of  claim 1  wherein the differentiation agent comprises one or more of the following: a PPARγ activator, modulator or inhibitor; a PPARα activator or modulator; a PPARδ activator or modulator; a dual PPARα and PPARδ activator or modulator; a pan-PPAR (α, δ, γ) activator or modulator; a PDE4 inhibitor; a PDE7 inhibitor; a NRIP1 (RIP140) inhibitor, a PTEN inhibitor; an α1-adrenergic full or partial agonist; an RXRα activator or modulator; a PGC-1α activator; a PGC-1β inhibitor or activator; adiponectin or an activator of adiponectin receptor AdipoR1 and/or AdipoR2; an NOS inhibitor or activator; a Rho kinase-ROCK inhibitor; BDNF; a monoamine oxidase (MAO) A inhibitor and/or a MAO B inhibitor; an activator of SRC, an inhibitor of EGFR; an inhibitor of FAAH; an inhibitor of MAPK 1 or 2 or 4 or 5 or 7 or 8; an inhibitor of CDK9; a TGR5 agonist; an AMPK activator; BMP-7, an mTOR inhibitor; an adenylate cyclase activator; or combinations of any of the foregoing. 
     
     
         3 . The method of  claim 1  wherein the differentiation agent comprises rosiglitazone. 
     
     
         4 . The method of  claim 1  wherein the brown adipocyte is characterized by one or more of the following: expression of UCP1 protein or mRNA, expression of mtTFA protein or mRNA, expression of PGC-1α protein or mRNA, uncoupled respiration, metabolic rate, glucose utilization rate, fatty acid oxidation rate, and a combination of any of the foregoing. 
     
     
         5 . The method of  claim 4  wherein the brown adipocyte contains UCP1 mRNA at a level higher than that of in vivo brown adipocytes in human skeletal muscle. 
     
     
         6 . The method of  claim 1  wherein the BAT progenitor cell is CD34 positive. 
     
     
         7 . The method of  claim 6  wherein the BAT progenitor cell is CD45 negative. 
     
     
         8 . The method of  claim 6  wherein the BAT progenitor cell is CD56 negative. 
     
     
         9 . The method of  claim 6  wherein the BAT progenitor cell is CD146 negative. 
     
     
         10 . The method of  claim 6  wherein the BAT progenitor cell is in primary culture. 
     
     
         11 . The method of  claim 6  wherein the BAT progenitor cell is expanded in culture for one or more passages. 
     
     
         12 . A method for inducing differentiation of brown adipose tissue (BAT) progenitor cells into brown adipocytes comprising:
 administering a differentiation agent to a patient having a metabolic disease; and   inducing differentiation of a BAT progenitor cell into a brown adipocyte in vivo;   wherein the BAT progenitor cell is in skeletal muscle of the patient.   
     
     
         13 . The method of  claim 12  wherein the differentiation agent comprises one or more of the following: a PPARγ activator, modulator or inhibitor; a PPARα activator or modulator; a PPARδ activator or modulator; a dual PPARα and PPARδ activator or modulator; a pan-PPAR (α, δ, γ) activator or modulator; a PDE4 inhibitor; a PDE7 inhibitor; a NRIP1 (RIP140) inhibitor, a PTEN inhibitor; an α1-adrenergic full or partial agonist; an RXRα activator or modulator; a PGC-1α activator; a PGC-1β inhibitor or activator; adiponectin or an activator of adiponectin receptor AdipoR1 and/or AdipoR2; an NOS inhibitor or activator; a Rho kinase-ROCK inhibitor; BDNF; a monoamine oxidase (MAO) A inhibitor and/or a MAO B inhibitor; an activator of SRC, an inhibitor of EGFR; an inhibitor of FAAH; an inhibitor of MAPK 1 or 2 or 4 or 5 or 7 or 8; an inhibitor of CDK9; a TGR5 agonist; an AMPK activator; BMP-7, an mTOR inhibitor; an adenylate cyclase activator; or combinations of any of the foregoing. 
     
     
         14 . The method of  claim 12  wherein the differentiation agent comprises rosiglitazone. 
     
     
         15 . The method of  claim 12  wherein the metabolic disease is one or more of the following: obesity, overweight, impaired glucose tolerance, insulin-resistance, type 2 diabetes, dyslipidemia, hypertension, cardiovascular disease, or metabolic syndrome. 
     
     
         16 . The method of  claim 12  wherein the metabolic disease is type 2 diabetes.