PNMT as a novel marker for progenitor cells

Abstract

In certain aspects, the present invention provides methods and compositions relating to a Pnmt-positive progenitor cell. In certain aspects, the present invention relates to methods for isolating and transplanting the subject progenitor cells, and methods for treating diseases such as myocardiac injuries and neurodegenerative disorders.

1 . A method of identifying a cardiac progenitor cell, comprising detecting expression of phenylethanolamine n-methyltransferase (Pnmt) in a cell, wherein if the cell expresses Pnmt and is capable of differentiating into a cardiomyocyte, the cell is identified as a cardiac progenitor cell. 
   
   
       2 . The method of  claim 1 , wherein the cardiomyocyte is selected from the group consisting of a pacemaker cell, a His bundle (HIS) cell, a Purkinje fiber (PUR) cell, an atrial working myocyte, and a ventricular working myocyte. 
   
   
       3 . The method of  claim 2 , wherein the pacemaker cell is selected from the group consisting of a sinoatrial node (SAN) cell and an atrioventricular node (AVN) cell. 
   
   
       4 . The method of  claim 1 , wherein the cell further expresses a marker selected from the group consisting of c-kit, Sca-1, and MDR1. 
   
   
       5 . The method of  claim 1 , wherein the cell is isolated from a tissue selected from the group consisting of a developing heart and an adult heart. 
   
   
       6 . The method of  claim 1 , wherein the cell is isolated from a cultured stem cell line. 
   
   
       7 . The method of  claim 1 , wherein the cell is a human cell. 
   
   
       8 . The method of  claim 1 , wherein the cell comprises a marker gene located at one or more loci of the Pnmt gene. 
   
   
       9 . The method of  claim 7 , wherein the marker gene is selected from the group consisting of GFP, beta-galactosidase, and luciferase. 
   
   
       10 . A method of inducing a Pnmt-positive cardiac progenitor cell to differentiate into a cardiomyocyte, comprising treating the Pnmt-positive cardiac progenitor cell with an effective amount of an agent that induces the progenitor cell to differentiate into a cardiomyocyte. 
   
   
       11 . The method of  claim 10 , wherein the progenitor cell is selected from the group consisting of a pacemaker cell, a His bundle (HIS) cell, a Purkinje fiber (PUR) cell, an atrial working myocyte, and a ventricular working myocyte. 
   
   
       12 . The method of  claim 11 , wherein the pacemaker cell is selected from the group consisting of a sinoatrial node (SAN) cell and an atrioventricular node (AVN) cell. 
   
   
       13 . The method of  claim 10 , wherein the progenitor cell is positive for a marker selected from the group consisting of c-kit, Sca-1, and MDR1. 
   
   
       14 . The method of  claim 10 , wherein the progenitor cell is isolated from a cultured stem cell line. 
   
   
       15 . The method of  claim 10 , wherein the progenitor cell is isolated from a tissue selected from the group consisting of a developing heart and an adult heart. 
   
   
       16 . The method of  claim 10 , wherein the progenitor cell is a human progenitor cell. 
   
   
       17 . The method of  claim 10 , wherein the agent is selected from the group consisting of catecholamine, a growth factor, a hormone, and an extracellular matrix protein. 
   
   
       18 . A method of treating an individual for a myocardial injury, comprising introducing Pnmt-positive cardiac progenitor cells into the individual, wherein the progenitor cell differentiates into a cardiomyocyte. 
   
   
       19 . The method of  claim 18 , wherein the progenitor cell is selected from the group consisting of a pacemaker cell, a His bundle (HIS) cell, a Purkinje fiber (PUR) cell, an atrial working myocyte, and a ventricular working myocyte. 
   
   
       20 . The method of  claim 18 , wherein the progenitor cell is positive for a marker selected from the group consisting of c-kit and c-kit, Sca-1, and MDR1. 
   
   
       21 . The method of  claim 18 , wherein the progenitor cell is isolated from a cultured stem cell line. 
   
   
       22 . The method of  claim 18 , wherein the progenitor cell is isolated from a tissue selected from the group consisting of a developing heart and an adult heart. 
   
   
       23 . The method of  claim 18 , wherein the progenitor cell is a human progenitor cell. 
   
   
       24 . The method of  claim 18 , wherein the myocardial injury is selected from the group consisting of myocardial infarction, cardiomyopathy and congenital heart disease. 
   
   
       25 . The method of  claim 18 , wherein the progenitor cell is introduced into a damaged cardiac region. 
   
   
       26 . The method of  claim 18 , further comprising expanding the progenitor cell ex vivo before transferring the progenitor cell into the patient. 
   
   
       27 . The method of  claim 18 , further comprising treating the individual with an immunosuppressive agent. 
   
   
       28 . The method of  claim 18 , further comprising inactivating expression of a major histocompatibility (MHC) gene in the Pnmt-positive progenitor cells prior to introducing the cells into the individual. 
   
   
       29 . The method of  claim 28 , wherein the immunosuppressive agent is selected from the group consisting of FK-506, cyclosporin, and GAD65 antibodies. 
   
   
       30 . A method of assessing a Pnmt-positive cardiac progenitor cell for its ability to regenerate cardiac tissues in vivo, comprising: (a) obtaining a Pnmt-positive cardiac progenitor cell; and (b) transferring the progenitor cell into a subject in need thereof; and (c) monitoring the transferred progenitor cell. 
   
   
       31 . The method of  claim 30 , wherein the transferred progenitor cell contains a marker. 
   
   
       32 . The method of  claim 31 , wherein the marker is selected from the group consisting of GFP, luciferase, beta-galactosidase. 
   
   
       33 . The method of  claim 30 , wherein the transferred progenitor cell is monitored by magnetic resonance imaging (MRI). 
   
   
       34 . The method of  claim 30 , wherein the transferred progenitor cell is monitored by bioluminescence imaging (BLI). 
   
   
       35 . An isolated Pnmt-positive mammalian progenitor cell. 
   
   
       36 . The isolated progenitor cell of  claim 35 , wherein the progenitor cell is a cardiac progenitor cell. 
   
   
       37 . The isolated progenitor cell of  claim 36 , wherein the progenitor cell is positive for a marker selected from the group consisting of c-kit, Sca-1, and MDR1. 
   
   
       38 . The isolated progenitor cell of  claim 36 , which differentiates into a cardiomyocyte. 
   
   
       39 . The isolated progenitor cell of  claim 38 , wherein the progenitor cell is selected from the group consisting of a pacemaker cell, a His bundle (HIS) cell, a Purkinje fiber (PUR) cell, an atrial working myocyte, and a ventricular working myocyte. 
   
   
       40 . The isolated progenitor cell of  claim 39 , wherein the pacemaker cell is selected from the group consisting of a sinoatrial node (SAN) cell and an atrioventricular node (AVN) cell. 
   
   
       41 . The isolated progenitor cell of  claim 35 , wherein the progenitor cell is a neural progenitor cell. 
   
   
       42 . The isolated progenitor cell of  claim 41 , wherein the progenitor cell is positive for nestin. 
   
   
       43 . The isolated progenitor cell of  claim 41  which differentiates into a brainstem neuronal cell. 
   
   
       44 . The isolated progenitor cell of  claim 41  which differentiates into a retinal neuronal cell. 
   
   
       45 . The isolated progenitor cell of  claim 35  which differentiates into an adrenergic cell. 
   
   
       46 . The isolated progenitor cell of  claim 35 , wherein the progenitor cell is isolated from a tissue selected from the group consisting of a developing heart and an adult heart. 
   
   
       47 . The isolated progenitor cell of  claim 35 , wherein the progenitor cell is isolated from a developing brain. 
   
   
       48 . The isolated progenitor cell of  claim 35 , wherein the progenitor cell is isolated from a cultured stem cell line. 
   
   
       49 . The isolated progenitor cell of  claim 35 , wherein the progenitor cell is a human progenitor cell. 
   
   
       50 . A transplant graft comprising isolated progenitor cells of  claim 35 . 
   
   
       51 . The transplant graft of  claim 50 , wherein the transplant graft does not cause graft versus host rejection when transplanted into an animal. 
   
   
       52 . A cell comprising a marker gene located at one or more loci of the Pnmt gene. 
   
   
       53 . The cell of  claim 52 , wherein the marker gene is selected from the group consisting of GFP, luciferase, beta-galactosidase. 
   
   
       54 . The cell of  claim 52 , wherein at least one copy of the Pnmt gene is deleted. 
   
   
       55 . The cell of  claim 52 , wherein the cell is a progenitor cell. 
   
   
       56 . The cell of  claim 52 , wherein the cell is a mammalian cell. 
   
   
       57 . A method of making a cell of  claim 52 . 
   
   
       58 . A non-human transgenic animal whose genome comprises a marker gene located at one or more loci of the Pnmt gene. 
   
   
       59 . The animal of  claim 58 , wherein the marker gene is selected from the group consisting of GFP, luciferase, and beta-galactosidase. 
   
   
       60 . The animal of  claim 58 , wherein at least one copy of the Pnmt gene is deleted. 
   
   
       61 . The animal of  claim 58 , wherein the animal is selected from the group consisting of a mouse, a rat, and a rabbit. 
   
   
       62 . A method of identifying a compound that increases expression of Pnmt in a cell, comprising: (a) contacting a cell comprising a marker located at one or more loci of the Pnmt gene with a test compound; (b) determining the expression level of the marker gene in the cell, wherein if expression level of the marker gene is higher in the presence of the test compound than in the absence of the test compound, the test compound is a compound that increases expression of Pnmt. 
   
   
       63 . The method of  claim 62 , further comprising assessing the ability of the test compound to increase expression of Pnmt in vivo. 
   
   
       64 . A method of delivering a scaffold graft in a target tissue, comprising:
 a) seeding Pnmt-positive progenitor cells onto a biocompatible scaffold, thereby forming a scaffold graft; and   b) implanting the scaffold graft from (a) in direct contact with, or adjacent to, a target tissue for a sufficient time,   wherein cells of the target tissue associate with the implanted scaffold graft, thereby to form new tissue.   
   
   
       65 . Use of an isolated Pnmt-positive human progenitor cell for treating an individual having a myocardial injury.