Methods and compositions for modulating angiogenesis and vasculogenesis

Abstract

Disclosed herein are methods and compositions for stimulating angiogenesis, using cells descended from marrow adherent stromal cells that have been transfected with sequences encoding a Notch intracellular domain. Applications of these methods and compositions include treatment of ischemic disorders such as stroke.

What is claimed is: 
     
         1 . A method for providing an angiogenic factor to a subject in need thereof, wherein the angiogenic factor is selected from the group consisting of one or more of angiogenin, angiopoietin-2, epidermal growth factor, basic fibroblast growth factor, leptin, platelet-derived growth factor-BB, and placental growth factor; the method comprising administering to the subject a therapeutically effective amount of cells, wherein the cells are made by a method comprising:
 (a) providing a culture of mesenchymal stem cells,   (b) contacting the cell culture of step (a) with a polynucleotide comprising sequences encoding a Notch intracellular domain (NICD) wherein said polynucleotide does not encode a full-length Notch protein,   (c) selecting cells that comprise the polynucleotide of step (b), and   (d) further culturing the selected cells of step (c) in the absence of selection and in the absence of trophic factors.   
     
     
         2 . The method of  claim 1 , wherein the one or more angiogenic factors promotes a process selected from the group consisting of one or more of prevention of endothelial cell death, enhancement of endothelial cell survival, stimulation of endothelial cell proliferation, and enhancement of blood vessel branching. 
     
     
         3 . A combination for use in stimulating angiogenesis in a subject in need thereof, wherein the combination comprises:
 (a) cells made by a method comprising:
 (i) providing a culture of mesenchymal stem cells, 
 (ii) contacting the cell culture of step (i) with a polynucleotide comprising sequences encoding a Notch intracellular domain (NICD) wherein said polynucleotide does not encode a full-length Notch protein, 
 (iii) selecting cells that comprise the polynucleotide of step (ii), and 
 (iv) further culturing the selected cells of step (iii) in the absence of selection and in the absence of trophic factors; and 
   (b) a second pro-angiogenic agent.   
     
     
         4 . The combination of  claim 3 , wherein the second pro-angiogenic agent is a polypeptide. 
     
     
         5 . The combination of  claim 4 , wherein the polypeptide is a transcription factor that activates transcription of a pro-angiogenic protein. 
     
     
         6 . The combination of  claim 5 , wherein the pro-angiogenic protein is vascular endothelial growth factor (VEGF). 
     
     
         7 . The combination of  claim 6 , wherein the transcription factor is a non-naturally-occurring zinc finger protein that activates transcription of the VEGF gene. 
     
     
         8 . The combination of  claim 3 , wherein the second pro-angiogenic agent is a nucleic acid. 
     
     
         9 . The combination of  claim 8 , wherein the nucleic acid encodes a transcription factor that activates transcription of a pro-angiogenic protein. 
     
     
         10 . The combination of  claim 9 , wherein the pro-angiogenic protein is vascular endothelial growth factor (VEGF). 
     
     
         11 . The combination of  claim 10 , wherein the transcription factor is a non-naturally-occurring zinc finger protein that activates transcription of the VEGF gene. 
     
     
         12 . A method for stimulating angiogenesis in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the combination of  claim 3 . 
     
     
         13 . The method of  claim 12 , wherein said administering results in promoting a process selected from the group consisting of one or more of prevention of endothelial cell death, enhancement of endothelial cell survival, stimulation of endothelial cell proliferation, and enhancement of blood vessel branching.