Methods and compositions for stem cell self-renewal

Abstract

The present invention relates to methods for expanding a stem cell population. More particularly, the invention relates, inter alia, to methods and compositions for expanding a stem cell population, particularly a hematopoietic stem cell population.

1 - 96 . (canceled) 
     
     
         97 . A composition for expanding a hematopoietic stem cell (HSC) population comprising:
 a PTEN inhibitor; and   a GSK-3β inhibitor;   wherein the composition is effective to enable expansion of the HSC population while maintaining a multilineage differentiation potential in the HSC population.   
     
     
         98 . The composition according to  claim 97 , wherein
 (i) the PTEN inhibitor is selected from the group consisting of a small molecule, a biologic, an antisense RNA, a small interfering RNA (siRNA), and combinations thereof; and   (ii) the GSK-3β inhibitor is selected from the group consisting of a small molecule, a biologic, an antisense RNA, a small interfering RNA (siRNA), and combinations thereof.   
     
     
         99 . The composition according to  claim 97 , wherein the PTEN inhibitor is selected from the group consisting of shikonin, a bisperoxovanadium compound, SF-1751 (Semafore Pharmaceuticals), pharmaceutical salts thereof, and combinations thereof, and the GSK-3β inhibitor is selected from the group consisting of Hymenialdisine, Flavopiridol, Kenpaullone, Alsterpaullone, Azakenpaullone, Indirubin-30-oxime, 6-Bromoindirubin-30-oxime (BIO), 6-Bromoindirubin-30-acetoxime, Aloisine A, Aloisine B, TDZD8, Compound 12, CHIR98014, CHIR99021 (CT99021), CT20026, Compound 1, SU9516, ARA014418, Staurosporine, Compound 5a, Compound 29, Compound 46, GF109203x (bisindolylmaleimide I), Ro318220 (bisindolylmaleimide IX), SB216763, SB415286, 15, CGP60474, Compound 8b, TWS119, Compound 1A, Compound 17, Lithium, Beryllium, Zinc, small molecule GSK-3β inhibitors (Vertex Pharmaceuticals), NP-12 (Neuropharma), GSK-3β inhibitors (Amphora), GSK-3β inhibitors (CrystalGenomics), SAR-502250 (Sanofi-Aventis), 3544 (Hoffmann-La Roche), GSK-3β inhibitors (Lundbeck), TDZD-8 (Cancer Center, University of Rochester), pharmaceutically acceptable salts thereof, and combinations thereof. 
     
     
         100 . The composition according to  claim 99 , wherein the bisperoxovanadium compound is selected from the group consisting of bpV(phen)2, bpV(pic), pharmaceutical salts thereof, and combinations thereof. 
     
     
         101 . The composition according to  claim 97 , wherein PTEN inhibitor is bpV(pic), and the GSK-3β inhibitor is CHIR99201. 
     
     
         102 . The composition according to  claim 97 , wherein the PTEN inhibitor is a reversible PTEN inhibitor. 
     
     
         103 . The composition according to  claim 102 , wherein the reversible PTEN inhibitor is selected from the group consisting of shikonin, a bisperoxovanadium compound, SF-1751 (Semafore Pharmaceuticals), pharmaceutical salts thereof, and combinations thereof. 
     
     
         104 . The composition according to  claim 103 , wherein the bisperoxovanadium compound is selected from the group consisting of bpV(phen)2, bpV(pic), pharmaceutical salts thereof, and combinations thereof. 
     
     
         105 . The composition according to  claim 97 , wherein the GSK-3β inhibitor is a reversible GSK-3β inhibitor. 
     
     
         106 . The composition according to  claim 105 , wherein the reversible GSK-3β inhibitor is selected from the group consisting of Hymenialdisine, Flavopiridol, Kenpaullone, Alsterpaullone, Azakenpaullone, Indirubin-30-oxime, 6-Bromoindirubin-30-oxime (BIO), 6-Bromoindirubin-30-acetoxime, Aloisine A, Aloisine B, TDZD8, Compound 12, CHIR98014, CHIR99021 (CT99021), CT20026, Compound 1, SU9516, ARA014418, Staurosporine, Compound 5a, Compound 29, Compound 46, GF109203x (bisindolylmaleimide I), Ro318220 (bisindolylmaleimide IX), SB216763, SB415286, I5, CGP60474, Compound 8b, TWS119, Compound 1A, Compound 17, Lithium, Beryllium, Zinc, small molecule GSK-3β inhibitors (Vertex Pharmaceuticals), NP-12 (Neuropharma), GSK-3β inhibitors (Amphora), GSK-3β inhibitors (CrystalGenomics), SAR-502250 (Sanofi-Aventis), 3544 (Hoffmann-La Roche), GSK-3β inhibitors (Lundbeck), TDZD-8 (Cancer Center, University of Rochester), pharmaceutically acceptable salts thereof, and combinations thereof. 
     
     
         107 . The composition according to  claim 106 , wherein the reversible GSK-3β inhibitor is CHIR99021. 
     
     
         108 . The composition according to  claim 97 , wherein the HSC population is obtained from a mammalian tissue selected from the group consisting of cord blood, peripheral blood, and bone marrow. 
     
     
         109 . The composition according to  claim 97 , wherein the expansion of the number of HSCs is by a factor of at least 40-fold. 
     
     
         110 . The composition according to  claim 97 , wherein the expansion of the number of HSCs is by a factor of at least 80-fold. 
     
     
         111 . The composition according to  claim 97 , wherein the expansion of the number of HSCs is by a factor of at least 150-fold. 
     
     
         112 . The composition according to  claim 97 , wherein the expansion of the number of HSCs is by a factor of at least 250-fold.