Methods for generating functional hematopoietic stem cells

Abstract

Described in the present application are methods for preparing populations of hematopoietic stem cells (HSCs), e.g., autologous and/or allogenic HSCs, using mechanical stretching or Trpv4 agonisists, and methods of use of the HSCs in transplantation. In some embodiments, the methods include providing a population comprising hemogenic endothelial (HE) cells, and (i) contacting the HE cells with an amount of an agonist of transient receptor potential cation channel-subfamily vanilloid member 4 (Trpv4); and/or (ii) subjecting the cells to cyclic 2-dimensional stretching, for a time and under conditions sufficient to stimulating endothelial-to-HSC transition. Also provided herein are methods for treating subjects who have, bone marrow, metabolic, and immune diseases; the methods include administering to the subject a therapeutically effective amount of hematopoietic stem cells (HSCs) obtained by a method described herein.

What is claimed is: 
     
         1 . A method of preparing a population of hematopoietic stem cells (HSC), the method comprising:
 providing a population comprising hemogenic endothelial (HE) cells, and   (i) contacting the HE cells with an amount of an agonist of transient receptor potential cation channel-subfamily vanilloid member 4 (Trpv4); and/or   (ii) subjecting the cells to cyclic 2-dimensional strectching, for a time and under conditions sufficient to stimulating endothelial-to-HSC transition.   
     
     
         2 . The method of  claim 1 , wherein the HE cells are obtained from iPSC. 
     
     
         3 . The method of  claim 1 , wherein the agonist of Trpv4 is selected from the group consisting of Arachidonic Acid, 5,6-EET, 8,9-EET, bisandrographolide A (BAA), Phorbol ester (e.g., 4α-PDD and 4α-PDH), RN-1747, substituted 1,4-diaminobutane or 1,3-diaminopropane analogues; and GSK10116790A. 
     
     
         4 . The method of  claim 1 , wherein the cells are obtained from a subject who has a blood, bone marrow, metabolic, or immune disease. 
     
     
         5 . The method of  claim 1 , wherein the subject does not have a hematological malignancy. 
     
     
         6 . A method of treating a subject who has a blood, bone marrow, metabolic, and immune diseases, the method comprising administering to the subject a therapeutically effective amount of hematopoietic stem cells (HSCs) obtained by a method comprising:
 providing a population comprising hemogenic endothelial (HE) cells, and   (i) contacting the HE cells with an amount of an agonist of transient receptor potential cation channel-subfamily vanilloid member 4 (Trpv4); and/or   (ii) subjecting the cells to cyclic 2-dimensional stretching, for a time and under conditions sufficient to stimulating endothelial-to-HSC transition.   
     
     
         7 . The method of  claim 6 , wherein the HE cells are obtained from iPSC. 
     
     
         8 . The method of  claim 6 , wherein the agonist of Trpv4 is selected from the group consisting of Arachidonic Acid, 5,6-EET, 8,9-EET, bisandrographolide A (BAA), Phorbol ester (e.g., 4α-PDD and 4α-PDH), RN-1747, substituted 1,4-diaminobutane or 1,3-diaminopropane analogues; and GSK10116790A. 
     
     
         9 . The method of  claim 6 , wherein the subject is a human. 
     
     
         10 . The method of  claim 6 , wherein the subject has multiple myeloma; non-Hodgkin lymphoma; Hodgkin disease; acute myeloid leukemia; neuroblastoma; a germ cell tumor; an autoimmune disorder (systemic lupus erythematosus (SLE) or systemic sclerosis); or amyloidosis.