Method of Enhancing Hematopoietic Cell Transplantation

Abstract

The invention relates to a method for enhancing the transplantation of hematopoietic cells to supplement or fully reconstitute the hematopoietic system, such as in myeloablated patients or patients otherwise deficient in hematopoietic cells. The method involves administering CD34 + cells having enhanced expression of one or more of Musashi-2, PTEN, phospho-GSK-3β, Mcl-1, Atg7, phospho-Akt(ser473), HoxB4, Bmi-1, UBC13, phospho-Akt(thr308), GATA2, cMyc, and E47 at levels that provide desirable therapeutically effective amounts of self-renewal of the administered cells and desirable therapeutically effective amounts of differentiation of the administered cells into the various progeny cells of the hematopoietic system (i.e., therapeutically effective amounts of hematopoietic reconstitution). To provide such cells to a subject, the invention relates to detecting such cells prior to or during treatment to ascertain whether such cells are present in clinically-relevant amounts. It may also relate to treating a subject so as to provide clinically-relevant numbers of such cells, as with specific mobilization agents.

What is claimed is: 
     
         1 . A method for assessing the capacity of a sample to therapeutically effect hematopoietic reconstitution in a subject, the method comprising: assessing CD34 +  cells for enhanced expression of one or more of Musashi-2, PTEN, phospho-GSK-3β, Mcl-1, Atg7, phospho-Akt(ser473), HoxB4, Bmi-1, UBC13, phospho-Akt(thr308), GATA2, cMyc, and E47, in individual cells in the sample and determining the number of such cells. 
     
     
         2 . The method of  claim 1  wherein the one or more of Musashi-2, PTEN, phospho-GSK-3β, Mcl-1, Atg7, phospho-Akt(ser473), HoxB4, Bmi-1, UBC13, phospho-Akt(thr308), GATA2, cMyc, and E47 are expressed at a level greater than the mean expression level in un-mobilized bone marrow. 
     
     
         3 . A method to prepare a subject to donate blood for hematopoietic-reconstituting cell (HRC) transplantation, the method comprising obtaining a blood sample containing hematopoietic cells from the subject; determining number of CD34 +  cells having enhanced expression of one or more of Musashi-2, PTEN, phospho-GSK-3β, Mcl-1, Atg7, phospho-Akt(ser473), HoxB4, Bmi-1, UBC13, phospho-Akt(thr308), GATA2, cMyc, and E47 in individual cells from the blood sample; and administering to the subject a mobilizing agent when the blood sample does not contain a desired therapeutically-effective amount of such CD34 +  cells. 
     
     
         4 . The method of  claim 3  wherein the one or more of Musashi-2, PTEN, phospho-GSK-3β, Mcl-1, Atg7, phospho-Akt(ser473), HoxB4, Bmi-1, UBC13, phospho-Akt(thr308), GATA2, cMyc, and E47 are expressed at a level greater than the mean expression level in un-mobilized bone marrow. 
     
     
         5 . The method of  claim 3 , further comprising the step of administering a mobilizing agent to the subject prior to the step of obtaining a blood sample. 
     
     
         6 . A method for transplanting hematopoietic-reconstituting cells in a subject in need thereof, the method comprising administering to the subject nucleated blood cells comprising a therapeutically effective amount of CD34 +  cells having enhanced expression of one or more of Musashi-2, PTEN, phospho-GSK-3β, Mcl-1, Atg7, phospho-Akt(ser473), HoxB4, Bmi-1, UBC13, phospho-Akt(thr308), GATA2, cMyc, and E47. 
     
     
         7 . The method of  claim 6  wherein the one or more of Musashi-2, PTEN, phospho-GSK-3β, Mcl-1, Atg7, phospho-Akt(ser473), HoxB4, Bmi-1, UBC13, phospho-Akt(thr308), GATA2, cMyc, and E47 are expressed at a level greater than the mean expression level in un-mobilized bone marrow. 
     
     
         8 . The method of  claim 6  wherein the CD34 +  cells expressing the one or more of Musashi-2, PTEN, phospho-GSK-3β, Mcl-1, Atg7, phospho-Akt(ser473), HoxB4, Bmi-1, UBC13, phospho-Akt(thr308), GATA2, cMyc, and E47 are isolated. 
     
     
         9 . The method of  claim 6  wherein the subject has a disorder treatable by hematopoietic stem cell transplantation. 
     
     
         10 . The method of  claim 9  wherein the disorder is a hematopoietic deficiency or malignancy. 
     
     
         11 . A method to identify a molecule, the expression of which is correlated with the hematopoietic reconstituting function, the method comprising assessing expression of a molecule in individual CD34 +  cells in samples having different levels of potency and identifying molecules, the expression of which correlates with potency, by correlating differences in expression of such molecules with the potency of the different samples. 
     
     
         12 . The method of  claim 11  wherein greater potency is associated with an increase in expression. 
     
     
         13 . The method of  claim 11  wherein greater potency is associated with a decrease in expression. 
     
     
         14 . The method of  claim 11  wherein the samples that are compared are un-mobilized bone marrow, mobilized peripheral blood from healthy subjects, and umbilical cord blood. 
     
     
         15 . The method of any of  claims 1 - 4 ,  6 ,  7 , and  11  wherein the expression that is assayed is selected from the group consisting of RNA, protein, and post-translational modification. 
     
     
         16 . A method to assess the potency of a sample for hematopoietic reconstitution function, the method comprising assessing the expression level of a molecule identified by the method in  claim 11  in CD34 +  cells in the sample.