Cell potency assay for therapeutic potential

Abstract

The invention provides cell potency assays for measuring, determining, identifying, confirming, or validating the therapeutic potential of a cell population. Cell potency assays may be performed with various types of cells, including stem or progenitor cells, such as, for example, hematopoietic stem or progenitor cells. Cell potency assays may also be performed on stem or progenitor cells that have been treated with one or more agents to enhance therapeutic potential. Hematopoietic cells having therapeutic potential are useful in downstream clinical applications for increasing engraftment, reconstitution, homing, and proliferation in vivo.

1 - 48 . (canceled) 
     
     
         49 . An in vitro or ex vivo method to determine the therapeutic potential of a population of cells comprising:
 a) measuring expression of a plurality of genes in a population of cells comprising hematopoietic cells;   b) identifying increased expression in at least two of the plurality of genes in the population of cells compared to a control population of cells;   wherein the hematopoietic cells have sufficient therapeutic potential if the increased expression in the at least two of the plurality of genes in the population of cells is at least two-fold.   
     
     
         50 . The method of  claim 49 , wherein the population of cells is bone marrow cells (BMCs), umbilical cord blood cells (UCBCs), placental blood cells, mobilized peripheral blood cells (MPBCs), hematopoietic stem cells (HSCs), hematopoietic progenitor cells (HPCs), or CD34+ cells. 
     
     
         51 . (canceled) 
     
     
         52 . The method of  claim 49 , wherein at least a portion of the population of cells is modulated ex vivo prior to measuring expression of the plurality of genes. 
     
     
         53 . The method of  claim 49 , wherein at least a portion of the population of cells is expanded ex vivo prior to measuring expression of the plurality of genes. 
     
     
         54 . The method of  claim 49 , wherein at least a portion of the population of cells is cryopreserved prior to measuring expression of the plurality of genes. 
     
     
         55 - 56 . (canceled) 
     
     
         57 . The method of  claim 49 , wherein the population of cells is modulated ex vivo prior to measuring expression of the plurality of genes by contacting the population of cells with at least one agent selected from the group consisting of a cAMP analogue or enhancer, a Ga-s activator, and a prostaglandin pathway agonist. 
     
     
         58 . The method of  claim 57 , wherein the prostaglandin pathway agonist selectively binds the PGE 2  EP 2  or PGE 2  EP 4  receptor. 
     
     
         59 . The method of  claim 57 , wherein the prostaglandin pathway agonist comprises PGE 2 , or a PGE 2  analogue or derivative. 
     
     
         60 . The method of  claim 57 , wherein the prostaglandin pathway agonist is selected from the group consisting of: PGE 2 , 16,16-dmPGE 2 , 15(S)-15-methyl PGE 2 , 20-ethyl PGE 2 , and 8-iso-16-cyclohexyl-tetranor PGE 2 . 
     
     
         61 . The method of  claim 57 , wherein the prostaglandin pathway agonist comprises 16,16-dmPGE 2 . 
     
     
         62 . The method of  claim 57 , wherein the population of cells is further contacted with a glucocorticoid. 
     
     
         63 - 68 . (canceled) 
     
     
         69 . The method of  claim 57 , wherein the population of cells has been contacted with the at least one agent for a time of at least about one hour to at least about 24 hours. 
     
     
         70 - 73 . (canceled) 
     
     
         74 . The method of  claim 57 , wherein the population of cells has been contacted with the at least one agent at a temperature of about 37° C. 
     
     
         75 - 78 . (canceled) 
     
     
         79 . The method of  claim 49 , wherein the plurality of genes comprises two or more genes selected from the group consisting of: hairy/enhancer-of-split related with YRPW motif 1 (HEYi), UL16 binding protein 2 (ULBP2), hyaluronan synthase 1 (HASI), GTP-binding protein GEM (GEM), renin (REN), collagen, type I, alpha 1 (COLIAI), cyclooxygenase 2 (COX-2), angiopoietin 1 (ANGPTI), chemokine (C-X-C motif) ligand 6 (CXCL6), prominin 1 (PROMI), bone morphogenetic protein 4 (BMP4), angiopoietin 2 (ANGPT2), inhibitor of kappaB kinase beta (IKBKB), platelet/endothelial cell adhesion molecule 1 (PECAMI), tyrosine kinase with immunoglobulin-like and EGF-like domains 1 (TIEI), amphiregulin (AREG), caspase 3 (CASP3), jagged 1 (JAGI), aryl hydrocarbon receptor nuclear translocator (ARNT), cAMP-responsive element modulator (CREM), connective tissue growth factor (CTGF), CD40 ligand (CD40L), BCL2-associated X protein (BAX), hepatocyte growth factor (HGF), superoxide dismutase 2 (SOD2), platelet derived growth factor B (PDGFB), thrombospondin 1 (THBSI), dual specificity protein phosphatase 4 (DUSP4), cysteine-rich protein 61 (CYR61), chemokine (C-X-C motif) ligand 1 (CXCLI), endothelial tyrosine kinase (TEK), CASP8 and FADD-like apoptosis regulator (CFLAR), insulin growth factor 2 (IGF2), chemokine (C-X-C motif) receptor 4 (CXCR4), matrix metalloprotease 2 (MMP2), fibroblast growth factor 2 (FGF2), prostaglandin endoperoxide synthase 2 (PTGS2), RAS-related C3 botulinum substrate 2 (RAC2), platelet derived growth factor receptor (PDGFR), nuclear receptor subfamily 4, group A, member 2 (NR4A2), nuclear receptor subfamily 4, group A, member 3 (NR4A3), telomerase reverse transcriptase (TERT), transforming growth factor beta 1 (TGFBI), matrix metalloprotease 9 (MMP9), CD40 antigen (CD40), CD44 antigen (CD44), high mobility group box 1 (HMGBI), nitrogen oxide synthase 3 (NOS3), kinase insert domain receptor (KDR), integrin beta 1 (ITGBI), catenin (cadherin-associated protein), beta 1 (CTNNBI), colony stimulating factor 3 (CSF3), interleukin 8 (IL8), plasminogen activator, urokinase receptor (PLAUR), B cell CLL/lymphoma 2 (BCL2), bone morphogenetic protein 2 (BMP2), colony stimulating factor 1 (CSFI), v-akt murine thymoma viral oncogene homolog 1 (AKTI), vascular endothelial growth factor A (VEGFA), intercellular adhesion molecule 1 (ICAMI), chemokine (C-X-C motif) ligand 3 (CXCL3), caspase 8 (CASP8), CD34 antigen (CD34), interleukin IA (IL1A), CD47 antigen (CD47), chemokine (C-C motif) ligand 7 (CCL7), hypoxia inducible factor IA (HIFIA), EDNI (endothelin 1), sphingosine-1-phosphate receptor 1 (SIPRI), chemokine (C-C motif) receptor 1 (CCRI), SMAD family member 4 (SMAD4), fms-related tyrosine kinase 1 (FLTI), CD151 antigen (CD151), placental growth factor (PGF), nuclear factor of kappa light polypeptide gene enhancer in B-cells 1 (NFKB 1), SMAD family member 2 (SMAD2), CXC chemokine receptor 7 (CXCR7), transforming growth factor beta 3 (TGFB3), chemokine (C-X-C motif) ligand 5 (CXCL5), cyclin DI (CCNDI), heparin-binding EGF-like growth factor (HBEGF), nuclear receptor subfamily 3, group C, member 1 (NR3C1), tumor necrosis factor (TNF), integrin alpha L (ITGAL), CXC chemokine receptor 2 (CXCR2), signal transducer and activator of transcription 1 (STATI), integrin alpha 4 (ITGA4), leukemia inhibitory factor (LIF), RAS p21 protein activator 1 (RASAI), cadherin 5 (CDH5), ephrin B2 (EFNB2), regulator of G-protein signaling 16 (RGS16), chemokine (C-X-C motif) ligand 2 (CXCL2), integrin alpha 5 (ITGA5), chemokine (C-X-C motif) ligand 12 (CXCL12), tissue inhibitor of metalloprotease 1 (TEMPI), Fos related antigen 2 (FOSL2), integrin beta 2 (ITGB2), and tissue inhibitor of metalloprotease 2 (TIMP2). 
     
     
         80 - 105 . (canceled) 
     
     
         106 . A method of increasing hematopoietic engraftment in a subject, comprising administering a population of hematopoietic cells having sufficient therapeutic potential according to  claim 49  to increase hematopoietic engraftment in the subject. 
     
     
         107 . (canceled) 
     
     
         108 . The method of  claim 106 , wherein the hematopoietic engraftment is increased about 10% by hematopoeitic cells that have therapeutic potential compared to the engraftment associated with control hematopoietic cells. 
     
     
         109 - 112 . (canceled) 
     
     
         113 . The method of  claim 106 , wherein the hematopoietic engraftment is increased about two-fold by hematopoeitic cells that have therapeutic potential compared to the engraftment associated with control hematopoietic cells. 
     
     
         114 - 115 . (canceled) 
     
     
         116 . A method of cell-based therapy, comprising administering a population of autologous hematopoietic cells having sufficient therapeutic potential according to  claim 49  to a subject in need thereof. 
     
     
         117 . A composition comprising a population of hematopoietic cells having therapeutic potential according  claim 49 . 
     
     
         118 . A gene expression panel for identifying a population of hematopoietic cells having therapeutic potential, comprising a plurality of genes selected from the group consisting of: HEYi, COX2, ULBP2, HASI, GEMi, REN, COLIAI, ANGPTI, CXCL6, PROMI, BMP4, ANGPT2, IKBKB, PECAMI, TIEI, AREG, CASP3, JAGI, ARNT, CREM, CTGF, CD40L, BAX, HGF, SOD2, PDGFB, THBSI, DUSP4, CYR61, CXCLI, TEK, CFLAR, IGF2, CXCR4, MMP2, FGF2, PTGS2, RAC2, PDGFR, NR4A2, NR4A3, TERT, TGFBI, MMP9, CD40, CD44, HMGBI, NOS3, KDR, ITGBI, CTNNBI, CSF3, IL8, PLAUR, BCL2, BMP2, CSFI, AKTI, VEGFA, ICAMI, CXCL3, CASP8, CD34, IL1A, CD47, CCL7, HIFIA, EDNI, SIPRI, CCRI, SMAD4, FLTI, CD151, PGF, NFKBI, SMAD2, CXCR7, TGFB3, CXCL5, CCNDI, HBEGF, NR3C1, TNF, ITGAL, CXCR2, STATI, ITGA4, LIF, RASAI, CDH5, EFNB2, RGS16, CXCL2, ITGA5, CXCL12, TIMPI, FOSL2, ITGB2, and TIMP2. 
     
     
         119 - 123 . (canceled)