Genetic markers for engraftment of human cardiac ventricular progenitor cells

Abstract

The present invention provides genetic markers for identifying engraftable human cardiac ventricular progenitor cells. The engraftment markers of the invention include angiogenic markers and extracellular matrix markers. Human ventricular progenitor cells expressing these markers are capable of forming ventricular tissue in vivo that is vascularized and supported by an extracellular matrix. Methods of engrafting human cardiac ventricular progenitor cells by transplanting into a subject progenitor cells that express the engraftment markers are also provided.

1 . A method of identifying engraftable human ventricular progenitor cells (HVPs), the method comprising:
 detecting expression of at least one engraftment marker in the HVPs to thereby identify engraftable HVPs, wherein:   the HVPs also express at least one surface marker selected from the group consisting of: JAG1, FZD4, LIFR, FGFR3 and/or TNFSF9.   
     
     
         2 . The method of  claim 1 , which comprises:
 culturing human cells containing cardiac progenitor cells under conditions causing differentiation into human ventricular progenitor cells (HVPs);   detecting expression on the HVPs of at least one surface marker selected from the group consisting of JAG1, FZD4, LIFR, FGFR3 and/or TNFSF9;   detecting expression in the HVPs of at least one engraftment marker; and   isolating HVPs that co-express the at least one surface marker and the at least one engraftment marker to thereby isolate engraftable HVPs.   
     
     
         3 . The method of  claim 1 , which comprises:
 transplanting engraftable human ventricular progenitor cells (HVPs) into the subject such that human ventricular tissue is engrafted in the subject,   wherein prior to transplantation, engraftable HVPs are identified by detecting expression of at least one engraftment marker in the HVPs; and   wherein the HVPs express at least one surface marker selected from the group consisting of: JAG1, FZD4, LIFR, FGFR3 and/or TNFSF9.   
     
     
         4 . The method of  claim 1 , which comprises:
 culturing human cells containing cardiac progenitor cells under conditions causing differentiation into human ventricular progenitor cells (HVPs);   detecting expression on the HVPs of at least one surface marker selected from the group consisting of JAG1, FZD4, LIFR, FGFR3 and/or TNFSF9;   detecting expression in the HVPs of at least one engraftment marker;   isolating HVPs that co-express the at least one surface marker and the at least one engraftment marker to thereby isolate engraftable HVPs; and   transplanting engraftable the HVPs into the subject such that human ventricular tissue is engrafted in the subject.   
     
     
         5 . The method of  claim 1 , which comprises:
 culturing human cells containing cardiac progenitor cells under conditions causing differentiation into human ventricular progenitor cells (HVPs) expressing at least one surface marker selected from the group consisting of JAG1, FZD4, LIFR, FGFR3 and/or TNFSF9;   isolating the HVPs expressing the at least one surface marker to form an HVP population;   detecting expression of at least one engraftment marker in a sample of the HVP population; and   transplanting the HVPs expressing the at least one surface marker and the at least one engraftment marker into the subject such that human ventricular tissue is engrafted in the subject.   
     
     
         6 . The method of  claim 1 , wherein the at least one engraftment marker detected is at least one positive angiogenic marker. 
     
     
         7 . The method of  claim 6 , wherein three or more positive angiogenic markers are detected. 
     
     
         8 . The method of  claim 6 , wherein ten or more positive angiogenic markers are detected. 
     
     
         9 . The method of  claim 1 , wherein the at least one engraftment marker detected is at least one positive extracellular matrix marker. 
     
     
         10 . The method of  claim 9 , wherein three or more positive extracellular matrix markers are detected. 
     
     
         11 . The method of  claim 9 , wherein ten or more positive extracellular matrix markers are detected. 
     
     
         12 . The method of  claim 1 , wherein the at least one engraftment marker detected is at least one positive angiogenic marker and at least one positive extracellular matrix marker. 
     
     
         13 . The method of  claim 6 , wherein the at least one positive angiogenic marker is selected from the group consisting of: FGF10, PRKD1, CCBE1, PDGFRA, EPHB2, GATA2, NTRK1, PTGIS, BMPER, BMP4, C1GALT1, MEIS1, TBX1, PKNOX1, ID1, TCF21, HEY1, HOXB3, HGF, IL6, GHRL, IHH, SRPK2, GATA6, HAND1, AMOT, NRP2, PTEN, SEMA3E, APOLD1, SETD2, DAB2IP, KDR, PGF, EMP2, TAL1, ACVR1, HIPK2, CSPG4, TNFAIP3, NRP1, NFATC4, CDC42, ANGPTL4, BCAS3, HIPK1, NRXN3, FZD5 and HHEX. 
     
     
         14 . The method of  claim 9 , wherein the at least one positive extracellular matrix marker is selected from the group consisting of: FGF10, SMOC1, CCBE1, COL6A6, ADAMTS12, COL19A1, LAMA1, BMP4, FBLN7, FBLN2, NDNF, HTRA1, HAPLN1, EMILIN1, SPOCK3, PODNL1, IHH, ACAN, NID2, COL4A6, LAMC1, FMOD, MUC4, EMID1, HMCN1, NID1, VCAN, CILP2, SOD3, ADAMTS3, ZP3, ANGPTL4, CRTAC1, LTBP4 and FREM1. 
     
     
         15 . The method of  claim 12 , wherein:
 the at least one positive angiogenic marker is selected from the group consisting of: FGF10, PRKD1, CCBE1, PDGFRA, EPHB2, GATA2, NTRK1, PTGIS, BMPER, BMP4, C1GALT1, MEIS1, TBX1, PKNOX1, ID1, TCF21, HEY1, HOXB3, HGF, IL6, GHRL, IHH, SRPK2, GATA6, HAND1, AMOT, NRP2, PTEN, SEMA3E, APOLD1, SETD2, DAB2IP, KDR, PGF, EMP2, TAL1, ACVR1, HIPK2, CSPG4, TNFAIP3, NRP1, NFATC4, CDC42, ANGPTL4, BCAS3, HIPK1, NRXN3, FZD5 and HHEX; and   the at least one positive extracellular matrix marker is selected from the group consisting of: FGF10, SMOC1, CCBE1, COL6A6, ADAMTS12, COL19A1, LAMA1, BMP4, FBLN7, FBLN2, NDNF, HTRA1, HAPLN1, EMILIN1, SPOCK3, PODNL1, IHH, ACAN, NID2, COL4A6, LAMC1, FMOD, MUC4, EMID1, HMCN1, NID1, VCAN, CILP2, SOD3, ADAMTS3, ZP3, ANGPTL4, CRTAC1, LTBP4 and FREM1.   
     
     
         16 . The method of  claim 1 , wherein the at least one engraftment marker detected is at least one negative angiogenic marker. 
     
     
         17 . The method of  claim 1 , wherein the at least one engraftment marker detected is at least one negative extracellular matrix marker. 
     
     
         18 . The method of  claim 1 , wherein the at least one engraftment marker detected is at least one negative angiogenic marker and at least one negative extracellular matrix marker. 
     
     
         19 . The method of  claim 1 , wherein detecting expression of at least one engraftment marker comprises detecting expression of mRNA encoding the at least one engraftment marker in the HVPs. 
     
     
         20 . The method of  claim 1 , wherein detecting expression of at least one engraftment marker comprises detecting lack of expression of mRNA encoding the at least one engraftment marker in the HVPs.