Expression of PROKR1 and PROKR2 in Human Enteric Neural Precursor Cells and Identification of Sequence Variants Suggest a Role in HSCR 英文参考文献.docVIP

ExpressionofPROKR1andPROKR2inHumanEnteric NeuralPrecursorCellsandIdentificationofSequence VariantsSuggestaRoleinHSCR MacarenaRuiz-Ferrer1,2,AnaTorroglosa1,2,Roc?′oNu′n?ez-Torres1,2,JuanCarlosdeAgust?′n3 ,Guillermo Antin?olo1,2,SaludBorrego1,2 * 1UnidaddeGestio′nCl?′nicadeGene′tica,Reproduccio′nyMedicinaFetal,InstitutodeBiomedicinadeSevilla,HospitalUniversitarioVirgendelRoc?′o/CSIC/Universidadde Sevilla,Sevilla,Spain,2CentrodeInvestigacio′nBiome′dicaenReddeEnfermedadesRaras(CIBERER),Sevilla,Spain,3UnidaddeGestio′nCl?′nicadeCirug?′aInfantil,Hospital UniversitarioVirgendelRoc?′o,Sevilla,Spain Abstract Background:Theentericnervoussystem(ENS)isentirelyderivedfromneuralcrestanditsnormaldevelopmentisregulated by specific molecular pathways. Failure in complete ENS formation results in aganglionic gut conditions such as Hirschsprung’sdisease(HSCR).Recently,PROKR1expressionhasbeendemonstratedinmouseentericneuralcrestderived cellsandProk-1wasshowntoworkcoordinatelywithGDNFinthedevelopmentoftheENS. PrincipalFindings:Inthepresentreport,ENSprogenitorswereisolatedandcharacterizedfromtheganglionicgutfrom children diagnosed with and without HSCR, and the expression of prokineticin receptors was examined. Immunocyto- chemical analysis of neurosphere-forming cells demonstrated that both PROKR1 and PROKR2 were present in human entericneuralcrestcells.Inaddition,wealsoperformedamutationalanalysisofPROKR1,PROKR2,PROK1andPROK2genes in a cohort of HSCR patients, evaluating themfor the first time as susceptibility genes for the disease. Several missense variants were detected, most of them affecting highly conserved amino acid residues of the protein and located in functionaldomainsofbothreceptors,whichsuggestsapossibledeleteriouseffectintheirbiologicalfunction. Conclusions:OurresultssuggestthatnotonlyPROKR1,butalsoPROKR2mightmediateacomplementarysignallingtotheRET/ GFRa1/GDNFpathwaysupportingproliferation/survivalanddifferentiationofprecursorcellsduringENSdevelopment.These findings,toge