title: Preventing Pluripotent Cell Teratoma in Regenerative Medicine Applied to Hematology Disorders.
abstract: Iatrogenic tumorigenesis is a major limitation for the use of human induced pluripotent stem cells (hiPSCs) in hematology. The teratoma risk comes from the persistence of hiPSCs in differentiated cell populations. Our goal was to evaluate the best system to purge residual hiPSCs before graft without compromising hematopoietic repopulation capability. Teratoma risk after systemic injection of hiPSCs expressing the reporter gene luciferase was assessed for the first time. Teratoma formation in immune-deficient mice was tracked by in vivo bioimaging. We observed that systemic injection of hiPSCs produced multisite teratoma as soon as 5 weeks after injection. To eliminate hiPSCs before grafting, we tested the embryonic-specific expression of suicide genes under the control of the pmiR-302/367 promoter. This promoter was highly active in hiPSCs but not in differentiated cells. The gene/prodrug inducible Caspase-9 (iCaspase-9)/AP20187 was more efficient and rapid than thymidine kinase/ganciclovir, fully specific, and without bystander effect. We observed that iCaspase-9-expressing hiPSCs died in a dose-dependent manner with AP20187, without reaching full eradication in vitro. Unexpectedly, nonspecific toxicity of AP20187 on iCaspase-9-negative hiPSCs and on CD34+cells was evidenced in vitro. This toxic effect strongly impaired CD34+-derived human hematopoiesis in adoptive transfers. Survivin inhibition is an alternative to the suicide gene approach because hiPSCs fully rely on survivin for survival. Survivin inhibitor YM155 was more efficient than AP20187/iCaspase-9 for killing hiPSCs, without toxicity on CD34+cells, in vitro and in adoptive transfers. hiPSC purge by survivin inhibitor fully eradicated teratoma formation in immune-deficient mice. This will be useful to improve the safety management for hiPSC-based medicine. Stem Cells Translational Medicine 2017;6:382-393.
authors.affiliation: Université de Bordeaux, Bordeaux, France.|INSERM U1035, Bordeaux, France.|Bordeaux University Hospital, Bordeaux, France.
authors.forename: Aurelie
authors.initials: A
authors.lastname: Bedel
authors.affiliation: Université de Bordeaux, Bordeaux, France.|INSERM U1035, Bordeaux, France.
authors.forename: François
authors.initials: F
authors.lastname: Beliveau
authors.affiliation: Université de Bordeaux, Bordeaux, France.|INSERM U1035, Bordeaux, France.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Lamrissi-Garcia
authors.affiliation: Université de Bordeaux, Bordeaux, France.|Bordeaux University Hospital, Bordeaux, France.
authors.forename: Benoit
authors.initials: B
authors.lastname: Rousseau
authors.affiliation: Université de Bordeaux, Bordeaux, France.|INSERM U1035, Bordeaux, France.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Moranvillier
authors.affiliation: Université de Bordeaux, Bordeaux, France.|INSERM U1035, Bordeaux, France.
authors.forename: Benoit
authors.initials: B
authors.lastname: Rucheton
authors.affiliation: Université de Bordeaux, Bordeaux, France.|Bordeaux University Hospital, Bordeaux, France.
authors.forename: Veronique
authors.initials: V
authors.lastname: Guyonnet-Dupérat
authors.affiliation: Université de Bordeaux, Bordeaux, France.|INSERM U1035, Bordeaux, France.
authors.forename: Bruno
authors.initials: B
authors.lastname: Cardinaud
authors.affiliation: Université de Bordeaux, Bordeaux, France.|INSERM U1035, Bordeaux, France.|Bordeaux University Hospital, Bordeaux, France.
authors.forename: Hubert
authors.initials: H
authors.lastname: de Verneuil
authors.affiliation: Université de Bordeaux, Bordeaux, France.|INSERM U1035, Bordeaux, France.|Bordeaux University Hospital, Bordeaux, France.|Plateforme de Vectorologie, Université de Bordeaux, Bordeaux, France.
authors.forename: François
authors.initials: F
authors.lastname: Moreau-Gaudry
authors.affiliation: Université de Bordeaux, Bordeaux, France.|INSERM U1035, Bordeaux, France.|Bordeaux University Hospital, Bordeaux, France.
authors.forename: Sandrine
authors.initials: S
authors.lastname: Dabernat
mesh_terms: D000818:Animals; D053453:Caspase 9 / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D049109:Cell Proliferation; D002471:Cell Transformation, Neoplastic / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D004305:Dose-Response Relationship, Drug; D015972:Gene Expression Regulation, Neoplastic; D041722:Genes, Transgenic, Suicide; D006402:Hematologic Diseases / Q000601:surgery*; D018380:Hematopoietic Stem Cell Transplantation / Q000009:adverse effects / Q000379:methods*; D006412:Hematopoietic Stem Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology*; D057026:Induced Pluripotent Stem Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology / Q000637:transplantation*; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D009285:Naphthoquinones / Q000494:pharmacology*; D010641:Phenotype; D044968:Regenerative Medicine / Q000379:methods*; D018570:Risk Assessment; D000077022:Survivin / Q000037:antagonists & inhibitors* / Q000378:metabolism; D016559:Tacrolimus / Q000031:analogs & derivatives* / Q000494:pharmacology; D013724:Teratoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D013997:Time Factors; D047368:Tumor Burden; D023041:Xenograft Model Antitumor Assays
chemical_list: C463061:AP20187; C107489:BIRC5 protein, human; D007093:Imidazoles; D009285:Naphthoquinones; D000077022:Survivin; D053453:Caspase 9; C523798:sepantronium; D016559:Tacrolimus
keywords: Hematology; Hematopoietic stem cell; Induced pluripotent stem cells; Regenerative medicine; Safety; Suicide gene; Survivin inhibitor; Teratoma; Thymidine kinase; iCaspase-9