title: Pre-clinical development of gene modification of haematopoietic stem cells with chimeric antigen receptors for cancer immunotherapy.
abstract: Patients with refractory or recurrent B-lineage hematologic malignancies have less than 50% of chance of cure despite intensive therapy and innovative approaches are needed. We hypothesize that gene modification of haematopoietic stem cells (HSC) with an anti-CD19 chimeric antigen receptor (CAR) will produce a multi-lineage, persistent immunotherapy against B-lineage malignancies that can be controlled by the HSVsr39TK suicide gene. High-titer third-generation self-inactivating lentiviral constructs were developed to deliver a second-generation CD19-specific CAR and the herpes simplex virus thymidine kinase HSVsr39TK to provide a suicide gene to allow ablation of gene-modified cells if necessary. Human HSC were transduced with such lentiviral vectors and evaluated for function of both CAR and HSVsr39TK. Satisfactory transduction efficiency was achieved; the addition of the suicide gene did not impair CAR expression or antigen-specific cytotoxicity, and determined marked cytotoxicity to ganciclovir. NSG mice transplanted with gene-modified human HSC showed CAR expression not significantly different between transduced cells with or without HSVsr39TK, and expression of anti-CD19 CAR conferred anti-tumor survival advantage. Treatment with ganciclovir led to significant ablation of gene-modified cells in mouse tissues. Haematopoietic stem cell transplantation is frequently part of the standard of care for patients with relapsed and refractory B cell malignancies; following HSC collection, a portion of the cells could be modified to express the CD19-specific CAR and give rise to a persistent, multi-cell lineage, HLA-independent immunotherapy, enhancing the graft-versus-malignancy activity.
authors.affiliation: a Department of Internal Medicine , David Geffen School of Medicine at UCLA , Los Angeles , CA , USA.
authors.forename: Sarah M
authors.initials: SM
authors.lastname: Larson
authors.affiliation: b Department of Pediatrics , David Geffen School of Medicine at UCLA , Los Angeles , CA , USA.
authors.forename: Laurel C
authors.initials: LC
authors.lastname: Truscott
authors.affiliation: b Department of Pediatrics , David Geffen School of Medicine at UCLA , Los Angeles , CA , USA.
authors.forename: Tzu-Ting
authors.initials: TT
authors.lastname: Chiou
authors.affiliation: c Western University of Health Sciences , Pomona , CA , USA.
authors.forename: Amie
authors.initials: A
authors.lastname: Patel
authors.affiliation: b Department of Pediatrics , David Geffen School of Medicine at UCLA , Los Angeles , CA , USA.
authors.forename: Roy
authors.initials: R
authors.lastname: Kao
authors.affiliation: b Department of Pediatrics , David Geffen School of Medicine at UCLA , Los Angeles , CA , USA.
authors.forename: Andy
authors.initials: A
authors.lastname: Tu
authors.affiliation: b Department of Pediatrics , David Geffen School of Medicine at UCLA , Los Angeles , CA , USA.
authors.forename: Tulika
authors.initials: T
authors.lastname: Tyagi
authors.affiliation: a Department of Internal Medicine , David Geffen School of Medicine at UCLA , Los Angeles , CA , USA.|d Clinical Translational Science Institute (CTSI), David Geffen School of Medicine at UCLA , Los Angeles , CA , USA.
authors.forename: Xiang
authors.initials: X
authors.lastname: Lu
authors.affiliation: a Department of Internal Medicine , David Geffen School of Medicine at UCLA , Los Angeles , CA , USA.|d Clinical Translational Science Institute (CTSI), David Geffen School of Medicine at UCLA , Los Angeles , CA , USA.
authors.forename: David
authors.initials: D
authors.lastname: Elashoff
authors.affiliation: b Department of Pediatrics , David Geffen School of Medicine at UCLA , Los Angeles , CA , USA.
authors.forename: Satiro N
authors.initials: SN
authors.lastname: De Oliveira
mesh_terms: D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology; D018106:CD28 Antigens / Q000276:immunology; D015774:Ganciclovir / Q000008:administration & dosage; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells; D006801:Humans; D007167:Immunotherapy; D019169:Jurkat Cells; D016086:Lentivirus / Q000235:genetics; D016393:Lymphoma, B-Cell / Q000276:immunology / Q000628:therapy*; D051379:Mice; D009364:Neoplasm Recurrence, Local / Q000628:therapy; D011948:Receptors, Antigen, T-Cell / Q000235:genetics* / Q000276:immunology* / Q000627:therapeutic use; D018139:Simplexvirus / Q000201:enzymology / Q000235:genetics; D013937:Thymidine Kinase / Q000235:genetics; D014161:Transduction, Genetic
chemical_list: C000605956:CD19-specific chimeric antigen receptor; D018106:CD28 Antigens; D011948:Receptors, Antigen, T-Cell; D013937:Thymidine Kinase; D015774:Ganciclovir
keywords: CAR; HSC; cancer immunotherapy; gene therapy; suicide gene