title: Toward eliminating HLA class I expression to generate universal cells from allogeneic donors.
abstract: Long-term engraftment of allogeneic cells necessitates eluding immune-mediated rejection, which is currently achieved by matching for human leukocyte antigen (HLA) expression, immunosuppression, and/or delivery of donor-derived cells to sanctuary sites. Genetic engineering provides an alternative approach to avoid clearance of cells that are recognized as "non-self" by the recipient. To this end, we developed designer zinc finger nucleases and employed a "hit-and-run" approach to genetic editing for selective elimination of HLA expression. Electro-transfer of mRNA species coding for these engineered nucleases completely disrupted expression of HLA-A on human T cells, including CD19-specific T cells. The HLA-A(neg) T-cell pools can be enriched and evade lysis by HLA-restricted cytotoxic T-cell clones. Recognition by natural killer cells of cells that had lost HLA expression was circumvented by enforced expression of nonclassical HLA molecules. Furthermore, we demonstrate that zinc finger nucleases can eliminate HLA-A expression from embryonic stem cells, which broadens the applicability of this strategy beyond infusing HLA-disparate immune cells. These findings establish that clinically appealing cell types derived from donors with disparate HLA expression can be genetically edited to evade an immune response and provide a foundation whereby cells from a single donor can be administered to multiple recipients.
authors.affiliation: Division of Pediatrics, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA.
authors.forename: Hiroki
authors.initials: H
authors.lastname: Torikai
authors.forename: Andreas
authors.initials: A
authors.lastname: Reik
authors.forename: Frank
authors.initials: F
authors.lastname: Soldner
authors.forename: Edus H
authors.initials: EH
authors.lastname: Warren
authors.forename: Carrie
authors.initials: C
authors.lastname: Yuen
authors.forename: Yuanyue
authors.initials: Y
authors.lastname: Zhou
authors.forename: Denise L
authors.initials: DL
authors.lastname: Crossland
authors.forename: Helen
authors.initials: H
authors.lastname: Huls
authors.forename: Nicholas
authors.initials: N
authors.lastname: Littman
authors.forename: Ziying
authors.initials: Z
authors.lastname: Zhang
authors.forename: Scott S
authors.initials: SS
authors.lastname: Tykodi
authors.forename: Partow
authors.initials: P
authors.lastname: Kebriaei
authors.forename: Dean A
authors.initials: DA
authors.lastname: Lee
authors.forename: Jeffrey C
authors.initials: JC
authors.lastname: Miller
authors.forename: Edward J
authors.initials: EJ
authors.lastname: Rebar
authors.forename: Michael C
authors.initials: MC
authors.lastname: Holmes
authors.forename: Rudolf
authors.initials: R
authors.lastname: Jaenisch
authors.forename: Richard E
authors.initials: RE
authors.lastname: Champlin
authors.forename: Philip D
authors.initials: PD
authors.lastname: Gregory
authors.forename: Laurence J N
authors.initials: LJ
authors.lastname: Cooper
mesh_terms: D018941:Antigens, CD19 / Q000378:metabolism; D001483:Base Sequence; D002454:Cell Differentiation; D003602:Cytotoxicity, Immunologic / Q000276:immunology; D003851:Deoxyribonucleases / Q000235:genetics*; D018274:Electroporation; D053595:Embryonic Stem Cells / Q000166:cytology; D018014:Gene Transfer Techniques; D057809:HEK293 Cells; D015395:Histocompatibility Antigens Class I / Q000378:metabolism*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000166:cytology; D008969:Molecular Sequence Data; D015202:Protein Engineering; D033581:Stem Cell Transplantation / Q000379:methods*; D013601:T-Lymphocytes / Q000276:immunology; D014184:Transplantation, Homologous; D016335:Zinc Fingers
chemical_list: D018941:Antigens, CD19; D015395:Histocompatibility Antigens Class I; D003851:Deoxyribonucleases