Toward eliminating HLA class I expression to generate universal cells from allogeneic donors

Abstract

Key Points Allogeneic-donor–derived cells can be genetically modified to eliminate expression of HLA-A. HLA-A disruption from donor cells is a step toward generating allogeneic cells as an off-the-shelf therapeutic.

Multiplex Genome-Edited T-cell Manufacturing Platform for “Off-the-Shelf” Adoptive T-cell Immunotherapies

“…On the basis of these analyses, we conclude that TALEN-mediated genome editing can be incorporated into a large-scale T-cell culture process as a practical and safe approach to the manufacturing of third-party CAR T cells for use in adoptive T-cell immunotherapies. Zinc finger nucleases have previously been applied for gene editing in human T cells, including inactivation of both the TCRa and TCRb genes for enabling allogeneic "off-the-shelf" and TCR replacement strategies and inactivation of an HLA class I gene (34)(35)(36). Our observations, regarding the functional capabilities of TRAC KO T cells, are consistent with the results described in the aforementioned publications; T cells lacking a TCR component lose CD3 expression and all capacity for activation via either the TCR or through the CD3 complex.…”

CD62L+ NKT cells have prolonged persistence and antitumor activity in vivo

“…Moreover, a recent report demonstrated that reconstitution of NKTs in peripheral blood is associated with long-term remission in pediatric leukemia patients receiving haploidentical stem cell transplantation (71,72). Hence, unlike donor-derived CAR-T cells, CAR-NKTs can be given early after HSCT when the disease burden is minimal and the curative potential of immunotherapy jci.org Volume 126 Number 6 June 2016 system as previously described (76). A clone with an intermediate level of CD1d expression induced the highest rate of NKT proliferation and was further modified with lentiviral vectors encoding CD86, 4-1BBL, or OX40L followed by single cell sorting and clonal expansion.…”

“…We rendered K562 cells HLA null by eliminating the HLA-C gene from the K562 cell genome using an HLA-Cspecific ZFN as previously described (76). Briefly, ZFNs containing 5 or 6 fingers were designed and assembled using an established archive of prevalidated 2-finger and 1-finger modules (Sangamo BioScience).…”

“…After electroporation, cells were immediately placed in prewarmed complete medium. HLA-C-negative clones were obtained using limiting dilution, and HLA deletion was confirmed by Surveyor nuclease assay (76). A clone of HLA null K562 cells was then electroporated with human CD1d cDNA using Sleeping Beauty transposon/transposase…”

T-cell–based Immunotherapy: Adoptive Cell Transfer and Checkpoint Inhibition

“…Regarding applicability of engineered T cells, strategies are being developed to generate "universal" T cells that could be manufactured from one donor and administered to multiple patients. For instance, this could be done by eliminating the expression of endogenous TCR to generate T cells devoid of alloreactive TCRs (27). Finally, ACT efficacy may be improved by generating and/or selecting T cells with enhanced proliferative and antitumor capacity.…”