Concurrent transposon engineering and CRISPR/Cas9 genome editing of primary CLL-1 chimeric antigen receptor-natural killer cells

被引：17

作者：

Gurney, Mark ^{[1
]}

O'Reilly, Eimear ^{[1
,2
]}

Corcoran, Sarah ^{[2
]}

Brophy, Sarah ^{[2
]}

Krawczyk, Janusz ^{[1
]}

Otto, Neil M. ^{[3
]}

Hermanson, David L. ^{[3
]}

Childs, Richard W. ^{[4
]}

Szegezdi, Eva ^{[1
]}

O'Dwyer, Michael E. ^{[1
,2
]}

机构：

[1] Natl Univ Ireland Galway, Galway, Ireland

[2] ONK Therapeut, Galway, Ireland

[3] Biotechne, Minneapolis, MN USA

[4] NHLBI, NIH, Bethesda, MD USA

来源：

CYTOTHERAPY | 2022年 / 24卷 / 11期

基金：

英国惠康基金; 爱尔兰科学基金会;

关键词：

CAR-NK cells; CRISPR/Cas9; DNA transposon; Genome editing; ACUTE MYELOID-LEUKEMIA; T-CELLS; NK CELLS; TRANSPLANTATION; CHEMOTHERAPY; SURVIVAL; CISH;

D O I：

10.1016/j.jcyt.2022.07.008

中图分类号：

Q813 [细胞工程];

学科分类号：

摘要：

Background: Natural killer (NK) cell genome editing promises to enhance the innate and alloreactive antitumor potential of NK cell adoptive transfer. DNA transposons are versatile non-viral gene vectors now being adapted to primary NK cells, representing important tools for research and clinical product development. Aims and Methods: We set out to generate donor-derived, primary chimeric antigen receptor (CAR)-NK cells by combining the TcBuster transposon system with Epstein-Barr virus-transformed lymphoblastoid feeder cell-mediated activation and expansion. Results: This approach allowed for clinically relevant NK-cell expansion capability and CAR expression, which was further enhanced by immunomagnetic selection based on binding to the CAR target protein.The resulting CAR-NK cells targeting the myeloid associated antigen CLL-1 efficiently targeted CLL-1-positive AML cell lines and primary AML populations, including a population enriched for leukemia stem cells. Subsequently, concurrent delivery of CRISPR/Cas9 cargo was applied to knockout the NK cell cytokine checkpoint cytokine-inducible SH2-containing protein (CIS, product of the CISH gene), resulting in enhanced cytotoxicity and an altered NK cell phenotype. Conclusions: This report contributes a promising application of transposon engineering to donor-derived NK cells and emphasizes the importance of feeder mediated NK cell activation and expansion to current protocols. (c) 2022 International Society for Cell & Gene Therapy. Published by Elsevier Inc. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/)

引用

页码：1087 / 1094

页数：8