CD38 knockout natural killer cells expressing an affinity optimized CD38 chimeric antigen receptor successfully target acute myeloid leukemia with reduced effector cell fratricide

There is a strong biological rationale for the augmentation of allogeneic natural killer (NK) cell therapies with a chimeric antigen receptor (CAR) to enhance acute myeloid leukemia (AML) targeting. CD38 is an established immunotherapeutic target in multiple myeloma and under investigation as a targ...

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Published inHaematologica (Roma) Vol. 107; no. 2; pp. 437 - 445
Main Authors Gurney, Mark, Stikvoort, Arwen, Nolan, Emma, Kirkham-McCarthy, Lucy, Khoruzhenko, Stanislav, Shivakumar, Rama, Zweegman, Sonja, Van de Donk, Niels W.C.J., Mutis, Tuna, Szegezdi, Eva, Sarkar, Subhashis, O’Dwyer, Michael
Format Journal Article
LanguageEnglish
Published Italy Fondazione Ferrata Storti 01.02.2022
Ferrata Storti Foundation
Subjects
ADP-ribosyl Cyclase 1
Cell Line, Tumor
Cytotoxicity, Immunologic
Gene Knockout Techniques
Humans
Immunotherapy, Adoptive
Killer Cells, Natural
Leukemia, Myeloid, Acute - therapy
Membrane Glycoproteins
Receptors, Chimeric Antigen - genetics
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Summary:There is a strong biological rationale for the augmentation of allogeneic natural killer (NK) cell therapies with a chimeric antigen receptor (CAR) to enhance acute myeloid leukemia (AML) targeting. CD38 is an established immunotherapeutic target in multiple myeloma and under investigation as a target antigen in AML. CD38 expression on NK cells and its further induction during ex vivo NK cell expansion represent barriers to the development of a CD38 CAR-NK cell therapy. We set out to develop a CD38 CAR-NK cell therapy for AML, first by using an NK cell line which has low baseline CD38 expression and subsequently NK cells expanded from healthy donors. To overcome anticipated fratricide due to NK cell CD38 expression when using primary expanded NK cells, we applied CRISPR/Cas9 genome editing to disrupt the CD38 gene during expansion, achieving a mean knockdown efficiency of 84%. The resulting CD38 knockdown expanded NK cells, after expression of an affinity optimized CD38 CAR, showed reduced NK-cell fratricide and an enhanced ability to target primary AML blasts. Furthermore, the cytotoxic potential of CD38 CAR-NK cells was augmented by pretreatment of the AML cells with all-trans retinoic acid which drove enhanced CD38 expression, offering a rational combination therapy. These findings support the further investigation of CD38 knockdown - CD38 CAR-NK cells as a viable immunotherapeutic approach to the treatment of AML.
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Disclosures
MG has received educational funding from Janssen Pharmaceuticals and Takeda. AS and SS have received research funding from ONK Therapeutics Limited. LKM is an employee of ONK Therapeutics Limited. SK and RS are employees of Maxcyte Inc. SZ has received research funding from Takeda, Celgene, and Janssen and is a member of the board of directors or an advisory committee for Takeda, Celgene, and Janssen. NWCJvdD has received research funding from Janssen Pharmaceuticals, Amgen, Celgene, Novartis, and BMS and has participated in advisory boards for Janssen Pharmaceuticals, Amgen, Celgene, BMS, Takeda, Roche, Novartis, Bayer, and Servier. TM has received research funding from Gilead, Celgene, Novartis, ONK Therapeutics Limited, Genmab, Janssen and has been a member of an advisory board for Janssen. ES has collaborated in research projects with Janssen, Roche, Celgene, and Takeda. MOD has received research funding from ONK Therapeutics Limited, BMS, Celgene, and Glycomimetics; is a member of the board of directors or an advisory committee for Janssen, Abbvie, and ONK Therapeutics Limited; and owns equity in ONK Therapeutics Limited. EN has no potential conflicts of interest to disclose.
SS and MOD conceived the research. EN, MG, AS and LKM performed functional assays. SK and RS contributed to electroporation optimization. ES contributed to acquisition of patients’ samples and the primary AML assay design. TM, SZ and NVD developed CD38 CAR-KHYG1 cells and associated functional assays. MG, SS and AS wrote the manuscript and prepared the figures. All authors contributed to editing and reviewing the final manuscript prior to submission.
Contributions
ISSN:0390-6078
1592-8721
1592-8721
DOI:10.3324/haematol.2020.271908