CAR-NK Cells Generated with mRNA-LNPs Kill Tumor Target Cells In Vitro and In Vivo

被引：27

作者：

Golubovskaya, Vita ^{[1
]}

Sienkiewicz, John ^{[1
]}

Sun, Jinying ^{[1
]}

Zhang, Shiming ^{[1
]}

Huang, Yanwei ^{[1
]}

Zhou, Hua ^{[1
]}

Harto, Hizkia ^{[1
]}

Xu, Shirley ^{[1
]}

Berahovich, Robert ^{[1
]}

Wu, Lijun ^{[1
,2
]}

机构：

[1] Promab Biotechnol, 2600 Hilltop Dr, Richmond, CA 94806 USA

[2] Forevertek Biotechnol, Changsha Hitech Ind Dev Zone, Lab Crit Qual Attributes Cell Therapy Prod, Janshan Rd, Changsha 410205, Peoples R China

来源：

INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES | 2023年 / 24卷 / 17期

关键词：

natural killer cells; chimeric antigen receptor; lipid nanoparticles; mRNA; leukemia; multiple myeloma; tumor; immunotherapy; T-CELLS; CANCER; INHIBITION; DELIVERY; THERAPY; GROWTH;

D O I：

10.3390/ijms241713364

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Natural killer (NK) cells are cytotoxic lymphocytes that are critical for the innate immune system. Engineering NK cells with chimeric antigen receptors (CARs) allows CAR-NK cells to target tumor antigens more effectively. In this report, we present novel CAR mRNA-LNP (lipid nanoparticle) technology to effectively transfect NK cells expanded from primary PBMCs and to generate functional CAR-NK cells. CD19-CAR mRNA and BCMA-CAR mRNA were embedded into LNPs that resulted in 78% and 95% CAR expression in NK cells, respectively. BCMA-CAR-NK cells after transfection with CAR mRNA-LNPs killed multiple myeloma RPMI8226 and MM1S cells and secreted IFN-gamma and Granzyme B in a dose-dependent manner in vitro. In addition, CD19-CAR-NK cells generated with CAR mRNA-LNPs killed Daudi and Nalm-6 cells and secreted IFN-gamma and Granzyme B in a dose-dependent manner. Both BCMA-CAR-NK and CD19-CAR-NK cells showed significantly higher cytotoxicity, IFN-gamma, and Granzyme B secretion compared with normal NK cells. Moreover, CD19-CAR-NK cells significantly blocked Nalm-6 tumor growth in vivo. Thus, non-viral delivery of CAR mRNA-LNPs can be used to generate functional CAR-NK cells with high anti-tumor activity.

引用

页数：13

共 33 条

[1] CAR-Ts: new perspectives in cancer therapy [J].

Abrantes, Rafaela ;

Duarte, Henrique O. ;

Gomes, Catarina ;

Walchli, Sebastien ;

Reis, Celso A. .

FEBS LETTERS, 2022, 596 (04) :403-416

[2] Targeting cardiac fibrosis with engineered T cells [J].

Aghajanian, Haig ;

Kimura, Toru ;

Rurik, Joel G. ;

Hancock, Aidan S. ;

Leibowitz, Michael S. ;

Li, Li ;

Scholler, John ;

Monslow, James ;

Lo, Albert ;

Han, Wei ;

Wang, Tao ;

Bedi, Kenneth ;

Morley, Michael P. ;

Saldana, Ricardo A. Linares ;

Bolar, Nikhita A. ;

McDaid, Kendra ;

Assenmacher, Charles-Antoine ;

Smith, Cheryl L. ;

Wirth, Dagmar ;

June, Carl H. ;

Margulies, Kenneth B. ;

Jain, Rajan ;

Pure, Ellen ;

Albelda, Steven M. ;

Epstein, Jonathan A. .

NATURE, 2019, 573 (7774) :430-+

[3] CAR T therapy beyond cancer: the evolution of a living drug [J].

Baker, Daniel J. ;

Arany, Zoltan ;

Baur, Joseph A. ;

Epstein, Jonathan A. ;

June, Carl H. .

NATURE, 2023, 619 (7971) :707-715

[4] CAR-T Cells Based on Novel BCMA Monoclonal Antibody Block Multiple Myeloma Cell Growth [J].

Berahovich, Robert ;

Zhou, Hua ;

Xu, Shirley ;

Wei, Yuehua ;

Guan, Jasper ;

Guan, Jian ;

Harto, Hizkia ;

Fu, Shuxiang ;

Yang, Kaihuai ;

Zhu, Shuying ;

Li, Le ;

Wu, Lijun ;

Golubovskaya, Vita .

CANCERS, 2018, 10 (09)

[5] FLAG-tagged CD19-specific CAR-T cells eliminate CD19-bearing solid tumor cells in vitro and in vivo [J].

Berahovich, Robert ;

Xu, Shirley ;

Zhou, Hua ;

Harto, Hizkia ;

Xu, Qumiao ;

Garcia, Andres ;

Liu, Fenyong ;

Golubovskaya, Vita ;

Wu, Lijun .

FRONTIERS IN BIOSCIENCE-LANDMARK, 2017, 22 :1644-1654

[6] Nonviral technologies can pave the way for CAR-NK cell therapy [J].

Bexte, Tobias ;

Reindl, Lisa Marie ;

Ullrich, Evelyn .

JOURNAL OF LEUKOCYTE BIOLOGY, 2023, 114 (05) :475-486

[7] CS1-specific chimeric antigen receptor (CAR)-engineered natural killer cells enhance in vitro and in vivo antitumor activity against human multiple myeloma [J].

Chu, J. ;

Deng, Y. ;

Benson, D. M. ;

He, S. ;

Hughes, T. ;

Zhang, J. ;

Peng, Y. ;

Mao, H. ;

Yi, L. ;

Ghoshal, K. ;

He, X. ;

Devine, S. M. ;

Zhang, X. ;

Caligiuri, M. A. ;

Hofmeister, C. C. ;

Yu, J. .

LEUKEMIA, 2014, 28 (04) :917-927

[8] CAR-NK cells: the next wave of cellular therapy for cancer [J].

Daher, May ;

Garcia, Luciana Melo ;

Li, Ye ;

Rezvani, Katayoun .

CLINICAL & TRANSLATIONAL IMMUNOLOGY, 2021, 10 (04)

[9] mRNA-Lipid Nanoparticle (LNP) Delivery of Humanized EpCAM-CD3 Bispecific Antibody Significantly Blocks Colorectal Cancer Tumor Growth [J].

Golubovskaya, Vita ;

Sienkiewicz, John ;

Sun, Jinying ;

Huang, Yanwei ;

Hu, Liang ;

Zhou, Hua ;

Harto, Hizkia ;

Xu, Shirley ;

Berahovich, Robert ;

Bodmer, Walter ;

Wu, Lijun .

CANCERS, 2023, 15 (10)

[10] Novel CS1 CAR-T Cells and Bispecific CS1-BCMA CAR-T Cells Effectively Target Multiple Myeloma [J].

Golubovskaya, Vita ;

Zhou, Hua ;

Li, Feng ;

Berahovich, Robert ;

Sun, Jinying ;

Valentine, Michael ;

Xu, Shirley ;

Harto, Hizkia ;

Sienkiewicz, John ;

Huang, Yanwei ;

Wu, Lijun .

BIOMEDICINES, 2021, 9 (10)

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