Engineering of potent CAR NK cells using non-viral Sleeping Beauty transposition from minimalistic DNA vectors

被引：19

作者：

Bexte, Tobias ^{[1
,2
,3
,4
,5
]}

Botezatu, Lacramioara ^{[6
,7
]}

Miskey, Csaba ^{[6
]}

Gierschek, Fenja ^{[1
,2
]}

Moter, Alina ^{[1
,2
]}

Wendel, Philipp ^{[1
,2
,8
,9
,10
]}

Reindl, Lisa Marie ^{[1
,2
]}

Campe, Julia ^{[1
,2
]}

Villena-Ossa, Jose Francisco ^{[11
,12
]}

Gebel, Veronika ^{[1
,2
,3
,4
]}

Stein, Katja ^{[1
,2
,3
]}

Cathomen, Toni ^{[11
,12
,13
,14
]}

Cremer, Anjali ^{[2
,3
,4
,8
,9
,15
]}

Wels, Winfried S. ^{[2
,8
,9
,16
]}

Hudecek, Michael ^{[17
,18
]}

Ivics, Zoltan ^{[6
,7
]}

Ullrich, Evelyn ^{[1
,2
,3
,4
,8
,9
]}

机构：

[1] Goethe Univ, Dept Pediat Expt Immunol & Cell Therapy, Theodor Stern Kai 7, D-60590 Frankfurt, Germany

[2] Goethe Univ, Frankfurt Canc Inst FCI, Frankfurt, Germany

[3] Univ Canc Ctr UCT Frankfurt, Frankfurt, Germany

[4] Hosp Goethe Univ Frankfurt, Mildred Scheel Career Ctr MSNZ, Frankfurt, Germany

[5] German Red Cross Blood Serv Baden Wurttemberg Hess, Inst Transfus Med & Immunohematol, Frankfurt, Germany

[6] Paul Ehrlich Inst, Res Ctr, Div Hematol Gene & Cell Therapy, Langen, Germany

[7] German Canc Consortium DKTK, Partner Site Heidelberg, Heidelberg, Germany

[8] German Canc Consortium DKTK, Partner Site Frankfurt Mainz, Heidelberg, Germany

[9] German Canc Res Ctr, Heidelberg, Germany

[10] Tech Univ Darmstadt, Inst Organ Chem & Biochem, Darmstadt, Germany

[11] Univ Freiburg, Inst Transfus Med & Gene Therapy, Med Ctr, Freiburg, Germany

[12] Univ Freiburg, Ctr Chron Immunodeficiency, Med Ctr, Freiburg, Germany

[13] Univ Freiburg, Fac Med, Freiburg, Germany

[14] German Canc Consortium DKTK, Partner Site Freiburg, Freiburg, Germany

[15] Univ Hosp Frankfurt, Dept Hematol Oncol, Frankfurt, Germany

[16] Inst Tumor Biol & Expt Therapy, Frankfurt, Germany

[17] Univ Hosp Wurzburg, Dept Med 2, Chaire Cellular Immunotherapy, Wurzburg, Germany

[18] Fraunhofer Inst Cell Therapy & Immunol, Cellular Immunotherapy Branch Site Wurzburg, Wurzburg, Germany

来源：

MOLECULAR THERAPY | 2024年 / 32卷 / 07期

关键词：

CHIMERIC ANTIGEN RECEPTOR; STABLE GENE-TRANSFER; NATURAL-KILLER-CELLS; MESSENGER-RNA; PIGGYBAC; STEM; TRANSFECTION; EXPRESSION; SELECTION; INSERTION;

D O I：

10.1016/j.ymthe.2024.05.022

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Natural killer (NK) cells have high intrinsic cytotoxic capacity, and clinical trials have demonstrated their safety and efficacy for adoptive cancer therapy. Expression of chimeric antigen receptors (CARs) enhances NK cell target specificity, with these cells applicable as off-the-shelf products generated from allogeneic donors. Here, we present for the first time an innovative approach for CAR NK cell engineering employing a non-viral Sleeping Beauty (SB) transposon/transposase-based system and minimized DNA vectors termed minicircles. SB-modified peripheral blood-derived primary NK cells displayed high and stable CAR expression and more frequent vector integration into genomic safe harbors than lentiviral vectors. Importantly, SB-generated CAR NK cells demonstrated enhanced cytotoxicity compared with non-transfected NK cells. A strong antileukemic potential was confirmed using established acute lymphocytic leukemia cells and patient-derived primary acute B cell leukemia and lymphoma samples as targets in vitro and in vivo in a xenograft leukemia mouse model. Our data suggest that the SB-transposon system is an efficient, safe, and costeffective approach to non-viral engineering of highly functional CAR NK cells, which may be suitable for cancer immunotherapy of leukemia as well as many other malignancies.

引用

页码：2357 / 2372

页数：16

共 71 条

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