High antileukemic efficiency of CD19-CAR NK cells engineered with
                        Sleeping Beauty transposon vectors

T Bexte; L Botezatu; C Miskey; LM Reindl; J Campe; S Mertlitz; V Gebel; R Schubert; A Cremer; E Rettinger; S Prommersberger; O Penack; WS Wels; M Hudecek; Z Ivics; E Ullrich

doi:10.1055/s-0042-1748751

Klinische Pädiatrie, Table of Contents

Klin Padiatr 2022; 234(03): 190-191
DOI: 10.1055/s-0042-1748751

Abstracts

High antileukemic efficiency of CD19-CAR NK cells engineered with Sleeping Beauty transposon vectors

T Bexte

¹Experimental Immunology, Department for Children and Adolescents Medicine, Hospital of the Goethe University Frankfurt, 60590, Germany

²Frankfurt Cancer Institute, Goethe University, Frankfurt am Main, Germany

³University Cancer Center (UCT) Frankfurt-Marburg

,

L Botezatu

⁴Division of Medical Biotechnology, Paul Ehrlich Institute, Langen, Germany

,

C Miskey

⁴Division of Medical Biotechnology, Paul Ehrlich Institute, Langen, Germany

,

LM Reindl

¹Experimental Immunology, Department for Children and Adolescents Medicine, Hospital of the Goethe University Frankfurt, 60590, Germany

,

J Campe

¹Experimental Immunology, Department for Children and Adolescents Medicine, Hospital of the Goethe University Frankfurt, 60590, Germany

,

S Mertlitz

⁵Charité Universitätsmedizin Berlin, Berlin, Germany

,

V Gebel

¹Experimental Immunology, Department for Children and Adolescents Medicine, Hospital of the Goethe University Frankfurt, 60590, Germany

²Frankfurt Cancer Institute, Goethe University, Frankfurt am Main, Germany

³University Cancer Center (UCT) Frankfurt-Marburg

,

R Schubert

⁶Department for Children and Adolescents, University Hospital, Goethe-University, Frankfurt, Germany

,

A Cremer

⁷Department of Hematology and Oncology, Johann Wolfgang Goethe University, Frankfurt, Germany

,

E Rettinger

⁸Division of Stem Cell Transplantation and Immunology, Department of Children and Adolescent Medicine, Hospital of the Goethe University Frankfurt, 60590, Germany

,

S Prommersberger

⁹Department of Internal Medicine II, University Hospital Würzburg, Würzburg, Germany

,

O Penack

⁵Charité Universitätsmedizin Berlin, Berlin, Germany

,

WS Wels

²Frankfurt Cancer Institute, Goethe University, Frankfurt am Main, Germany

¹⁰Georg-Speyer-Haus, Institute for Tumor Biology and Experimental Therapy, Frankfurt am Main, Germany

¹¹German Cancer Consortium (DKTK) partner site Frankfurt/Mainz, Frankfurt am Main, Germany

,

M Hudecek

⁹Department of Internal Medicine II, University Hospital Würzburg, Würzburg, Germany

,

Z Ivics

⁴Division of Medical Biotechnology, Paul Ehrlich Institute, Langen, Germany

,

E Ullrich

¹Experimental Immunology, Department for Children and Adolescents Medicine, Hospital of the Goethe University Frankfurt, 60590, Germany

²Frankfurt Cancer Institute, Goethe University, Frankfurt am Main, Germany

³University Cancer Center (UCT) Frankfurt-Marburg

¹¹German Cancer Consortium (DKTK) partner site Frankfurt/Mainz, Frankfurt am Main, Germany

› Author Affiliations

Abstract

Full Text

Expression of chimeric antigen receptors (CAR) constitutes a promising approach to enhance therapeutic efficacy of NK cells. Here, we demonstrate that primary CAR NK cells can be generated using the non-viral Sleeping Beauty (SB) transposon/transposase.

SB transposons vectorized as minicircles, encoding a CD19-CAR were transfected together with the hyperactive SB100X transposase. SB-modified NK cells displayed long-lasting CD19-CAR expression during ex-vivo expansion. Assessment of vector integration revealed a significantly higher frequency of insertion into genomic safe harbors for SB-CAR-NK cells compared to lentivirally-transduced CAR-NK cells. SB-CD19-CAR NK cells demonstrated significantly higher cytotoxicity compared to non-modified NK cells in vitro. Enhanced antitumor potential was confirmed in a systemic CD19-expressing leukemia xenograft model (NSG-NALM-6/Luc) in vivo. In addition, enhanced SB-CD19-CAR NK cell-mediated killing capacity was also observed against primary patient-derived B-ALL blasts.

The Sleeping Beauty transposon system represents a safe and cost-effective platform for CAR NK cells generation and can be suitable also for other cancer immunotherapies.