AAV-mediated in vivo CAR gene therapy for targeting human T-cell leukemia

Waqas Nawaz,Bilian Huang,Linjing Zhu,Zhiwei Wu,Yanlei Li,Hu Yiqiao,Shijie Xu,Xilin Wu

doi:10.1038/s41408-021-00508-1

Waqas Nawaz, Bilian Huang + Show 6 more

Open Access

https://doi.org/10.1038/s41408-021-00508-1

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Abstract

Chimeric antigen receptor (CAR) T-cell therapy is the most active field in immuno-oncology and brings substantial benefit to patients with B cell malignancies. However, the complex procedure for CAR T-cell generation hampers its widespread applications. Here, we describe a novel approach in which human CAR T cells can be generated within the host upon injecting an Adeno-associated virus (AAV) vector carrying the CAR gene, which we call AAV delivering CAR gene therapy (ACG). Upon single infusion into a humanized NOD.Cg-Prkdcscid Il2rgem26/Nju tumor mouse model of human T-cell leukemia, AAV generates sufficient numbers of potent in vivo CAR cells, resulting in tumor regression; these in vivo-generated CAR cells produce antitumor immunological characteristics. This instantaneous generation of in vivo CAR T cells may bypass the need for patient lymphodepletion, as well as the β processes of traditional CAR T-cell production, which may make CAR therapy simpler and less expensive. It may allow the development of intricate, individualized treatments in the form of on-demand and diverse therapies.

Highlights

Chimeric antigen receptor (CAR) T cells have achieved significant clinical recognition with remarkable responses documented for autologous CD19 redirected T cells for the treatment of patients with relapsed or refractory B cell malignancies [1]
We furnished our associated virus (AAV)-CD4CAR with the scFv region of the Hu5A8 resulted in the production of enough CD4-CAR T cells, which antibody specific for CD4, which we reported previously for HIV circulate in the host body for weeks (Fig. 3B)
We evaluated the functional ability of AAVCD4CAR to eliminate CD4+ cells in Peripheral blood mononuclear cells (PBMCs) from various donors (n = 10) during T-cell expansion following AAV-CD4CAR transducbiodistribution of AAV-CD4CAR T cells in different organs after injecting AAV-CD4CAR into NCG-HuPBL mice

Summary

Introduction

Chimeric antigen receptor (CAR) T cells have achieved significant clinical recognition with remarkable responses documented for autologous CD19 (a B cell-specific antigen) redirected T cells for the treatment of patients with relapsed or refractory B cell malignancies [1]. Current clinical-scale manufacturing of T lymphocytes involves complex processes of isolation, genetic modification, and selective expansion of redirected cells ex vivo prior to their infusion into patients. This makes the manufacturing process complicated, costly, and associated with clinical hazards [6]. The most crucial step to overcome these limitations is to create novel strategies for making this complicated and risky cell therapy affordable, safer and customizable. This may in turn enable CAR T cells to outcompete chemotherapy as front-line therapy

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