Human CD3+ T-Cells with the Anti-ERBB2 Chimeric Antigen Receptor Exhibit Efficient Targeting and Induce Apoptosis in ERBB2 Overexpressing Breast Cancer Cells.

Rusheni Munisvaradass,Chandramohan Govindasamy,Pooi Mok,Khalid Alnumair,Suresh Kumar

doi:10.3390/ijms18091797

Abstract

Breast cancer is a common malignancy among women. The innate and adaptive immune responses failed to be activated owing to immune modulation in the tumour microenvironment. Decades of scientific study links the overexpression of human epidermal growth factor receptor 2 (ERBB2) antigen with aggressive tumours. The Chimeric Antigen Receptor (CAR) coding for specific tumour-associated antigens could initiate intrinsic T-cell signalling, inducing T-cell activation, and cytotoxic activity without the need for major histocompatibility complex recognition. This renders CAR as a potentially universal immunotherapeutic option. Herein, we aimed to establish CAR in CD3+ T-cells, isolated from human peripheral blood mononucleated cells that could subsequently target and induce apoptosis in the ERBB2 overexpressing human breast cancer cell line, SKBR3. Constructed CAR was inserted into a lentiviral plasmid containing a green fluorescent protein tag and produced as lentiviral particles that were used to transduce activated T-cells. Transduced CAR-T cells were then primed with SKBR3 cells to evaluate their functionality. Results showed increased apoptosis in SKBR3 cells co-cultured with CAR-T cells compared to the control (non–transduced T-cells). This study demonstrates that CAR introduction helps overcome the innate limitations of native T-cells leading to cancer cell apoptosis. We recommend future studies should focus on in vivo cytotoxicity of CAR-T cells against ERBB2 expressing tumours.

Highlights

Cancer has become an endemic disease that is prevalent in the modern world
We successfully showed that these genetically modified CD3+ cells were able to target and induce apoptosis in the ERBB2 overexpressing breast cancer cell line, SKBR3
The lentivirus was packaged by 293FT cells once the presence of Chimeric Antigen Receptor (CAR) gene within the lentiviral expression transfer plasmid was confirmed (Figure S1)

Summary

Introduction

Cancer has become an endemic disease that is prevalent in the modern world. The occurrence of cancer is commonly linked to genetic code malfunctions, leading to abnormal expression of human epidermal growth factor receptor 2 (ERBB2), progesterone [4], oestrogen receptors [5], tumour protein P53 [6], and others. ERBB2 amplification and overexpression have been shown to possess a prevalence of 15–25% in breast cancer patients [8]. This overexpression, in turn, lowers the efficiency of chemotherapy and endocrine therapy, resulting in an earlier relapse with poor prognosis. Current conventional therapy to target ERBB2 antigen in breast cancer includes treatment with trastuzumab, a humanized monoclonal antibody that binds to the extracellular domain of the receptor [13]. The efficiency of this targeted therapy is shown to be only 12–34% [15,16]

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