Overcoming the challenge of transduction of human T-cells with chimeric antigen receptor (CAR) specific for ERBB2 antigen

Breast cancer is one of the most common malignancies among woman. Decades of scientific study have linked the overexpression of ERBB2 antigen to aggressive tumors. To target aggressive breast cancer, chimeric antigen receptor (CAR) technology can be utilized. For this, human T-cells are transduced w...

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Main Authors:	Rusheni Munisvaradass, Lee, Shirley Ding Suet, Koh, Avin Ee Hwan, Suresh Kumar, Lim, Moon Nian, Shalini Vellasamy, Syahril Abdullah, Alarfaj, Abdullah A., Ling, Mok Pooi
Format:	Article
Language:	English
Published:	Penerbit Universiti Kebangsaan Malaysia 2017
Online Access:	http://journalarticle.ukm.my/11542/ http://journalarticle.ukm.my/11542/ http://journalarticle.ukm.my/11542/1/21%20Rusheni%20Munisvaradass.pdf

id	ukm-11542
recordtype	eprints
spelling	ukm-115422018-04-10T09:56:14Z http://journalarticle.ukm.my/11542/ Overcoming the challenge of transduction of human T-cells with chimeric antigen receptor (CAR) specific for ERBB2 antigen Rusheni Munisvaradass, Lee, Shirley Ding Suet Koh, Avin Ee Hwan Suresh Kumar, Lim, Moon Nian Shalini Vellasamy, Syahril Abdullah, Alarfaj, Abdullah A. Ling, Mok Pooi Breast cancer is one of the most common malignancies among woman. Decades of scientific study have linked the overexpression of ERBB2 antigen to aggressive tumors. To target aggressive breast cancer, chimeric antigen receptor (CAR) technology can be utilized. For this, human T-cells are transduced with a gene sequence encoding a CAR that is specific for tumor-associated antigens (TAAs). These genetically-engineered CAR transduced T-cells (CAR-T cells) are able to target the tumor antigen without the need for major histocompatibility complex (MHC) recognition, rendering it a potentially universal immunotherapeutic option. However, efficient transduction of therapeutic gene into human T-cells and further cell expansion are challenging. In this study, we reported a successful optimization of a transduction protocol using spinoculation on CD3+ T-cells with different concentrations of lentiviral plasmid encoding the CAR gene. CD3+T-cells were isolated from the peripheral blood mononuclear cells (PBMCs). The constructed CAR gene was inserted into a lentiviral plasmid containing the green fluorescent protein (GFP) tag and lentiviral particles were produced. These lentiviral particles were used to transduce activated T-cells by spinoculation. T-cells were activated using Dynabead-conjugated CD3/CD28 human T-cell activator and interleukin-2 (IL-2) before transduction. CD3+ T-cells were selected and GFP expression, which indicated transduction, was observed. Future studies will focus on in vitro and in vivo models to determine the efficiency of CAR-T cells in specifically targeting ERBB2-expressing cells. Penerbit Universiti Kebangsaan Malaysia 2017-10 Article PeerReviewed application/pdf en http://journalarticle.ukm.my/11542/1/21%20Rusheni%20Munisvaradass.pdf Rusheni Munisvaradass, and Lee, Shirley Ding Suet and Koh, Avin Ee Hwan and Suresh Kumar, and Lim, Moon Nian and Shalini Vellasamy, and Syahril Abdullah, and Alarfaj, Abdullah A. and Ling, Mok Pooi (2017) Overcoming the challenge of transduction of human T-cells with chimeric antigen receptor (CAR) specific for ERBB2 antigen. Sains Malaysiana, 46 (10). pp. 1831-1838. ISSN 0126-6039 http://www.ukm.my/jsm/english_journals/vol46num10_2017/contentsVol46num10_2017.html
repository_type	Digital Repository
institution_category	Local University
institution	Universiti Kebangasaan Malaysia
building	UKM Institutional Repository
collection	Online Access
language	English
description	Breast cancer is one of the most common malignancies among woman. Decades of scientific study have linked the overexpression of ERBB2 antigen to aggressive tumors. To target aggressive breast cancer, chimeric antigen receptor (CAR) technology can be utilized. For this, human T-cells are transduced with a gene sequence encoding a CAR that is specific for tumor-associated antigens (TAAs). These genetically-engineered CAR transduced T-cells (CAR-T cells) are able to target the tumor antigen without the need for major histocompatibility complex (MHC) recognition, rendering it a potentially universal immunotherapeutic option. However, efficient transduction of therapeutic gene into human T-cells and further cell expansion are challenging. In this study, we reported a successful optimization of a transduction protocol using spinoculation on CD3+ T-cells with different concentrations of lentiviral plasmid encoding the CAR gene. CD3+T-cells were isolated from the peripheral blood mononuclear cells (PBMCs). The constructed CAR gene was inserted into a lentiviral plasmid containing the green fluorescent protein (GFP) tag and lentiviral particles were produced. These lentiviral particles were used to transduce activated T-cells by spinoculation. T-cells were activated using Dynabead-conjugated CD3/CD28 human T-cell activator and interleukin-2 (IL-2) before transduction. CD3+ T-cells were selected and GFP expression, which indicated transduction, was observed. Future studies will focus on in vitro and in vivo models to determine the efficiency of CAR-T cells in specifically targeting ERBB2-expressing cells.
format	Article
author	Rusheni Munisvaradass, Lee, Shirley Ding Suet Koh, Avin Ee Hwan Suresh Kumar, Lim, Moon Nian Shalini Vellasamy, Syahril Abdullah, Alarfaj, Abdullah A. Ling, Mok Pooi
spellingShingle	Rusheni Munisvaradass, Lee, Shirley Ding Suet Koh, Avin Ee Hwan Suresh Kumar, Lim, Moon Nian Shalini Vellasamy, Syahril Abdullah, Alarfaj, Abdullah A. Ling, Mok Pooi Overcoming the challenge of transduction of human T-cells with chimeric antigen receptor (CAR) specific for ERBB2 antigen
author_facet	Rusheni Munisvaradass, Lee, Shirley Ding Suet Koh, Avin Ee Hwan Suresh Kumar, Lim, Moon Nian Shalini Vellasamy, Syahril Abdullah, Alarfaj, Abdullah A. Ling, Mok Pooi
author_sort	Rusheni Munisvaradass,
title	Overcoming the challenge of transduction of human T-cells with chimeric antigen receptor (CAR) specific for ERBB2 antigen
title_short	Overcoming the challenge of transduction of human T-cells with chimeric antigen receptor (CAR) specific for ERBB2 antigen
title_full	Overcoming the challenge of transduction of human T-cells with chimeric antigen receptor (CAR) specific for ERBB2 antigen
title_fullStr	Overcoming the challenge of transduction of human T-cells with chimeric antigen receptor (CAR) specific for ERBB2 antigen
title_full_unstemmed	Overcoming the challenge of transduction of human T-cells with chimeric antigen receptor (CAR) specific for ERBB2 antigen
title_sort	overcoming the challenge of transduction of human t-cells with chimeric antigen receptor (car) specific for erbb2 antigen
publisher	Penerbit Universiti Kebangsaan Malaysia
publishDate	2017
url	http://journalarticle.ukm.my/11542/ http://journalarticle.ukm.my/11542/ http://journalarticle.ukm.my/11542/1/21%20Rusheni%20Munisvaradass.pdf
first_indexed	2023-09-18T20:00:35Z
last_indexed	2023-09-18T20:00:35Z
_version_	1777406821481316352

Overcoming the challenge of transduction of human T-cells with chimeric antigen receptor (CAR) specific for ERBB2 antigen

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