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...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Penerbit Universiti Kebangsaan Malaysia
2017
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| Online Access: | http://psasir.upm.edu.my/id/eprint/60480/ http://psasir.upm.edu.my/id/eprint/60480/1/Overcoming%20the%20challenge%20of%20transduction%20of%20human%20T-cells%20with%20chimeric%20antigen%20receptor%20%28CAR%29%20specific%20for%20ERBB2%20antigen.pdf |
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| author | Munisvaradass, Rusheni Ding, Shirley Suet Lee Koh, Avin Ee Hwan Subbiah, Suresh Kumar Lim, Moon Nian Vellasamy, Shalini Abdullah, Syahril Alarfaj, Abdullah A. Mok, Pooi Ling |
| author_facet | Munisvaradass, Rusheni Ding, Shirley Suet Lee Koh, Avin Ee Hwan Subbiah, Suresh Kumar Lim, Moon Nian Vellasamy, Shalini Abdullah, Syahril Alarfaj, Abdullah A. Mok, Pooi Ling |
| author_sort | Munisvaradass, Rusheni |
| building | UPM Institutional Repository |
| collection | Online Access |
| 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. |
| first_indexed | 2025-11-15T11:06:11Z |
| format | Article |
| id | upm-60480 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T11:06:11Z |
| publishDate | 2017 |
| publisher | Penerbit Universiti Kebangsaan Malaysia |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-604802018-05-21T03:52:18Z http://psasir.upm.edu.my/id/eprint/60480/ Overcoming the challenge of transduction of human T-cells with chimeric antigen receptor (CAR) specific for ERBB2 antigen Munisvaradass, Rusheni Ding, Shirley Suet Lee Koh, Avin Ee Hwan Subbiah, Suresh Kumar Lim, Moon Nian Vellasamy, Shalini Abdullah, Syahril Alarfaj, Abdullah A. Mok, Pooi Ling 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 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/60480/1/Overcoming%20the%20challenge%20of%20transduction%20of%20human%20T-cells%20with%20chimeric%20antigen%20receptor%20%28CAR%29%20specific%20for%20ERBB2%20antigen.pdf Munisvaradass, Rusheni and Ding, Shirley Suet Lee and Koh, Avin Ee Hwan and Subbiah, Suresh Kumar and Lim, Moon Nian and Vellasamy, Shalini and Abdullah, Syahril and Alarfaj, Abdullah A. and Mok, Pooi Ling (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 10.17576/jsm-2017-4610-21 |
| spellingShingle | Munisvaradass, Rusheni Ding, Shirley Suet Lee Koh, Avin Ee Hwan Subbiah, Suresh Kumar Lim, Moon Nian Vellasamy, Shalini Abdullah, Syahril Alarfaj, Abdullah A. Mok, Pooi Ling Overcoming the challenge of transduction of human T-cells with chimeric antigen receptor (CAR) specific for ERBB2 antigen |
| title | 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_short | 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 |
| url | http://psasir.upm.edu.my/id/eprint/60480/ http://psasir.upm.edu.my/id/eprint/60480/ http://psasir.upm.edu.my/id/eprint/60480/ http://psasir.upm.edu.my/id/eprint/60480/1/Overcoming%20the%20challenge%20of%20transduction%20of%20human%20T-cells%20with%20chimeric%20antigen%20receptor%20%28CAR%29%20specific%20for%20ERBB2%20antigen.pdf |