Anti-tumour effect of the fourth-generation chimeric antigen receptor T cells targeting CD133 against cholangiocarcinoma cells

Thanich Sangsuwannukul; Kamonlapat Supimon; Jatuporn Sujjitjoon; Nattaporn Phanthaphol; Thaweesak Chieochansin; Naravat Poungvarin; Sopit Wongkham; Mutita Junking; Pa-Thai Yenchitsomanus

doi:10.1016/j.intimp.2020.107069

Anti-tumour effect of the fourth-generation chimeric antigen receptor T cells targeting CD133 against cholangiocarcinoma cells

Int Immunopharmacol. 2020 Dec;89(Pt B):107069. doi: 10.1016/j.intimp.2020.107069. Epub 2020 Nov 24.

Authors

Thanich Sangsuwannukul¹, Kamonlapat Supimon¹, Jatuporn Sujjitjoon², Nattaporn Phanthaphol¹, Thaweesak Chieochansin², Naravat Poungvarin³, Sopit Wongkham⁴, Mutita Junking², Pa-Thai Yenchitsomanus⁵

Affiliations

¹ Siriraj Center of Research Excellence for Cancer Immunotherapy (SiCORE-CIT), Research Department, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand; Graduate Program in Immunology, Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
² Siriraj Center of Research Excellence for Cancer Immunotherapy (SiCORE-CIT), Research Department, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand; Division of Molecular Medicine, Research Department, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
³ Department of Clinical Pathology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
⁴ Department of Biochemistry, and Center for Translational Medicine, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand; Cholangiocarcinoma Research Institute, Khon Kaen University, Khon Kaen, Thailand.
⁵ Siriraj Center of Research Excellence for Cancer Immunotherapy (SiCORE-CIT), Research Department, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand; Division of Molecular Medicine, Research Department, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand. Electronic address: pathai.yen@mahidol.edu.

PMID: 33242709
DOI: 10.1016/j.intimp.2020.107069

Abstract

Current treatment of cholangiocarcinoma (CCA) - a lethal bile duct cancer - is ineffective because the disease is usually diagnosed at late and advanced stage. Thus, a novel therapeutic modality is urgently required. Fourth-generation chimeric antigen receptor (CAR4) T cells was created to target CD133, a well-known cancer stem cell marker, that is highly expressed and associates with cancer progression. The anti-CD133-CAR4 T cells showed high efficacy against CD133-expressing CCA cells. Tumour cell lysis occurred in a dose- and CD133 antigen-dependent manner, and significantly higher, up to 57.59% ± 9.62 at effector to target ratio of 5:1 in a CCA cell line - KKU-213A cells, compared to mock control (p = 0.008). Similarly, significant IFN-γ (p = 0.011) and TNF-α (p = 0.002) upregulation was observed upon tumour treatment. The effectiveness of our anti-CD133-CAR4 T cells will be beneficial not only for CD133-expressing CCA, but also for other CD133-expressing tumours. This study may guide future in vivo study and clinical trials.

Keywords: Adoptive T cell therapy; CAR T cell; CD133; Cancer stem cell; Chimeric antigen receptor; Cholangiocarcinoma.

MeSH terms

AC133 Antigen / immunology
AC133 Antigen / metabolism*
Bile Duct Neoplasms / immunology
Bile Duct Neoplasms / metabolism
Bile Duct Neoplasms / pathology
Bile Duct Neoplasms / therapy*
CD28 Antigens / genetics
CD28 Antigens / immunology
CD28 Antigens / metabolism
CD3 Complex / genetics
CD3 Complex / immunology
CD3 Complex / metabolism
Cell Line, Tumor
Cholangiocarcinoma / immunology
Cholangiocarcinoma / metabolism
Cholangiocarcinoma / pathology
Cholangiocarcinoma / therapy*
Coculture Techniques
Cytotoxicity, Immunologic
Humans
Immunotherapy, Adoptive*
Interferon-gamma / metabolism
Neoplastic Stem Cells / immunology
Neoplastic Stem Cells / metabolism*
Neoplastic Stem Cells / pathology
Single-Chain Antibodies / genetics
Single-Chain Antibodies / immunology
Single-Chain Antibodies / metabolism*
Spheroids, Cellular
T-Lymphocytes / immunology
T-Lymphocytes / metabolism*
T-Lymphocytes / transplantation
Tumor Necrosis Factor Receptor Superfamily, Member 7 / genetics
Tumor Necrosis Factor Receptor Superfamily, Member 7 / immunology
Tumor Necrosis Factor Receptor Superfamily, Member 7 / metabolism
Tumor Necrosis Factor Receptor Superfamily, Member 9 / genetics
Tumor Necrosis Factor Receptor Superfamily, Member 9 / immunology
Tumor Necrosis Factor Receptor Superfamily, Member 9 / metabolism
Tumor Necrosis Factor-alpha / metabolism

Substances

AC133 Antigen
CD28 Antigens
CD3 Complex
IFNG protein, human
PROM1 protein, human
Single-Chain Antibodies
TNF protein, human
TNFRSF9 protein, human
Tumor Necrosis Factor Receptor Superfamily, Member 7
Tumor Necrosis Factor Receptor Superfamily, Member 9
Tumor Necrosis Factor-alpha
Interferon-gamma