Table 3.

Summary table of preclinical and clinical research studies focused on chimeric antigen receptor (CAR) T cell therapies for CCA. Abbreviations: chimeric antigen receptor 4th generation (CAR4 T), epidermal growth factor receptor (EGFR), interleukin (IL), intrahepatic cholangiocarcinoma (ICC), mucin 1 (MUC1), natural killer (NK), programmed cell death protein (PD-1), T cell immunoglobulin and ITIM domain (Tigit), T cell immunoglobulin and mucin domain-containing-3 (TIM-3), transforming growth factor-β (TGF-β).

		Pre-Clinical
Author	Title	Date	Summary	Phase, Enrollment Status	Results	Ref.
Sangsuwannuku T et al.	Anti-tumor effect of the fourth-generation chimeric antigen receptor T cells targeting CD133 against CCA cells	2020	Anti-CD133-CAR4 T cells can be used to target CD133-expressing cancers as an alternative cellular immunotherapy in CD133-positive CCA, and may also be beneficial for treating other CD133-expressing cancers.			[52]
Supimon K et al.	Anti-mucin 1 chimeric antigen receptor T cells for adoptive T cell therapy of CCA	2021	Anti-MUC1-CAR4 T cells could effectively disrupt KKU-213A spheroids.			[66]
Phanthaphol N et al.	Chimeric Antigen Receptor T Cells Targeting Integrin αvβ6 Expressed on CCA Cells	2021	A20-4G CAR T cells had lower levels of cytokine production, but with higher proliferation rates; represents a promising potential adoptive T cell therapy for integrin αvβ6-positive CCA.			[115]
Supimon K et al.	Cytotoxic activity of anti-mucin 1 chimeric antigen receptor T cells expressing PD-1-CD28 switch receptor against CCA cells	2023	The cytotoxic function of aM.CAR/SR T cells was enhanced over the aM.CAR T cells, which have potential to be further tested for CCA treatment.			[125]
Mao L et al.	Development of Engineered CAR T-cells Targeting Tumor-Associated Glycoforms of MUC1 for the Treatment of Intrahepatic CCA	2023	Tn-MUC1 may be a potential therapeutic target for ICC, and its expression level was positively correlated with poor prognosis of ICC patients.			[9]
Qiao Y et al.	Enhancement of CAR T-cell activity against CCA by simultaneous knockdown of six inhibitory membrane proteins	2023	PTG-T16R-scFV-CAR T cells with knockdown of sextuplet inhibitory molecules (PD-1, Tim-3, Tigit, TGFβR, IL-10R, IL-6R) exhibited strong anti-tumor effect against CCA and long-term efficacy both in vitro and in vivo.			[131]
Chiawpanit C et al.	Precision immunotherapy for cholangiocarcinoma: Pioneering the use of human-derived anti-cMET single chain variable fragment in anti-cMET chimeric antigen receptor (CAR) NK cells	2024	Anti-cMET CAR-NK cells were developed using a human-derived ScFv to target cMET. These engineered NK cells effectively killed cMET-expressing CCA cells, highlighting their potential as a promising therapy for CCA.			[132]
		Animal Model
Nebbia M et al. (Abstract)	B7-h3 targeted CAR T-cell immunotherapy for primary and metastatic/multi-focal intrahepatic CCA	2022	B7-H3 CAR T cell therapy is effective in eradicating both primary and multi- focal disease and ICC metastases established in NSG mice and in prolonging their survival.			[133]
		Clinical
Feng K et al.	Cocktail treatment with EGFR-specific and CD133-specific chimeric antigen receptor modified T cells in a patient with advanced CCA	2017	CAR T cocktail immunotherapy may be feasible for the treatment of CCA as well as other solid malignancies; however, the toxicities, especially the epidermal/endothelial damages, require further investigation.	Phase I N = 1 4-day successive infusion of 2.2 × 106/kg total (CART-EGFR) 2nd cycle: 2.1 × 106/kg CART-EGFR 1 cycle of CART-EGFR + 2 cycles nivolumab 1.22 × 106/kg CD133- CART	8.5 month PR from CART-EGFR, 4.5 month PR from CART133	[65]
Feng K et al.	Phase I study of chimeric antigen receptor modified T cells in treating HER2-positive advanced biliary tract cancers and pancreatic cancers	2018	Data from this study demonstrated the safety and feasibility of CART-HER2 immunotherapy and showed encouraging signals of clinical activity.	Phase I N = 4 pCCA, 4 iCCA, 1–2 cycles (median dose: 2.45 × 106/kg)	1 partial response (4.5 months PFS), 3 stable disease, 4 progressive disease Median PFS: 3.25 months (range, 1.5–5 months)	[99]
Guo Y et al.	Phase I Study of Chimeric Antigen Receptor–Modified T Cells in Patients with EGFR-Positive Advanced Biliary Tract Cancers	2018	The CART-EGFR cell immunotherapy was a safe and active strategy for EGFR-positive advanced BTCs. The enrichment of Tcm in the infused CART-EGFR cells could predict clinical response.	Phase I N = 14 CCA 1–3 cycles (median dose, 2.65 × 106/kg; range, 0.8–4.1 × 106/kg) within 6 months	1 complete response, 10 stable disease Median PFS: 4 months (range, 2.5–22 months)	[90]