靶向甲胎蛋白-磷脂酰肌醇蛋白聚糖3的嵌合抗原受体T细胞介导肝癌抗肿瘤疗效增强。
Chimeric antigen receptor-T cells targeting AFP-GPC3 mediate increased antitumor efficacy in hepatocellular carcinoma.
作者信息
Li Mingxing, Chen Tailin, Huang Rongshi, Cen Yanhui, Zhao Feilan, Fan Rong, He Guozhen
机构信息
Embryo Formation Teaching and Research Section, Guangxi University of Chinese Medicine, No.13 Wuhe Avenue, Nanning 530200, Guangxi, China.
Embryo Formation Teaching and Research Section, Guangxi University of Chinese Medicine, No.13 Wuhe Avenue, Nanning 530200, Guangxi, China.
出版信息
Arab J Gastroenterol. 2025 Feb;26(1):84-93. doi: 10.1016/j.ajg.2024.12.002. Epub 2025 Jan 4.
BACKGROUND AND STUDY AIMS
As a novel immunotherapy, chimeric antigen receptor T (CAR-T) cell technology is successful in treating hematologic malignancies, and exhibits potential benefits in partial solid tumors. Therapies targeting one antigen have some weaknesses, and dual-targeted CAR-T cells may be a better option. Alpha-fetoprotein (AFP) and glypican-3 (GPC3) are both highly expressed in hepatocellular carcinoma (HCC) and serve as important markers. Our study aimed to compare the cytotoxicity effect of AFP and GPC3 dual-targeted CAR-T cells on HCC cells in vitro and its therapeutic effects on a SCID xenograft model with those of single-targeted CAR-T cells.
MATERIALS AND METHODS
pLVX lentivirus vectors loaded with AFP CAR, GPC3 CAR, or AFP-GPC3 CAR constructs were transfected into human T lymphocytes. Control T, AFP CAR-T, GPC3 CAR-T, and AFP-GPC3 CAR-T cells were used as effector cells, and HLE (AFPGPC3), Sh-GPC3-Huh-7 (AFP), Sh-AFP-Huh-7 (GPC3), and Huh-7 (AFPGPC3) cells were used as target cells. After their co-culture for 6 h, the LDH cytotoxicity assay was employed to estimate the cell-killing effects of CAR-T cells on the target HCC cells. SCID mice bearing Huh-7 cell-derived neoplasms were injected with CAR-T cells, after which the pathological changes, CD3ζ expression, and IL-2 and IFN-γ levels in mouse tumor tissues were determined.
RESULTS
AFP and GPC3 were highly expressed in Huh-7 cells. AFP-GPC3 CAR-T cells exerted significant cell-killing effects on the HCC cells that expressed specific targeting antigen molecules (AFP and GPC3). Besides, AFP-GPC3 CAR-T cells better promoted Th cytokine secretion by Huh-7 cells than AFP CAR-T and GPC3 CAR-T cells. In vivo results suggested that AFP-GPC3 CAR-T cells better inhibited the growth of Huh-7 cell (AFPGPC3)-derived neoplasms than AFP CAR-T and GPC3 CAR-T cells.
CONCLUSION
AFP and GPC3 dual-targeted CAR-T cells showed better anti-tumor effects in HCC than AFP or GPC3 single-targeted CAR-T cells.
背景与研究目的
嵌合抗原受体T（CAR-T）细胞技术作为一种新型免疫疗法，在治疗血液系统恶性肿瘤方面取得了成功，并在部分实体瘤中显示出潜在益处。靶向单一抗原的疗法存在一些弱点，双靶点CAR-T细胞可能是更好的选择。甲胎蛋白（AFP）和磷脂酰肌醇蛋白聚糖-3（GPC3）在肝细胞癌（HCC）中均高表达，且均为重要标志物。本研究旨在比较AFP和GPC3双靶点CAR-T细胞对HCC细胞的体外细胞毒性作用及其对SCID异种移植模型的治疗效果与单靶点CAR-T细胞的差异。
材料与方法
将装载有AFP CAR、GPC3 CAR或AFP-GPC3 CAR构建体的pLVX慢病毒载体转染至人T淋巴细胞中。将对照T细胞、AFP CAR-T细胞、GPC3 CAR-T细胞和AFP-GPC3 CAR-T细胞用作效应细胞，将HLE（AFPGPC3）、Sh-GPC3-Huh-7（AFP）、Sh-AFP-Huh-7（GPC3）和Huh-7（AFPGPC3）细胞用作靶细胞。共培养6小时后，采用乳酸脱氢酶（LDH）细胞毒性测定法评估CAR-T细胞对靶HCC细胞的杀伤作用。对携带Huh-7细胞来源肿瘤的SCID小鼠注射CAR-T细胞，之后测定小鼠肿瘤组织中的病理变化、CD3ζ表达以及白细胞介素-2（IL-2）和干扰素-γ（IFN-γ）水平。
结果
AFP和GPC3在Huh-7细胞中高表达。AFP-GPC3 CAR-T细胞对表达特异性靶向抗原分子（AFP和GPC3）的HCC细胞具有显著的杀伤作用。此外，AFP-GPC3 CAR-T细胞比AFP CAR-T细胞和GPC3 CAR-T细胞能更好地促进Huh-7细胞分泌Th细胞因子。体内结果表明，AFP-GPC3 CAR-T细胞比AFP CAR-T细胞和GPC3 CAR-T细胞能更好地抑制Huh-7细胞（AFPGPC3）来源肿瘤的生长。
结论
AFP和GPC3双靶点CAR-T细胞在HCC中显示出比AFP或GPC3单靶点CAR-T细胞更好的抗肿瘤效果。
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