1. Guangdong Cardiovsacular Institute, Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Science, Guangzhou 510080, China 2. The Second Clinical Medical College, Guangzhou University of Chinese Medicine, Department of Laboratory Medicine/State Key Laboratory of Traditional Chinese Medicine Syndrome, Guangdong Provincial Hospital of Chinese Medicine, Guangzhou 510120, China 3. Department of Medical Oncology, The First Affiliated Hospital of Guangxi Medical University, Nanning 530021, China 4. Department of Stomatology, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, China 5. Medical Research Institute, Guangdong Provincial People’s Hospital (Guangdong Academy of Medical Science), Southern Medical University, Guangzhou 510080, China 6. School of Food and Drug, Shenzhen Polytechnic University, Shenzhen 518055, China 7. Shenzhen Second People’s Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen 518055, China
CD39 serves as a crucial biomarker for neoantigen-specific CD8+ T cells and is associated with antitumor activity and exhaustion. However, the relationship between CD39 expression levels and the function of chimeric antigen receptor T (CAR-T) cells remains controversial. This study aimed to investigate the role of CD39 in the functional performance of CAR-T cells against hepatocellular carcinoma (HCC) and explore the therapeutic potential of CD39 modulators, such as mitochondrial division inhibitor-1 (mdivi-1), or knockdown CD39 through short hairpin RNA. Our findings demonstrated that glypican-3-CAR-T cells with moderate CD39 expression exhibited a strong antitumor activity, while high and low levels of CD39 led to an impaired cellular function. Methods modulating the proportion of CD39 intermediate (CD39int)-phenotype CAR-T cells such as mdivi-1 and CD39 knockdown enhanced and impaired T cell function, respectively. The combination of mdivi-1 and CD39 knockdown in CAR-T cells yielded the highest proportion of infiltrated CD39int CAR-T cells and demonstrated a robust antitumor activity in vivo. In conclusion, this study revealed the crucial role of CD39 in CAR-T cell function, demonstrated the potential therapeutic efficacy of combining mdivi-1 with CD39 knockdown in HCC, and provided a novel treatment strategy for HCC patients in the field of cellular immunotherapy.
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