PDF(Pubmed)
Abstract：
BACKGROUND: Pancreatic cancer is one of the most lethal malignancies and the lack of treatment options makes it more deadly. Chimeric Antigen Receptor T-cell (CAR-T) immunotherapy has revolutionized cancer treatment and made great breakthroughs in treating hematological malignancies, however its success in treating solid cancers remains limited mainly due to the lack of tumor-specific antigens. On the other hand, the prolonged traditional manufacturing process poses challenges, taking 2 to 6 weeks and impacting patient outcomes. CD276 has recently emerged as a potential therapeutic target for anti-solid cancer therapy. Here, we investigated the efficacy of CD276 CAR-T and rapidly-manufactured CAR-T against pancreatic cancer.
METHODS: In the present study, CD276 CAR-T was prepared by CAR structure carrying 376.96 scFv sequence, CD8 hinge and transmembrane domain, 4-1BB and CD3ζ intracellular domains. Additionally, CD276 rapidly-manufactured CAR-T (named CD276 Dash CAR-T) was innovatively developed by shortening the duration of ex vitro culture to reduce CAR-T manufacturing time. We evaluated the anti-tumor efficacy of CD276 CAR-T and further compared the functional assessment of Dash CAR-T and conventional CAR-T in vitro and in vivo by detecting the immunophenotypes, killing ability, expansion capacity and tumor-eradicating effect of CAR-T.
RESULTS: We found that CD276 was strongly expressed in multiple solid cancer cell lines and that CD276 CAR-T could efficiently kill these solid cancer cells. Moreover, Dash CAR-T was successfully manufactured within 48-72 h and the functional validation was carried out subsequently. In vitro, CD276 Dash CAR-T possessed a less-differentiated phenotype and robust proliferative ability compared to conventional CAR-T. In vivo xenograft mouse model, CD276 Dash CAR-T showed enhanced anti-pancreatic cancer efficacy and T cell expansion. Besides, except for the high-dose group, the body weight of mice was maintained stable, and the state of mice was normal.
CONCLUSIONS: In this study, we proved CD276 CAR-T exhibited powerful activity against pancreatic cancer cells in vitro and in vivo. More importantly, we demonstrated the manufacturing feasibility, acceptable safety and superior anti-tumor efficacy of CD276 Dash CAR-T generated with reduced time. The results of the above studies indicated that CD276 Dash CAR-T immunotherapy might be a novel and promising strategy for pancreatic cancer treatment.
METHODS: In the present study, CD276 CAR-T was prepared by CAR structure carrying 376.96 scFv sequence, CD8 hinge and transmembrane domain, 4-1BB and CD3ζ intracellular domains. Additionally, CD276 rapidly-manufactured CAR-T (named CD276 Dash CAR-T) was innovatively developed by shortening the duration of ex vitro culture to reduce CAR-T manufacturing time. We evaluated the anti-tumor efficacy of CD276 CAR-T and further compared the functional assessment of Dash CAR-T and conventional CAR-T in vitro and in vivo by detecting the immunophenotypes, killing ability, expansion capacity and tumor-eradicating effect of CAR-T.
RESULTS: We found that CD276 was strongly expressed in multiple solid cancer cell lines and that CD276 CAR-T could efficiently kill these solid cancer cells. Moreover, Dash CAR-T was successfully manufactured within 48-72 h and the functional validation was carried out subsequently. In vitro, CD276 Dash CAR-T possessed a less-differentiated phenotype and robust proliferative ability compared to conventional CAR-T. In vivo xenograft mouse model, CD276 Dash CAR-T showed enhanced anti-pancreatic cancer efficacy and T cell expansion. Besides, except for the high-dose group, the body weight of mice was maintained stable, and the state of mice was normal.
CONCLUSIONS: In this study, we proved CD276 CAR-T exhibited powerful activity against pancreatic cancer cells in vitro and in vivo. More importantly, we demonstrated the manufacturing feasibility, acceptable safety and superior anti-tumor efficacy of CD276 Dash CAR-T generated with reduced time. The results of the above studies indicated that CD276 Dash CAR-T immunotherapy might be a novel and promising strategy for pancreatic cancer treatment.
摘要：
背景：胰腺癌是最致命的恶性肿瘤之一，缺乏治疗选择使其更加致命。嵌合抗原受体T细胞（CAR-T）免疫疗法彻底改变了癌症治疗，并在治疗血液恶性肿瘤方面取得了重大突破，然而，它在治疗实体癌方面的成功仍然有限，主要是由于缺乏肿瘤特异性抗原。另一方面,延长的传统制造过程带来了挑战，服用2至6周，影响患者预后。CD276最近已成为抗实体癌治疗的潜在治疗靶标。这里,我们研究了CD276CAR-T和快速制造的CAR-T对胰腺癌的疗效.
方法：在本研究中，CD276CAR-T是通过CAR结构制备的，携带376.96scFv序列，CD8铰链和跨膜结构域，4-1BB和CD3ζ胞内结构域。此外,CD276快速制造的CAR-T（名为CD276DashCAR-T）是通过缩短体外培养时间以减少CAR-T制造时间而创新开发的。我们评估了CD276CAR-T的抗肿瘤功效，并通过检测免疫表型进一步比较了DashCAR-T和常规CAR-T在体外和体内的功能评估。杀伤能力,CAR-T的扩增能力和肿瘤根除作用
结果：我们发现CD276在多种实体癌细胞系中强烈表达，CD276CAR-T可以有效杀死这些实体癌细胞。此外，DashCAR-T在48-72小时内成功制造，随后进行了功能验证。体外，与常规CAR-T相比，CD276DashCAR-T具有较低分化的表型和强大的增殖能力。体内异种移植小鼠模型,CD276DashCAR-T显示出增强的抗胰腺癌功效和T细胞扩增。此外,除了高剂量组,小鼠体重保持稳定,老鼠的状态正常。
结论：在这项研究中，我们证明了CD276CAR-T在体外和体内对胰腺癌细胞表现出强大的活性。更重要的是,我们证明了制造的可行性，以减少的时间产生的CD276DashCAR-T具有可接受的安全性和优异的抗肿瘤功效。上述研究结果表明，CD276DashCAR-T免疫疗法可能是一种新颖且有前途的胰腺癌治疗策略。
方法：在本研究中，CD276CAR-T是通过CAR结构制备的，携带376.96scFv序列，CD8铰链和跨膜结构域，4-1BB和CD3ζ胞内结构域。此外,CD276快速制造的CAR-T（名为CD276DashCAR-T）是通过缩短体外培养时间以减少CAR-T制造时间而创新开发的。我们评估了CD276CAR-T的抗肿瘤功效，并通过检测免疫表型进一步比较了DashCAR-T和常规CAR-T在体外和体内的功能评估。杀伤能力,CAR-T的扩增能力和肿瘤根除作用
结果：我们发现CD276在多种实体癌细胞系中强烈表达，CD276CAR-T可以有效杀死这些实体癌细胞。此外，DashCAR-T在48-72小时内成功制造，随后进行了功能验证。体外，与常规CAR-T相比，CD276DashCAR-T具有较低分化的表型和强大的增殖能力。体内异种移植小鼠模型,CD276DashCAR-T显示出增强的抗胰腺癌功效和T细胞扩增。此外,除了高剂量组,小鼠体重保持稳定,老鼠的状态正常。
结论：在这项研究中，我们证明了CD276CAR-T在体外和体内对胰腺癌细胞表现出强大的活性。更重要的是,我们证明了制造的可行性，以减少的时间产生的CD276DashCAR-T具有可接受的安全性和优异的抗肿瘤功效。上述研究结果表明，CD276DashCAR-T免疫疗法可能是一种新颖且有前途的胰腺癌治疗策略。
