PO.CL05.08 · 临床研究
利用多功能纳米衔接器在人源化肾癌中直接体内激活并功能重编程人TILs
Direct in vivo activation and functional reprogramming of human TILs using a multifunctional nanoengager in humanized renal carcinoma
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
引言：肿瘤浸润淋巴细胞（TILs）疗法在黑色素瘤中取得了显著的临床成功。然而，其在其他癌症中的应用受到高制造成本和毒性预处理方案的限制。为克服这些局限，我们旨在开发一种利用多功能纳米衔接器（TIL-NPs）在体内直接激活并扩增TILs的新策略。在本研究中，我们报道了使用癌症小鼠模型（包括一个高度重现人类免疫-肿瘤相互作用的人源化肾癌模型）对TIL-NPs的验证。
方法：TIL-NPs通过将叠氮基功能化的PEG-PLGA纳米颗粒与DBCO修饰的抗CD3、抗4-1BB和IL-2偶联而生成。使用MC38和786-O模型评估其激活和扩增TILs的能力。在MC38研究中，携带双侧肿瘤的小鼠仅在右侧腹肿瘤内接受TIL-NPs，并使用分离的MC38 TILs与MC38细胞体外共培养进一步评估TIL激活。为评估人类相关性，使用肾细胞癌（RCC）患者样本测试了TIL-NP介导的人TILs扩增，并通过将786-O细胞接种到植入人PBMCs的NSG小鼠中建立了人源化RCC模型。
结果：我们构建了展示抗CD3、抗4-1BB和IL-2的多功能纳米颗粒（TIL-NPs），尺寸约126 nm，每mg NP偶联约50 μg免疫激动剂。体外，用TIL-NPs刺激的MC38肿瘤来源TILs对MC38细胞显示出显著增强的细胞毒性，并产生了相比游离抗体或未刺激TILs最高的IFN-gamma水平。体内，在双侧MC38模型中瘤内递送TIL-NPs诱导所有注射肿瘤完全消退，并产生强烈的全身反应，对侧肿瘤显著消退且生存改善。使用RCC患者来源的TILs，TIL-NPs相比游离抗体也增强了对786-O和ACHN细胞的细胞毒性。在人源化RCC模型中，TIL-NPs使治疗肿瘤近乎完全消退，并显著抑制对侧肿瘤生长。免疫组化证实TIL-NP治疗的肿瘤中CD8⁺T细胞浸润密集，表明细胞毒性T细胞的激活和募集增强。总体而言，TIL-NPs通过受体聚集增强T细胞信号传导，延长生存，并引发强效的局部和全身抗肿瘤反应。
结论：TIL-NPs可在体内强效激活并重编程TILs，增强细胞毒性和全身抗肿瘤免疫，凸显了其在实体瘤过继性T细胞治疗中的转化潜力。
查看英文原文 English abstract
INTRODUCTION: Tumor-infiltrating lymphocytes (TILs) therapy has demonstrated remarkable clinical success in melanoma. However, its application in other cancers has been limited by high manufacturing cost and toxic conditioning regimen. To overcome these limitations, we aimed to develop a novel strategy to directly activate and expand TILs in vivo using a multifunctional nanoengager (TIL-NPs). In this study, we report the validation of TIL-NPs using mouse models of cancer, including a humanized renal carcinoma model that closely recapitulates human immune-tumor interactions.
METHODS: TIL-NPs were generated by conjugating azide-functionalized PEG-PLGA nanoparticles with DBCO-modified anti-CD3, anti-4-1BB, and IL-2. Their ability to activate and expand TILs was assessed using both MC38 and 786-O models. For the MC38 study, mice bearing bilateral tumors received intratumoral TIL-NPs in the right flank tumor only, and TIL activation was further evaluated using isolated MC38 TILs co-cultured with MC38 cells in vitro. To assess human relevance, TIL-NP-mediated expansion of human TILs was tested using renal cell carcinoma (RCC) patient samples, and a humanized RCC model was established by inoculating 786-O cells into NSG mice engrafted with human PBMCs.
RESULTS: We engineered multifunctional nanoparticles (TIL-NPs) displaying anti-CD3, anti-4-1BB, and IL-2, measuring ~126 nm with ~50 μg of immune agonists conjugated per mg NP. In vitro, MC38 tumor-derived TILs stimulated with TIL-NPs showed markedly enhanced cytotoxicity against MC38 cells and produced the highest IFN-gamma levels compared with free antibodies or unstimulated TILs. In vivo, intratumoral delivery of TIL-NPs in a bilateral MC38 model induced complete regression of all injected tumors and generated a strong systemic response, with substantial regression of contralateral tumors and improved survival. Using RCC patient-derived TILs, TIL-NPs also enhanced cytotoxicity against 786-O and ACHN cells relative to free antibodies. In a humanized RCC model, TIL-NPs produced near-complete regression of treated tumors and markedly suppressed contralateral tumor growth. Immunohistochemistry confirmed dense CD8⁺ T-cell infiltration in TIL-NP-treated tumors, indicating enhanced activation and recruitment of cytotoxic T cells. Overall, TIL-NPs strengthened T-cell signaling through receptor clustering, prolonged survival, and elicited potent local and systemic antitumor responses.
CONCLUSION: TIL-NPs potently activate and reprogram TILs in vivo, enhancing cytotoxicity and systemic antitumor immunity, highlighting their translational potential for adoptive T-cell therapy in solid tumors.
利益披露 Disclosure
M. Wang, None..
L. Ruan, None..
X. Wen, None..
T. Zhang, None.