免疫疗法已成为一种有前景的癌症治疗方法，溶瘤腺病毒显示出作为免疫治疗剂的功效。在这项研究中，我们研究了表达 CXCL9、CXCL10 或 IL-15 的腺病毒构建体在透明细胞肾细胞癌 (ccRCC) 肿瘤模型中的免疫治疗潜力。我们的结果表明病毒治疗后细胞因子分泌强劲，表明转基因表达有效。随后使用基于电阻的 Transwell 迁移和微流控芯片分析进行的分析表明，在 2D 和 3D 细胞模型中，受感染细胞分泌趋化因子后，T 细胞迁移增加。流式细胞术分析显示，CXCR3 受体在 T 细胞亚群中表达，其中 CD8 T 细胞的百分比最高，强调了它们在免疫细胞迁移中的关键作用。除了 T 细胞外，我们还在用编码细胞因子的腺病毒治疗的免疫受损小鼠的肿瘤中检测到 NK 细胞。此外，我们还发现了潜在的免疫原性抗原，可以增强我们的武装溶瘤腺病毒在 ccRCC 中的功效和特异性。总体而言，我们使用 ccRCC 细胞系、体内人源化小鼠、2D 生理相关 PDC 和 3D 患者源性类器官 (PDO) 的研究结果表明，带有趋化因子的腺病毒有望增强 ccRCC 中的 T 细胞迁移并改善免疫治疗结果。我们的研究通过阐明肿瘤微环境（TME）内的免疫细胞浸润和激活机制，有助于开发更有效的 ccRCC 治疗策略，并强调 PDO 在预测临床相关性和验证新型免疫治疗方法方面的有用性。总体而言，我们的研究为 ccRCC 病毒免疫疗法的合理设计和优化提供了见解。© 2024 作者。经泰勒许可出版

Immunotherapy has emerged as a promising approach for cancer treatment, with oncolytic adenoviruses showing power as immunotherapeutic agents. In this study, we investigated the immunotherapeutic potential of an adenovirus construct expressing CXCL9, CXCL10, or IL-15 in clear cell renal cell carcinoma (ccRCC) tumor models. Our results demonstrated robust cytokine secretion upon viral treatment, suggesting effective transgene expression. Subsequent analysis using resistance-based transwell migration and microfluidic chip assays demonstrated increased T-cell migration in response to chemokine secretion by infected cells in both 2D and 3D cell models. Flow cytometry analysis revealed CXCR3 receptor expression across T-cell subsets, with the highest percentage found on CD8+ T-cells, underscoring their key role in immune cell migration. Alongside T-cells, we also detected NK-cells in the tumors of immunocompromised mice treated with cytokine-encoding adenoviruses. Furthermore, we identified potential immunogenic antigens that may enhance the efficacy and specificity of our armed oncolytic adenoviruses in ccRCC. Overall, our findings using ccRCC cell line, in vivo humanized mice, physiologically relevant PDCs in 2D and patient-derived organoids (PDOs) in 3D suggest that chemokine-armed adenoviruses hold promise for enhancing T-cell migration and improving immunotherapy outcomes in ccRCC. Our study contributes to the development of more effective ccRCC treatment strategies by elucidating immune cell infiltration and activation mechanisms within the tumor microenvironment (TME) and highlights the usefulness of PDOs for predicting clinical relevance and validating novel immunotherapeutic approaches. Overall, our research offers insights into the rational design and optimization of viral-based immunotherapies for ccRCC.© 2024 The Author(s). Published with license by Taylor & Francis Group, LLC.