Zhao Shushu, Chen Yeqing, Bhojnagarwala Pratik S, Livingston Cory, Jose Joshua, Gao Yangcheng, Park Daniel H, Herlyn Meenhard, Weiner David B
Wistar Institute of Anatomy and Biology, Philadelphia, Pennsylvania, USA.
University of Pennsylvania, Philadelphia, Pennsylvania, USA.
J Immunother Cancer. 2025 Jun 8;13(6):e011073. doi: 10.1136/jitc-2024-011073.
Melanoma is a highly aggressive skin cancer, especially in advanced stages. While current treatments such as targeted therapies and immunotherapies have made significant progress, challenges like drug resistance and limited effectiveness in some patients persist. Therefore, ongoing development of novel therapies, particularly for late-stage melanoma, is crucial.
In this study, we explored the expression of interleukin-13 receptor subunit alpha-2 (IL13Rα2) in melanoma patient-derived xenograft models. We investigated IL13Rα2 as a potential target for melanoma treatment by employing an IL13Rα2-CD3 bispecific T-cell engager (BTE). We tested the effect of IL13Rα2-CD3 BTE on T cell activity by flow cytometry. We studied the potency of IL13Rα2-CD3 BTE in tumor killing assay in vitro. For in vivo studies, we administered DNA expression cassettes encoding IL13Rα2-CD3 BTE (IL13Rα2-CD3 DNA encoding BTE (dBTE)) into immunodeficient mice for direct in vivo expression. The mice were challenged with A375 cells and then treated with IL-13Rα2-CD3 dBTE versus control and reconstituted with human peripheral blood mononuclear cells (PBMCs) or T cells. Tumor development was monitored, and T cell infiltration in the tumor was analyzed throughflow cytometry.
Our findings revealed heterogeneous expression of IL-13Rα2, particularly in samples from advanced stages of melanoma. The IL13Rα2-CD3 BTE facilitated T-cell activation and proliferation by bridging melanoma cells and T cells. We also observed the ability of IL13Rα2-CD3 BTE to direct T cells to kill multiple melanoma patient-derived cell lines through xCELLigence assay in vitro, including those with various mutations associated with late-stage metastatic melanoma. IL13Rα2-CD3 dBTE expressed in vivo led to notable tumor regression through inducing increased T-cell infiltration and activation within the tumor microenvironment.
These promising findings underscore the potential of targeting IL13Rα2 as a relevant target for the development of biologics including dBTE aimed at treating specific subsets of melanoma.
黑色素瘤是一种侵袭性很强的皮肤癌，尤其是在晚期。虽然目前的治疗方法，如靶向治疗和免疫治疗已经取得了显著进展，但耐药性和对某些患者疗效有限等挑战仍然存在。因此，持续开发新的治疗方法，特别是针对晚期黑色素瘤的治疗方法至关重要。
在本研究中，我们在黑色素瘤患者来源的异种移植模型中探索了白细胞介素-13受体亚基α-2（IL13Rα2）的表达。我们通过使用IL13Rα2-CD3双特异性T细胞衔接器（BTE）研究IL13Rα2作为黑色素瘤治疗的潜在靶点。我们通过流式细胞术测试了IL13Rα2-CD3 BTE对T细胞活性的影响。我们在体外肿瘤杀伤试验中研究了IL13Rα2-CD3 BTE的效力。对于体内研究，我们将编码IL13Rα2-CD3 BTE的DNA表达盒（IL13Rα2-CD3 DNA编码BTE（dBTE））注射到免疫缺陷小鼠体内以进行直接体内表达。用A375细胞攻击小鼠，然后用IL-13Rα2-CD3 dBTE与对照进行处理，并用人类外周血单核细胞（PBMC）或T细胞进行重建。监测肿瘤发展，并通过流式细胞术分析肿瘤中的T细胞浸润情况。
我们的研究结果揭示了IL-13Rα2的异质性表达，特别是在黑色素瘤晚期样本中。IL13Rα2-CD3 BTE通过连接黑色素瘤细胞和T细胞促进T细胞活化和增殖。我们还通过体外xCELLigence分析观察到IL13Rα2-CD3 BTE能够引导T细胞杀死多种黑色素瘤患者来源的细胞系，包括那些与晚期转移性黑色素瘤相关的各种突变细胞系。体内表达的IL13Rα2-CD3 dBTE通过诱导肿瘤微环境中T细胞浸润和活化增加，导致显著的肿瘤消退。
这些有前景的发现强调了将IL13Rα2作为开发包括dBTE在内的生物制剂的相关靶点的潜力，这些生物制剂旨在治疗特定亚型的黑色素瘤。
