Giudice Anna Maria, Roth Sydney L, Matlaga Stephanie, Cresswell-Clay Evan, Mishra Pamela, Schürch Patrick M, Boateng-Antwi Kwame Attah M, Samanta Minu, Pascual-Pasto Guillem, Zecchino Vincent, Spear Timothy T, McIntyre Brendan, Chada Neil C, Wang Tingting, Liu Lingling, Wang Ruoning, Wilson John T, Wolpaw Adam J, Bosse Kristopher R
Division of Oncology and Center for Childhood Cancer Research, Children's Hospital of Philadelphia, Philadelphia, PA 19104, USA.
Division of Oncology and Center for Childhood Cancer Research, Children's Hospital of Philadelphia, Philadelphia, PA 19104, USA; Department of Biomedical and Health Informatics, Children's Hospital of Philadelphia, Philadelphia, PA 19104, USA.
Mol Ther. 2025 Sep 3;33(9):4552-4569. doi: 10.1016/j.ymthe.2025.05.025. Epub 2025 May 27.
Poor tumor trafficking and the immunosuppressive tumor microenvironment (TME) limit chimeric antigen receptor (CAR) T cell efficacy in solid tumors, such as neuroblastoma. We previously optimized GPC2 CARs in human neuroblastoma xenografts leading to clinical translation; however, there have not been preclinical studies using immunocompetent models. Thus, here we generated murine GPC2 CAR T cells using the D3-GPC2-targeting single-chain variable fragment being utilized clinically (NCT05650749) and tested them in neuroblastoma syngeneic allografts. Immune-profiling of GPC2 CAR T cell-treated tumors revealed significant reprogramming of the TME, most notably poor intra-tumor CAR T cell persistence being associated with increased recruitment of myeloid-derived suppressor cells (MDSCs), along with MDSC-recruiting CXCL1/2 chemokines. These tumor-infiltrating MDSCs directly inhibited GPC2 CAR T cell activation, proliferation, and cytotoxicity ex vivo. To both capitalize on this chemokine gradient and mitigate MDSC-tumor trafficking, we engineered GPC2 CAR T cells to express the CXCL1/2 receptor, CXCR2. CXCR2-armored GPC2 CAR T cells migrated toward CXCL1/2 gradients, enhanced anti-neuroblastoma efficacy, and reduced the level of MDSCs in the TME. Together, these findings suggest CAR T cell studies in immunocompetent models are imperative to define mechanisms of solid tumor immune escape and rationally design armoring strategies that will lead to durable clinical efficacy.
肿瘤归巢能力差以及免疫抑制性肿瘤微环境（TME）限制了嵌合抗原受体（CAR）T细胞在实体瘤（如神经母细胞瘤）中的疗效。我们之前在人神经母细胞瘤异种移植模型中优化了GPC2 CAR，从而推动了其临床转化；然而，尚未有使用免疫活性模型的临床前研究。因此，我们在此使用临床上正在使用的靶向D3-GPC2的单链可变片段生成了小鼠GPC2 CAR T细胞，并在神经母细胞瘤同基因异体移植模型中对其进行了测试。对接受GPC2 CAR T细胞治疗的肿瘤进行免疫分析发现，TME发生了显著的重编程，最明显的是肿瘤内CAR T细胞持久性差与髓系来源抑制细胞（MDSC）募集增加以及MDSC募集趋化因子CXCL1/2有关。这些肿瘤浸润性MDSC在体外直接抑制GPC2 CAR T细胞的活化、增殖和细胞毒性。为了利用这种趋化因子梯度并减轻MDSC向肿瘤的归巢，我们对GPC2 CAR T细胞进行基因工程改造，使其表达CXCL1/2受体CXCR2。CXCR2武装的GPC2 CAR T细胞向CXCL1/2梯度迁移，增强了抗神经母细胞瘤的疗效，并降低了TME中MDSC的水平。总之，这些发现表明，在免疫活性模型中进行CAR T细胞研究对于确定实体瘤免疫逃逸机制并合理设计能带来持久临床疗效的武装策略至关重要。
