Cancer-associated fibroblasts and extracellular matrix remodeling in pediatric neuroblastoma

项目总结 细胞外基质(ECM)重塑是实体瘤微环境中的一个重要过程。癌症-- 相关成纤维细胞(CAF)是基质蛋白和重塑酶的主要生产者，这些蛋白和重塑酶可交叉 连接和降解基质蛋白质。CAF是一个谜，因为它们可以促进或阻碍肿瘤的生长， 暗示着不完全理解的异质性。CAF异质性是如何超过肿瘤的 进展和这种演变如何影响实体肿瘤内ECM重塑仍然是一个悬而未决的问题。 在癌症生物学方面。最近的研究主要集中在CAF在成人癌症模型中的作用，但很少有人关注。 旨在了解儿童癌症中CAF的异质性和ECM重塑，这是 是病因不同的疾病。 神经母细胞瘤是一种儿童神经内分泌细胞癌，占儿童癌症的15%。 死亡。高危NB的5年总体存活率保持在50%，突显出需要更深层次的 了解推动这类患者肿瘤发生的分子和免疫学机制。 与其他实体肿瘤一样，NB肿瘤微环境包括恶性细胞、CAF和免疫细胞。我们的 实验室使用高危Nb的渗透性自发模型(TH-MYCN，AlkF1178L/+)研究Nb 免疫活性小鼠，允许体内研究肿瘤免疫微环境 从肿瘤发生到进行性疾病。使用这个模型，我们以前展示了促肿瘤发生的作用 通过显示巨噬细胞和CD4+T细胞的耗尽显著减少肿瘤的形成和 延长存活期。 应用10倍铬基因表达分析新生大鼠骨髓基质细胞的初步实验 成熟肿瘤发现了两个不同的CAF亚群，其特征是α-平滑肌肌动蛋白(Acta2)和 Wilms肿瘤1(Wt1)的表达。有趣的是，只有Wt1 CAF表达趋化因子和细胞因子参与 髓样细胞和T细胞的募集。WT1 CAF在早期肿瘤微环境中丰富并表达 编码ADAMTS酶的基因与蛋白多糖的降解有关。我们的预赛 研究结果表明，Wt1 CAF可能是我们模型中促肿瘤免疫细胞的关键早期募集者。 神经母细胞瘤。建议的研究将评估CAF的异质性如何影响ECM的重塑。 神经母细胞瘤的进展，检测肿瘤-CAF和CAF-免疫信号，并测试扰动的效果 细胞外基质沉积和降解对肿瘤形成和发展的影响。总之，这些研究将产生新的 和令人兴奋的信息，关于肿瘤微环境和潜在的脆弱性，可在儿科 实体瘤。

PROJECT SUMMARY Extracellular matrix (ECM) remodeling is a critical process within the solid tumor microenvironment. Cancer- associated fibroblasts (CAFs) are the primary producers of matrix proteins and remodeling enzymes that cross- link and degrade matrix proteins. CAFs are enigmatic in that they can promote or impede tumor growth, suggesting heterogeneity that is not completely understood. How CAF heterogeneity evolves over tumor progression and how this evolution impacts ECM remodeling within solid tumors remain an outstanding question in cancer biology. Recent studies have focused on CAF function in models of adult cancer, but little attention has been directed toward understanding CAF heterogeneity and ECM remodeling in childhood cancers, which are etiologically different diseases. Neuroblastoma (NB) is a childhood cancer of neuroendocrine cells that accounts for 15% of pediatric cancer deaths. The 5-year overall survival rate for high-risk NB remains at 50%, highlighting the need for a deeper understanding of the molecular and immunological mechanisms driving tumorigenesis in this subset of patients. Like other solid tumors, the NB tumor microenvironment includes malignant cells, CAFs, and immune cells. Our laboratory studies NB using a penetrant, spontaneous model of high-risk NB (TH-MYCN, AlkF1178L/+) in immunocompetent mice, allowing in vivo investigation of the tumor immune microenvironment from tumorigenesis to progressive disease. Using this model, we previously demonstrated the pro-tumorigenic role of macrophages and CD4+ T cells by showing that their depletion significantly reduced the formation of tumors and extended survival. My preliminary experiments utilizing 10X Chromium gene expression analysis of stromal cells from nascent and mature tumors identified two distinct subsets of CAFs characterized by alpha smooth muscle actin (Acta2) and Wilms tumor 1 (Wt1) expression. Interestingly, only Wt1 CAFs expressed chemokines and cytokines involved in recruitment of myeloid and T cells. Wt1 CAFs were enriched in the early tumor microenvironment and expressed genes encoding ADAMTS enzymes involved in degradation of the proteoglycan versican. Our preliminary findings indicate that Wt1 CAFs may be a crucial early recruiter of pro-tumorigenic immune cells in our model of neuroblastoma. The studies proposed will assess how CAF heterogeneity affects ECM remodeling over neuroblastoma progression, examine tumor-CAF and CAF-immune signaling, and test the effect of perturbing ECM deposition and degradation on tumor formation and progression. Together, these studies will generate new and exciting information about the tumor microenvironment and potential vulnerabilities to exploit in pediatric solid tumors.