Preclinical Models for Cancer Therapeutic Development

项目摘要/摘要 摘要胰腺导管腺癌(Pda)是一种高度侵袭性和致命性的疾病，其疗效较差。 目前的治疗方法。因此，我的研究重点是开发用于识别的临床前模型。 更好的治疗策略。PDA的两个主要特征是KRAS突变频率高 这对靶向治疗和广泛的促结缔组织增生性肿瘤微环境(TME)反应不佳。 由致密的细胞外基质(ECM)和多个非肿瘤细胞组成，ECM充当治疗的屏障 类型包括癌症相关成纤维细胞(CAF)、内皮细胞和免疫细胞。这两个突出的 PDA的特点使其难以满足当前的护理标准，需要量身定做的靶向治疗 以提高患者的存活率。 正如我们之前所报道的，在PDA发育过程中，致癌Kras的激活会导致 氧化还原动态平衡和有丝分裂吞噬途径，为支持氧化还原靶向方法提供了证据。这就做 采用基因工程小鼠模型(GEMM)、有机类化合物和有机类化合物移植模型 PDA测试氧化还原疗法的潜在疗效，特别是线粒体抑制剂或ROS诱导剂 与Meki(Kras信令的下游组件)组合。 我们之前的工作也发现了癌症相关成纤维细胞群体的异质性。 (CAF)，每个CAF都有自己独特的功能和活跃的途径。这些成纤维细胞包括肌成纤维细胞 (MyCAF)、炎症性(ICAF)和抗原提呈(ApCAF)CAF。了解潜在的 它们的活性通路的机制对于开发治疗肿瘤的策略是必要的。 特别促进成纤维细胞。我们报告说，Jaki将CAF亚型转变为myCAF和 抑制肿瘤生长。我继续通过IL1R拮抗作用靶向其他活跃的ICAF信号通路 或使用我们的GEMM模型将抗LIF抗体与免疫治疗相结合。理解 不同类型的CAF如何促进肿瘤生长将为开发消融策略提供新的途径 促进癌细胞的CAF。为此，我们将在我们的 基于单细胞RNA测序方法的新型导管内植入类人器官模型。这就做 用患者衍生的有机化合物建立一系列IGO模型，并用这些小鼠来测试联合治疗的效果。 通过应用scRNA-seq分析的结果，靶向癌细胞和促进癌症的CAF。 最后，我将开发病毒诱导的PDA GEMM，作为一个快速平台来研究 在我们的转录组或蛋白质组数据集中识别的候选基因的重要性来自我们的有机化合物 和老鼠模型。综上所述，我将采用这些多种方法来研究PDA，它的主要驱动因素 癌基因和异常改变的通路，以及周围的微环境将阐明关键通路 癌细胞需要有潜力的这些通路作为新的治疗靶点。

PROJECT SUMMARY/ABSTRACT Pancreatic ductal adenocarcinoma (PDA) is a highly aggressive and lethal disease due to the poor efficacy of current therapies. Therefore, my research focuses on development of preclinical models for the identification of better therapeutic strategies. Two main distinct features of PDA are the high frequency of KRAS mutations that is poorly responsive to targeted therapies and an extensive desmoplastic tumor microenvironment (TME) composed of a dense extracellular matrix (ECM), acting as a barrier to therapy, and multiple non-neoplastic cell types including cancer-associated fibroblasts (CAF), endothelial cells, and immune cells. These two prominent features of PDA contribute to its intractability to current standard-of-care, calling for tailored targeted therapies to improve patients’ survival. As we previously reported, activation of oncogenic Kras during PDA development results in alterations to redox homeostasis and mitophagy pathways, providing evidence to support a redox-targeting approach. I will employ genetically engineered mouse models (GEMMs), organoids, and organoid transplantation models of PDA to test the potential efficacy of redox therapies, in particular mitochondrial inhibitors or ROS inducers in combination with MEKi (downstream component of Kras signaling). Our prior work has also identified heterogeneity within the population of cancer-associated fibroblasts (CAFs), each with their own distinct functions and active pathways. These fibroblasts include myofibroblastic (myCAFs), inflammatory (iCAFs) and antigen-presenting (apCAFs) CAFs. Understanding the underlying mechanisms of their active pathways is necessary for the development of therapeutic strategies to ablate tumor- promoting fibroblasts specifically. We reported that JAKi shifted the CAF subtypes towards myCAFs and suppressed tumor growth. I continue to target other active iCAF-signaling pathways through IL1R antagonism or delivery of anti-LIF antibodies in combination with immunotherapy using our GEMM models. Understanding how different types of CAFs contribute to tumor growth will provide a new avenue to develop strategies to ablate the cancer cell-promoting CAFs. To this end, we will uncover the identities and functions of these CAFs in our novel intraductally engrafted human organoid (IGO) model using a single-cell RNA sequencing approach. I will establish a series of IGO models with patient-derived organoids and use these mice to test the efficacy of co- targeting cancer cells and cancer-promoting CAFs by applying the findings from scRNA-seq analysis. Lastly, I will develop viral-induced GEMMs of PDA that can serve as a rapid platform to investigate the importance of candidate genes identified in our transcriptomic or proteomic datasets derived from our organoid and mouse models. Taken together, these multiple approaches I will employ to studying PDA, its primary driving oncogene and aberrantly altered pathways, and the surrounding microenvironment will elucidate key pathways the cancer cells require with the potential of these pathways acting as new therapeutic targets.