Preclinical Models for Cancer Therapeutic Development

项目概要/摘要 胰腺导管腺癌（Pancreatic ductal adenocarcinoma，PDA）是一种侵袭性强、致死率高的恶性肿瘤 目前的疗法。因此，我的研究重点是开发临床前模型， 更好的治疗策略。PDA的两个主要特征是KRAS突变的高频率 对靶向治疗反应差和广泛的促结缔组织增生性肿瘤微环境（TME） 由作为治疗屏障的致密细胞外基质（ECM）和多种非肿瘤细胞组成 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的特征导致其难以达到当前的标准治疗，需要定制的靶向治疗 来提高病人的存活率 正如我们以前报道的，在PDA发展过程中致癌Kras的激活导致了 氧化还原稳态和线粒体自噬途径，提供证据支持氧化还原靶向方法。我会 采用基因工程小鼠模型（GEMM）、类器官和类器官移植模型， PDA测试氧化还原疗法的潜在功效，特别是线粒体抑制剂或ROS诱导剂， 与MEKi（Kras信号传导的下游组分）组合。 我们先前的工作也确定了癌症相关成纤维细胞群体的异质性 （CAFs），每个都有自己独特的功能和活性途径。这些成纤维细胞包括肌纤维母细胞 （myCAF）、炎性（iCAF）和抗原呈递（apCAF）CAF。了解潜在的 其活性途径的机制对于开发消融肿瘤的治疗策略是必要的， 促进成纤维细胞特异性。我们报道了JAKI将CAF亚型向myCAF转移， 抑制肿瘤生长。我继续通过IL 1 R拮抗作用靶向其他活性iCAF信号通路 或使用我们的GEMM模型与免疫疗法组合递送抗LIF抗体。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， 促进癌细胞生长的CAF为此，我们将在我们的 使用单细胞RNA测序方法的新型导管内移植人类器官（IGO）模型。我会 建立一系列具有患者源性类器官的IGO模型，并使用这些小鼠来测试共 通过应用scRNA-seq分析的结果靶向癌细胞和促癌CAF。 最后，我将开发病毒诱导的PDA GEMM，它可以作为一个快速的平台来研究 在我们的转录组学或蛋白质组学数据集中鉴定的候选基因的重要性 和小鼠模型。总之，我将采用这些多种方法来研究PDA，它的主要驱动力 癌基因和异常改变的途径，以及周围的微环境将阐明关键途径 癌细胞需要这些途径作为新的治疗靶点的潜力。

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.