CSHL-JAX Patient-Derived Models of Pancreatic Cancer as Systems for Investigating Tumor Heterogeneity

项目总结 摘要胰腺导管腺癌(Pda)是一种高致命性的恶性肿瘤，其五年存活率较低。 而不是8%。这些严峻的统计数据在一定程度上归因于PDA对当前疗法的深刻抵制。更多 有效的治疗策略将为PDA患者带来巨大的好处。新出现的证据表明 表观遗传和遗传异质性同时存在于肿瘤和非肿瘤间隔中。 胰腺肿瘤，这种异质性可能会影响治疗反应。我们自己的研究已经 鉴定了PDA中存在的癌症相关成纤维细胞(CAF)的两个亚群 微环境，表达肌成纤维细胞标志物的癌近端群体和癌远端 表达炎症标志物的人群，如细胞因子IL-6。然而，全额的 PDA肿瘤内存在的细胞群及其可能通过的机制 对肿瘤生物学的影响尚不清楚。解决这些问题需要准确概括的模型 人体掌上电脑。患者来源的异种移植物(PDX)、类器官培养、类器官-基质共培养和类器官 异种移植物已经成为“下一代”模型，它能更好地模拟存在于 掌上电脑的微环境。然而，每个模型在多大程度上保留了在 人类肿瘤仍不清楚。为了解决这个问题，我们建议生成一系列匹配 我们将用来描述和扰动肿瘤亚群的器官和异种移植模型 细胞和CAF。我们将生成并表征五组配对的器官培养和PDX模型，其中 配对是从相同的未鉴定的患者肿瘤标本中提取的。此外，我们还将开发一种新的模式， 在胰腺癌中，患者衍生的PDA有机类化合物通过注射到小鼠体内而原位传递给小鼠。 主胰管，比其他异种移植更能概括人PDA的发展轨迹 接近了。我们将使用单细胞RNA和DNA测序以及条形码有机系来 描述和扰乱我们模型中存在的肿瘤和CAF人群。白介素6在脑出血中的作用研究 更直接地，我们将使用生化和遗传策略来干扰CAF介导的IL-6信号转导。 我们的模特。其中，我们将利用一种经过改造的小鼠模型来表达人IL-6作为宿主 我们的PDA异种移植，恢复了基质中产生的IL-6向移植的人发出信号的能力 肿瘤细胞。最终，我们的研究将阐明不同的肿瘤细胞亚群和 CAF存在于PDA微环境中，并有助于识别这些亚群中的哪些发挥功能 促进PDA肿瘤的侵袭性特征。我们从学习中学到的知识将会 对肿瘤微环境的本质提供有价值的见解。这样的洞察力应该为小说 有效治疗PDA的战略想法，这是一个具有迫切未满足需求的临床目标。

PROJECT SUMMARY Pancreatic ductal adenocarcinoma (PDA) is a highly lethal malignancy with a five-year survival rate of less than 8%. These grim statistics owe, in part, to the profound resistance of PDA to current therapies. More effective treatment strategies would offer tremendous benefit to PDA patients. Emerging evidence suggests that epigenetic and genetic heterogeneity exist in both the neoplastic and non-neoplastic compartments of pancreatic tumors, and that this heterogeneity may impact response to treatment. Our own studies have identified two subpopulations of cancer-associated fibroblasts (CAFs) present within the PDA microenvironment, a cancer-proximal population that expresses myofibroblast markers and a cancer-distal population that expresses inflammatory markers such as the cytokine IL-6. However, the full complement of cell populations present within the PDA tumors and the mechanisms through which each population might impact tumor biology remain unclear. Tackling these questions requires models that accurately recapitulate human PDA. Patient-derived xenografts (PDXs), organoid cultures, organoid-stromal co-cultures, and organoid xenografts have emerged as “next-generation” models that better mimic the complex interactions present in the PDA microenvironment. Yet, the extent to which each model preserves the diverse populations found in human tumors remains unclear. To address this question, we propose to generate a series of matching organoid and xenograft models which we will use to characterize and perturb subpopulations of neoplastic cells and CAFs. We will generate and characterize five sets of paired organoid culture and PDX models, where pairs are derived the same de-identified patient tumor specimens. In addition, we will develop a new model for pancreatic cancer in which patient-derived PDA organoids are delivered to mice in situ via injection into the main pancreatic duct, better recapitulating the developmental trajectory of human PDA than other xenograft approaches. We will use single-cell RNA and DNA sequencing as well as barcoded organoid lines to characterize and perturb the neoplastic and CAF populations present in our models. To study to role of IL-6 in PDA more directly, we will use biochemical and genetic strategies to perturb CAF-mediated IL-6 signalling in our models. Among these, we will make use of a murine model engineered to express human IL-6 as a host for our PDA xenografts, restoring the ability for IL-6 generated in the stroma to signal to the transplanted neoplastic cells. Ultimately, our studies will shed light on the distinct subpopulations of neoplastic cells and CAFs present in the PDA microenvironment and help to identify which of those subpopulations function to promote the aggressive traits characteristic of PDA tumors. The knowledge gained from our studies will provide valuable insights into the nature of the tumor microenvironment. Such insights should inform novel ideas for strategies to effectively treat PDA, a clinical goal with urgent unmet need.