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

CSHL-JAX 患者衍生的胰腺癌模型作为研究肿瘤异质性的系统

基本信息

批准号：
10116310
负责人：
Paul Robson
金额：
$ 84.5万
依托单位：
COLD SPRING HARBOR LABORATORY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-03-06 至 2023-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10116310
关键词：
Address Affect Bar Codes Biochemical Biochemical Genetics Biological Models Biology Cell physiology Cells Characteristics Clinical Coculture Techniques Complement Complex DNA sequencing Development Disease Distal Disulfiram Engineering Environment Epigenetic Process Fibroblasts Genetic Genetic Heterogeneity Goals Growth Head of pancreas Heterogeneity Human Implant In Situ In Vitro Individual Injections Interleukin-6 Intraepithelial Neoplasia Investigation Knowledge Laboratories Lesion Light Main pancreatic duct Malignant Neoplasms Malignant neoplasm of pancreas Measures Mediating Methods Modeling Mus Myofibroblast Nature Organoids Pancreas Pancreatic Ductal Adenocarcinoma Pancreatic duct Pathway interactions Patients Population Population Heterogeneity Primary Neoplasm Research Resistance Role Series Signal Transduction Simvastatin Specimen Survival Rate System Therapeutic Transgenic Mice Transplantation Treatment Efficacy Tumor Biology Xenograft Model Xenograft procedure cancer cell cell type cytokine design effective therapy experimental study genetic approach genetic signature in vitro Model in vivo inflammatory marker inhibitor/antagonist insight model development mouse model neoplastic neoplastic cell next generation novel novel strategies pancreatic cancer model pancreatic ductal adenocarcinoma cell pancreatic ductal adenocarcinoma model pancreatic neoplasm pancreatic tumorigenesis patient derived xenograft model preservation programs recruit response single cell analysis single cell mRNA sequencing single-cell RNA sequencing standard of care statistics therapy resistant trait transcriptome treatment response treatment strategy tumor tumor heterogeneity tumor microenvironment tumor xenograft tumorigenesis

项目摘要

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.

项目摘要胰腺导管腺癌（PDA）是一种高度致命的恶性肿瘤，五年生存率较小比8％。这些严峻的统计数据部分归功于PDA对当前疗法的深刻抵抗。更多的有效的治疗策略将为PDA患者带来巨大的好处。新兴证据表明这种表观遗传和遗传异质性都存在于肿瘤和非塑性室中胰腺肿瘤，这种异质性可能会影响对治疗的反应。我们自己的研究有确定了PDA中存在的两个癌症相关成纤维细胞（CAF）的亚群微环境，一种表达肌纤维细胞标记和癌症抗癌症的癌症蛋白环境表达炎症标志物（例如细胞因子IL-6）的种群。但是，完整完成 PDA肿瘤中存在的细胞群体以及每个人群可能的机制影响肿瘤生物学尚不清楚。解决这些问题需要准确概括的模型人PDA。患者衍生的Xenographictic（PDXS），器官培养物，类带有质培养和器官培养异种移植物已成为“下一代”模型，可以更好地模仿存在 PDA微环境。然而，每个模型在多大程度上保留了在人类肿瘤仍然不清楚。为了解决这个问题，我们建议生成一系列匹配器官和异种移植模型我们将用于表征和扰动肿瘤的亚群细胞和CAF。我们将生成和表征五组配对的器官培养和PDX模型，其中对得出相同的去识别患者肿瘤标本。此外，我们将为胰腺癌通过注射将患者衍生的PDA器官在原位输送到小鼠主胰管，比其他异种移植物更好地概括人类PDA的发育轨迹方法。我们将使用单细胞RNA和DNA测序以及条形码的类符号线表征和扰动我们模型中存在的肿瘤和CAF种群。研究IL-6在 PDA更直接地，我们将使用生化和遗传策略来扰动CAF CAF介导的IL-6信号传导我们的模型。其中，我们将利用设计的鼠模型来表达人类IL-6作为主机我们的PDA Xenographtics，恢复了基质中产生的IL-6的能力，以发出移植的信号肿瘤细胞。最终，我们的研究将阐明肿瘤细胞和 PDA微环境中存在的CAF，并有助于确定哪些亚群的功能促进PDA肿瘤的侵略性特征。从我们的研究中获得的知识将对肿瘤微环境的性质提供宝贵的见解。这样的见解应该告知小说有效治疗PDA的策略的想法，这是一个未满足的临床目标。