Novel model to study PDAC using normal human pancreatic tissue

使用正常人胰腺组织研究 PDAC 的新模型

• 批准号: 9502935
• 负责人: Pei Wang
• 金额: $ 16.58万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-06-09 至 2020-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9502935
• 关键词: Acinar Cell; Adenocarcinoma Cell; Animal Model; Area; Biological Models; Cancer Model; Cancer cell line; Cancerous; Cell Line; Cells; Clinical; Clinical Trials; Common Neoplasm; Data; Desmoplastic; Development; Disease; Duct (organ) structure; Ductal; Ductal Epithelial Cell; Early Diagnosis; Early identification; Engineering; Epidermal Growth Factor Receptor; Essential Genes; Event; Exploratory/Developmental Grant; Genes; Genetic; Genetically Engineered Mouse; Genome; Genomic Instability; Grant; Growth; Guide RNA; Human; Human Engineering; In Vitro; Islets of Langerhans Transplantation; KRAS2 gene; Knock-out; Lentivirus Vector; Ligands; MADH4 gene; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of pancreas; Metaplasia; Metastatic Neoplasm to the Liver; Methods; Modeling; Modification; Molecular; Mus; Mutate; Mutation; NOD/SCID mouse; Neoplasm Metastasis; Neoplasms; Normal Cell; Oncogenes; Oncogenic; Organ; Organoids; Pancreas; Pancreatic Ductal Adenocarcinoma; Pathway interactions; Patients; Pharmaceutical Preparations; Publishing; Relapse; Research; Sampling; Signal Pathway; Signal Transduction; Study models; Survival Rate; System; TP53 gene; Testing; Time; Tissues; Transforming Growth Factor beta; Transplantation; Tumor stage; Validation; Xenograft procedure; actionable mutation; cancer diagnosis; clinically relevant; design; diagnostic biomarker; early detection biomarkers; effective therapy; genetically modified cells; human disease; human model; improved; in vivo; method development; model development; mutant; novel; precision medicine; rare cancer; success; three-dimensional modeling; treatment strategy; tumor; tumor growth; tumorigenesis; vector

Novel model for studying PDAC using normal human pancreatic tissue For the past twenty years, the survival rate for many cancers has improved, but survival for pancreatic ductal adenocarcinoma (PDAC) has not, due to lack of both early detection methods and effective treatments. Human PADC cancer cell lines have been widely used. However, such cell lines are far from original organ tissue, and their highly unstable genome complicates interpretation. On the other hand, while animal models overcome many of the limitations of cell lines, genetically engineered mouse models (GEMMs) may overlook some profound differences between human and mouse cells. Indeed, nearly 90% of drugs with proven efficacy in animal models are ineffective in clinical trials. In our previous studies, we have successfully induced ADM in primary human normal acinar cells and found distinct features of human cells. We further engineered these acinar-derived ductal like (AD) cells to PDAC cells by specifically introducing the landscape mutation combination (KRAS/P16/TP53/SMAD4, KPTS) of human PDAC. Importantly, when transplanted into mice, these genetically engineered cells gave rise to invasive desmoplastic tumors, recapitulating the clinical features of PDAC in human patients, highlighting the power of this novel system to modeling human PDAC development. Given the fact that PDAC gains mutations during progression, we propose to use our novel system to generate engineered cells carrying fewer mutations, which may give rise to non-invasive neoplasia when orthotropically transplanted into NOD-SCID mice. This model will not only help us to understand the mechanisms of human PDAC development, it may also be a valuable platform to identify biomarkers for early diagnosis. Since our engineered AD cells give rise to tumors recapitulating clinical features of human PDAC, they are a good model to identify key targets to stop the growth of fully established tumors in vivo. To test this concept, we will utilize an inducible gRNA expressing vector to specifically knockout oncogenic KRAS in vivo to induce tumor size reduction. Success with this method would provide a powerful model to identify other essential genes supporting human PDAC progression and to reveal possible pathways for human PDAC relapse after KRAS knockout. The proposed novel models have great clinical potential on precision medicine, identification of early diagnosis methods, and development of new treatment strategies for human PDAC.

利用正常人胰腺组织研究PDAC的新模型 在过去的20年里，许多癌症的存活率有所提高，但胰腺癌的存活率仍然很低。 由于缺乏早期检测方法和有效治疗，导管腺癌（PDAC）还没有。 人PADC癌细胞系已被广泛使用。然而，这样的细胞系远离原始器官， 组织，它们高度不稳定的基因组使解释复杂化。另一方面，虽然动物模型 为了克服细胞系的许多局限性，基因工程小鼠模型（GEMM）可能会忽视 人类和小鼠细胞之间的一些深刻差异。事实上，近90%的已证实有效的药物 在临床试验中是无效的。在我们以前的研究中，我们已经成功地诱导ADM在 原代人正常腺泡细胞，发现了人细胞的独特特征。我们进一步设计了这些 腺泡衍生的导管样（AD）细胞转化为PDAC细胞， 在一些实施方案中，使用人PDAC的组合（KRAS/P16/TP 53/SMAD 4，KPTS）。重要的是，当移植到小鼠体内时， 这些基因工程细胞引起了侵袭性促结缔组织增生性肿瘤， PDAC在人类患者中的特征，突出了这种新型系统对人类PDAC建模的能力 发展鉴于PDAC在进展过程中获得突变的事实，我们建议使用我们的新的 系统产生携带较少突变的工程细胞，这可能会导致非侵入性瘤形成 当移植到NOD-SCID小鼠时。这个模型不仅可以帮助我们理解 尽管它可能是人类PDAC发展机制的重要组成部分，但它也可能是一个有价值的平台，用于识别早期PDAC的生物标志物。 诊断.由于我们的工程化AD细胞产生重现人PDAC临床特征的肿瘤， 它们是鉴定关键靶点以阻止体内完全建立的肿瘤生长的良好模型。为了验证这一 根据这一概念，我们将利用诱导型gRNA表达载体在体内特异性敲除致癌KRAS， 诱导肿瘤尺寸减小。这种方法的成功将提供一个强大的模型来识别其他 支持人类PDAC进展的必需基因，并揭示人类PDAC的可能途径 KRAS敲除后复发。所提出的新模型在精准医疗方面具有巨大的临床潜力， 早期诊断方法的鉴定和人PDAC新治疗策略的开发。