Project 1: Elucidating the genetics and cell of origin of pancreatic cancer initiation

项目1：阐明胰腺癌发生的遗传学和细胞起源

• 批准号: 10704080
• 负责人: LAURA D ATTARDI
• 金额: $ 40.95万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10704080
• 关键词: Acinar Cell; Affect; Aging; Alleles; Bar Codes; CDKN2A gene; CRISPR/Cas technology; Cells; Chromosome Mapping; Clinical; Clinical Management; Clonal Expansion; Data; Development; Disease; Duct (organ) structure; Ductal Epithelial Cell; Early Diagnosis; Early treatment; Epithelial Cells; Event; Evolution; Gene Expression; Gene Expression Profile; Genetic; Genetic Determinism; Genetically Engineered Mouse; Genotype; High-Throughput Nucleotide Sequencing; Homeostasis; Human; Immune; Intervention; KRAS oncogenesis; KRAS2 gene; KRASG12D; Knowledge; Lesion; MADH4 gene; Malignant Neoplasms; Malignant neoplasm of pancreas; Measures; Metaplasia; Modeling; Molecular; Mus; Mutate; Mutation; Oncogenic; Pancreas; Pancreatic Ductal Adenocarcinoma; Pancreatic Intraepithelial Neoplasia; Pancreatitis; Pathway interactions; Positioning Attribute; Process; Recurrence; Research; Research Personnel; Risk; Risk Factors; Role; Route; System; TP53 gene; Testing; Tumor Suppressor Genes; Tumor-Suppressor Gene Inactivation; Variant; Viral; cancer care; cancer cell; cancer genetics; cancer initiation; carcinogenesis; cell type; chronic pancreatitis; design; genome editing; genome sequencing; immunoregulation; improved; in vivo; innovation; insight; mortality; mouse model; mutant; neoplastic; novel; premalignant; reduce symptoms; regenerative; repaired; single-cell RNA sequencing; somatic cell gene editing; synergism; transcriptome; transdifferentiation; tumor; tumor initiation; tumor microenvironment

ABSTRACT (Project 1) Pancreatic ductal adenocarcinoma (PDAC) is a prevalent and almost uniformly fatal malignancy. Human PDAC genome sequencing has identified recurrent mutations in Kras and several major tumor suppressor genes (TSGs), but the impact of these alterations in the initiation and evolution of human PDAC development remain poorly understood at the molecular level. Understanding the basis of tumor initiation and development has critical implications for improving early diagnosis and therapy. While PDACs were originally thought to arise from ductal cells, evidence from mouse models suggests that acinar cells can give rise to PDACs through a regenerative, transdifferentiation process known as Acinar to Ductal Metaplasia (ADM). ADM is stimulated by chronic pancreatitis, a known risk factor for PDAC, and generates premalignant Pancreatic Intraepithelial Neoplasias (PanINs), which lead to PDAC. Our preliminary data suggest that the p53 TSG can inhibit ADM and that p53 inactivation in acinar cells expressing oncogenic KrasG12D drives PDAC. However, expression of oncogenic KrasG12D and p53 inactivation in ductal cells can also induce PDAC, suggesting alternate routes of PDAC carcinogenesis. Interestingly, we find that acinar cell-derived and ductal cell-derived tumors resemble the classical and basal-like subtypes of human PDAC, respectively. These results underscore the unique value of mouse models in uncovering the determinants that dictate different paths of PDAC evolution to provide insight into human PDAC. The roles of other common PDAC TSGs in early stage pre-neoplastic lesions, including whether they normally function in acinar cells and/or ductal cells, however, remains unclear. Furthermore, oncogenic Kras mutations are diverse in PDAC, yet it is unknown whether different mutations have distinct impacts on PDAC development from different pancreatic epithelial cell types. We hypothesize that the cell type of origin and tumor genotype cooperate to drive different paths of PDAC development, and to test this idea, we will systematically inactivate several major PDAC TSGs and express different Kras alleles in both putative cell types of origin. We will characterize the transcriptomes and the cellular milieu of early lesions and tumors and employ innovative approaches like somatic genome editing and molecular barcoding. We will compare our studies in tractable mouse models to studies of human PDAC development. Together, these transformative studies will establish how genetic determinants and cell-of-origin influence the molecular pathways and the cellular microenvironment during PDAC evolution in mice and humans, critical knowledge for better clinical management of this deadly disease.

摘要(项目1) 胰腺导管腺癌(PDAC)是一种常见且几乎一致致命的恶性肿瘤。人类PDAC 基因组测序发现了Kras和几个主要肿瘤抑制基因的反复突变 (TSG)，但这些变化在人类PDAC发育的启动和演变中的影响仍然存在 在分子水平上知之甚少。了解肿瘤发生和发展的基础至关重要 对改进早期诊断和治疗的启示。而PDAC最初被认为起源于导管 细胞，来自小鼠模型的证据表明腺泡细胞可以通过再生的， 转分化过程称为腺泡到导管的化生(ADM)。ADM受慢性刺激 胰腺炎，已知的PDAC的危险因素，并导致癌前胰腺上皮内肿瘤 (Panins)，这导致了PDAC。我们的初步数据表明，P53 TSG可以抑制ADM，而P53 表达致癌基因KrasG12D的腺泡细胞失活可驱动PDAC。然而，致癌基因的表达 导管细胞中KrasG12D和P53的失活也可以诱导PDAC，提示PDAC的替代途径 致癌。有趣的是，我们发现腺泡细胞来源的肿瘤和导管细胞来源的肿瘤类似于 人PDAC的典型亚型和类基底细胞亚型。这些结果突显了 小鼠模型揭示决定PDAC进化不同路径的决定因素以提供洞察力 变成人类的PDAC。其他常见的PDAC TSG在早期癌前病变中的作用，包括 然而，它们是否在腺泡细胞和/或导管细胞中正常发挥功能仍不清楚。此外， 致癌的kras基因突变在pDAC中是多样的，但不同的突变是否有不同的差异尚不清楚。 不同类型胰腺上皮细胞对PDAC发育的影响。我们假设细胞类型 起源和肿瘤基因型共同驱动了PDAC发展的不同途径，为了检验这一想法，我们 将系统地灭活几个主要的PDAC TSG，并在两个假定的细胞中表达不同的Kras等位基因 原产地类型。我们将描述早期病变和肿瘤的转录本和细胞环境，以及 使用创新的方法，如体细胞基因组编辑和分子条形码。我们将比较我们的 从易驯化的小鼠模型研究人类PDAC发育。总而言之，这些变革性的 研究将确定遗传决定因素和起源细胞如何影响分子途径和 小鼠和人类PDAC进化过程中的细胞微环境--改善临床的关键知识 对这种致命疾病的管理。