Pancreatic cancer stem cells: PD2-mediated novel mechanistic link and metabolomic alterations

胰腺癌干细胞：PD2介导的新机制联系和代谢组学改变

• 批准号: 10306412
• 负责人: Saswati Karmakar
• 金额: $ 8.82万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-13 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10306412
• 关键词: 3-Dimensional; Area; Binding Sites; CRISPR/Cas technology; Cancer Biology; Cell Maintenance; Cells; ChIP-seq; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; Data; Derivation procedure; Development; Disease; Drug resistance; Epigenetic Process; Equilibrium; Exhibits; Foundations; Funding; Genes; Genetic; Genomics; Glycolysis; Goals; Human; Implant; Infrastructure; Knock-out; Knockout Mice; Knowledge; Link; Maintenance; Malignant Neoplasms; Malignant neoplasm of ovary; Malignant neoplasm of pancreas; Mass Spectrum Analysis; Mediating; Mentors; Messenger RNA; Metabolic; Metabolism; Metastatic to; Mitochondria; Modeling; Molecular; Molecular Biology; Mus; Neoplasm Metastasis; Nude Mice; Oncoproteins; Organoids; Oxidative Phosphorylation; Pancreas; Pathogenesis; Pathway interactions; Phase; Phenotype; Play; Polymerase; Population; Postdoctoral Fellow; Protein Dynamics; Proteins; Proteomics; Pyruvate; RNA Polymerase II; Recurrence; Research; Research Personnel; Research Project Grants; Research Proposals; Resistance development; Role; Techniques; Therapeutic; Tissues; Training; Transcription Elongation; Transgenic Mice; Tumor Burden; Tumor Tissue; United States; Work; base; biomarker identification; cancer cell; cancer cell differentiation; cancer drug resistance; cancer stem cell; career; cell type; combinatorial; conventional therapy; effective therapy; embryonic stem cell; experimental study; improved; in vivo; islet stem cells; knock-down; knockout gene; lipidomics; metabolic phenotype; metabolic profile; metabolomics; neoplastic; neoplastic cell; novel; overexpression; oxidation; pancreatic cancer cells; pancreatic cancer model; pancreatic differentiation 2 protein; pancreatic tumorigenesis; post-doctoral training; pre-doctoral; programs; self-renewal; skills; skills training; stem; stem cell biomarkers; stem cell differentiation; stem cell self renewal; stem cells; stem-like cell; stemness; targeted treatment; therapeutic target; transcription factor; transcriptome sequencing; tumor heterogeneity; tumor metabolism; tumor progression; tumorigenesis

PROJECT SUMMARY / ABSTRACT This research proposal is intended to provide predoctoral and postdoctoral training to develop the necessary skills for a career as an independent investigator in cancer biology. The long-term research focus is development of effective therapy for pancreatic cancer (PC) through 1) determining the contribution of cancer stem cells (CSCs) to PC progression and metastasis, 2) understanding the mechanism of CSC maintenance and CSC- mediated drug resistance, 3) identifying genetic, epigenetic, and metabolic factors essential for CSC maintenance with the aim of identifying novel epigenetic and metabolic targets that can be exploited for combinatorial therapy against PC. The objective of my dissertation research (F99 phase) is to define the role of Pancreatic Differentiation 2 (PD2) in pancreatic CSCs and CSC-mediated PC progression, with the goal of deciphering the mechanism of PD2-dependent CSC maintenance. PD2 is a ubiquitous multifunctional protein, a core component of human RNA Polymerase II-Associated Factor 1 complex (PAF1C) that functions in transcription elongation and mRNA processing. We discovered that PD2 is a novel pancreatic CSC marker and mediates drug resistance of CSCs. Knowledge of the molecular mechanism of PD2-dependent CSC maintenance and drug resistance is critical. We have recently made several discoveries relevant to this concept. First, knockdown of PD2 significantly reduces the levels of established CSC and self-renewal markers. Importantly, PD2 depletion significantly reduces tumor burden in vivo. Moreover, RNA-sequencing and transcription factor PCR array analyses revealed that several stemness and metastasis genes were significantly downregulated following PD2 depletion. Based on aforementioned information and additional data, we hypothesize that PD2 functions as a master-regulator of stem cell maintenance and thereby mediates PC progression. Our research will utilize high throughput genomic techniques such as chromatin immunoprecipitation sequencing and RNA-sequencing in presence and absence of PD2 to define downstream targets of PD2, and identify the pathway for PD2-dependent maintenance of pancreatic CSCs. We will also determine the role of PD2 in CSC-mediated PC progression using a novel CRISPR- based PD2 knockout model crossed with KPC model of PC progression. To expand upon the future research direction and to build the foundation for independence, I will pursue postdoctoral training in cancer metabolism. The goal of the proposed K00 postdoctoral training is to gain expertise on :1) the current metabolomic strategies; 2) understand the mechanisms that promote acquisition of different metabolic programs by CSCs and differentiated tumor cells; 3) contribution of metabolism to CSC-mediated drug resistance; and 4) development of novel combinatorial therapeutics based on metabolic targeting to treat cancer. Ultimately, the proposed F99/K00 training will provide a strong intellectual foundation for R01 funding that will establish my independence and will provide professional training in the skills required to be an effective PI and mentor.

项目概要/摘要 该研究计划旨在提供博士前和博士后培训，以开发必要的 作为癌症生物学独立研究者的职业技能。长期研究重点是发展 通过 1) 确定癌症干细胞的贡献来确定胰腺癌 (PC) 的有效治疗方法 (CSC) 到 PC 进展和转移，2) 了解 CSC 维持和 CSC- 的机制 介导的耐药性，3) 识别 CSC 必需的遗传、表观遗传和代谢因素 维护的目的是识别新的表观遗传和代谢目标，可用于 针对 PC 的联合治疗。我的论文研究（F99 阶段）的目标是定义角色 胰腺 CSC 中的胰腺分化 2 (PD2) 和 CSC 介导的 PC 进展，目标是 破译PD2依赖的CSC维持机制。 PD2是一种普遍存在的多功能蛋白 人类 RNA 聚合酶 II 相关因子 1 复合物 (PAF1C) 的核心成分，其功能 转录延伸和 mRNA 加工。我们发现 PD2 是一种新型胰腺 CSC 标志物 介导 CSC 的耐药性。了解PD2依赖性CSC的分子机制 维持和耐药性至关重要。我们最近有了一些与这个概念相关的发现。 首先，PD2 的敲低显着降低了已建立的 CSC 和自我更新标记的水平。 重要的是，PD2 消耗显着降低了体内肿瘤负荷。此外，RNA测序和 转录因子 PCR 阵列分析显示，几个干性和转移基因显着 PD2 耗尽后下调。根据上述信息和附加数据，我们 假设 PD2 作为干细胞维持的主调节因子发挥作用，从而介导 电脑进步。我们的研究将利用高通量基因组技术，例如染色质 在存在和不存在 PD2 的情况下进行免疫沉淀测序和 RNA 测序以确定下游 PD2 的靶点，并确定胰腺 CSC 的 PD2 依赖性维持途径。我们还将 使用基于 CRISPR 的新型 PD2 敲除模型确定 PD2 在 CSC 介导的 PC 进展中的作用 与 PC 进展的 KPC 模型交叉。拓展未来的研究方向并建立 为独立奠定基础，我将从事癌症代谢方面的博士后培训。拟议的目标 K00博士后培训旨在获得以下方面的专业知识：1）当前的代谢组学策略； 2）了解 促进 CSC 和分化肿瘤细胞获得不同代谢程序的机制； 3） 代谢对 CSC 介导的耐药性的贡献； 4）新型组合的开发 基于代谢靶向治疗癌症的疗法。最终，拟议的 F99/K00 培训将提供 为 R01 资助奠定坚实的智力基础，这将建立我的独立性并提供专业的 培训成为有效的 PI 和导师所需的技能。