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: 10535431
• 负责人: Saswati Karmakar
• 金额: $ 9.26万
• 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/10535431
• 关键词: 3-Dimensional; Area; Binding Sites; CRISPR/Cas technology; Cancer Biology; Cell Maintenance; Cell Separation; 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; KPC model; 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; Mitochondria; Modeling; Molecular; Molecular Biology; Mus; Neoplasm Metastasis; Nude Mice; Oncogenes; 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; Role; Techniques; Therapeutic; Tissues; Training; Transcription Elongation; Transgenic Mice; Tumor Burden; Tumor Tissue; United States; Work; 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; stem cell biomarkers; 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.

项目总结/摘要 本研究建议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。长期的研究重点是发展 胰腺癌（PC）的有效治疗方法，通过1）确定癌症干细胞的贡献 （2）了解CSC维持和CSC-转移的机制， 介导的耐药性，3）鉴定CSC所必需的遗传、表观遗传和代谢因素 维持，目的是鉴定新的表观遗传和代谢靶点， 联合治疗PC。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（PD 2）和CSC介导的PC进展，目标是 解译PD 2依赖的CSC维持的机制。PD 2是一种普遍存在的多功能蛋白， 人RNA聚合酶II-相关因子1复合物（PAF 1C）的核心组分，在 转录延伸和mRNA加工。我们发现PD 2是一种新的胰腺CSC标志物， 介导CSC的耐药性。PD 2依赖性CSC的分子机制 维持和耐药性至关重要。我们最近有几个发现与这个概念有关。 首先，PD 2的敲低显著降低了已建立的CSC和自我更新标志物的水平。 重要的是，PD 2消耗显著降低体内肿瘤负荷。此外，RNA测序和 转录因子PCR阵列分析显示，几个干细胞和转移基因， 在PD 2耗尽后下调。根据上述信息和其他数据，我们 假设PD 2作为干细胞维持的主调节因子发挥作用，从而介导 PC进展。我们的研究将利用高通量基因组技术，如染色质 在存在和不存在PD 2的情况下进行免疫沉淀测序和RNA测序，以确定下游 PD 2的靶点，并确定胰腺CSC的PD 2依赖性维持途径。我们还将 使用基于CRISPR的新型PD 2敲除模型确定PD 2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，构建 为独立奠定基础，我将继续进行癌症代谢方面的博士后培训。建议的目标 K 00博士后培训是为了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和分化的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;和4）开发新的组合药物， 基于代谢靶向的治疗方法来治疗癌症。最终，拟议的F99/K 00培训将提供 R 01资金的强大知识基础，将建立我的独立性，并将提供专业的 培训成为有效PI和导师所需的技能。