Mechanisms of Dynamic Transcriptional Reprogramming in Metastasis Stem Cells

转移干细胞动态转录重编程的机制

基本信息

批准号：
10340384
负责人：
Karuna Ganesh
金额：
$ 40.49万
依托单位：
SLOAN-KETTERING INST CAN RESEARCH
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-03-01 至 2027-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10340384
关键词：
3&apos Untranslated Regions Adherens Junction Advanced Malignant Neoplasm Aging Binding Binding Proteins Biochemical Biological Assay Biological Models Cancer Etiology Cell Cycle Cells Cessation of life Chromatin Clinical Colitis Colon Colorectal Cancer Complex Cytoplasmic Granules Data Development Disseminated Malignant Neoplasm Distant E-Cadherin Engineering Epigenetic Process Epithelial Epithelial Cells Family Fluorescence Gene Expression Genes Genetic Genetic Transcription Homeostasis Immunofluorescence Immunologic Immunoprecipitation Impairment Injections Intestinal Neoplasms Intestines Knock-in LGR5 gene Maintenance Malignant Neoplasms Mediating Mediator of activation protein Messenger RNA Modeling Molecular Mutation Natural regeneration Neoplasm Metastasis Neural Cell Adhesion Molecule L1 Neurons Neurosecretory Systems Oncogenes Organ Organoids Outcome Output Patient-Focused Outcomes Patients Phase Phenotype Physiologic pulse Physiological Primary Neoplasm Proliferating Proteins RNA RNA-Binding Proteins Recurrence Regulation Reporter Resistance Role Sampling Stress Structure Structure of splenic vein Time Tissues Untranslated Regions Xenograft procedure base cancer cell cancer stem cell colorectal cancer metastasis crosslink in vivo insight intestinal epithelium knock-down mRNA Transcript Degradation member mouse model neoplastic neuroendocrine differentiation novel overexpression progenitor programs protein complex rectal regenerative regenerative cell repaired response restoration self-renewal single cell mRNA sequencing stem cell self renewal stem cells stemness therapeutic target therapy outcome transcriptional reprogramming transcriptome sequencing transcriptomics tumor tumor initiation tumor progression wound healing

项目摘要

Metastasis causes >90% of cancer death. The persistence and lethality of metastasis is driven by cells capable of self-renewal, slow cell-cycling, tumor re-initiation, and therapy resistance, termed metastasis stem cells (MetSCs). Development of effective strategies for eliminating metastasis requires a better understanding of the mechanisms that MetSCs exploit for survival. We recently demonstrated that (1) disseminating colorectal cancers (CRC) undergo a dynamic phenotypic switch from an LGR5+ tumor-initiating cancer stem cell (CSC) state to a distinct LGR5lowL1CAM+ state required for metastasis. (2) This phenotypic plasticity of MetSCs is retained in ex vivo patient derived organoids, which can be used to dissect mechanisms of plasticity. (3) L1CAM+ MetSCs are functionally distinct from intestinal tumor-initiating LGR5+ CSCs: L1CAM is required for organoid formation, the regeneration of intestinal epithelium after colitis, and tumor formation after metastatic dissemination. But unlike LGR5, it is dispensable for epithelial homeostasis or intestinal tumor initiation. In contrast to tumor initiation, where homeostatic stem cells undergo oncogene-driven hyper proliferation in intact tissues, metastasis subverts a regenerative mechanism to survive and regrow outside an intact epithelial niche. (4) We have shown that the principal driver of L1CAM expression is loss of epithelial integrity itself, acting via loss of E-cadherin intercellular adherens junctions to transiently displace the transcriptional silencer REST/NSRF from chromatin in quiescent MetSCs, in turn derepressing expression of L1CAM and other genes required for tissue regeneration1. Proliferation, restoration of epithelial structures, and macrometastatic outgrowth, on the other hand, require high REST levels. Our evidence suggests that MetSCs cells are regenerative stem cells that emerge directly in response to loss of epithelial integrity to drive repair, a phenotype of physiological wound healing that is redeployed in MetSCs. In this proposal, we will define the molecular mechanism by which REST chromatin binding is dynamically regulated in MetSCs, and how this in turn enables cell fate plasticity from stemness to proliferation. Our preliminary data implicates the mRNA binding protein ZFP36L1/2 in REST- mediated metastatic plasticity. Project hypothesis: The ZFP36L1/2REST axis is a master regulator of cell fate plasticity in intestinal epithelial progenitors. Aim 1: Define the function of the ZFP36L1/2-REST axis in normal and neoplastic intestinal stem cell self-renewal, differentiation, and proliferation. Aim 2: Dissect the molecular mechanism of ZFP36L1/2-mediated cell fate plasticity. Aim 3: Determine the functional consequences of ZFP36L1/2-REST dynamic regulation in metastatic seeding and colonization. Results will define mechanisms of cancer progression and identify putative therapeutic targets to limit regenerative plasticity, with potential to impact clinical outcomes.

转移导致>90%的癌症死亡。转移的持久性和致死性是由有能力的细胞驱动的自我更新、缓慢的细胞周期、肿瘤重新启动和治疗抵抗，称为转移干细胞（气象SC）。制定消除转移的有效策略需要更好地了解 MetSC 赖以生存的机制。我们最近证明 (1) 传播结直肠癌癌症 (CRC) 经历 LGR5+ 肿瘤起始癌症干细胞 (CSC) 的动态表型转变状态转变为转移所需的不同 LGR5lowL1CAM+ 状态。 (2) MetSCs 的表型可塑性是保留在体外患者衍生的类器官中，可用于剖析可塑性机制。 (3)L1CAM+ MetSC 在功能上不同于肠道肿瘤启动 LGR5+ CSC：类器官需要 L1CAM 结肠炎后肠上皮的形成、再生以及转移后肿瘤的形成传播。但与 LGR5 不同的是，它对于上皮稳态或肠道肿瘤的发生是可有可无的。在与肿瘤起始相反，在肿瘤起始中，稳态干细胞会完整地经历癌基因驱动的过度增殖。在组织中，转移破坏了再生机制，使其在完整的上皮微环境之外生存和再生。 (4) 我们已经表明，L1CAM 表达的主要驱动因素是上皮完整性本身的丧失，通过 E-钙粘蛋白细胞间粘附连接的丢失暂时取代转录沉默子 REST/NSRF 来自静态 MetSC 中的染色质，进而抑制 L1CAM 和其他基因的表达组织再生1.增殖、上皮结构的恢复和巨转移生长，另一方面，需要高休息水平。我们的证据表明 MetSC 细胞是再生干细胞直接响应上皮完整性的丧失而出现，以驱动修复，这是生理伤口的一种表型在 MetSC 中重新部署的治疗。在本提案中，我们将定义 REST 的分子机制染色质结合在 MetSC 中受到动态调节，以及这如何反过来使细胞命运可塑性从干性增殖。我们的初步数据表明 REST-中的 mRNA 结合蛋白 ZFP36L1/2 介导的转移可塑性。项目假设：ZFP36L1/2REST 轴是细胞命运的主要调节因子肠上皮祖细胞的可塑性。目标 1：定义 ZFP36L1/2-REST 轴正常情况下的功能以及肿瘤性肠干细胞的自我更新、分化和增殖。目标 2：剖析分子 ZFP36L1/2介导的细胞命运可塑性机制。目标 3：确定功能后果 ZFP36L1/2-REST 在转移播种和定植中的动态调节。结果将定义机制癌症进展并确定限制再生可塑性的假定治疗靶点，有可能影响临床结果。