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：
10577727
负责人：
Karuna Ganesh
金额：
$ 39.68万
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/10577727
关键词：
3&apos Untranslated Regions Acceleration Adherens Junction Advanced Malignant Neoplasm 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 Elements Engineering Epigenetic Process Epithelial Cells Epithelium Family Fluorescence Gene Expression Genes Genetic Genetic Transcription Homeostasis Immunofluorescence Immunologic Immunoprecipitation Impairment Injections Intestinal Neoplasms Intestines Invaded Knock-in LGR5 gene Maintenance Malignant Neoplasms Mediating Mediator 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 Rectum Recurrence Regulation Reporter Resistance Role Sampling Stress Structure Structure of splenic vein Time Tissues Tumor Suppressor Proteins Untranslated Regions Work Xenograft procedure cancer cell cancer initiation cancer stem cell colorectal cancer metastasis crosslink derepression in vivo insight intestinal epithelium knock-down mRNA Transcript Degradation member mouse model neoplastic neuroendocrine differentiation novel overexpression posttranscriptional 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 tissue regeneration transcriptional reprogramming transcriptome sequencing transcriptomics tumor tumor initiation tumor progression whole genome 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由能够细胞驱动自我更新，缓慢的细胞循环，肿瘤再生和耐药性，称为转移干细胞（METSC）。制定消除转移的有效策略需要更好地理解 Metscs利用生存的机制。我们最近证明了（1）传播大肠癌症（CRC）从LGR5+肿瘤发射癌症干细胞（CSC）进行动态表型转换状态到转移所需的不同LGR5LOWL1CAM+状态。（2）METSC的这种表型可塑性是保留在离体患者衍生的类器官中，可用于剖析可塑性机制。（3）L1CAM+ METSC在功能上与肠道肿瘤发射LGR5+ CSC不同：类器官需要L1CAM 形成，结肠炎后肠上皮的再生以及转移后的肿瘤形成传播。但是与LGR5不同，它对于上皮稳态或肠道肿瘤的启动是可分配的。在与肿瘤的起始形成鲜明对比，肿瘤的开始稳态干细胞在完整中经历癌基因驱动的超增殖组织，转移会颠覆一种再生机制，使其在完整的上皮生态裂市场外生存和再生。（4）我们已经表明，L1CAM表达的主要驱动力是上皮完整性本身的丧失，通过 E-钙黏着蛋白间的粘附蛋白连接处的损失会瞬时移位转录消音器静止/NSRF 从静态METSC中的染色质，反过组织再生1。增殖，上皮结构的恢复和大量转移的产物另一方面，需要较高的休息水平。我们的证据表明，METSCS细胞是再生干细胞直接响应上皮完整性以驱动修复的出现，这是生理伤口的表型在METSC中重新部署的康复。在此提案中，我们将定义分子机制染色质结合在METSC中动态调节，这又如何使细胞命运可塑性从茎向增殖。我们的初步数据暗示mRNA结合蛋白ZFP36L1/2在静止介导的转移性可塑性。项目假设：ZFP36L1/2REST轴是细胞命运的主调节器肠上皮祖细胞中的可塑性。 AIM 1：在正常中定义ZFP36L1/2-RES轴的功能和肿瘤性肠道干细胞自我更新，分化和增殖。目标2：剖析分子 ZFP36L1/2介导的细胞命运可塑性的机理。目标3：确定功能后果 ZFP36L1/2-rest动态调节转移性播种和定殖。结果将定义癌症进展并确定推定的治疗靶标，以限制再生可塑性，并有可能影响临床结果。