Investigating microRNA Function in Homeostasis, Regeneration and Cancer

研究 microRNA 在稳态、再生和癌症中的功能

基本信息

批准号：
10407066
负责人：
Andrea Ventura
金额：
$ 50.92万
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：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10407066
关键词：
Acute Address Adult Affect Aging Alleles Applications Grants Binding Biogenesis Biological Process Cancer cell line Cells Chronic Code Complex Cues Development Doxycycline Ensure Gene Expression Gene Expression Regulation Gene Fusion Genes Genetic Transcription Genetically Engineered Mouse Goals Homeostasis In Vitro Infection Inflammation Injury Lead Liver Maintenance Malignant Neoplasms Mammals Mediating Messenger RNA MicroRNAs Mouse Strains Mus Natural regeneration Normal tissue morphology Nucleotides Partner in relationship Pathway interactions Phenotype Population Process Protein Family Proteins Repression Research Resolution Ribonucleoproteins Role Series Site Testing Time Tissues Transgenic Mice Transgenic Organisms Untranslated RNA Work anti-cancer base cancer cell cell type experimental study gain of function gene repression human cancer mouse model in vivo innovation insight intestinal epithelium loss of function member novel novel strategies prevent protein degradation recruit small molecule inhibitor stem cell population tissue regeneration tumor tumor initiation tumor progression tumorigenesis

项目摘要

ABSTRACT Precise temporal and spatial control of gene expression is essential to ensure normal development and homeostasis and is achieved through multiple regulatory mechanisms acting at every step from gene transcription to protein degradation. MicroRNAs (miRNAs), constitute a large class of highly conserved regulatory short non-coding RNAs that modulate gene expression at the post-transcriptional level. Mechanistically, miRNAs repress their targets as part of a large multicomponent ribonucleoprotein known as the miRNA-Induced Silencing Complex (miRISC), which include Argonaute proteins (AGO) and members of the TNRC6 family of proteins. How miRISC assembly and activity are regulated in vivo remains poorly understood and until recently it was tacitly assumed that miRNAs are constitutively active in all cell types. Recent work from several groups, including ours, has challenged this assumption, revealing that assembly of the miRISC is tightly regulated in vivo, and suggesting the possibility that in many adult tissues and in quiescent cells the bulk of miRNA-bound AGO proteins are not bound to target mRNAs, and are not engaged in their repression. Why many adult tissues and quiescent cells contain high levels of functionally inactive miRNAs and whether, in addition to mitogenic cues, other perturbations—including cancer, aging, infection, and inflammation—regulate miRISC assembly and activity are two major unanswered questions in the field and the main focus of this grant proposal. As a first step to address these two questions we propose a series of experiment that take advantage of a novel genetically engineered mouse strain we have recently generated that allows to control miRISC assembly in vivo in a temporally and spatially controlled fashion. We will use this novel mouse strain to determine the requirement for miRISC activity during normal tissue homeostasis, in development, and during tissue regeneration (Aim 1). In Aim 2, we will use it to directly test the hypothesis that miRISC activity is required for tumor progression and tumor maintenance in vivo and we will determine the potential of miRISC inhibition as a novel anticancer strategy. Finally, in Aim 3 we will take advantage of a novel mouse strain we have developed to gain mechanistic insights into how miRNAs control these essential processes. Successful completion of this project will greatly advance our understanding of the role of miRNA-mediated gene repression in mammals and could lead to the development of novel anti-cancer strategies.

抽象的基因表达的精确时空控制对于确保正常发育和发育至关重要体内平衡是通过作用于基因每一步的多重调控机制来实现的转录到蛋白质降解的微小RNA（miRNA）构成了一大类高度保守的蛋白质。在转录后水平调节基因表达的调节性短非编码RNA。从机制上讲，miRNA 作为大型多组分核糖核蛋白（称为 miRNA 诱导沉默复合物 (miRISC)，其中包括 Argonaute 蛋白 (AGO) 和 TNRC6 蛋白家族。 miRISC 组装和活性如何在体内调节仍然知之甚少，直到最近才被人们所了解。默示地假设 miRNA 在所有细胞类型中都具有组成性活性，包括几个小组的最新工作。我们的研究挑战了这一假设，揭示了 miRISC 的组装在体内受到严格调控，并且表明在许多成体组织和静止细胞中，大量 miRNA 结合的 AGO 可能存在蛋白质不与靶标 mRNA 结合，也不参与其抑制。为什么许多成体组织和静止细胞含有高水平的功能失活的 miRNA 和除了促有丝分裂线索之外，是否还存在其他干扰——包括癌症、衰老、感染和炎症——调节 miRISC 的组装和活性是该领域尚未解答的两个主要问题以及本次拨款提案的主要重点。作为解决这两个问题的第一步，我们提出了一系列利用新颖的实验我们最近培育出的基因工程小鼠品系可以在体内控制 miRISC 组装我们将使用这种新型小鼠品系以时间和空间控制的方式来确定需求。正常组织稳态、发育和组织再生期间的 miRISC 活性（目标 1）。在目标 2 中，我们将用它来直接检验肿瘤进展需要 miRISC 活性的假设，以及体内肿瘤维持，我们将确定 miRISC 抑制作为一种新型抗癌策略的潜力。最后，在目标 3 中，我们将利用我们开发的新型小鼠品系来获得机制见解研究 miRNA 如何控制这些重要过程。该项目的成功完成将极大地增进我们对miRNA介导基因作用的理解哺乳动物的抑郁症，并可能导致新的抗癌策略的发展。