The role of long noncoding RNA CRNDE in normal physiology and cancer

长链非编码RNA CRNDE在正常生理和癌症中的作用

基本信息

批准号：
10715065
负责人：
Joshua T Mendell
金额：
$ 48.96万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-06-01 至 2028-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10715065
关键词：
Alleles Alternative Splicing Animal Model Animals Binding Biogenesis Biology Bone Marrow Transplantation Breeding CRISPR interference Cancer Biology Cancer Patient Cause of Death Cell Line Cell Nucleolus Cell Proliferation Cell model Characteristics Code Collaborations Colorectal Neoplasms Constitution Constitutional Custom Cytoplasm Data Development Diagnosis Disease Elements Genes Genetic Engineering Genomics Growth Hematopoietic Hematopoietic Neoplasms Human Impairment Knockout Mice Knowledge Length Libraries Malignant - descriptor Malignant Neoplasms Maps Mediating Molecular Molecular Mechanisms of Action Mus Nucleotides Pancreas Pathogenesis Pathway interactions Patients Phenocopy Phenotype Physiological Physiology Play Predisposition Production Proliferating Proteins Proteomics RNA Renal Cell Carcinoma Renal carcinoma Research Resolution Resources Ribosomal RNA Ribosomes Role Shwachman-Diamond syndrome Testing Tissues Transcript Translations Tumor Promotion United States Untranslated RNA Variant cancer cell cancer type differential expression effective therapy experience experimental study in vivo knock-down loss of function mouse model novel novel therapeutic intervention overexpression patient derived xenograft model premature prevent regeneration model skeletal abnormality therapeutic target tissue injury tissue regeneration transcriptome sequencing tumor tumor growth tumor progression urologic

项目摘要

PROJECT SUMMARY A growing body of evidence indicates that long noncoding RNAs (lncRNAs), a diverse class of non-protein- coding transcripts >200 nucleotides in length, play important roles in the initiation and progression of cancer. LncRNAs have been proposed to regulate all cancer hallmarks, but, in the vast majority of cases, their molecular mechanisms of action remain poorly understood. This knowledge gap is a major impediment towards realizing the potential of lncRNAs as therapeutic targets in cancer and other diseases. As in many human malignancies, lncRNAs are frequently dysregulated in renal cell carcinoma (RCC). RCC is the most common type of kidney cancer and the most lethal malignant urological tumor, with approximately 70,000 new cases diagnosed annually in the United States. To date, most genomic studies of RCC (and other cancers) have focused on identifying disease-associated alterations of protein-coding genes. Our understanding of the molecular pathways regulated by lncRNAs in RCC, and the functional roles of these transcripts in this malignancy, remains limited. We mined RNA-seq data from RCC patients to identify a set of 805 lncRNAs that are commonly overexpressed in this tumor type. We generated a custom CRISPR interference (CRISPRi) library targeting these lncRNAs and performed screens in multiple RCC cell lines to identify lncRNAs that are essential for RCC cell proliferation. These experiments revealed that the lncRNA Colorectal neoplasia differentially expressed (CRNDE) is required for growth of all tested RCC cell lines. Although this lncRNA has been shown to be overexpressed and is associated with poor patient survival in RCC and other types of cancer, its molecular function remains unclear. We identified a critical region of CRNDE that is necessary for RCC cell proliferation and we identified proteins that interact with this sequence. We also generated novel genetically-engineered alleles in mice that enable constitutional or conditional deletion of critical Crnde sequences. In this proposal, we will leverage our new understanding of this lncRNA, and the novel resources we have generated, in order to dissect the molecular function of CRNDE and define its role in normal physiology and in RCC pathogenesis in vivo. These experiments will take advantage of our extensive experience, and that of our collaborators, in evaluating noncoding RNA functions and RCC biology using cellular and animal models. Successful completion of the proposed research will address two major knowledge gaps in the fields of RNA biology and cancer biology: 1) our limited understanding of the molecular mechanisms-of-action of lncRNAs; and 2) how these mechanisms are co-opted by cancer cells to promote tumor growth, particularly in RCC. We anticipate that the principles revealed by these studies will be broadly applicable to our understanding of the roles of other lncRNAs in cancer cells and may set the stage for developing therapeutics that target CRNDE or the pathways it controls.

项目摘要越来越多的证据表明，长期不编码的RNA（LNCRNA）是一类多种非蛋白质 - 编码转录物>长度为200个核苷酸，在癌症的起始和进展中起重要作用。已提议lncrnas规范所有癌症的标志，但在绝大多数情况下，作用的分子机制仍然很少理解。这个知识差距是主要障碍要实现LNCRNA作为癌症和其他疾病的治疗靶标的潜力。就像许多人一样人类恶性肿瘤，LNCRNA在肾细胞癌（RCC）中经常失调。 RCC是最大的常见的肾癌和最致命的恶性泌尿科肿瘤，约有70,000个新的每年在美国诊断的病例。迄今为止，大多数RCC（和其他癌症）的基因组研究已经专注于鉴定蛋白质编码基因的疾病相关改变。我们对由LNCRNA在RCC中调节的分子途径，这些转录本在此中的功能作用恶性，仍然有限。我们从RCC患者那里挖掘了RNA-Seq数据，以识别一组805个LNCRNA 在这种肿瘤类型中通常过表达。我们产生了自定义CRISPR干扰（CRISPRI）针对这些LNCRNA并在多个RCC细胞系中进行屏幕的库，以识别LNCRNA RCC细胞增殖必不可少的。这些实验表明LNCRNA结直肠肿瘤所有测试的RCC细胞系生长需要差异表达（CRNDE）。虽然这个lncrna具有被证明过表达，与RCC和其他类型的患者生存不良癌症，其分子功能尚不清楚。我们确定了Crnde的关键区域 RCC细胞增殖和我们确定了与该序列相互作用的蛋白质。我们还产生了小说小鼠中的基因工程等位基因，使临界CRNDE的宪法或条件缺失序列。在此提案中，我们将利用我们对这种lncrna的新理解和新颖的资源为了剖析crnde的分子功能并定义了其在正常情况下的作用，我们已经产生了生理学和体内RCC发病机理。这些实验将利用我们的广泛经验以及我们的合作者的经验，用于评估非编码RNA功能和RCC生物学的经验细胞和动物模型。成功完成拟议的研究将解决两个主要知识 RNA生物学和癌症生物学领域的差距：1）我们对分子的有限理解 LNCRNA的作用机制； 2）这些机制如何被癌细胞选中以促进肿瘤生长，特别是在RCC中。我们预计这些研究所揭示的原则将是广泛的适用于我们理解其他LNCRNA在癌细胞中的作用，并可能为开发针对CRNDE或它控制的途径的治疗剂。

项目成果

期刊论文数量（2）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Target-directed microRNA degradation regulates developmental microRNA expression and embryonic growth in mammals.

DOI：
10.1101/gad.350906.123
发表时间：
2023-07-01
期刊：
GENES & DEVELOPMENT
影响因子：
10.5
作者：
Jones, Benjamin T.;Han, Jaeil;Zhang, He;Hammer, Robert E.;Evers, Bret M.;Rakheja, Dinesh;Acharya, Asha;Mendell, Joshua T.
通讯作者：
Mendell, Joshua T.