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.

项目概要越来越多的证据表明，长非编码 RNA (lncRNA) 是一类不同类型的非蛋白质- 编码转录本长度>200个核苷酸，在癌症的发生和进展中发挥重要作用。 LncRNA 已被提议调节所有癌症标志，但在绝大多数情况下，它们的分子作用机制仍知之甚少。这种知识差距是一个主要障碍实现lncRNA作为癌症和其他疾病治疗靶点的潜力。正如许多在人类恶性肿瘤中，lncRNA 在肾细胞癌 (RCC) 中经常失调。 RCC 是最肾癌的常见类型和最致命的泌尿系统恶性肿瘤，约有 70,000 例新发肾癌美国每年诊断出的病例数。迄今为止，大多数肾细胞癌（和其他癌症）的基因组研究专注于识别与疾病相关的蛋白质编码基因的改变。我们的理解 RCC 中 lncRNA 调节的分子途径，以及这些转录本在此过程中的功能作用恶性程度仍然有限。我们挖掘了 RCC 患者的 RNA-seq 数据，以确定一组 805 个 lncRNA，在这种肿瘤类型中通常过度表达。我们生成了定制的 CRISPR 干扰 (CRISPRi) 文库针对这些 lncRNA，并在多个 RCC 细胞系中进行筛选，以鉴定对于 RCC 细胞增殖至关重要。这些实验表明，lncRNA 结直肠肿瘤所有测试的 RCC 细胞系的生长都需要差异表达 (CRNDE)。虽然这个lncRNA有已被证明过度表达，并且与 RCC 和其他类型的患者生存率较差有关癌症，其分子功能仍不清楚。我们确定了 CRNDE 的一个关键区域，这是 RCC 细胞增殖，我们鉴定了与该序列相互作用的蛋白质。我们还生成了小说小鼠体内的基因工程等位基因能够实现关键 Crnde 的宪法性或条件性删除序列。在本提案中，我们将利用我们对这种 lncRNA 的新理解以及新颖的资源我们已经生成，以便剖析 CRNDE 的分子功能并定义其在正常情况下的作用生理学和 RCC 体内发病机制。这些实验将利用我们广泛的以及我们的合作者在评估非编码 RNA 功能和 RCC 生物学方面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）我们对分子生物学的了解有限 lncRNA的作用机制； 2）癌细胞如何利用这些机制来促进肿瘤生长，尤其是肾细胞癌。我们预计这些研究揭示的原则将得到广泛的应用适用于我们对其他 lncRNA 在癌细胞中的作用的理解，并可能为开发针对 CRNDE 或其控制途径的疗法。

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.