Project 3

项目3

基本信息

批准号：
8744319
负责人：
DAVID L SPECTOR
金额：
$ 42.22万
依托单位：
COLD SPRING HARBOR LABORATORY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-01-01 至 2016-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8744319
关键词：
Address Alleles Alternative Splicing Biochemical Biological Breast Breast Cancer Model Cancer Biology Cells Chromatin Code Complex Development Diagnostic Disease Embryo Etiology Eukaryota Exhibits Fibroblasts Functional RNA Future Gene Expression Gene Expression Regulation Genes Genetic Transcription Genome Genomics Goals Human Human Genome Knock-out Knockout Mice Laboratories Lead Liver Lung Malignant Neoplasms Mammary Neoplasms Modeling Molecular Mus NCI Center for Cancer Research Neoplasm Metastasis Neoplasm Transplantation Nonmetastatic Nuclear Nucleic Acid Regulatory Sequences Organism Outcome Pathway interactions Pattern Play Polycomb Property Prostate Proteins Proteome RNA RNA Sequence Analysis RNA Splicing Ribonucleoproteins Role Sampling Series Site Spliced Genes Tetanus Helper Peptide Tissue-Specific Gene Expression Tissues Transcript Transgenic Organisms cancer initiation cancer therapy cell type gain of function genome sequencing human disease in vivo innovation insight interest loss of function mRNA Precursor malignant breast neoplasm mammary gland development mouse development mouse genome mouse model next generation overexpression therapeutic target tumor tumor progression tumorigenesis

项目摘要

Long nuclear retained non-coding RNAs (IncRNAs) represent a large and relatively unmined class of RNAs that are likely to play critical roles in gene regulation and disease etiology. A major challenge is to understand the molecular functions of specific IncRNAs both at the cellular level and within the context of an organism. The long-term goal of this project is to identify and characterize IncRNAs that play a critical role in mammary gland development and the initiation and progression of breast cancer. Here, a series of Aims are presented to dissect out the role of Malati, an abundant IncRNA localized to nuclear speckles and focally amplified in a significant number of metastatic breast cancers. Studies are proposed to develop innovative loss-of-function and gain-of-function mouse models combined with cell biological approaches to assess the function of Malati in normal development and in breast cancer initiation and metastasis. The impact of alterations in the level of Malati on tissue organization will be examined, and its effect on alternative splicing in a tissue-specific manner will be pursued by next-generation RNA-sequencing analyses. Complementary cell biological studies will delve into the role that Malati plays in nuclear organization and its impact on the dynamics of pre-mRNA splicing factors enriched in nuclear speckles. The Malati RNP will be purified using an RNA-tagging strategy and its proteome will be characterized in order to identify proteins responsible for its nuclear retention and to provide additional insight into its function. In the final Aim a series of newly identified IncRNAs, that are misregulated in breast cancer, will be prioritized and several will be selected for functional analyses, to identify genes and pathways that they target, and to elucidate the mechanisms by which they contribute to breast cancer tumorigenesis. Together, the proposed studies will provide important insights into the role of several IncRNAs in normal development and cancer and will lead to new opportunities for the identification and characterization of a new and exciting class of potential therapeutic targets.

长核保留的非编码RNA（ERCRNA）代表了大型且相对未固定的RNA类，它们可能在基因调节和疾病病因学中起关键作用。一个主要的挑战是在细胞水平和生物体的背景下了解特定增量的分子功能。该项目的长期目标是识别和表征在乳腺发育以及乳腺癌的启动和进展中起关键作用的增量。在这里，提出了一系列目标，以阐明马拉蒂的作用，马拉蒂的作用是一种局部到核斑点的大量增量，并在大量的转移性乳腺癌中局部扩增。提出了研究以开发创新的功能丧失和功能获取小鼠模型，并结合细胞生物学方法来评估Malati在正常发育以及乳腺癌的起始和转移中的功能。将检查Malati水平的变化对组织组织的影响，并将通过下一代RNA序列分析来追求其对组织特异性剪接的影响。互补的细胞生物学研究将研究马拉蒂在核组织中所起的作用及其对富含核斑点的MRNA剪接因子动态的影响。 Malati RNP将使用RNA的策略进行纯化，其蛋白质组将被表征，以鉴定负责其核保留率的蛋白质，并提供对其功能的更多见解。在最终目标中，将优先考虑一系列在乳腺癌中被误导的新鉴定的增量，并将选择一些用于功能分析，以识别其靶向的基因和途径，并阐明它们对乳腺癌肿瘤造成贡献的机制。拟议的研究将共同提供有关几个增量在正常发育和癌症中作用的重要见解，并将为鉴定和表征一系列新的潜在治疗靶标的新机会。