Characterization of long non-coding RNAs in the DNA damage response

描述（由申请人提供）：拟议的研究旨在研究长链非编码rna （lncRNAs）在DNA损伤反应（DDR）和乳腺癌中的作用。作为早期肿瘤发生的主要抗癌屏障，DDR涉及一个复杂而纠缠的过程网络，该过程感知和修复DNA损伤以维持基因组完整性，其中lncRNAs作为一类新的调控RNA分子可能发挥重要作用。在本研究中，我发现了强有力的证据，证明lncrna受ATM的调控，ATM是DDR中的一种必需激酶。其中两种lncrna， JANDA （Jade1相关ncRNA DNA损伤激活）和MANDS （Mfhas1相关ncRNA DNA损伤抑制），在染色质重塑和乳腺肿瘤发生和转移中具有重要影响。我认为DDR诱导的JANDA是连接DDR与组蛋白H4乙酰化的关键功能环节，DDR抑制的mands通过靶向Mfhas1作为肿瘤抑制因子，Mfhas1是一种候选癌基因之外的ROCO蛋白，JANDA的失调可能有助于乳腺癌的进展和转移。在Aim 1中，我将探讨JANDA和MANDS在DDR中的分子机制。我还将确定JANDA和MANDS对基因表达的全球调控及其在DDR和DNA修复中的作用。我的初步数据显示，JANDA在人类乳腺肿瘤中过度表达，可能在乳腺癌起始过程中具有促癌特性，因此在Aim 2中，我将评估JANDA和H4乙酰化在乳腺癌组织中的表达水平以及对乳腺细胞致癌应激的反应。JANDA可能是一种新的阴性预后因子，在HER2型肿瘤中表现出促转移活性，因此在Aim 3中，我将评估JANDA在乳腺癌中作为转移促进因子的作用，并检测沉默JANDA在曲妥珠单抗耐药HER2型肿瘤中的作用。最后，我将评估JANDA作为乳腺癌的潜在治疗靶点。这次申请的学习将为我提供最好的机会，开始我独立的学术生涯。我的长期目标是探索非编码rna在健康和疾病中的作用。为此，我将继续提高我在分子生物学、癌症生物学、表观遗传学、小鼠工程和操作方面的技术技能，并扩大我在癌症基因组学、干细胞生物学和转化生物学方面的专业知识。为了我的职业发展，我将学习提高自己的技能，管理实验室，指导博士后和学生，科学写作和演讲，建立新的合作。MD安德森癌症中心的癌症生物系在转化性癌症研究领域有着悠久的历史，我能够将最先进的研究成果转化为临床应用。在K99/R00的支持下，我将能够1)完成拟议的研究，并探索从初步研究中产生的新想法，2)寻找新的策略，使人类乳腺癌细胞在化疗和放疗中敏感化，3)建立自己的独立研究团队，并将我的网络扩展到DNA损伤，非编码RNA和乳腺癌领域。

DESCRIPTION (provided by applicant): The proposed study is to investigate the roles of long non-coding RNAs (lncRNAs) in the DNA damage response (DDR) and breast cancer. As a primary anti-cancer barrier in early tumorigenesis, the DDR involves a complicated and entangled network of processes that sense and repair DNA damage to maintain genomic integrity, in which lncRNAs, a novel class of regulatory RNA molecules, may play important roles. In this proposed study, I found strong evidence that lncRNAs were regulated by ATM, an essential kinase in the DDR. Two of such lncRNAs, termed JANDA (Jade1 associated ncRNA DNA damage activated) and MANDS (Mfhas1 associated ncRNA DNA damage suppressed), have significant impacts in chromatin remodeling and breast tumorigenesis and metastasis. I propose that DDR induced-JANDA is a key functional link that connects the DDR to histone H4 acetylation, and DDR suppressed-MANDS serves as a tumor suppressor by targeting Mfhas1, an ROCO protein beyond a candidate oncogene, and dysregulation of JANDA may contribute to breast tumor progression and metastasis. In Aim 1, I will explore the molecular mechanism of JANDA and MANDS in the DDR. I will also determine global regulation of gene expression by JANDA and MANDS and their roles in the DDR and DNA repair. My preliminary data showed JANDA was overexpressed in human breast tumors and may have pro-oncogenic property in breast cancer initiation, so in Aim 2 I will evaluate the expression level of JANDA and H4 acetylation in breast cancer tissues and in response to oncogenic stress in mammary cells. JANDA may serve as a novel negative prognostic factor and exhibit pro-metastatic activity in HER2 type tumors, so in Aim 3, I will assess the role of JANDA as a metastasis promoter in breast cancer and detect the effects of silencing JANDA in trastuzumab-resistant HER2 type tumors. Lastly, I will evaluate JANDA as a potential therapeutic target in breast cancer. This proposed study will provide me with the best opportunity to start up my independent career in academia. My long term goal is to explore the contribution of non-coding RNAs in health and disease. To that end, I will continue enhancing my technical skills in molecular biology, cancer biology, epigenetics, mouse engineering and manipulation, and expanding my expertise in cancer genomics, stem cell biology and translational biology. For my career development, I will learn to improve my skills on managing laboratory, mentoring postdoctoral fellows and students, scientific writing and presentation and establishing new collaborations. The Department of Cancer Biology at MD Anderson Cancer Center has a long history of being a leader in the field of translational cancer research, where I am able to translate state-of-art studies into clinical applications. With the support of K99/R00, I will be able to 1) complete the proposed studies and explore novel ideas that arise from preliminary studies, 2) search for novel strategies to sensitize human breast cancer cells in chemotherapy and radiotherapy, 3) establish my independent research team and expand my network into DNA damage, non-coding RNA and breast cancer fields.