Role of Splicing Factors in Breast Cancer

描述（由申请人提供）：癌细胞经常表现出异常的选择性剪接，导致蛋白质异构体的产生，可以增加细胞增殖、迁移和凋亡抵抗。我的长期目标是建立一个独立的研究实验室，在那里我将阐明通过改变各种癌基因和肿瘤抑制基因的表达，选择性剪接错误调节在癌症中发挥作用的分子机制。重要的是，这些发现将转化为新的治疗策略的发展。K99/R00职业奖将有助于实现这一目标，在我的主要导师Adrian Krainer博士的指导下，通过推进我在反义寡核苷酸技术方面的培训；Michael Schatz博士共同指导下的下一代测序；和转移性乳腺癌模型在Mikala Egeblad博士的共同指导下。这次培训将补充我之前在乳腺癌研究和RNA剪接机制方面的专业知识。冷泉港实验室非常刺激的科学环境不仅将为我提供完成这个项目的指导阶段所需的专业知识和设施，而且还将为我顺利过渡到一个独立的教师职位做好准备。在K99指导阶段，我将定义剪接因子和剪接失调在乳腺癌中的作用。在随后的R00独立阶段，我将确定可能构成治疗靶点的致癌剪接事件

DESCRIPTION (provided by applicant): Cancer cells often display aberrant profiles of alternative splicing, leading to the production of protein isoforms that can increase cell proliferation, migration, and apoptotic resistance. My long-term goal is to establish an independent research lab, where I will elucidate molecular mechanisms by which alternative splicing misregulation plays a role in cancer by altering the expression of various oncogenes and tumor-suppressor genes. Importantly, these findings will translate into the development of novel therapeutic strategies. The K99/R00 career award will help in achieving this goal by advancing my training in: antisense oligonucleotide technologies under the guidance of my primary mentor Dr. Adrian Krainer; next-generation sequencing under the co-mentorship of Dr. Michael Schatz; and metastatic breast cancer models under the co-mentorship of Dr. Mikala Egeblad. This training will complement my previous expertise in breast cancer research and RNA splicing mechanisms. The very stimulating scientific environment at Cold Spring Harbor Laboratory will not only provide me with the expertise and facilities necessary for the completion of the mentored phase of this project, but will also prepare me to transition smoothly into an independent faculty position. During the K99 mentored phase, I will define the role of splicing factors and splicing misregulation in breast cancer. In the subsequent R00 independent phase, I will indentify oncogenic splicing events that could constitute therapeutic targets to pursue during my future independent research. We have previously demonstrated that overexpression of the splicing factor SRSF1 can transform human mammary epithelial cells in vitro and in vivo. We have also shown that SRFS1 levels are directly regulated by the MYC oncoprotein. However, additional splicing factors are also overexpressed in human breast tumors, suggesting that they may also play a role in breast cancer. In Aim 1, during the K99 phase, I will determine the role of specific splicing factors in breast cancer using relevant cell and animal models that mimic the biological context in which the tumors arise. This includes: (i) identifying changes in alternative splicing events underlying splicing-factor- mediated transformation by next-generation RNA-sequencing in 3-D cultures of human mammary epithelial cells; and (ii) defining the metastatic potential of these oncogenic splicing factors using in vivo mouse models. In Aim 2, during the K99 phase, I will determine the role of MYC in the regulation of alternative splicing in breast cancer, by identifying changes in both splicing-factor expression and in alternative splicing profiles by next-generation RNA-sequencing in a MYC-inducible cell culture system. In Aim 3, during the R00 phase, I will determine the role of splicing factors in acquisition of resistance to tyrosine kinase inhibitors in breast cancer, by identifying changes in splicing-factor levels and i alternative splicing events. Finally, starting in the K99 phase and leading into the R00 phase, I will determine the therapeutic potential of using antisense oligonucleotides to modulate specific oncogenic alternative splicing events identified in Aims 1-3. The proposed research will identify not only splicing factors involved in breast cancer but also their regulators and specific targets. This plan will establish the basis for my independent research program, in which I plan to contribute to the development of new cancer therapies based on modulating the expression and activity of splicing factors or their targets.