Role of Splicing Factors in Breast Cancer

剪接因子在乳腺癌中的作用

• 批准号: 8568241
• 负责人: OLGA ANCZUKOW-CAMARDA
• 金额: $ 10.71万
• 依托单位: COLD SPRING HARBOR LABORATORY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8568241
• 关键词: 3-Dimensional; Address; Alternative Splicing; Animal Model; Antisense Oligonucleotides; Apoptotic; Award; BCL2L11 gene; Biological; Breast; Breast Cancer Cell; Breast Cancer Model; Cancer cell line; Candidate Disease Gene; Cell Culture System; Cell Culture Techniques; Cell Line; Cell Proliferation; Cell model; Cells; Complement; Critiques; Data; Databases; Development; Environment; Epithelial; Epithelial Cells; Event; Exhibits; Faculty; Future; Gene Expression; Goals; Human; In Vitro; Individual; Laboratories; Lead; Literature; Malignant Neoplasms; Mammary Neoplasms; Mammary gland; Mediating; Mentors; Mentorship; Molecular; Monitor; Neoplasm Metastasis; Oncogene Proteins; Oncogenic; Phase; Phenotype; Play; Positioning Attribute; Process; Production; Protein Family; Protein Isoforms; RNA Sequences; RNA Splicing; Regulation; Reporting; Research; Resistance; Role; Serine; Technology; Therapeutic; Training; Translating; Tumor Suppressor Genes; Tyrosine Kinase Inhibitor; Writing; anticancer research; base; cancer cell; cancer initiation; cancer therapy; career; cell transformation; human ARMET protein; in vivo; malignant breast neoplasm; meetings; member; migration; mouse model; next generation; next generation sequencing; novel; novel therapeutics; overexpression; prevent; programs; public health relevance; therapeutic target; transcriptome sequencing; tumor; vector

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.

描述（由申请人提供）：癌细胞通常显示出替代剪接的异常谱，从而导致产生蛋白质同工型，这些蛋白质可以增加细胞增殖，迁移和凋亡耐药性。我的长期目标是建立一个独立的研究实验室，在该实验室中，我将通过改变各种癌基因和肿瘤抑制基因的表达来阐明分子机制在癌症中发挥作用。重要的是，这些发现将转化为新型治疗策略的发展。 K99/R00职业奖将通过在我的主要导师Adrian Krainer博士的指导下推进我的培训来帮助实现这一目标；在迈克尔·沙茨（Michael Schatz）博士的联合学下进行下一代测序；以及Mikala Egeblad博士的同学下的转移性乳腺癌模型。该培训将补充我以前在乳腺癌研究和RNA剪接机制方面的专业知识。冷泉港实验室的非常刺激的科学环境不仅会为我提供完成该项目的指导阶段所需的专业知识和设施，而且还将使我能够使我平稳地过渡到独立的教师职位。在K99的指导阶段，我将定义剪接因子的作用和剪接不调节在乳腺癌中的作用。在随后的R00独立阶段，我将缩减可能构成治疗目标的致癌剪接事件 我未来的独立研究。我们先前已经证明，剪接因子SRSF1的过表达可以在体外和体内转化人类乳腺上皮细胞。我们还表明，SRFS1水平由MYC癌蛋白直接调节。但是，其他剪接因子在人乳腺肿瘤中也过表达，这表明它们也可能在乳腺癌中起作用。在AIM 1中，在K99阶段，我将确定 使用相关细胞和动物模型模仿肿瘤出现的生物学环境的特定剪接因子。这包括：（i）识别替代方案的变化 剪接事件是通过在人类乳腺上皮细胞的3-D培养物中进行的下一代RNA测序的剪接因子介导的转化的； （ii）使用体内小鼠模型定义这些致癌剪接因子的转移潜力。在AIM 2中，在K99阶段，我将通过鉴定剪接因子表达的变化以及在MYC诱导的细胞培养系统中通过下一代RNA序列确定剪接因子表达和替代剪接曲线的变化，从而确定MYC在调节乳腺癌中的作用。在AIM 3中，在R00阶段，我将确定剪接因子在获得抵抗力中的作用 乳腺癌中的酪氨酸激酶抑制剂通过鉴定剪接因子水平的变化和我的替代剪接事件。最后，从K99阶段开始，进入R00阶段，我将确定使用反义寡核苷酸来调节AIMS 1-3中确定的特定过量替代剪接事件的治疗潜力。拟议的研究不仅会确定乳腺癌涉及的剪接因子，还可以确定其调节剂和特定目标。 该计划将建立我的独立研究计划的基础，我计划基于调节剪接因素的表达和活性或目标，为新的癌症疗法开发。