Nuclear body assembly and function in telomerase-free cancer cells

我的整个职业目标是成为一名老牌的癌细胞生物学家，运用多学科 确定相变如何用于促进癌细胞生长和发展的方法 针对癌症治疗的这些过程的策略。我对生物物质自组装的兴趣 来自于我在脂质双层分子动力学方面的博士工作和在相态方面的博士后工作 RNA颗粒的分离。在宾夕法尼亚大学物理科学肿瘤学中心(PSOC)的支持下，我开始 自2015年以来，我在宾夕法尼亚大学从事癌症方面的博士后工作，并对细胞和 癌症的分子物理学。我决定将我的独立研究集中在这一领域，并发起了一个项目 我会把它带到我未来的实验室。我的项目是关于端粒酶阴性的癌细胞 另一种延长端粒(ALT)途径以保持端粒长度的方法。ALT的存在- 相关早幼粒细胞白血病核体(APB)是ALT的一个独特特征，用于 诊断。APB对于ALT的端粒维持是必不可少的，但它们的形成和功能都是如此 端粒延长是ALT癌症表型的关键部分，但目前尚不清楚。我观察到 端粒DNA损伤诱导的APB表现出液相凝聚的行为特征， 这让我假设ALT细胞中端粒的缩短导致APB的成核 凝聚物作为端粒延长的一种机制。APBS的液体性质将促进 APB聚集成簇的端粒，这是ALT细胞的另一个特征。同时， APB液滴的凝聚可以浓缩DNA修复因子，为端粒提供使用机会 彼此作为修复模板以在APB内伸长。我开发了一种光遗传方法，并诱导了 APB的新组装，表现出液体行为并结合以驱动端粒聚集。在这K22里，我会 验证我对APB功能的假设，为以APB为靶点的癌症治疗提供依据。目标1侧重于 从APB的形成如何驱动端粒看APB功能与材料性质的关系 目标2通过测试，重点研究APB函数与其化学组成的依赖关系 APB的形成是否足以合成端粒。我正在洽谈我对……职位的报价 助理教授，计划明年早些时候开始我的独立职业生涯。通过我的多学科训练 背景和令人兴奋的初步数据支持我的假设，我处于有利地位，可以实现我的 作为一名独立的癌症研究人员的职业目标。然而，为了过渡到癌症研究， 顺利和成功地进行进一步培训，以获得癌症生物学方面的知识和技术专长， 特别是在DNA修复和端粒生物学方面，是非常需要的。因此，由此产生的额外保护时间 K22对我来说是至关重要的，我要专注于研究，沉浸在癌症生物学中，并培养与专业知识相衔接的技能 来自多个学科，以确立我作为癌症液体冷凝方面的专家的地位。

My overall career goal is to become an established cancer cell biologist who employs multidisciplinary approaches to determine how phase transition is used to promote cancer cell growth and to develop strategies targeting these processes for cancer therapy. My interest in self-assembly of biological matter originated from my Ph.D. work on molecular dynamics in lipid bilayers and postdoctoral work on phase separation of RNA granules. Supported by the Physical Science Oncology Center (PSOC) at Penn, I started working in cancer in my current postdoctoral position at Penn since 2015 and became interested in cell and molecular physics of cancer. I decided to focus my independent research in this area and initiated a project that I will bring with me to my future laboratory. My project concerns telomerase-negative cancer cells that rely on an alternative lengthening of telomeres (ALT) pathway to maintain telomere length. The presence of ALT- associated promyelocytic leukemia nuclear bodies (APBs) is a unique characteristic of ALT and is used for diagnosis. APBs are essential for telomere maintenance in ALT, but both how they form and how they function in telomere lengthening, which is a crucial part of the ALT cancer phenotype, are unknown. I observed that APBs induced by DNA damage at telomeres exhibit behavior characteristic of liquid phase condensation, leading me to hypothesize that telomere shortening in ALT cells induces nucleation of APB condensates as a mechanism for telomere elongation. The liquid nature of APBs would promote coalescence of APBs to cluster telomeres within APBs, another characteristic of ALT cells. Meanwhile, condensation of APB droplets can concentrate DNA repair factors, providing opportunities for telomeres to use one another as repair templates to elongate within APBs. I developed an optogenetic approach and induced de novo assembly of APBs that exhibit liquid behavior and coalesce to drive telomere clustering. In this K22, I will test my hypothesis for APB function to provide a basis for cancer therapy targeting APBs. Aim 1 focuses on the dependence of APB function on their material properties by asking how APB formation drives telomere clustering, and Aim 2 focuses on the dependence of APB function on their chemical composition by testing whether APB formation is sufficient for telomere synthesis. I am negotiating my offers for the position of Assistant Professor and plan to start my independent career earlier next year. With my multidisciplinary training background and exciting preliminary data that supports my hypothesis, I am well positioned to achieve my career objectives as an independent cancer researcher. However, to make the transition into cancer research smooth and successful, further training to gain knowledge and technical expertise in cancer biology, particularly on DNA repair and telomere biology, is much needed. Therefore, additional protected time from this K22 is crucial for me to focus on research, immerse in cancer biology, and develop skills bridging expertise from multiple disciplines to establish myself as an expert on liquid phase condensation in cancer.