Translational studies of cellular senescence as a regulator of doxorubicin-mediated arterial dysfunction

项目摘要/摘要 这个K99/R00应用程序的目的是为Zachary Clayton博士提供支持，他是一位有前途的职位- 道格拉斯·海尔斯博士实验室的博士生。这项额外的研究和培训将使他能够 成功转型为转化性心血管(CV)领域的独立研究者 生理学。作为他提议的K99训练计划的一部分，他将学习新的技术技能，提高他的智力 和专业技能，并参加各种职业发展活动，包括那些有助于 使他成为心脏肿瘤学和细胞衰老领域的领导者。他提出的项目旨在 探讨细胞衰老在调节动脉功能障碍(即主要危险因素)中的作用 对于心血管疾病[CVD])使用常用化疗药物阿霉素(Doxo)，首先在小鼠身上(K99) 后来翻译到老鼠和人类身上(R00)。Doxo后，衰老细胞在CV系统中积累 而细胞衰老可能会加剧动脉功能障碍的上游调节因素(即炎症和 氧化应激)。在强大的初步数据的指导下，克莱顿博士将首先(目标1)确认Doxo导致 利用互补的小鼠模型通过细胞衰老导致动脉功能障碍 系统清除衰老细胞。有来自国际的技术技能的指导和培训 公认的细胞衰老专家，他将(目标2)进行创新的体外细胞培养 在血管细胞中确定细胞衰老在关键表型中的作用的实验 Doxo处理的小鼠的血浆引起的潜在的动脉功能障碍。在过渡到教职员工之后 接下来，克莱顿博士将(Aim 3；R00)首先将他的发现翻译到老鼠身上，进行老鼠对老鼠的实验 血浆交换实验确定Doxo介导的动脉功能障碍是否可以转移 以及这种反应是否依赖于细胞衰老的激活。接下来，他会 通过确定服用阿昔洛韦的癌症患者的血浆在人类中的作用，将他的发现扩展到人类 促进培养的血管细胞的动脉功能障碍相关表型。总体而言，拟议的研究 有潜力解决NHLBI的两个战略研究重点：1)研究新的病理生物学 心血管疾病发生的重要机制；2)确定降低癌症血管发病率的策略 幸存者。拟议的项目将为未来的可资助研究提供机会，最终将达到 在该奖项的第4-5年期间提交小说R01。Seals博士(主要导师)是一位国际专家 美国国立卫生研究院资助的科学家，在翻译简历研究方面有很强的成功指导历史。 在他的指导和共同导师朱迪思·坎皮西博士的指导下，顾问团成员杨霍博士 Bae、Thomas LaRocca、Saul Villeda、Lavanya Kondapalli、Kamali Kimdar和生物统计学家尤志颖博士， 克莱顿博士将能够成功完成拟议的研究和培训计划，并过渡到 一家顶级研究机构(R1)的独立、由外部资金资助的终身教职。

PROJECT SUMMARY/ABSTRACT The purpose of this K99/R00 application is to provide support for Dr. Zachary Clayton, a promising post- doctoral fellow in the laboratory of Dr. Douglas Seals. This additional research and training will allow him to successfully transition into an independent investigator in the field of translational cardiovascular (CV) physiology. As part of his proposed K99 training plan, he will learn new technical skills, enhance his intellectual and professional skills and participate in various career development activities, including those that will help establish him as a leader in the fields of cardio-oncology and cellular senescence. His proposed project seeks to investigate the role of cellular senescence in regulating arterial dysfunction (i.e. the primary risk factor for CV diseases [CVD]) with the common chemotherapeutic agent Doxorubicin (DOXO), first in mice (K99) and later translating to mice and humans (R00). Senescent cells accumulate in the CV system following DOXO and cellular senescence may exacerbate upstream regulators of arterial dysfunction (i.e. inflammation and oxidative stress). Guided by strong preliminary data, Dr. Clayton will first (Aim 1) confirm that DOXO causes arterial dysfunction via cellular senescence by utilizing complementary mouse models in which he can systemically clear senescent cells. With guidance and training in technical skills from an internationally recognized expert in cellular senescence, he will then (Aim 2) conduct innovative ex vivo cell culture experiments in vascular cells to determine the role of cellular senescence in mediating key phenotypes underlying arterial dysfunction, evoked by plasma from DOXO-treated mice. After transitioning to a faculty position, Dr. Clayton will next (Aim 3; R00) translate his findings first to mice, by performing mouse-to-mouse plasma exchange experiments to determine whether DOXO-mediated arterial dysfunction can be transferred via the circulation and whether this response is dependent upon cellular senescence activation. Next, he will extend his findings to humans by determining the role of plasma from DOXO-treated cancer patients in facilitating arterial dysfunction-related phenotypes in cultured vascular cells. Overall, the proposed research has the potential to address 2 strategic research priorities of NHLBI: 1) investigate new pathobiological mechanisms important to the onset of CVD; 2) determine strategies for reducing vascular morbidity in cancer survivors. The proposed projects will provide opportunities for future fundable research, culminating in submission of a novel R01 during years 4-5 of this award. Dr. Seals (primary mentor) is an internationally recognized and NIH funded scientist with a strong history of successful mentoring in translational CV research. With his guidance and the guidance of co-mentor Dr. Judith Campisi, advisory team members Drs. Youngho Bae, Thomas LaRocca, Saul Villeda, Lavanya Kondapalli, Kamali Kimdar and biostatistician Dr. Zhiying You, Dr. Clayton will be able to successfully complete the proposed research and training plan and transition to an independent, extramurally-funded tenure-track position at a top-tier (R1) research institution.