Characterizing TLR Function of Human Dendritic Cells in the Elderly

描述(由申请人提供)： 候选人：候选人是一名内科和传染病专业的医生，是肺部和重症监护医学的研究员，也是耶鲁大学医学院研究医学项目的博士候选人。在他的临床培训期间，他意识到了老年患者护理方面的复杂挑战。因此，他决定将研究重点放在了解人类免疫反应中与年龄相关的变化上，重点放在先天免疫上。他的长期职业目标是在一家学术机构获得终身教职，他的大部分时间都致力于基础科学研究。申请者将在两位高素质导师(阿尔伯特·肖和理查德·布卡拉)的监督下，在由耶鲁大学免疫学(Peter Cresswell和Ruslan Medzhitov)、老年病学(Mary Tinetti)和临床流行病学和生物统计学(Heather Allore)领导人组成的导师咨询委员会的指导下，实施拟议的研究计划，以实现这一目标。申请者提出了一个有重点的教学课程和参加研究研讨会的计划，这些课程将补充他完成的研究生课程。最终，当候选人以终身教职的方式建立自己的独立研究实验室时，他将整合自己的临床和研究培训，以进行关于肺部衰老和先天免疫反应的转化性研究，这一决定得到了他的导师的支持。 环境：耶鲁大学的环境非常适合从事人类翻译研究。这位候选人和他的导师与耶鲁大学老龄化和胡椒中心项目有着长期的合作关系，该项目为人类研究提供招聘、信息学和生物统计学支持，并与耶鲁大学免疫生物学系的领导人保持着长期的合作关系，后者位于同一栋建筑内。 研究：这位候选人最近发表的研究表明，与年轻人(21-30岁)相比，年龄较大(65岁以上)的树突状细胞(DC)群体中的Toll样受体(TLR)功能普遍存在缺陷。在这里，重点是了解在老年人中观察到的与年龄相关的TLR1表面表达下降的基础，这似乎是由翻译后机制引起的。PI将使用包括脉冲追逐实验、共聚焦显微镜和荧光共振能量转移在内的细胞生物学技术来评估TLR1表面表达、细胞内分布和与TLR2异源二聚的动力学随年龄的变化。他还将评估TLR伴侣蛋白的贡献，如PRAT4a，已发现其在老年人细胞中的表达减少。最后，由于候选人计划建立他的独立研究计划，将对年轻人和老年人的支气管肺泡灌洗液中肺来源的肺泡巨噬细胞的TLR功能进行初步研究，以获得初步数据。综上所述，拟议的研究、职业发展和指导活动将有助于候选人作为一名多产的内科科学家进行培训。 公共卫生相关性：老龄化与免疫功能(免疫衰老)的进行性下降有关，导致对病毒和细菌感染的易感性增加，对疫苗的反应降低。虽然合并症很可能是导致观察到的死亡率增加的原因，但很明显，与衰老相关的宿主防御系统受损也导致了发病率和死亡率的增加。我们的研究重点是老年人先天免疫力的缺陷，最终，我们希望我们的工作将有助于解释老年人免疫力下降的原因，并有助于合理开发专门针对老年人的新疗法和疫苗。

DESCRIPTION (provided by applicant): Candidate: The candidate is a physician trained in Internal Medicine and Infectious Diseases who is a fellow in Pulmonary and Critical Care Medicine and a Ph.D. candidate in the Investigative Medicine Program at the Yale University School of Medicine. During his clinical training, he became aware of the complex challenges in the care of patients in the geriatric age group. As a result, he decided to focus his research on understanding age-associated changes in human immune responses, with an emphasis on innate immunity. His long-term career goal is to obtain a tenure-track position at an academic institution with a majority of his time dedicated to basic scientific investigation. The applicant will achieve this objective by carrying out the proposed research plan under the supervision of two highly qualified mentors (Albert Shaw and Richard Bucala), with guidance by a Mentorship Advisory Committee composed of leaders in immunology (Peter Cresswell and Ruslan Medzhitov), geriatrics (Mary Tinetti), and clinical epidemiology and biostatistics (Heather Allore) at Yale. The applicant proposes a focused program of didactic courses and participation in research seminars that will complement his completed graduate curriculum. Ultimately, when the candidate establishes his own independent research laboratory in a tenure-track faculty appointment, he will integrate his clinical and research training to pursue translational research on aging and innate immune responses in the lung, a decision supported by his mentors. Environment: The environment at Yale is ideal for pursuing human translational research. The candidate and his mentors have a longstanding collaboration with the Program on Aging and Pepper Center at Yale, which provide recruitment, informatics and biostatistical support for human studies, and have longstanding relationships with leaders in the Department of Immunobiology at Yale, who are located in the same building. Research: The candidate's published work recently demonstrated a generalized defect in Toll-like Receptor (TLR) function in dendritic cell (DC) populations from older (over 65 years), compared to young (21-30 years) individuals. Here, the focus was on understanding the basis for the age-associated decrease in TLR1 surface expression observed in older individuals, which appears to arise from post-translational mechanisms. The PI will employ cell biology techniques including pulse-chase experiments, confocal microscopy and fluorescence resonance energy transfer to evaluate age-associated changes in the kinetics of TLR1 surface expression, intracellular distribution and heterodimerization with TLR2. He will also evaluate the contribution of TLR chaperone proteins such as PRAT4a, the expression of which has been found to be decreased in cells from older individuals. Finally, in anticipation of the candidate's plans for establishing his independent research program, a pilot study of TLR function in lung-derived alveolar macrophages in bronchoalveolar lavage fluid from young and older individuals will be carried out to obtain pilot data. Taken together, the proposed research, career development and mentoring activities will facilitate the candidate's training as a productive physician- scientist. Public Health Relevance: Aging is associated with a progressive decline in immune function (immunosenescence) resulting in increased susceptibility to viral and bacterial infections and decreased response to vaccines. While it is likely that comorbid conditions contribute to the observed increase in mortality, it is clear that impaired host defenses associated with aging also contribute to increased morbidity and mortality. Our research focuses on defects of the innate immunity arm in older adults and, ultimately, we hope that our work will help explain the deterioration of immunity seen in older adults and aid in the rational development of novel treatments and vaccines geared specifically towards older adults.