Characterization of CDKL5 function in host antiviral defense

CDKL5 在宿主抗病毒防御中的功能表征

• 批准号: 10282404
• 负责人: Josephine Wanjiru Thinwa
• 金额: $ 19.66万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-18 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10282404
• 关键词: Antiviral Agents; Antiviral Response; Autophagocytosis; Autophagosome; Biological Assay; Brain; Capsid; Capsid Proteins; Cell Death; Cells; Central Nervous System Infections; Co-Immunoprecipitations; Communicable Diseases; Complement; Cyclin-Dependent Kinases; Data; Defect; Defense Mechanisms; Doctor of Philosophy; Double Stranded DNA Virus; Generations; Genes; Goals; Health; Herpesvirus 1; Host Defense; Human; Infection; Inflammatory; Inflammatory Response; Internal Medicine; Interphase; Knockout Mice; Leadership; Learning; Life Cycle Stages; Lysosomes; Mentors; Mentorship; Molecular; Morphogenesis; Mus; Mutation; Natural Immunity; Neurodevelopmental Disorder; Neurons; Nonstructural Protein; Pathway interactions; Physicians; Physiological; Play; Positioning Attribute; Postdoctoral Fellow; Process; Production; Proteins; RNA; RNA Viruses; Research; Research Personnel; Resources; Role; Scientist; Sindbis Virus; Small Interfering RNA; Structural Genes; Structural Protein; Technical Expertise; Terminal Disease; Testing; Tissues; Training; Translations; Viral; Viral Load result; Viral Pathogenesis; Viral Proteins; Viral Structural Proteins; Virion; Virus; Virus Diseases; Virus Replication; Work; antiviral immunity; base; biomedical scientist; brain tissue; career; career development; cell type; cytokine; experience; genome-wide; imaging modality; in vivo; mutant; neuron loss; neurotropic virus; novel; particle; pathogen; pathogenic virus; programs; reconstitution; skills

Project Summary The goal of this K08 application is to provide a rigorous 5-year scientific and careered development training plan that will facilitate Dr. Josephine Thinwa to transition from a post-doctoral fellow to an independent investigator. The candidate completed her MD/PhD degree at UT Health San Antonio where she focused on innate immunity. After completing internal medicine and infectious diseases training at UT Southwestern, she joined the lab of Dr. Beth Levine, a renowned expert in autophagy, to pursue a project on the interface between autophagy and antiviral immunity. Autophagy is known to function in antiviral innate immunity by specifically targeting intracellular viral components for lysosomal degradation, a process called virophagy. However, how virophagy is activated and regulated is still poorly understood. Based on an unbiased screen for host genes necessary for autophagy induction during infection with two prototypic neurotropic viruses, Sindbis virus (SINV) and Herpes Simplex Virus-1 (HSV-1), her preliminary work identified CDKL5 as a candidate novel regulator of virophagy. She demonstrated that cells deficient in CDKL5 accumulated high levels of SINV capsid protein. Additionally, she determined that CDKL5KO mice were more susceptible to lethal CNS infections with SINV and HSV-1, suggesting that CDKL5 is a critical host antiviral factor. After learning of her mentor’s likely terminal illness, Dr. Thinwa worked with Dr. Levine to establish a mentoring team of accomplished scientists who would provide the expertise and training necessary for her to successfully elucidate the function of CDKL5 in antiviral immunity and become an independent investigator. Her mentorship transitioned smoothly after Dr. Levine passed away to Dr. Michael Shiloh, her primary co-mentor who is a physician scientist and expert in innate immunity and autophagy, and Dr. Julie Pfeiffer, a renowned scientist in viral pathogenesis. They have successfully trained numerous postdoctoral fellows to achieve scientific independence. This mentoring team is perfectly complemented for the proposed studies at the interface between autophagy and antiviral immunity by her outside co-mentor Dr. Hebert “Skip” Virgin. He is a physician scientist and an undisputed leader in the field of autophagy and innate antiviral immunity. He also has a very strong track record of mentoring postdoctoral trainees. A committee of advisors and collaborators consisting of three outstanding scientists will provide additional scientific expertise to enhance her technical skills and offer guidance in scientific and career matters. In her proposed research, the applicant will aim to 1) define the function of CDKL5 in virophagy, 2) delineate which step(s) of the SINV life cycle are modulated by CDKL5, and 3) determine the role of CDKL5 in host antiviral response in vivo. Overall, the proposed studies are likely to expand our understanding of how host cells mitigate the toxic overproduction of viral proteins during infection. Together with access to excellent institutional resources, and training from didactic courses on leadership, grantsmanship and technical skills, she is positioned to become a successful physician- scientist focused on antiviral innate immunity.

项目摘要 此K08应用程序的目标是提供严格的5年科学和职业发展培训计划 这将有助于Josephine Thinwa博士从博士后研究员转变为独立研究员。 这位候选人在德克萨斯大学圣安东尼奥分校完成了医学/博士学位，在那里她专注于先天免疫。 在德克萨斯大学西南分校完成内科和传染病培训后，她加入了博士的实验室。 贝丝·莱文，一位著名的自噬专家，致力于研究自噬和自噬之间的接口。 抗病毒免疫。已知自噬通过特异性靶向在抗病毒的先天免疫中发挥作用。 溶酶体降解的细胞内病毒成分，这一过程称为病毒吞噬。然而，如何吞噬病毒 是否被激活和监管仍然知之甚少。基于对宿主基因的无偏见筛选 辛德比斯病毒(SINV)和疱疹病毒感染自噬诱导 单纯疱疹病毒1型(HSV-1)，她的初步工作确定CDKL5是一个候选的新的病毒吞噬调节因子。 她证明，CDKL5缺陷的细胞积累了高水平的SINV衣壳蛋白。另外， 她确定CDKL5KO小鼠更容易感染SINV和HSV-1的致命性中枢神经系统， 提示CDKL5是一种重要的宿主抗病毒因子。在得知她的导师可能得了绝症后。 Thinwa与Levine博士合作建立了一个由有成就的科学家组成的指导团队，他们将为 她需要的专业知识和培训，以成功地阐明CDKL5在抗病毒免疫和 成为一名独立调查员。在莱文博士去世后，她的导师生涯顺利过渡。 迈克尔·希洛是她的主要合作导师，是一名内科科学家，也是先天性免疫和自噬方面的专家， 以及著名的病毒发病机制科学家朱莉·法伊弗博士。他们已经成功地培训了许多 博士后研究员实现科学独立。这一指导团队与 她的外部合作导师赫伯特博士提议对自噬和抗病毒免疫之间的接口进行研究 “跳过”维珍。他是一位内科科学家，也是自噬和先天抗病毒领域无可争议的领导者。 豁免权。他在指导博士后实习生方面也有着非常出色的记录。顾问委员会 由三位杰出科学家组成的合作者将提供额外的科学专业知识，以增强 她的技术技能，并在科学和职业方面提供指导。在她提出的研究中，申请者 将旨在1)确定CDKL5在病毒吞噬中的功能，2)描述SINV生命周期的哪一步(S) 3)确定CDKL5在体内宿主抗病毒反应中的作用。总体而言， 拟议中的研究可能会扩大我们对宿主细胞如何减轻有毒的过量生产的理解 感染过程中的病毒蛋白。再加上获得优秀的机构资源，以及从教育中获得培训 关于领导力、资质和技术技能的课程，她有能力成为一名成功的内科医生- 科学家专注于抗病毒的先天免疫。