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: 10434124
• 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/10434124
• 关键词: 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K 08应用程序的目标是提供一个严格的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过渡到独立研究员。 该候选人在UT Health圣安东尼奥完成了她的医学博士/博士学位，在那里她专注于先天免疫。 在UT西南完成内科和传染病培训后，她加入了博士的实验室。 Beth Levine，一位著名的自噬专家，继续一个关于自噬和 抗病毒免疫已知自噬通过特异性靶向抗病毒先天免疫而发挥作用 细胞内的病毒组分用于溶酶体降解，这一过程称为病毒吞噬。然而， 是如何被激活和调节的，我们仍然知之甚少。基于对宿主基因的无偏筛选， 辛德毕斯病毒和疱疹病毒感染时的自噬诱导 单纯疱疹病毒-1（HSV-1），她的初步工作确定了CDKL 5作为候选的新型噬病毒调节剂。 她证明了CDKL 5缺陷的细胞积累了高水平的SINV衣壳蛋白。此外，本发明还 她确定CDKL 5 KO小鼠对SINV和HSV-1的致死性CNS感染更敏感， 提示CDKL 5是一个重要的宿主抗病毒因子。在得知她的导师可能患上了绝症后， 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， 她成功阐明CDKL 5在抗病毒免疫中的功能所需的专业知识和培训， 成为独立调查员。她的导师顺利过渡后，莱文博士去世，博士。 迈克尔夏伊洛，她的主要共同导师谁是医生科学家和专家在先天免疫和自噬， 以及著名的病毒致病科学家朱莉·菲佛博士他们成功地训练了许多 博士后研究员实现科学独立。这个指导团队是完美的补充 她的外部共同导师赫伯特博士提出了关于自噬和抗病毒免疫之间界面的研究 “跳过”处女他是一位内科科学家，也是自噬和先天性抗病毒领域无可争议的领导者。 免疫力他在指导博士后学员方面也有很好的记录。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， 她的技术技能，并在科学和职业问题上提供指导。在她提出的研究中，申请人 将致力于1）定义CDKL 5在病毒吞噬中的功能，2）描述SINV生命周期的哪些步骤是 受CDKL 5调节，和3）确定CDKL 5在体内宿主抗病毒应答中的作用。总体看 拟议的研究可能会扩大我们对宿主细胞如何减轻有毒的过度生产的理解。 病毒感染时的蛋白质再加上获得优秀的机构资源， 关于领导力，专业知识和技术技能的课程，她被定位为成为一名成功的医生- 科学家专注于抗病毒先天免疫。