The effect of HPV on apoptosis and cellular proliferation in laryngeal papillomas

HPV对喉乳头状瘤细胞凋亡和增殖的影响

• 批准号: 8490338
• 负责人: Michael P Underbrink
• 金额: $ 22.13万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8490338
• 关键词: Address; Adjuvant; Adult; Affect; Apoptosis; Apoptotic; Area; Binding; Biological Assay; Biometry; Caspase; Cell Communication; Cell Culture Techniques; Cell Cycle; Cell Proliferation; Cell physiology; Cells; Child; Clinical; Confocal Microscopy; Cytochromes; DNA Damage; Data; Debridement; Disease; Epidemiology; Epithelium; Functional disorder; Funding; Genes; Genotoxic Stress; Goals; HIV; HPV-High Risk; Hoarseness; Human; Human Papillomavirus; Human papilloma virus infection; Human papillomavirus 11; Human papillomavirus 6; Immunoblotting; Immunohistochemistry; Impairment; Individual; Infection; K-Series Research Career Programs; Knowledge; Larynx; Lead; Learning; Life Cycle Stages; Low risk HPV; Malignant neoplasm of cervix uteri; Maps; Mediating; Mediator of activation protein; Medical; Mentors; Modeling; Molecular; Molecular Analysis; Molecular Biology; Operative Surgical Procedures; Otolaryngology; Outcome; Papilloma; Papillomavirus Transforming Protein E6; Pathway interactions; Patients; Physicians; Positioning Attribute; Process; Proliferating; Property; Proteins; Recurrence; Research; Research Design; Research Project Grants; Research Training; Residencies; Residual state; Risk; Scientist; Skin; Squamous Papilloma of the Larynx; Supervision; Surgeon; Techniques; Testing; Therapeutic; Therapeutic Agents; Training; Undifferentiated; Upper respiratory tract; Virus; Virus Diseases; Voice; Voice Disorders; Writing; airway obstruction; bak protein; career; clinical efficacy; clinically relevant; cost; effective therapy; experience; high risk; human RCN2 protein; human UBE3A protein; improved; innovation; insight; interest; keratinocyte; meetings; molecular pathology; novel; novel therapeutics; post-doctoral training; prevent; respiratory; response; responsible research conduct; skills; small hairpin RNA; success; treatment effect; tumor; tumor growth; ubiquitin ligase; ubiquitin-protein ligase; virology; vocal cord

DESCRIPTION (provided by applicant): The overall goal of this Mentored Clinical Scientist Research Career Development Award proposal is to assist in establishing the independent research career of the candidate in the study of virus-host cell interactions, specifically of the cellular processes contributing to vocal cord dysfunction and upper airway obstruction caused by human papillomavirus (HPV) infections that lead to recurrent respiratory papillomatosis (RRP). The candidate is a board-certified otolaryngology surgeon with a focus on the management of voice disorders and difficult airways. Besides his clinical residency in otolaryngology, the candidate received postdoctoral training in the molecular biology of genus beta HPVs and the mechanisms by which they induce cell proliferation and block apoptosis after DNA damage - issues similar to those to be studied in the research component of this project, focused on the RPP-related genus alpha HPVs. The training and research components of the project are under the supervision of the co-mentors, Dr. Stephen Tyring and Dr. Thomas Albrecht. In addition to the mentored research component, the training plan calls for didactic coursework in such areas as virology, biostatistics, scientific writing, and several aspects of the responsible conduct of research, as well as attending relevant local and national meetings. The research project focuses on elucidating the mechanisms by which RPP-related HPVs block apoptosis and induce cellular proliferation in human keratinocytes. In high-risk HPV infections, the E6 viral protein targets the pro-apoptotic cellular Bak protein (as well as p53) for degradation, and so prevents the normal process of apoptosis from being induced by genotoxic stress. The candidate's preliminary data show that the RRP-related HPVs also retain the ability to target Bak for degradation. Aim 1 of the research project is to use immunoblotting and shRNA knockdown to determine the ability of HPV6 and HPV11 E6 proteins to blunt the apoptotic response, and also to determine the mechanism for Bak degradation. Aim 2 is to use a combination of protein pull down assays to identify conserved (Bak, E6AP) and novel binding partners of the 6E6 and 11E6 proteins. Finally, Aim 3 is to use immunostaining techniques to determine the effects of treatments targeted towards the induction of the extrinsic and/or intrinsic apototis pathways in both undifferentiated and differentiated E6-expressing cell cultures. Upon successful completion of this project, there will be a much better understanding of the molecular biology and pathogenic mechanisms of the "low- risk," RPP-related HPVs; this information will be invaluable in developing effective, function-sparing treatments for this presently incurable disease process. Further, the candidate will have obtained the preliminary data needed to compete successfully for R01 funding, and the skill set necessary for him to achieve his long-range goal - success as an established, extramurally funded surgeon-scientist focused on effective treatments for RPP and other laryngeal-related diseases.

描述（由申请人提供）：该指导临床科学家研究职业发展奖提案的总体目标是协助建立候选人在病毒-宿主细胞相互作用研究中的独立研究职业生涯，特别是导致声带功能障碍和上呼吸道阻塞的细胞过程，由人乳头瘤病毒（HPV）感染导致复发性呼吸道乳头瘤病（RRP）。候选人是一名委员会认证的耳鼻喉科外科医生，专注于语音障碍和困难气道的管理。除了他在耳鼻喉科的临床住院医师外，该候选人还接受了β HPV属分子生物学的博士后培训，以及它们在DNA损伤后诱导细胞增殖和阻断细胞凋亡的机制-与本项目研究部分中研究的问题相似，重点是RPP相关的α HPV属。该项目的培训和研究部分由共同导师Stephen Tyring博士和托马斯Albrecht博士监督。除了指导研究部分外，培训计划还要求在病毒学、生物统计学、科学写作和负责任地开展研究的若干方面等领域开展教学课程，并参加有关的地方和国家会议。该研究项目的重点是阐明RPP相关HPV阻断人类角质形成细胞凋亡和诱导细胞增殖的机制。在高风险HPV感染中，E6病毒蛋白靶向促凋亡细胞巴克蛋白（以及p53）进行降解，从而防止基因毒性应激诱导的正常凋亡过程。候选物的初步数据显示，与RPR相关的HPV也保留了靶向巴克降解的能力。本研究的第一个目的是利用免疫印迹和shRNA敲低来确定HPV 6和HPV 11 E6蛋白减弱凋亡反应的能力，并确定巴克降解的机制。目的2是使用蛋白质下拉测定的组合来鉴定6 E6和11 E6蛋白的保守（巴克，E6 AP）和新的结合配偶体。最后，目的3是使用免疫染色技术，以确定治疗的效果，针对诱导的外在和/或内在apototis途径在未分化和分化的E6表达细胞培养物。在成功完成该项目后，将更好地了解“低风险”RPP相关HPV的分子生物学和致病机制;这些信息对于开发针对这种目前无法治愈的疾病过程的有效的功能保留治疗方法将是非常宝贵的。此外，候选人将获得成功竞争R 01资金所需的初步数据，以及他实现长期目标所需的技能-成功作为一个既定的，壁外资助的外科医生科学家，专注于RPP和其他喉相关疾病的有效治疗。