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: 8190228
• 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/8190228
• 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. PUBLIC HEALTH RELEVANCE: Recurrent respiratory papillomatosis (RRP) is characterized by the growth of tumors in the upper respiratory tract, caused by infection with the human papilloma virus (HPV). Understanding the effects of HPV6 and HPV11 infection in the airway cells (keratinocytes) will clarify how RRP tumors proliferate in the upper respiratory tract and should lead to the discovery of novel, targeted therapies that can be used for the safe, voice-preserving treatment of this disease. Additionally, principles learned through the molecular study of RRP-associated HPV-host interactions will undoubtedly impact the study and treatment of other virus-induced diseases.

描述（由申请人提供）：这项受过指导的临床科学家研究职业发展奖的总体目标是帮助建立候选人在病毒宿主相互作用的研究中的独立研究职业，特别是针对人类Papillomavirus（HPV）引起的诉讼的细胞过程，导致声带功能障碍和上部气道阻塞，导致了诉讼（RESS）。该候选人是一名经过董事会认证的耳鼻喉科医生，重点是对语音障碍和难度的管理。除了他在耳鼻喉科方面的临床居留外，该候选人还接受了βHPV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凋亡的机制 - 与该项目的研究成分相似的问题，重点是RPP -RPP -RPP -RPPENUS ALPHAHPVS。该项目的培训和研究组成部分是在联席会员斯蒂芬·泰林（Stephen Tyring）和托马斯·阿尔布雷希特（Thomas Albrecht）博士的监督下。除了指导的研究部分外，培训计划还要求在病毒学，生物统计学，科学写作以及负责任的研究行为的几个方面以及参加相关的地方和国家会议等领域进行教学课程。该研究项目的重点是阐明与RPP相关的HPV阻断凋亡的机制，并诱导人角质形成细胞中的细胞增殖。在高风险的HPV感染中，E6病毒蛋白靶向降解促凋亡的细胞BAK蛋白（以及p53），因此可以防止正常的凋亡过程是由遗传毒性应激诱导的。候选人的初步数据表明，与RRP相关的HPV还保留了靶向BAK降解的能力。研究项目的目标1是使用免疫印迹和shRNA敲低来确定HPV6和HPV11 E6蛋白钝化凋亡反应的能力，并确定BAK降解的机制。 AIM 2是使用蛋白质下拉测定法的组合来鉴定6E6和11E6蛋白的保守（BAK，E6AP）和新型结合伙伴。最后，目标3是使用免疫染色技术来确定针对诱导外在和/或固有的apotis途径的治疗的影响，在未分化和分化的E6表达E6细胞培养物中。成功完成该项目后，将对“低风险”的分子生物学和致病机制有更好的了解，与RPP相关的HPV；对于目前无法治愈的疾病过程，为开发有效的功能性治疗而言，此信息将是无价的。此外，候选人将获得成功竞争R01资金所需的初步数据，以及他实现长期目标所必需的技能 - 成功，作为一个既定的，外部资助的外科医生，专注于RPP和其他喉相关疾病的有效治疗方法。 公共卫生相关性：复发性呼吸乳头状瘤病（RRP）的特征是由人类乳头状瘤病毒（HPV）感染引起的上呼吸道中肿瘤的生长。了解HPV6和HPV11感染对气道细胞（角质形成细胞）的影响将阐明RRP肿瘤在上呼吸道中如何增殖，并应导致发现可用于这种疾病的安全，语音治疗的新型，有针对性的疗法。此外，通过对RRP相关的HPV-HOST相互作用的分子研究学到的原则无疑会影响其他病毒诱导疾病的研究和治疗。