Roles of RhoA/Rho-kinase Signaling in Cavernous Neurons Following Axonal Injury

描述（由申请人提供）：我提议的指导临床科学家研究职业发展（K 08）奖将集中在一个研究课题，具有巨大的潜力，影响神经泌尿学领域。前列腺癌是美国最常见的癌症。前列腺癌手术治疗的两个主要后遗症是术后尿失禁和勃起功能障碍（艾德）。根治性膀胱切除术（RP）后的尿失禁已大大减少，然而，“保留神经”RP后的神经源性ED仍然存在。神经泌尿学领域的进展主要集中在制定和实施策略，以保留RP后阴茎勃起的神经基础和海绵体神经的功能完整性，从而最大限度地提高术后勃起功能结局。本申请中提出的实验将海绵体神经中RhoA/ROCK信号传导的分子生物学和神经解剖学研究与体内神经生理学研究联合收割机结合。我们假设变性和神经元细胞凋亡是由于海绵体神经损伤后主要盆神经节中RhoA/ROCK的表达/活性增加。生理学研究的选择性RhoA和ROCK抑制剂的神经调节勃起功能和轴突再生的背景下提出海绵体神经损伤后的影响。了解损伤后RhoA/ROCK信号在海绵体神经中的有害作用及其对阴茎神经功能和修复的生理学影响，可以预期推进艾德的治疗策略，特别是那些与神经损伤以及其他周围神经病变相关的治疗策略。 我获得了杜兰大学分子药理学的医学博士/博士学位。我的博士论文主要研究糖尿病等全身性疾病对内皮细胞生物学的影响，因为它与心血管疾病和泌尿生殖道功能有关。在约翰霍普金斯大学接受泌尿外科培训期间，我获得了美国泌尿外科协会基金会的MD/PhD奖学金，研究镰状细胞病相关性阴茎异常勃起中内皮细胞功能障碍的病理生理学。在我的住院医师和博士后研究期间，我在临床和基础科学研究方面获得了宝贵的经验，这些研究侧重于确定问题并将传播疾病开始和进展的分子和病理事件联系起来。我的职业发展奖将侧重于进一步了解疾病的推定分子机制，从而设计新的疾病特异性药物治疗，以预防和治疗泌尿生殖道疾病。更重要的是，它将使我在一个新的研究领域获得额外的专业知识，并显着提高我的研究能力，努力获得经验和指导，这将导致一个独立和富有成效的研究生涯。 K 08奖将为我提供一个重要的受保护的时间块，致力于我的研究计划。对于临床活跃的科学家来说，这种保护的重要性再强调也不过分。病人护理的需求难以预测，而且还在不断扩大。此外，虽然许多早期职业赠款涵盖2年或3年，但K 08涵盖5年。较长的获奖期增强了我作为一名调查员对问题进行深入和严格研究的能力。布雷迪泌尿研究所非常适合让我成为一名独立的外科医生/泌尿科学家。约翰霍普金斯大学在培养独立科学家方面有着丰富的历史，并拥有开展我的K 08资助计划中的研究所需的所有资源。我的导师，亚瑟伯内特博士，是一个NIH支持的研究员，非常适合导航我的职业发展，并为我提供职业建议，同时帮助我平衡一个活跃的研究实验室和临床实践。 我在未来五年的目标是获得最好的指导，作为一个独立的外科医生科学家成功整合神经科学，细胞信号传导和疾病发病机制的神经生物学领域。我希望开发一个研究项目，重点是识别和表征神经损伤后促进轴突再生的新途径。我提出了一系列的实验，利用海绵体神经损伤的动物模型，这是一个建立的模型，以研究根治性切除术后ED的病理生理。我的职业发展奖的培训目标包括：1。了解周围神经系统功能的分子基础，包括神经发育，突触传递和神经病理学; 2。了解神经细胞的轴突和许旺细胞成分的基本解剖组织和组织学。此外，获得技能，以识别和区分正常，受伤和再生神经细胞的各种状态;和3。提高我对遗传学的理解，并通过专注于基因表达谱和生物信息学的研究来补充这一点，因为它与疾病病理生物学有关。我将接受的宝贵教育将使我了解轴突损伤的病理生理学，并使我能够设计疾病特异性分子治疗方法来保护下尿路功能。我的长期职业目标包括在R系列中获得额外的NIH资金，并希望有机会参与联合收割机结合多学科方法治疗泌尿生殖系统疾病过程的项目赠款。我希望有一天能指导其他年轻的外科医生/科学家，并为他们提供我的部门和K 08资金将为我提供的手段和机会。

DESCRIPTION (provided by applicant): My proposed Mentored Clinical Scientist Research Career Development (K08) Award will focus on a research topic which has tremendous potential to impact the field of Neurourology. Prostate cancer is the most commonly diagnosed cancer in the United States. Two major sequelae from the surgical management of prostate cancer are post-operative incontinence and erectile dysfunction (ED). Urinary incontinence following radical prostatectomy (RP) has been greatly reduced; however, neurogenic-ED following "nerve-sparing" RP persists. Advances in the field of neurourology have focused on the development and implementation of strategies for preserving the neurogenic basis of penile erection and functional integrity of the cavernous nerves following RP and thus maximizing postoperative erectile function outcomes. The experiments proposed in this application combine molecular biologic and neuroanatomic investigation of RhoA/ROCK signaling in the cavernous nerve with in vivo neurophysiological studies. We hypothesize that degeneration and neuronal cell apoptosis is a result of increased expression/activity of RhoA/ROCK in the major pelvic ganglion after cavernous nerve injury. Physiologic studies of selective RhoA and ROCK inhibitors effects in the context of neuroregulated erectile function and axonal regeneration are proposed after cavernous nerve injury. An understanding of the detrimental effects of RhoA/ROCK signaling in the cavernous nerve after injury and its influence on physiology of penile nerve function and repair can be expected to advance therapeutic strategies for ED particularly those related to nerve injury as well as other peripheral nerve neuropathies. I obtained my MD/PhD from Tulane University in Molecular Pharmacology. My PhD thesis focused on the influence of systemic disease states such as diabetes on endothelial cell biology, as it relates to cardiovascular disease and genitourinary tract function. During my urology training at Johns Hopkins, I was awarded a MD/PhD Fellowship from the American Urological Association Foundation to study the pathophysiology of endothelial cell dysfunction in sickle cell disease-associated priapism. During my residency and post-doctoral fellowship, I gained invaluable experience in both clinical and basic science investigations focusing on identifying a problem and linking molecular and pathological events which propagate disease initiation and progression. My career development award will focus on furthering my education about identifying putative molecular mechanisms of disease and thus design new disease-specific pharmacotherapies for the prevention and treatment of genitourinary tract disorders. More importantly, it will allow me to gain additional expertise in a new research area and significantly enhance my research capabilities in an effort to gain experience and mentorship that will lead to an independent and productive research career. The K08 award will afford me a significant block of protected time to devote to my research program. For a clinically active scientist the importance of this protection cannot be over-emphasized. The demands of patient care are difficult to predict and ever-expanding. Furthermore, while many early career grants cover 2 or 3 years, the K08 covers 5 years. The longer award period enhances my ability as an investigator to perform in-depth and rigorous study of a problem. The Brady Urological Institute is ideally suited to allow me to become an independent surgeon/scientist in Urology. Johns Hopkins has a rich history of training independent scientists and has all the resources necessary to carry out the proposed studies in my K08 grant. My mentor, Dr. Arthur Burnett, is a NIH supported investigator and is ideally suited to navigate my career development and provide me with career advice while helping me balance an active research lab and clinical practice. My goal over the next five year period is to obtain the best mentorship possible to succeed as an independent surgeon scientist integrating the fields of neuroscience, cell signaling, and neurobiology of disease pathogenesis. I hope to develop a research program focused on the identification and characterization of novel pathways involved in the promotion of axonal regeneration in response to nerve injury. I have proposed a series of experiments that utilize an animal model of cavernous nerve injury which is an established model to study the pathophysiology of post-radical prostatectomy ED. My career development award training goals include: 1. Understanding the molecular basis of peripheral nervous system function including neural development, synaptic transmission, and neuropathology; 2. Understanding the fundamental anatomical organization and histology of the axon and Schwann cell components of a nerve cell. Additionally acquire the skills to identify and differentiate the various states of normal, injured, and regenerating nerve cells; and 3. Improve my understanding of genetics and complement this with studies focused on gene expression profiling and bioinformatics as it relates to disease pathobiology. The invaluable education I will receive would allow me to understand the pathophysiology of axonal injury and allow me to design disease-specific molecular-based therapies to preserve lower urinary tract function. My long term career goals include obtaining additional NIH funding in the R-series and hopefully have the opportunity to participate in program project grants which combine a multi-disciplinary approach to genitourinary disease processes. I hope to one day mentor other young surgeon/scientist and provide them with the means and opportunity that my department and K08 funding will afford me.