Hypothalamic Orexin Role in Menopause-Associated Hot Flashes and Mood/Sleep Disru

描述（由申请人提供）：目前，我在以下方面接受了广泛的培训：1）在我的硕士论文期间，女性性激素和行为的神经内分泌学; 2）在我的博士论文期间，压力反应的神经内分泌学和功能性神经解剖学; 3）在我的博士后培训期间，翻译动物模型。该培训集中于整个动物的生理和行为，结合使用微透析的体内神经化学测量;药理学和道追踪化合物的立体定位递送，以及使用免疫组织化学检测立即早期基因蛋白的离体功能性脑成像。2008年和2010年，我分别晋升为研究助理教授，然后晋升为精神病学系助理教授。在此期间，我发表了第一作者自然医学文章，其中主要发现是下丘脑食欲素系统在公认的惊恐障碍大鼠模型中起着关键作用，并且惊恐障碍患者的中枢食欲素水平升高。我继续确定是否有其他病理与过度活跃的食欲素系统有关。正是在这里，我在临床前研究中注意到雌激素抑制食欲素活性，最近的一项临床研究表明，绝经期雌激素的急剧丧失导致食欲素中枢水平增加300%，而雌激素替代治疗则逆转了这一趋势。这一证据，结合食欲素在体温调节中的已知作用，以及食欲素系统位于富含两种雌激素受体的下丘脑区域，导致我目前的假设，即“食欲素系统在更年期相关症状中起着关键作用，如潮热，情绪/睡眠中断”。大约70%的接近更年期的妇女会受到诸如潮热之类的更年期不良症状的影响，北美更年期协会认为每天有6,000多名美国妇女达到更年期。此外，63%的50-64岁妇女目前有工作（美国人口普查局，2003年）。因此，不利的更年期症状通过损失工作日和医疗服务的直接保健费用对工作场所产生重大的经济影响。这个K 01培训补助金代表了我职业生涯中一个重要但合乎逻辑的方向，需要IU专家的额外指导和培训：1）丹尼尔Rusyniak医学博士的体温调节啮齿动物研究; 2）Kathryn Jones博士的性激素神经内分泌学; 3）人类受试者的更年期相关“潮热”和Janet Carpenter博士，RN，FAAN教授的更年期课程;和4）转化研究方法和动物模型由Anantha Shekhar，MD/PhD。我的培训还将包括托德Skaar博士针对目标1的药理学培训; IUSM的William Truitt博士和Kenneth Cornetta博士针对目标2和3的分子生物学课程和培训;以及IUSM的Robert Bies博士针对目标3b的昼夜节律分析培训。在Rusyniak和Carpenter博士的最初指导下，我获得了内部资助的CTSI项目开发补助金和2011年转化研究的KL-2 CTSI青年研究者基础科学奖，以获得该补助金的初步数据。这里概述的目标和研究将有助于从“路线图”K奖过渡到“路线图”R奖。我在2011年的美国神经精神药理学学院和2012年的转化科学会议上提出了初步结果，并分别入选了“数据闪电战”会议;并获得了学者摘要奖。我还在准备一份关于我们的“潮热”脆弱性的新模型的手稿。本研究具有创新性，原因如下：1）据我们所知，这是首次尝试确定增食欲素的作用 绝经期不良症状截至2013年3月，在PubMed上搜索“食欲素”和“潮热”（这是主要的更年期症状）没有结果; 2）这项拟议的研究是基于关于食欲素和受体生理学的雌激素调节的临床前信息，食欲素在睡眠觉醒周期中的作用，以及我们最近将过度活跃的食欲素系统与焦虑和温度失调联系起来的临床前和临床数据; 3）我们将进行动物研究以阐明食欲素诱导更年期症状的潜在机制，据我们所知，这在以前没有报道过; 4）我已经开发了新的潮热脆弱性动物模型来测试我的假设; 5）我将应用药理学、急性基因和慢性慢病毒siRNA基因沉默技术来理解机制; 6）我将阐明调节潮热的神经机制，重点是下丘脑食欲素系统; 7）我将提供机制研究，这将导致翻译临床方案，发现食欲素受体拮抗剂的新用途，提交FDA批准用于失眠。我最近接受了IUSM解剖学和细胞生物学系的终身助理教授职位。我的实验室（湿实验室，手术区和生理/行为室）在Truitt博士的大厅对面，Rusyniak博士的一层楼，步行即可到达我的其他导师，这使其成为理想的指导和协作环境。因此，这是这个培训补助金的最终目标，帮助我成为一个独立的调查员，广泛了解如何戏剧性的损失雌激素导致扰乱体温调节。这将使我能够对我们对导致潮热的神经和神经化学机制的知识做出重大贡献，并确定潜在的新的非激素治疗靶点。

DESCRIPTION (provided by applicant): Currently I have had extensive training in: 1) neuroendocrinology of female sex hormones and behavior during my Masters with Thesis; 2) neuroendocrinology and functional neuroanatomy of stress responses during my PhD dissertation; and 3) translational animal modeling during my postdoctoral training. This training was collectively focused on whole animal physiology and behavior, combined with in vivo neurochemistry measures using microdialysis; stereotaxic delivery of pharmacological and tract tracing compounds, and ex vivo functional brain imaging using immunohistochemistry to detect immediate early gene proteins. In 2008 and 2010, I was respectively promoted to Research Assistant Professor, and then to Assistant Professor in the Department of Psychiatry. During this time I published a 1st author Nature Medicine article, where the main discovery was that a hypothalamic orexin system plays a critical role panic vulnerability in an accepted rat model of panic disorder and that central orexin levels are elevated in patients with panic disorder. I proceeded to determine if other pathologies were associated with a hyperactive orexin system. It was here that I noted in preclinical studies that estrogens suppress orexin activity and that ina recent clinical study dramatic loss of estrogen during menopause leads to a 300% increase in central levels of orexin, which was reversed with estrogen replacement. This evidence, combined with orexin's known role in thermoregulation, and that the orexin system is located in a hypothalamic area enriched in both estrogen receptors, led to my current hypothesis that "the orexin system plays a critical role in menopause-related symptoms such as hot flashes, and mood/sleep disruption". Adverse menopausal symptoms such as hot flashes affect about 70% of women approaching menopause and The North American Menopause Society posits that over 6,000 American women reach menopause daily. In addition, 63% of women ages 50-64 years are currently employed (U.S. Bureau of the Census, 2003). Thus, adverse menopausal symptoms have a significant financial impact in the workplace through lost work days and direct health care costs for medical services. This K01 training grant represents a significant, but logical, redirection in my career that requires additional mentoring and training by experts at IU in: 1) rodent studies in thermoregulation by Daniel Rusyniak MD; 2) sex hormone neuroendocrinology by Kathryn Jones PhD; 3) menopause-related "hot flashes" in human subjects and a Menopause course taught by Janet Carpenter, PhD, RN, FAAN; and 4) translational research approaches and animal modeling by Anantha Shekhar, MD/PhD. My training will also involve pharmacology training by Todd Skaar PhD for aim 1; molecular biology coursework and training by William Truitt, PhD, and Kenneth Cornetta MD at IUSM for aims 2 and 3; and training in circadian analyses by Robert Bies PhD at IUSM in aim 3b. Under the initial mentorship of Drs. Rusyniak and Carpenter, I have obtained an internally funded CTSI project development grant and a KL-2 CTSI Young Investigator Basic Science Award in Translational Research in 2011 to gain preliminary data for this grant. The aims and studies outlined here will aid in the transition from a 'Roadmap' K award to 'Roadmap' R awards. I have presented preliminary results at the American College of Neuropsychopharmacology in 2011 and at the Translational Science meeting in 2012, where I was respectively selected for the "Data Blitz" session; and won a Scholar's Abstract Award. I am also preparing a manuscript on our novel models of "hot flash" vulnerability. The proposed research outlined here is innovative for the following reasons: 1) to our knowledge this is the first attempt to determine orexin's role in adverse menopausal symptoms. A PubMed search for "orexin" and "hot flash" which is the primary menopause symptom yields no results as of March 2013; 2) this proposed research is based on preclinical information about estrogen regulation of orexin and receptor physiology, orexin's role in sleep wakes cycles, and our recent preclinical and clinical data linking a hyperactive orexin system to anxiety and temperature dysregulation; 3) we will conduct animal studies to elucidate the mechanisms underlying orexin induction of menopausal symptoms, which to our knowledge, has not been previously reported; 4) I have developed novel animal models of hot flash vulnerability to test my hypotheses; 5) I will apply pharmacology, acute gene and chronic lentiviral siRNA gene silencing techniques to understand the mechanism; 6) I will elucidate the neurologic mechanisms that regulate hot flashes with emphasis on the hypothalamic orexin system; and 7) I will provide mechanistic studies that will lead to a translational clinical protocol finding a novel use of an orexin receptor antagonist submitted for FDA approval for insomnia. I recently accepted a tenure track Assistant Professor position in the Department of Anatomy & Cell Biology at IUSM. My laboratory (a wetlab, surgical area and physiology/behavior room) is across the hall from Dr. Truitt and one floor up from Dr. Rusyniak, and within walking distance of my other mentors, making this an ideal mentoring and collaborative setting. Therefore, it is the ultimate goal of this training grant to help me become an independent investigator with an extensive understanding of how dramatic loss of estrogens lead to disrupted thermoregulation. This will allow me to make significant contributions to our knowledge of the neural and neurochemical mechanisms that lead to hot flashes and to identify potential novel nonhormonal targets for treatment.