The unique role of B-Raf in murine cardiac development and postnatal function

B-Raf 在小鼠心脏发育和产后功能中的独特作用

• 批准号: 8700504
• 负责人: Natasha N Chattergoon
• 金额: $ 12万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8700504
• 关键词: Address; Adrenergic Agents; Adult; Affect; Animal Model; Animals; Apoptosis; Appointment; Area; BRAF gene; Biochemical; Birth; Blood Vessels; Bone Marrow; Calcitonin Gene-Related Peptide; Cardiac; Cardiac Myocytes; Cardiology; Cardiomegaly; Cardiovascular Diseases; Cell Cycle Regulation; Chronic; Clinical Research; Communication; Complement; Consultations; Cutaneous; Cyclic AMP; Development; Doctor of Philosophy; Educational process of instructing; Environment; Exhibits; Faculty; Fetal Development; Fetal Heart; Fetus; Funding; Future; Genes; Genetic; Genetic Models; Goals; Grant; Growth; Growth Factor; Health; Health Sciences; Heart; Heart Research; Human; Hypertrophy; Hypoxia; In Vitro; Infusion procedures; Institutes; Institution; Insulin-Like Growth Factor I; Interdisciplinary Study; Isoproterenol; Knockout Mice; Knowledge; Laboratories; Leadership; Learning; Life; Link; Low Mitotic Activity; Lung; Malignant Neoplasms; Malignant neoplasm of thyroid; Mammals; Manuscripts; Measurement; Measures; Medical center; Mentorship; Methods; Mitogen-Activated Protein Kinases; Modeling; Modification; Molecular; Mus; Muscle Cells; Mutate; Mutation; Myocardium; Newborn Infant; Operative Surgical Procedures; Oregon; Pathway interactions; Performance; Perinatal; Pharmacology; Phase; Phenylephrine; Physiology; Population; Positioning Attribute; Preparation; Primary Cell Cultures; Process; Proliferating; Protein Analysis; Protein Isoforms; Ras/Raf; Rattus; Recording of previous events; Regulation; Renaissance; Renal carcinoma; Reperfusion Injury; Research; Research Activity; Research Personnel; Research Project Grants; Resources; Risk; Role; Scientist; Sheep; Signal Pathway; Signal Transduction; Signaling Molecule; Smooth Muscle Myocytes; Staging; Stress; Stromal Cells; Syndrome; Techniques; Testing; Time; Tissues; Training; Transfection; Transgenic Model; United States Department of Veterans Affairs; Universities; Vascular Smooth Muscle; Vasodilator Agents; Western Blotting; Wild Type Mouse; Work; abstracting; adrenergic; base; cardiogenesis; cardiovascular imaging; career; career development; cell growth; experience; fetal; fetal programming; heart function; hemodynamics; human FRAP1 protein; human disease; in utero; in vivo; inhibitor/antagonist; insight; interest; melanoma; mouse model; post-doctoral training; postnatal; pressure; prevent; primary pulmonary hypertension; professor; programs; raf Kinases; regenerative; response; stressor; vascular smooth muscle cell proliferation

DESCRIPTION (provided by applicant): Candidate: Dr. Chattergoon, M.S., Ph.D. My long-term career objective is to build a strong multidisciplinary research program, utilizing appropriat developmental models that focus on understanding the risks of future cardiovascular disease. My five-year career goal is to become an independent investigator in the area of cardiac maturation in the perinatal period and especially understanding the signaling mechanisms that govern this phase of heart development. I completed my M.S. and Ph.D. degrees at Tulane University in the Department of Pharmacology in August 2000 and August 2004, respectively. I studied vascular smooth muscle growth in the context of primary pulmonary hypertension for my doctoral dissertation. This area of research offered training in primary cell culture of aortic and pulmonary vascular smooth muscle cells (VSMC), protein analysis by western blot, immunostaining, adenoviral gene transfection, and primary bone marrow stromal cell characterization. We determined that calcitonin gene-related peptide, a powerful vasodilator, is potent inhibitor of VSMC proliferation in vitro by stimulating cAMP pathways that affect cell cycle regulation. I began my postdoctoral training in Dr. Kent L. Thornburg's laboratory at Oregon Health and Science University (OHSU) in August of 2004. I joined this laboratory to train in fetal heart development and how changes to the fetal environment dramatically alter heart growth. I have invested the several years required to become facile with the fetal sheep model used in Dr. Thornburg's laboratory. With that came extensive teaching by our collaborators, Drs. G Giraud, S Louey, S Jonker, D Andersen, L Davis, and J Faber. With this group of scientists I have learned the surgical techniques required for chronic fetal studies, methods for continuous hemodynamic measurements, and the general methods of chronic large animal in vivo studies, as well as primary culture of fetal cardiomyocytes. I have also gained great experience in grant and manuscript preparation. I plan to apply what I have learned in large mammal heart development to how genetic perturbations in utero may have detrimental consequences for post-natal life under the new mentorship of Drs. Stork and Van Winkle. My proposed studies will allow me generate and utilize transgenic models in a way that helps to marry my interests in both whole animal physiology as well as molecular signaling in the heart. Also, by using echocardiographic techniques to determine cardiac function in the mouse model I will introduce a new layer to the work already under way Dr. Stork's lab. I will also use various genetic models for my future studies. Attending, presenting and interacting with leaders in cardiac development and signaling are also important aspects of my training. These research activities are essential to my career development in this field. As I transition to my new appointment of Research Assistant Professor I feel I am in the perfect position and at the ideal institution to bridge the gaps of knowledge we have in understanding fetal programming with reference to cardiac development. The funding for this project will also be the basis of setting my long-term goals. Environment: Oregon Health and Science University (OHSU) is especially fertile for developing my research program. Through this proposal I will interact with research teams at Portland Veterans Administration Medical Center, the Vollum Institute, the Biomedical Information Communications Center, the Heart Research Center, and the Cardiovascular Imaging lab. The Division of Cardiology has a strong history of clinical and research excellence and had undergone a renaissance under the leadership of Dr. Sanjiv Kaul. Dr. Kaul has an unwavering commitment to academics and a history of supporting junior faculty scientists. The university has roughly 12 shared Core resources facilities that offer consultation on study analyses and I will be working with 5 of them for this project. Research Project: The fetal period is a critical stage for heart development. In larger mammals, like the sheep, cardiomyocytes gradually cease dividing and become primarily binucleated (terminal differentiation) just prior to birth. Once cardiac myocytes terminally differentiate they can no longer divide but they retain a remarkable capacity to enlarge. In the mouse, cardiomyocytes continue to proliferate and terminally differentiate over the first week of postnatal life. There is a large gap in our understanding of the signaling pathways that yield the proper number of cardiomyocytes at birth. This proposal will address one major portion of that gap. The mitogen activated protein kinase (MAPK) is a major proliferative pathway that is influenced by Ras/Raf-1 signaling. This proposal will examine the role of B-Raf blockade in both genetic models and pharmacological models in the mouse. B- Raf has not been studied in the developing heart. We hypothesize that blockade of B-Raf control of MAPK in cardiomyocytes will alter the proliferative and hypertrophic signals and will provide an animal model with pathophysiological consequences of altered cardiac cell growth. A positive finding would confirm the role of this important signaling molecule and will provide new potential avenues of growth therapy for under grown hearts. We will couple the results of B-Raf disruption to morphological, biochemical, and functional parameters of cardiac performance in the newborn and adult using echocardiographic measures. (End of Abstract)

描述（由申请人提供）：候选人：Dr. Chattergoon，M.S.，博士 我的长期职业目标是建立一个强大的多学科研究计划，利用适当的发展模型，重点了解未来心血管疾病的风险。我的五年职业目标是成为围产期心脏成熟领域的独立研究者，特别是了解控制心脏发育这一阶段的信号机制。我完成了我的MS。和博士分别于2000年8月和2004年8月在杜兰大学药理学系获得学位。我在博士论文中研究了原发性肺动脉高压背景下的血管平滑肌生长。这一研究领域提供了主动脉原代细胞培养的培训， 肺血管平滑肌细胞（VSMC），蛋白质印迹分析，免疫染色，腺病毒基因转染和原代骨髓基质细胞表征。我们确定降钙素基因相关肽是一种强有力的血管扩张剂，通过刺激影响细胞周期的cAMP途径，在体外是VSMC增殖的有效抑制剂 调控我在肯特L博士那里开始了我的博士后训练。2004年8月，索恩伯格在俄勒冈州健康与科学大学（OHSU）的实验室。我加入这个实验室是为了训练胎儿心脏的发育，以及胎儿环境的变化如何极大地改变心脏的发育。我已经投入了几年的时间来熟悉索恩伯格博士实验室使用的胎羊模型。随之而来的是我们的合作者G Giraud博士、S Louey博士、S Jonker博士、D Andersen博士、L Davis博士和J Faber博士的广泛教学。与这群科学家一起，我学习了慢性胎儿研究所需的手术技术、连续血流动力学测量方法、慢性大动物体内研究的一般方法以及胎儿心肌细胞的原代培养。我还获得了丰富的经验，在赠款和手稿的准备。 我计划将我在大型哺乳动物心脏发育方面所学到的知识应用于子宫内的遗传扰动如何对产后生活产生有害影响，这是在斯托克博士和货车温克尔博士的新指导下进行的。我所提出的研究将使我能够以一种有助于结合我对整个动物生理学和心脏分子信号传导的兴趣的方式生成和利用转基因模型。此外，通过使用超声心动图技术来确定小鼠模型的心脏功能，我将为斯托克博士实验室已经进行的工作引入一个新的层次。我也将在未来的研究中使用各种遗传模型。参加，展示和与心脏发育和信号传导方面的领导者互动也是我培训的重要方面。这些研究活动对我在这个领域的职业发展至关重要。当我过渡到我的研究助理教授的新任命，我觉得我在完美的位置，并在理想的机构，以弥合知识的差距，我们在了解胎儿编程与心脏发育。这个项目的资金也将是我制定长期目标的基础。环境：俄勒冈州健康与科学大学（OHSU）特别适合发展我的研究项目。通过这个提议，我将与波特兰退伍军人管理局医疗中心、Vollum研究所、生物医学信息通信中心、心脏研究中心和心血管成像实验室的研究团队进行互动。心脏病科拥有卓越的临床和研究历史，并在Sanjiv Kaul博士的领导下经历了复兴。考尔有一个坚定的承诺，以学术和支持初级教师科学家的历史。该大学大约有12个共享的核心资源设施，为研究分析提供咨询，我将与其中5个合作完成这个项目。研究项目：胎儿期是心脏发育的关键阶段。在较大的哺乳动物中，如绵羊，心肌细胞逐渐停止分裂，并在出生前主要变成双核（终末分化）。一旦心肌细胞终末分化，它们就不能再分裂，但它们保留了显著的扩大能力。在小鼠中，心肌细胞在出生后的第一周内继续增殖和终末分化。我们对出生时产生适当数量心肌细胞的信号通路的理解存在很大差距。这项建议将解决这一差距的一个主要部分。丝裂原活化蛋白激酶（MAPK）是受Ras/Raf-1信号影响的主要增殖途径。该提案将研究B-Raf阻断在小鼠遗传模型和药理学模型中的作用。尚未在发育中的心脏中研究过B- Raf。我们假设阻断心肌细胞中MAPK的B-Raf控制将改变增殖和肥大信号，并将提供一种具有改变心脏细胞生长的病理生理学后果的动物模型。一个积极的发现将证实这一重要的信号分子的作用 并将为生长不足的心脏提供新的潜在的生长治疗途径。我们将使用超声心动图测量新生儿和成人的B-Raf干扰结果与心脏性能的形态、生化和功能参数相结合。(End摘要）