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: 8506964
• 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/8506964
• 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)

候选人：Chattergoon博士，医学硕士，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。我五年的职业目标是成为围产期心脏成熟领域的独立研究员，特别是了解控制这一心脏发育阶段的信号机制。我分别于2000年8月和2004年8月在杜兰大学药理学系获得硕士和博士学位。在我的博士论文中，我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和主动脉原代细胞培养方面的培训。 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，以及原代骨髓基质细胞鉴定。我们确定降钙素基因相关肽是一种强大的血管扩张剂，通过刺激影响细胞周期的cAMP途径，在体外对VSMC的增殖起到有效的抑制作用。 监管。2004年8月，我开始在俄勒冈健康与科学大学(OHSU)肯特·L·桑伯格博士的实验室进行博士后培训。我加入这个实验室是为了培训胎儿心脏发育，以及胎儿环境的变化如何戏剧性地改变心脏发育。为了熟悉桑伯格博士实验室中使用的胎羊模型，我花了几年的时间。随之而来的是我们的合作者G·吉罗博士、S·娄伊博士、S·容克博士、D·安德森博士、L·戴维斯博士和J·费伯博士的广泛教学。和这群科学家一起，我学习了慢性胎儿研究所需的外科技术，连续血流动力学测量的方法，慢性大动物活体研究的一般方法，以及胎儿心肌细胞的原代培养。我还在赠款和手稿准备方面积累了丰富的经验。在斯托克博士和范·温克尔博士的新指导下，我计划将我在大型哺乳动物心脏发育中学到的知识应用到子宫内的遗传扰动如何对出生后的生活产生有害后果的问题上。我提议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以及心脏分子信号的兴趣的方式来产生和利用转基因模型。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，我将为斯托克博士的实验室已经进行的工作引入一个新的层面。我还将在未来的研究中使用各种遗传模型。参加、介绍心脏发育和信号方面的领导并与他们互动也是我培训的重要方面。这些研究活动对我在这一领域的职业发展至关重要。当我过渡到我新任命的研究助理教授时，我觉得我处于一个完美的位置，是一个理想的机构，可以弥合我们在理解心脏发育方面的胎儿编程方面的知识差距。这个项目的资金也将是我制定长期目标的基础。环境：俄勒冈健康与科学大学(OHSU)特别适合发展我的研究项目。通过这项提案，我将与波特兰退伍军人管理医疗中心、Vollum研究所、生物医学信息交流中心、心脏研究中心和心血管成像实验室的研究团队进行互动。心脏病科在临床和研究方面有着卓越的历史，并在桑吉夫·考尔博士的领导下经历了复兴。考尔博士坚定不移地致力于学术研究，并有支持初级教职科学家的历史。该大学大约有12个共享的核心资源设施，提供研究分析方面的咨询，我将与其中5个机构合作完成这个项目。研究项目：胎儿期是心脏发育的关键时期。在较大的哺乳动物中，如绵羊，心肌细胞在出生前逐渐停止分裂，成为主要的双核细胞(终末分化)。一旦心肌细胞终末分化，它们就不能再分裂了，但它们保留了非凡的放大能力。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。我们对在出生时产生适当数量的心肌细胞的信号通路的理解有很大的差距。这项提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。这项提案将研究B-Raf阻断在小鼠遗传模型和药理模型中的作用。B-Raf在发育中的心脏尚未被研究过。我们假设，阻断B-Raf对心肌细胞MAPK的控制将改变心肌细胞的增殖和肥大信号，并将提供一种改变心肌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种重要的信号分子的作用。 并将为未成年心脏的生长治疗提供新的潜在途径。我们将通过超声心动图测量，将B-Raf干扰的结果与新生儿和成人心脏功能的形态、生化和功能参数相结合。(摘要结束)