Mapping the Angiotensin II-TGFB-Integrin signaling triad to reveal therapeutic targets in aortic aneurysm

 描述（由申请人提供）：项目总结主动脉瘤是一种常见疾病，其定义为主动脉过度生长和中壁重塑，可导致致命性夹层和破裂。动脉瘤的有效药物治疗方法很少，这在很大程度上是由于对疾病机制的不完全理解。本提案的目的是更全面地了解与胸主动脉瘤进展不相符的分子事件，并在此过程中识别和验证新型药物治疗的潜在靶点。初步体外和体内数据 表明血管紧张素II、整合素和转化生长因子β（TGFβ）信号传导之间相互作用是动脉瘤发病机制的关键分子要素。主要研究者Sarah帕克博士使用马凡氏综合征（MFS）的基因敲入小鼠模型研究导致胸主动脉瘤中TGFβ信号失调的背景依赖性分子机制。在本提案的指导阶段，帕克博士将使用体外技术评估整合素β3（ITGβ3）过表达（如MFS中发生的）如何影响已知在主动脉瘤中发生改变的血管平滑肌细胞生理学方面，并使用新型质谱技术（数据独立采集MS）鉴定ITGβ3下游驱动VSMC生理学改变的致病性信号传导组分（目的1）。在向独立阶段的过渡中，帕克博士将确定另一个信号传导网络，即血管紧张素II 1型受体（AT 1 R）的β-抑制蛋白2（β ARR 2）偏置信号传导，如何促成MFS中动脉瘤期间发生的TGFβ信号传导失调和VSMC机械特性改变（目的2）。最后，帕克博士将整合目标1和2中的发现，以检验一个统一的假设，即ITGβ3改变的基质感知有助于MFS小鼠体外和体内AT 1 R的β ARR 2偏倚信号传导，并进一步确定ITGβ3和/或β ARR 2偏倚信号传导的药理学操作是否可以减缓MFS中的动脉瘤进展（目标3）。帕克博士获得了博士学位。威斯康星州医学院（MCW）的生理学。随后，她在约翰霍普金斯大学完成了前三年半的博士后研究，并得到了著名临床医生和结缔组织疾病和主动脉瘤医学遗传学专家Harry（Hal）Dietz博士和临床心血管蛋白质组学首席专家Jennifer货车Eyk博士的合作指导。基于她在心血管生理学和基于质谱的蛋白质组学技术方面的成熟专业知识，这个K99/R 00奖项将使帕克博士能够（1）发展分析和解释复杂分子数据集的信息学和计算技能，（2）与南加州大学的梅根·麦凯恩博士合作，开发一个体外模型来独立修改基质成分，平滑肌细胞类型和可溶性细胞外因子，以研究平滑肌细胞的收缩生理学，（3）继续建立在主动脉血管生物学的专业知识和（4）加强她的沟通，指导，管理和领导技能，为成功的独立生物医学研究人员做好准备。帕克博士将在雪松西奈医学中心（CSMC）完成该奖项的指导阶段，她的主要导师货车Eyk博士最近将她的实验室搬到了高级临床生物系统研究所。帕克博士已经招募了一个令人印象深刻的导师和顾问团队，包括雪松西奈当地（Jennifer货车Eyk博士，Moshe Arditi博士，Ben Berman博士，Ken伯恩斯坦博士）和外部机构（Hal Dietz博士，John Yates博士和Megan McCain博士），以促进她的科学和个人发展。帕克博士接受培训的机构（约翰霍普金斯，MCW）和将继续她的培训（CSMC）所培养的临床研究环境提供了理想的环境，以促进她的长期职业目标，即阐明原位的背景依赖性病理信号事件，并将其与疾病发病机制的细胞，组织和器官生理学特征改变联系起来。帕克博士将首先把她的方法集中在驱动升主动脉瘤的特定病因机制上，并打算最终将她的研究扩展到心血管生物学的其他领域，在这些领域中，必须理解外部和内部分子背景的全部复杂性，以便最好地预测细胞信号传导和病理生理学之间的因果关系。为了实现这一目标，帕克博士打算将基于质谱的发现工作流程与仔细的生物学验证和用于治疗特定病理的新型候选治疗药物的临床前测试联系起来。该奖项将是支持帕克博士建立一个研究计划的框架，将实现她的职业目标的基础。此外，通过完成本提案的目标，帕克博士将为预防主动脉瘤衰弱后果的新治疗方法的开发做出重大贡献。

 DESCRIPTION (provided by applicant): PROJECT SUMMARY Aortic aneurysm is a prevalent condition defined by excessive aortic growth and medial wall remodeling that can result in lethal dissection and rupture. Few effective pharmacological treatments exist for aneurysm, due in large part to an incomplete understanding of the mechanisms that underlie the disease. The goal of this proposal is to derive a more comprehensive understanding of the molecular events that belie thoracic aortic aneurysm progression, and in so doing identify and validate potential targets for novel pharmacological therapy. Preliminary in vitro and in vivo data indicate that interactions between angiotensin II, integrin, and transforming growth factor β (TGFβ) signaling are key molecular elements of aneurysm pathogenesis. The principal investigator, Dr. Sarah Parker, uses a genetic knock-in mouse model of Marfan syndrome (MFS) to study the context-dependent molecular mechanisms leading to dysregulated TGFβ signaling in thoracic aortic aneurysm. In the mentored phase of this proposal, Dr. Parker will use in vitro techniques to assess how of Integrin β3 (ITGβ3) overexpression, as occurs in MFS, impacts aspects of vascular smooth muscle cell physiology known to be altered in aortic aneurysm, and use novel mass spectrometry technologies (data independent acquisition MS) to identify pathogenic signaling components downstream of ITGβ3 that drive altered VSMC physiology (Aim 1). In the transition to the independent phase, Dr. Parker will identify how another signaling network, β-Arrestin 2 (βARR2) biased signaling by the Angiotensin II Type 1 Receptor (AT1R), contributes to dysregulated TGFβ signaling and altered mechanical properties of VSMCs that occur during aneurysm in MFS (Aim 2). Finally, Dr. Parker will integrate the findings in Aims 1 and 2 to test a unifying hypothesis that altered matrix sensing by ITGβ3 contributes to βARR2 biased signaling by AT1R both in vitro as well as in vivo in MFS mice, and further determine whether pharmacological manipulation of ITGβ3 and/or βARR2-biased signaling can attenuate aneurysm progression in MFS (Aim 3). Dr. Parker received her Ph.D. in Physiology from the Medical College of Wisconsin (MCW). She has subsequently completed the first three and a half years of her Post Doctoral fellowship at Johns Hopkins University under the collaborative mentorship of Dr. Harry (Hal) Dietz, a renowned clinician and expert in the medical genetics of connective tissue disorders and aortic aneurysm, and Dr. Jennifer Van Eyk, a premier expert in clinical cardiovascular proteomics. Building upon her established expertise in cardiovascular physiology and mass spectrometry-based proteomic techniques, this K99/R00 award will allow Dr. Parker to (1) develop informatics and computational skills for the analysis and interpretation of complex molecular data sets, (2) in collaboration with Dr. Megan McCain at the University of Southern California, develop an in vitro model to independently modify matrix components, smooth muscle cell types, and soluble extracellular factors in order to study contractile physiology in smooth muscle cells, (3) continue to build expertise in the vascular biology of the aorta and (4) strengthen her communication, mentoring, management, and leadership skills to prepare for success as an independent biomedical researcher. Dr. Parker will complete the mentored phase of this award at Cedars-Sinai Medical Center (CSMC), where her primary mentor, Dr. Van Eyk, has recently moved her laboratory to become the director of the Advanced Clinical Biosystems Research Institute. Dr. Parker has enlisted an impressive team of mentors and advisors both local to Cedars Sinai (Dr. Jennifer Van Eyk, Dr. Moshe Arditi, Dr. Ben Berman, Dr. Ken Bernstein) and at external institutions (Dr. Hal Dietz, Dr. John Yates, and Dr. Megan McCain) to facilitate her scientific and personal development. The clinical research environments fostered by the institutions where Dr. Parker has been trained (Johns Hopkins, MCW) and will continue her training (CSMC) provide ideal settings to facilitate her long-term career goal to elucidate context-dependent, pathological signaling events in situ and connect them with the altered cellular, tissue, and organ physiology characteristic of disease pathogenesis. Dr. Parker will first focus her approach on the specific etiological mechanisms that drive ascending aortic aneurysm, and intends to eventually expand her research into other areas of cardiovascular biology where the full complexity of external and internal molecular context must be understood in order to best predict the cause-and-effect relationships between cell signaling and pathophysiology. To achieve this goal, Dr. Parker intends to bridge focused mass spectrometry-based discovery workflows with careful biological validation and the pre-clinical testing of novel therapeutic candidates that will be used to treat specific pathologies. This award will be fundamental in supporting Dr. Parker to build the framework for a research program that will achieve her career goals. Furthermore, by completing the aims of this proposal Dr. Parker will make a significant contribution toward the development of new treatments that will prevent the debilitating consequences of aortic aneurysm.