Vermont Center for Cardiovascular and Brain Health

项目总结： 大脑血流量得到精确控制，以满足神经元的代谢需求。活跃的神经元向 血管系统通过多种神经血管耦合机制增加局部血流量的现象 称为功能性充血。充血反应增加了血液流动施加的摩擦力。 微动脉和毛细血管内皮细胞。我们最近证明了Piezo1频道是一种 脑毛细血管内皮细胞中的关键机械传感器，并介导钙信号对 机械刺激。此前的研究主要集中在内皮细胞Piezo1下游的作用机制 但内皮细胞质膜流动性如何控制其活性仍不清楚。作为回应 美国国立卫生研究院特别关注通知(非-GM-23-034)“IDEA的行政补充 为资助团队科学发展项目提供令人信服的初步证据 富含胆固醇和肌醇磷脂的结构域存在于中枢神经系统毛细血管网络中。 此外，我们提出并提供了脂类影响Piezo1活性的初步结果。家长奖， 在NIGMS(P20GM135007)的支持下，提供了一个建立在 早期职业教师研究社会面临的重大健康问题的非凡潜力：心血管 疾病、中风和认知障碍。本团队科学提案旨在奠定基础和框架 哈拉兹博士(现任项目总监，P20GM135007)和冈萨雷斯博士(前项目总监，P20GM130459) 以他们过去富有成效的合作为基础，并将他们的专业知识和技术应用于 影响深远的生物学问题。在此，我们假设脂类胆固醇和磷脂酰肌醇-4，5- 二磷酸(PIP2)在毛细血管网络中形成结构域，决定着毛细血管内皮细胞膜的流动性 和Piezo1活性。为了验证这一假设，被提议的团队科学项目将使用多样化的、最先进的 方法，包括膜片钳电生理技术和工程小鼠模型(Harraz Lab) 以及高分辨率血管成像(Gonzales Lab)，以表征膜脂对 内皮细胞Piezo1活性。该项目的完成将促进两个独立的 研究团队，并将支持Piezo1活性由脂质微环境决定的概念。这些 研究将进一步奠定坚实的基础，从而导致未来的独立资助(NIH R01申请)， 这将促进我们作为独立科学家的职业生涯。

Project Summary: Cerebral blood flow is precisely controlled to satisfy neuronal metabolic demands. Active neurons signal to the vasculature via multiple neurovascular coupling mechanisms to increase regional blood flow in a phenomenon known as functional hyperemia. The hyperemic response increases the frictional forces imposed by blood flow onto endothelial cells of arterioles and capillaries. We have recently demonstrated that the Piezo1 channel is a crucial mechanosensor in brain capillary endothelial cells, and that it mediates Ca2+ signals in response to mechanical stimuli. Previous research has focused on mechanisms engaged downstream of endothelial Piezo1 activation, but how endothelial cell plasma membrane fluidity controls its activity remains unknown. In response to the NIH Notice of Special Interest (NOT-GM-23-034) “Availability of Administrative Supplements to IDeA Awards to Fund Team Science Development Projects,” we provide compelling preliminary evidence that lipid domains enriched in cholesterol and phosphoinositides exist in central nervous system capillary networks. Further, we propose and provide preliminary results that lipids affect Piezo1 activity. The parent award, supported by NIGMS (P20GM135007), provides a platform to build sustainable research programs built on the exceptional potential of early career faculty to study the vital health problems facing society: cardiovascular disease, stroke and cognitive impairment. This Team Science proposal aims to lay the foundation and framework for Drs. Harraz (current Project Director, P20GM135007) and Gonzales (former Project Director, P20GM130459) to build upon their past productive collaborations and bring their expertise and techniques to synergistic work on impactful biological questions. Herein, we hypothesize that the lipids cholesterol and phosphatidylinositol-4,5- bisphosphate (PIP2) form domains in capillary networks, which determine capillary endothelial membrane fluidity and Piezo1 activity. To test this hypothesis, the proposed Team Science project will use diverse, state-of-the-art approaches, including patch-clamp electrophysiological techniques and engineered mouse models (Harraz Lab) along with high-resolution vascular imaging (Gonzales Lab) to characterize the impact of membrane lipids on endothelial Piezo1 activity. Completion of this project will foster collaboration between the two independent research teams and will support the concept that Piezo1 activity is dictated by lipid microenvironment. These studies will further establish a solid foundation that will lead to future independent funding (NIH R01 application), which will further our careers as independent scientists.