The acute effects of snake venom CRiSP toxins on blood and lymphatic endothelial cell permeability: new insights into the pathophysiology of snakebite

这个项目的总体目标是让本科生和研究生参与创新研究。 在分子毒理学中，有可能对我们的研究做出重要贡献的研究 对蛇咬伤的病理生理学的认识。蛇咬伤是一个巨大的全球健康问题，造成严重的 270万男子、妇女和儿童受伤，估计每年夺走12.5万人的生命。尽管它有 作为一个全球性的健康问题，人们对蛇咬伤的病理生理知之甚少。科学的 我们研究的前提是一个特定的蛇毒毒素家族，富含半胱氨酸的分泌蛋白 (SvCRiSPs)，通过靶向咬伤区域的血管和淋巴管，在蛇咬伤中发挥关键作用 并迅速而显著地增加血管通透性。我们假设svCRiSPs能增加血液 淋巴管内皮细胞通透性。减少引起的间质液体流量的增加 内皮细胞屏障功能，加速毒液从其在咬合处的沉积部位进入 体循环。一旦进入体循环，毒液中的大量毒素就会获得 接触到它们的目标组织，导致蛇的猎物迅速固定和死亡。测试我们的 假设我们将解决两个截然不同但相互补充的具体目标： 特异性目标1：研究svCRiSPs对血管通透性和内皮细胞的影响 功能。参加研究课程的本科生团队将净化和特色化 SvCRISP的毒液是一些最具医学意义的毒蛇物种，居住在我们的 收集。鉴定将包括血液和淋巴管内皮细胞屏障功能的体外分析 以及体内血管通透性的测定。 具体目标2.为了表征Hellerin作用的细胞和分子基础， 新发现的Crotalid svCRiSP，对血液和淋巴管内皮细胞功能的影响。进阶 本科生和研究生将在分离的svCRiSP中研究Heller的作用机制 来自南太平洋响尾蛇的毒液。学生们将测试Hellerin的活性 血液和淋巴管内皮细胞的增殖和屏障功能，并将成为细胞信号的特征 介导毒素对内皮细胞功能影响的途径和膜受体。 拟议的研究将通过为本科生提供支持我们项目的教育目标 和具有实践研究经验的研究生，这些经验提供了机会的入门 适用于生物医学研究的职业。这项研究计划也将提高学术水平 通过推动一项重要的全球健康研究计划，改善了我们大学的环境。知识 从这些研究中获得的结果将为深入了解这些影响的分子机制提供帮助 SvCRiSPs对血管功能的影响，有助于我们理解蛇咬伤的病理生理机制。

The overall goal of this project is to engage undergraduate and graduate students in innovative studies in molecular toxinology, studies that have the potential to contribute to important advances in our understanding of the pathophysiology of snakebite. Snakebite is a huge global health problem, causing serious injury to 2.7 million men, women and children and claiming an estimated 125,000 lives annually. In spite of its importance as a global health issue, very little is known of the pathophysiology of snakebite. The scientific premise for our study is that a specific family of snake venom toxins, the Cysteine-Rich Secretory Proteins (svCRiSPs), plays a critical role in snakebite by targeting blood and lymphatic vessels in the region of the bite and rapidly and dramatically increasing vascular permeability. We hypothesize that svCRiSPs increase blood and lymphatic endothelial cell permeability. The increase in interstitial fluid flow that results from decreased endothelial cell barrier function, accelerates the transit of the venom from its site of deposition in the bite into the systemic circulation. Once delivered into the systemic circulation, the numerous toxins in the venom gain access to their target tissues, causing the rapid immobilization and death of the snake's prey. To test our hypothesis we will address two distinct but complementary Specific Aims: Specific Aim 1: To profile the effect of svCRiSPs on vascular permeability and endothelial cell function. Teams of undergraduate students enrolled in research courses will purify and characterize svCRISP's from the venom some of the most medically significant species of venomous snakes housed in our collection. Characterization will include in vitro assays of blood and lymphatic endothelial cell barrier function and in vivo assays of vascular permeability. Specific Aim 2. To characterize the cellular and molecular basis for the effects of Hellerin, a newly identified crotalid svCRiSP, on the function of blood and lymphatic endothelial cells. Advanced undergraduate and graduate students will investigate the mechanism of action of Hellerin an svCRiSP isolated from the venom of the Southern Pacific Rattlesnake. Students will test the activity of Hellerin on the proliferation and barrier function of blood and lymphatic endothelial cells and will characterize the cell signaling pathways and membrane receptors that mediate the effects of the toxin on endothelial cell function. The proposed studies will support the educational goals of our program by providing undergraduate and graduate students with hands-on research experience that provides an introduction to the opportunities available for careers in biomedical research. This research program will also enhance the academic environment of our university by promoting an important program of global health research. Knowledge gained from these studies will provide insights into the molecular mechanisms that underlie the effects of svCRiSPs on vascular function and will contribute to our understanding of the pathophysiology of snakebite.