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，有可能有助于我们的重要进展的研究 了解蛇咬伤的病理生理学。蛇咬伤是一个巨大的全球性健康问题， 270万男子、妇女和儿童受伤，估计每年夺去125 000人的生命。尽管其 尽管蛇咬伤作为一个全球性的健康问题非常重要，但人们对蛇咬伤的病理生理学知之甚少。科学 我们研究的前提是，蛇毒毒素的一个特殊家族，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起着关键作用 并迅速和显著地增加血管渗透性。我们假设svCRiSPs增加血液 和淋巴管内皮细胞通透性。减少的组织间液流量增加 内皮细胞屏障功能，加速毒液从其沉积部位进入 体循环一旦进入体循环，毒液中的大量毒素 进入它们的目标组织，导致蛇的猎物迅速固定和死亡。来测试我们 假设我们将讨论两个不同但互补的具体目标： 具体目的1：描述svCRiSPs对血管通透性和内皮细胞增殖的影响。 功能参加研究课程的本科生团队将净化和表征 svCRISP是从毒液中提取的，这些毒液是我们国家最具医学意义的毒蛇种类之一。 收藏.表征将包括血液和淋巴管内皮细胞屏障功能的体外试验 和血管渗透性的体内测定。 具体目标2。为了表征Hellerin作用的细胞和分子基础， 新鉴定的crotalid svCRiSP，对血液和淋巴管内皮细胞的功能。先进 本科生和研究生将研究Hellerin的作用机制， 从南太平洋响尾蛇的毒液中分离出来学生们将测试Hellerin的活动， 血液和淋巴管内皮细胞的增殖和屏障功能，并将表征细胞信号传导 介导毒素对内皮细胞功能的作用的途径和膜受体。 拟议的研究将支持我们的计划的教育目标，提供本科 和研究生与实践研究经验，提供了一个介绍的机会， 可用于生物医学研究的职业生涯。这项研究计划也将提高学术 通过促进全球健康研究的重要计划，我们的大学环境。知识 从这些研究中获得的信息将有助于深入了解 svCRiSPs对血管功能的影响，将有助于我们理解蛇咬伤的病理生理学。

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.