RAPID: How do vertebrate endolysosomal Ca2+ channels control coronaviral tropism?

这项研究促进了国家繁荣和生物经济，增强了我们对冠状病毒在物种间传播机制的理解。本研究结果可能是鉴定SARS-CoV-2 （COVID-19或COVID）疾病严重程度或传播的生物标志物的基础。这个项目将有助于理解为什么一些通常会感染特定动物（如鸟类和蝙蝠）的冠状病毒菌株有时也会感染人类，而其他菌株则不会。这一知识对于理解病毒如何打破物种壁垒，在新宿主中定居并引发广泛感染至关重要，正如正在进行的COVID大流行期间所经历的那样。冠状病毒通过与特定的细胞表面受体结合并通过称为内体和溶酶体的细胞器进入细胞内部来感染细胞。该项目将研究这些细胞结构内的环境如何调节病毒感染的强度。通过在不同脊椎动物物种中使用比较生物学，目标将是确定内体和溶酶体中促进特定生物体感染的条件。这项研究可能是确定疾病严重程度或COVID传播的生物标志物的基础。这项研究还将提供工具，使广泛的研究社区能够研究COVID和其他病毒。该提案还使用资金来培训研究生和本科生。因此，这笔资金正在培养下一代科学领袖。这项工作的预期结果将有助于识别和控制COVID在人类和其他动物中的传播。冠状病毒科（cov）是一类单链RNA病毒，可感染三种脊椎动物：哺乳动物、鸟类和鱼类。理解支持这种向性的分子决定因素，即为什么冠状病毒自然有效地感染这些物种，并针对这些动物体内的特定组织，是一个基本的生物学概念。因此，定义限制冠状病毒趋向性的机制对于我们理解冠状病毒如何打破物种壁垒并在新宿主中定居至关重要。在本RAPID提案中，将研究不同脊椎动物中不同冠状病毒的内溶酶体系统的腔内微环境在决定冠状病毒感染效率中的作用。核心假设是，在低等哺乳动物和人类中，居住在核内体和溶酶体中的称为离子通道的细胞蛋白质的存在是不同的。此外，这些离子通道在低等哺乳动物的内溶酶体和溶酶体内创造了独特的环境，这对支持冠状病毒感染至关重要，组织和物种之间内溶酶体离子通道的差异表达调节了冠状病毒感染的有效性。这一假设将通过多种细胞模型、成像方法和感染分析进行验证。这项研究得到了美国国家科学基金会“共生、防御和自我认知”项目的支持。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

This research advances the national prosperity and the bioeconomy and enhances our understanding of the mechanisms that allow the spread of corona viruses between species. Results of this research may be the basis for identifying biomarkers of severity of disease or spread of SARS-CoV-2 (COVID-19 or COVID). This project will produce an understanding of why some coronavirus strains that normally infect specific animals (such as birds and bats) sometimes also infect humans, whereas other strains do not. This knowledge is critical for understanding how viruses break species barriers to colonize new hosts and precipitate widespread infections, as experienced during the ongoing COVID pandemic. Coronaviruses infect cells by binding to specific cell surface receptors and passing through organelles known as endosomes and lysosomes to access the cell interior. This project will investigate how the environment within these cellular structures regulates the intensity of viral infection. Through the use of comparative biology across different vertebrate species, the goal will be to identify conditions in endosomes and lysosomes that facilitate infection in particular organisms. This research may be the basis for identifying biomarkers of severity of disease or COVID spread. This research will also provide tools to enable a broad research community working on COVID and other viruses. This proposal also uses funding to train graduate and undergraduate students. As such this funding is training the next generation of leaders in science. Results expected from this work will facilitate identification and control of COVID spread in humans and other animals. The coronoaviridae (CoVs) are a family of single-stranded RNA viruses that infect three vertebrate groups: mammals, birds and fish. Understanding the molecular determinants underpinning this tropism, namely why CoVs naturally and efficiently infect these species, and target specific tissues within these animals, is a fundamental biological concept. Definition of the mechanisms restricting CoV tropism is therefore critical for our understanding of how CoVs break species barriers and colonize new hosts. In this RAPID proposal, the role of the luminal microenvironment of the endolysosomal system in determining the efficiency of CoV infection will be studied for different CoVs in different vertebrate species. The central hypothesis is that the cellular proteins called ion channels resident in endosomes and lysosomes are differentially present in lower mammals and humans. Furthermore, these ion channels uniquely create environments within endosomes and lysosomes in lower mammals that are critical for supporting coronaviral infection and the differential expression of endolysosomal ion channels between tissues and species regulates the effectiveness of CoV infection. This hypothesis will be tested using a variety of cell models, imaging approaches and infection assays. This research is supported by the Symbiosis, Defense and Self-recognition program of the National Science Foundation.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.