The role of SLC11A1 in neutrophil antimicrobial response

SLC11A1 在中性粒细胞抗菌反应中的作用

• 批准号: 10545065
• 负责人: Annica Stull-Lane
• 金额: $ 3.68万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-01-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10545065
• 关键词: Address; Agriculture; Amino Acid Sequence; Area; Autoimmune Diseases; Bacteremia; Basic Science; Biological; Biological Assay; Biological Sciences; Biology; Biomedical Research; Carrier Proteins; Cell Culture Techniques; Cell Line; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Code; Communicable Diseases; Core Facility; Cre lox recombination system; Data; Disease; Divalent Cations; Doctor of Philosophy; Education; Environment; Environmental Science; Faculty; Generations; Genes; Genetic; Genetic Polymorphism; Goals; Health; Host Defense; Human; Immune; Immune response; Immunology; Impairment; Inbred Mouse; Inbred Strains Mice; Infection; Iron; Knowledge; Link; Macrophage; Manganese; Mediating; Membrane; Mentors; Mentorship; Metals; Microbiology; Missense Mutation; Mouse Strains; Mus; Names; Nutritional Immunity; Pathology; Patients; Phagosomes; Phenotype; Predisposition; Protein Family; Proteins; Pump; Research; Research Personnel; Role; Salmonella infections; Salmonella typhimurium; Structure; Technology; Testing; Training; Translations; Vacuole; Veterinary Medicine; Veterinary Schools; Vitamin A Deficiency; Work; antimicrobial; antiporter; bactericide; clinically relevant; cofactor; college; conditional knockout; expectation; experience; experimental study; graduate school; improved; in vitro Model; in vivo; innovation; insight; medical schools; member; mouse genetics; mouse model; mutant; natural resistance-associated macrophage protein 1; neutrophil; novel; pathogen; response; solute; tool; translational study

PROJECT SUMMARY For decades, NRAMP1 (for natural resistance-associated macrophage protein 1, renamed SLC11A1 for solute carrier protein family 11, member 1) has been characterized as a divalent cation/H+ antiporter in the phagosomal membrane of macrophages that pumps iron and manganese out of the vacuole. This mechanism contributes to nutritional immunity against intracellular pathogens by depleting the phagosomal environment of essential metal cofactors. Mouse macrophages expressing mutant Slc11a1 are functionally compromised in control of several intracellular pathogens, including Salmonella enterica serovar Typhimurium (S. Typhimurium). Further, human polymorphisms in SLC11A1 are associated with susceptibility to both autoimmune and infectious disease. However, until now the role of SLC11A1 has been assumed to be macrophage-specific. Remarkably, during studies on vitamin A deficiency, our lab has uncovered an unsuspected role for SLC11A1 in control of systemic S. Typhimurium infection by neutrophils. Our preliminary results show that increased susceptibility of vitamin A-deficient mice to disseminated S. Typhimurium infection is dependent on synthesis of SLC11A1 by neutrophils, which challenges conventional wisdom that SLC11A1-dependent host defenses are associated exclusively with macrophages. However, strong evidence for a role of SLC11A1 as essential in neutrophil antimicrobial response is needed to establish this novel concept. Our central hypothesis is that SLC11A1 function promotes the bactericidal activity of neutrophils. The objective of this application is to investigate the role of SLC11A1 in neutrophil control of S. Typhimurium infection. To test our hypothesis, we will use a triangulated approach assessing neutrophil function and response to infection by utilizing human neutrophils in cell culture, mouse neutrophils ex vivo and a systemic S. Typhimurium infection in vivo mouse model. We are drawing on collaborative expertise to employ CRISPR for generation of a SLC11A1-/- cell line and utilize the Cre-lox system to create a mouse strain that has a conditional knockout of Slc11a1 only in neutrophils. Our rigorous studies of SLC11A1 in neutrophil response to infection will provide fundamental knowledge in the fields of microbiology and immunology, thereby advancing biomedical research. Further, this project will have broad implications for translation to improve human health, as polymorphisms in SLC11A1 make certain patients more susceptible to disease. The integrated training plan outlined in this application will provide an in-depth research experience, training in cutting-edge technologies like CRISPR, expertise from core facilities, and education from faculty in the School of Veterinary Medicine, the School of Medicine, the College of Agricultural and Environmental Sciences, and the College of Biological Sciences. Importantly, this plan embodies the structured dual-degree mentorship at UC Davis with a supportive sponsor and lab that addresses clinically relevant microbiological questions, a clinical mentor, a graduate school adviser, medical school advisers and an MD/PhD adviser.

项目摘要 几十年 溶质载体蛋白家族11，成员1）被描述为二价阳离子/h+抗腹剂 巨噬细胞的吞噬膜，将铁和锰从液泡中抽出。这种机制 通过耗尽吞噬环境，有助于对细胞内病原体的营养免疫力 必需的金属辅因子。表达突变体SLC11A1的小鼠巨噬细胞在功能上被妥协 控制了几种细胞内病原体，包括沙门氏菌血清鼠伤寒（S. typhimurium）。 此外，SLC11A1中的人类多态性与自身免疫性和感染性的敏感性有关 疾病。但是，直到现在，SLC11A1的作用被认为是巨噬细胞特异性的。 值得注意的是，在研究维生素A缺乏症的过程中，我们的实验室发现了无引起的作用 SLC11A1控制中性粒细胞的全身性孢子虫感染。我们的初步结果表明 维生素A缺陷小鼠对散布伤寒孢菌感染的敏感性增加取决于 中性粒细胞对SLC11A1的合成，这挑战了Slc11a1依赖宿主的传统智慧 防御与巨噬细胞仅相关。但是，有力证明SLC11A1的作用 需要在中性粒细胞抗菌反应中建立这种新颖的概念。 我们的中心假设是SLC11A1功能促进了中性粒细胞的杀菌活性。这 该应用的目的是研究SLC11A1在伤寒链霉菌感染的中性粒细胞控制中的作用。 为了检验我们的假设，我们将使用评估中性粒细胞功能的三角方法和对 通过在细胞培养中利用人类嗜中性粒细胞，小鼠嗜中性粒细胞和全身性孢子虫的感染感染 体内小鼠模型感染。我们正在利用协作专业知识来利用CRISPR来产生 SLC11A1 - / - 单元线并利用Cre-Lox系统创建一个有条件敲除的鼠标菌株 SLC11A1仅在中性粒细胞中。我们对SLC11A1在中性粒细胞对感染反应中的严格研究将提供 微生物学和免疫学领域的基本知识，从而进步生物医学研究。 此外，该项目将对改善人类健康的翻译具有广泛的影响，因为多态性 SLC11A1使某些患者更容易受到疾病的影响。 本应用程序中概述的综合培训计划将提供深入的研究经验， CRISPR等尖端技术的培训，核心设施的专业知识以及教师的教育 兽医学院，医学院，农业与环境学院 科学和生物科学学院。重要的是，该计划体现了结构化的双度 在加州大学戴维斯分校的指导下，由支持临床相关的微生物学的支持者和实验室 问题，临床导师，研究生顾问，医学院顾问和医学博士/博士顾问。

项目成果

Vitamin A supplementation boosts control of antibiotic-resistant Salmonella infection in malnourished mice.

• DOI: 10.1371/journal.pntd.0008737
• 发表时间: 2020-10
• 期刊: PLoS neglected tropical diseases
• 影响因子: 3.8
• 作者: Stull-Lane AR;Lokken-Toyli KL;Diaz-Ochoa VE;Walker GT;Cevallos SA;Winter ALN;Muñoz ADH;Yang GG;Velazquez EM;Wu CY;Tsolis RM
• 通讯作者: Tsolis RM