The Function of Small Proline Rich Proteins in Cutaneous Host Defense

富含脯氨酸的小蛋白在皮肤宿主防御中的功能

• 批准号: 10553219
• 负责人: Tamia Alisha Harris-Tryon
• 金额: $ 15.96万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-10 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10553219
• 关键词: 16S ribosomal RNA sequencing; Animals; Antibiotic Resistance; Award; Bacteria; Binding; Biochemical; Biochemistry; Cell Death; Cell Nucleus; Cell Wall; Cells; Cellulitis; Cutaneous; Data; Defense Mechanisms; Dermatology; Development; Doctor of Medicine; Doctor of Philosophy; Endoplasmic Reticulum; Enterococcus faecalis; Environment; Epithelium; Family; Fluorescent in Situ Hybridization; Gastrointestinal tract structure; Gene Family; Gene Proteins; Genes; Genetic Engineering; Genomics; Gland; Goals; Gram-Negative Bacteria; Gram-Positive Bacteria; Home; Host Defense; Host Defense Mechanism; Human; Immune; Immune system; Immunity; Immunologics; In Vitro; Infection; Infectious Skin Diseases; Inflammatory; International; Intestines; Invaded; Lipopolysaccharides; Membrane; Mentorship; Microbe; Modeling; Mus; Names; Oils; Pathway interactions; Persons; Physiological; Play; Postdoctoral Fellow; Predisposition; Proline; Protein Secretion; Proteins; Public Health; Research; Research Proposals; Role; SPRR1A; SPRR1B; Scientist; Sebaceous Glands; Shapes; Skin; Skin Tissue; Soft Tissue Infections; Staphylococcus aureus; Staphylococcus epidermidis; Streptococcus; Streptococcus pyogenes; Surface; Techniques; Testing; Training; Virus; Visualization; Work; Wound Infection; antimicrobial; burden of illness; career; experimental study; fighting; fungus; in vivo; infectious disease treatment; insight; keratinocyte; member; methicillin resistant Staphylococcus aureus; microbial host; microbiome; microbiota; microorganism; mouse model; novel therapeutics; pathogen; pathogenic bacteria; programs; proline-rich proteins; protein expression; recruit; resistant strain; resistin; response; skills; skin barrier; skin disorder; skin microbiome; trafficking; transcriptome sequencing

Project Summary/Abstract: Skin and soft tissue infections are a major public health threat. Exacerbating the problem has been the development of antibiotic resistant strains of bacteria. Like the hide of an animal, human skin creates a physical barrier between a person and its environment. The skin is also home to cells and molecules of the immune system that create an immunological barricade to infection. A critical component of the skin barrier are antimicrobial proteins- evolutionarily ancient immune effectors that maintain mutually-beneficial host-microbial relationships at multiple epithelial surfaces, such as the skin and the intestine. Recently, we discovered that proteins in the Resistin family (human Resistin and mouse RELMα) are antimicrobial and expressed by keratinocytes and by sebaceous glands - the oil producing glands of the skin. In addition to RELMα, there are innumerable proteins elaborated by sebaceous glands with unknown biochemical function. Little is understood about the role that bacteria play in regulating the expression of these proteins. We thus wanted to identify other genes expressed by the sebaceous gland that might, 1) play a role in host defense and 2) modulate the skin microbiome. Using whole transcriptome sequencing (RNA-seq), we compared the expression profile of sebocytes treated with lipopolysaccharide (LPS) to sebocytes treated with a vehicle control. We were surprised to find that 4 members of the small proline rich family (gene name SPRR) were upregulated by LPS. SPRR proteins also have structural features similar to other described antimicrobial proteins. Our preliminary data indicate that SPRR1A and SPRR2A proteins can kill skin commensals in vitro. In this proposal we will utilize an array of in vitro and in vivo techniques to: (Aim 1) determine whether SPRR1A can kill bacterial pathogens, (Aim 2) delineate how SPRR1 proteins are secreted from skin, and (Aim 3) determine the physiologic function of SPRR2A in skin. These studies will help us understand how the sebaceous gland contributes to cutaneous host defense. My long-term career goal is to develop an independent research program that investigates host defense at the skin surface. After completing MD/PhD and Dermatology training at Johns Hopkins, I was recruited to UT Southwestern to train as a post-doctoral fellow in the lab of Lora Hooper, Ph.D, an internationally recognized leader in mechanistic studies of the host/microbe interface and the characterization of antimicrobial proteins in the gastrointestinal tract. During my tenure in the Hooper lab, I have expanded the focus of the lab to include skin. This career develop award will support my final years in this unique training environment and give me the opportunity to gain new skills in: 1) cellular trafficking biochemistry and 2) bacterial genomics. I have created a mentorship team of scientists to help fill these gaps in my training, Sandra Schmid Ph.D., Heidi Kong M.D., and Julia Segre Ph.D. Through the completion of these aims, I will create a fully independent research platform, able to generate novel therapeutic solutions for skin infections and inflammatory skin diseases.

皮肤和软组织感染是一个主要的公共卫生威胁。 抗生素耐药菌株的发展使问题更加严重。像 动物的皮，人的皮，在人和环境之间形成了一个物理屏障。的 皮肤也是免疫系统的细胞和分子的家园，它们产生免疫屏障， 感染皮肤屏障的一个关键组成部分是抗菌蛋白-进化古老的免疫 在多个上皮表面维持互利的宿主-微生物关系的效应物，例如 皮肤和肠道一样。最近，我们发现抵抗素家族（人抵抗素）中的蛋白质 和小鼠RELMα）是抗微生物的，并由角质形成细胞和皮脂腺表达-油 产生皮肤的腺体。除了RELMα，还有无数的蛋白质由 皮脂腺的生化功能未知。人们对细菌所起的作用知之甚少 调节这些蛋白质的表达。因此，我们希望鉴定由这些细胞表达的其他基因。 皮脂腺可能，1）在宿主防御中发挥作用，2）调节皮肤微生物组。使用 全转录组测序（RNA-seq），我们比较了用 脂多糖（LPS）对用载体对照处理的皮脂细胞的作用。我们惊讶地发现， 富含脯氨酸的小家族成员（基因名SPRR）被LPS上调。SPRR蛋白 也具有与其它所述抗微生物蛋白相似的结构特征。我们的初步数据显示 SPRR 1A和SPRR 2A蛋白可以在体外杀死皮肤寄生虫。在本提案中，我们将利用 一系列体外和体内技术来：（目的1）确定SPRR 1A是否可以杀死细菌病原体， (Aim 2）描述SPRR 1蛋白如何从皮肤分泌，并且（目的3）确定SPRR 1蛋白的生理学特性。 SPRR 2A在皮肤中的功能这些研究将帮助我们了解皮脂腺如何贡献 皮肤宿主防御 我的长期职业目标是发展一个独立的研究项目， 皮肤表面的防御。在约翰霍普金斯完成医学博士/博士学位和皮肤病学培训后，我 招募到UT西南培训作为一个博士后研究员在实验室的劳拉胡珀，博士， 在宿主/微生物界面的机理研究和 在胃肠道中的抗微生物蛋白的表征。我在胡珀实验室任职期间， 我把实验室的重点扩大到皮肤。这个职业发展奖将支持我的最后几年 在这个独特的培训环境，让我有机会获得新的技能：1）细胞贩运 生物化学和2）细菌基因组学。我创建了一个科学家导师团队， Sandra Schmid博士在我的训练中的空白，Heidi Kong医学博士，和朱莉娅塞格雷博士通过完成 为了实现这些目标，我将创建一个完全独立的研究平台，能够产生新的治疗方法， 皮肤感染和炎症性皮肤病的解决方案。