PROJECT SUMMARY Parent award. The initial parent award (P20-GM103625) allowed the University of Arkansas for Medical Sciences (UAMS) to establish the Center for Microbial Pathogenesis and Host Inflammatory Responses (CMPHIR). The scientific focus of the CMPHIR is on understanding infectious disease from the perspective of diverse microbial pathogens and their impact on the host immunological and inflammatory responses influences the disease process. This theme is based on the premise that understanding the complex interplay between diverse pathogens and their common human host is a prerequisite to maximizing opportunities to manipulate one or both sides of this equation in favor of the desired therapeutic outcome. The project is currently in Phase III (P30-GM145393) with the goal of continuing to support and enhance the critical core facilities to support the growing number of CMPHIR investigators, continue to provide administrative support to promote further growth of the CMPHIR and the career development of its investigators, and promote the transition to a self-sustained Center of Biomedical Research Excellence with the expertise and resources required to address existing and emerging problems in infectious disease. The current proposal for a Team Science Administrative Supplement application is directly in line with the goals of the CMPHIR and that it takes a comprehensive, team-based approach to understanding how the crosstalk between microbiota and the host during antibiotic-induced dysbiosis, so that new therapeutic strategies can be implemented to maintain host homeostasis and mitigate disease risk. It is also a direct reflection of the growth of the CMPHIR in that it brings together three investigators (one current CMPHIR P30 pilot recipient and 2 investigators with various other past COBRE and INBRE ties) with distinct areas of expertise that can collectively take advantage of their specific strengths and cutting edge technologies to comprehensively investigate fundamental disease-related processes of highly consequential bacterial imbalance to an extent that otherwise would not be possible. Research question. Antibiotic (ABX) use has significantly increased by more than 30% in recent years. Although ABX fight infections, they also disrupt commensal microbe communities that are crucial for maintaining homeostasis of various local and distal processes. Microbial imbalance induced by ABX, termed herein as dysbiosis, has many systemic and long-lasting effects on the host including, but not limited to, increased risk of vascular pathologies, e.g., solid tumors. Thus, there is a pressing need to reach a deeper understanding of the crosstalk between microbiota and the host during dysbiosis, so that new therapeutic strategies can be implemented to maintain host homeostasis and mitigate disease risk. The long-term overarching goal of this supplement is to initiate a new line of investigation that brings together multiple scientific disciplines for a common goal of understanding how dysbiosis influences vascular development. The overall objective of this application is to rigorously and precisely define the molecular composition and cellular functions of small extracellular vesicles (sEVs) from independent perspectives of an endothelial cell biology and innate immunity in the setting of angiogenesis. Attaining this objective is critical for future deductive approaches that will guide development of more effective vascular therapies in patients with acute and chronic dysbiosis. Our central hypothesis is that independent lipid, nucleic acid and proteomic compositions of dysbiotic sEVs drives homing to vascular beds of activated vascular endothelial cells and local activation of neutrophils that propel angiogenesis. Team science approach. The work proposed here relies on the synergy created by a team of investigators, each of whom brings a unique expertise and/or resource to the project. This truly is a team science approach, as each of the laboratories bring distinct expertise, skillsets, resources and equipment with virtually no overlap but shared interest in vascular biology. Our team is a collaboration between a tumor angiogenesis biologist (Dings), an expert in small RNA and neutrophil biology (Allen), and a renowned expert in lipid metabolism and lipidomics (Morris) at UAMS and the Arkansas Children’s Nutrition Research Center at the Arkansas Children’s Hospital Research Institute (ACNC/ACRI). The Dings lab is an expert on ABX-induced dysbiosis and tumor angiogenesis and will contribute murine models, endothelial cell lines and overall vasculature development expertise necessary for the proposed studies. The Allen lab brings experience and expertise in extracellular small RNAs transported by sEVs and lipoproteins, as well as innate immune activity within the vasculature. The Morris lab is a renowned authority in lipid metabolomics and lipidomics. This supplement will facilitate the development of a strong collaborative team with complementary skillsets and experimental approaches, thus allowing for a unique and comprehensive experimental approach that would not otherwise be possible. Teams will meet at least twice monthly to discuss results and strategize. The data generated will be used for subsequent grant applications focused on understanding at the molecular and cellular level how dysbiosis-induced sEVs modulate host vasculature and innate immune responses. Moreover, the data, tools, protocols, and experimental approach will be applicable to any number of other diseases involving the microbiota. Ultimately, the work proposed will further enable integration of lipidomics into the parent COBRE and will allow to lay the foundation of establishing new research infrastructure and resources at the sponsoring institution to be able to support investigators of other local and national COBRE/INBRE/CTRs in the future.

Project Narrative The current proposal for a Team Science Administrative Supplement is associated and directly in line with the goals of the Center for Microbial Pathogenesis and Host Inflammatory Responses (CMPHIR; P30 GM145393; PD: Mark Smeltzer, Ph.D.), in that it takes a comprehensive, team-based approach to understanding how the crosstalk between microbiota and the host during antibiotic-induced dysbiosis, so that new therapeutic strategies can be implemented to maintain host homeostasis and mitigate disease risk. This supplement will facilitate the development of a strong collaborative team with complementary skillsets and experimental approaches, thus allowing for a unique and comprehensive experimental approach that would not otherwise be possible.