PROJECT SUMMARY/ABSTRACT It is unknown why some patients regain their pre-morbid health following appropriate treatment for Lyme disease while others progress to develop post-treatment Lyme disease (PTLD). Significant prior published and preliminary data suggest that variability in clinical, immunologic, and metabolic factors at the onset of infection with B. burgdorferi contribute to the development of divergent post-treatment outcomes, including PTLD. In this study, we will draw upon unique cohorts of patients with well-defined PTLD, healthy controls without a history of Lyme disease, and patients with early Lyme disease followed longitudinally up to 1 year after the end of treatment. Combining advanced, innovative statistical and laboratory methods, the objective of this grant is to identify these factors and examine how they relate to underlying symptom phenotypes among patients with PTLD. To accomplish this goal, we will identify clinically-relevant risk factors associated with post-treatment outcomes in Lyme disease over time. These factors will then be used to develop an assessment score to identify patients at increased risk of developing PTLD in the clinical setting (AIM 1). A longitudinal, multivariate, cross- domain analysis of this sort with the goal of direct translation of findings to clinical practice and the study of fundamental disease pathways in PTLD has not previously been performed. We will also identify novel auto- antibodies that are associated with PTLD through protein array, and examine their relationship with underlying clinical phenotypes (AIM 2). While an autoimmune process has been suspected to underlie the pathogenesis of PTLD, at least for some individuals, adaptive immune recognition of self-antigens is only beginning to be described. Discovery of autoantibodies in PTLD may reveal patient subsets marked by specific clinical features, such as predominant musculoskeletal pain or neurologic symptoms, and identify individuals whose disease is driven by an autoimmune process. Finally, we will study the longitudinal immunometabolic profile of patients with differing clinical outcomes using a novel, high-dimensional flow cytometry approach, and hypothesize that pharmacologic manipulation of perturbed pathways in vitro may be able to normalize a PTLD-associated metabolic signature (AIM 3). The ability of immunometabolism to globally report on the acute cellular environment suggests that interrogating immune cell metabolism in individuals develop PTLD in the future could provide fundamental insights into disease mechanism. Collectively, our studies represent a rigorous approach towards uncovering determinants that are associated with the outcome of PTLD and its underlying clinical phenotypes. The long-term goal of this project is to generate fundamental knowledge to inform the development of innovative therapeutic strategies as well as novel interventions to prevent or reduce the often significant symptom burden and functio...