Research Projects

Endometriosis, a debilitating disease affecting millions of women, often leads to menstrual pain and reduced quality of life. Although the lesions may be a primary cause of endometriosis-induced pain, surgical removal of the lesions does not provide respite from pain in all cases. This stark disconnect underscores a critical, unaddressed gap in our understanding that the pain of endometriosis is not merely local, but profoundly influenced by complex, systemic mechanisms. We propose a paradigm shift with neuroinflammation, specifically involving activated microglia, and the intricate interplay with the gut microbiome, are pivotal drivers of chronic endometriosis pain. Compelling evidence points to glial activation as a hallmark of chronic pain states, and growing research highlights altered gut microbiota in endometriosis patients. Our central hypothesis is twofold: 1. Endometriosis drives distinct, pathological microglial activation leading to distinct Disease-Associated Microglial(DAM) patterns in the CNS. 2. Targeted intervention in these microglial populations, through depletion and subsequent repopulation, can effectively mitigate neuroinflammation and alleviate endometriosis-associated pain. This project is a multi-pronged investigation: • Objective 1: Deciphering the Microglial Pain Signature in Endometriosis. We will establish a robust syngeneic murine endometriosis model, precisely characterizing chronic pain behavior using advanced machine learning. We will then map neuroinflammation by characterizing ectopic lesions and spinal columns for innervation, microglial presence (CD68, CD11b), and inflammatory cytokines (IL-6, TNF) via immunohistochemistry. Crucially, we will isolate brain microglia and perform targeted RNA sequencing (NanoString ) to identify unique DAM patterns specific to endometriosis. • Objective 2: Translating Findings to Human Pathophysiology. We will bridge the translational gap by generating patient-specific Induced Microglia-Like cells (iMGs) from the peripheral blood of women with surgically confirmed endometriosis. These human iMGs will be subjected to RNA sequencing to validate and identify human-specific DAM patterns. • Objective 3: Pioneering Microglial Replacement as a Therapeutic Strategy. Leveraging our murine model, we will rigorously test the therapeutic efficacy of pharmacologically induced microglial depletion (using PLX5622-formulated chow at high/low doses) followed by repopulation. Pain responses will be comprehensively evaluated before and after the intervention. The extent of microglial modulation will be validated via RNA sequencing to elucidate underlying mechanisms. • Objective 4: Unveiling the Microbiota-Microglia-Pain Axis. This objective will uncover the influence of the gut microbiome on microglial activation. We will delineate endometriosis-specific microbiome-metabolome signatures through 16S sequencing and unbiased metabolomics of patient stool samples. Identified metabolites will be validated in vitro on iMGs and in vivo in our murine model. Transformative impact: The concept of DAM in endometriosis represents a ground breaking understanding of chronic pelvic pain that moves beyond anatomical lesions. Identifying these DAM patterns will unveil novel, non-hormonal therapeutic targets. Our innovative use of patient-derived iMGs offers an unprecedented, non-invasive human platform for studying neuroinflammation and developing sorely needed diagnostic biomarkers. Crucially, generating proof-of-concept for microglial replacement therapy presents a highly innovative, non-hormonal, non-surgical strategy for long-term pain relief. Finally, establishing the causal link between gut microbiota and microglial activation will open entirely new avenues for dietary and microbial interventions, offering personalized and sustainable solutions for millions suffering from endometriosis.