Stratification of ME/CFS by mycotoxin exposure

Myalgic Encephalomyelitis/Chronic Fatigue Syndrome (ME/CFS) is a disabling, multisystem disease affecting up to 3.3 million Americans, predominantly women. Despite a significant public health burden, biological mechanisms remain poorly understood and no diagnostic biomarkers or targeted therapies exist. Mounting evidence suggests that environmental exposures, particularly to fungal mycotoxins, may contribute to ME/CFS pathogenesis by promoting immune dysfunction. However, there are no peer-reviewed published studies examining the molecular effects of mycotoxins on the immune system of ME/CFS patients. This proposal aims to conduct the first study on the molecular level of how mycotoxin exposure contributes to ME/CFS pathophysiology. We hypothesize that mycotoxins induce transcriptomic reprogramming in immune cells and extracellular vesicles (EVs), leading to immune exhaustion and chronic inflammation. Preliminary data from our group shows enrichment of fungal-response pathways, altered immune cell subsets, increased pro-inflammatory EV miRNAs, and exhaustion-associated signatures in ME/CFS patients with detectable urinary mycotoxins. We will recruit 150 ME/CFS patients and 50 matched sedentary healthy controls (HC), stratify ME/CFS cases by mycotoxin exposure, and pursue four specific aims: Aim 1: Quantify urinary mycotoxins and assess associations of mycotoxin levels and combinations with symptom severity and plasma cytokines. Aim 2: Use single-cell RNA sequencing to define mycotoxin contribution to transcriptomic alterations in PBMC subsets in ME/CFS patients. Aim 3: Identify long and small RNA signatures in plasma-derived EVs associated with mycotoxin exposure. Aim 4: Integrate multi-omics data using machine learning to identify mechanistic correlations between mycotoxin exposure, immune dysregulation, and symptom severity. We will use a validation cohort of 20 ME/CFS patients and 10 HC to confirm our results. Our approach is innovative in combining environmental exposure data with transcriptomics and EV RNA profiling to define a biologically and clinically distinct ME/CFS subgroup. The integration of scRNA-seq, EV-RNA profiles, and urinary mycotoxins will enable the discovery of robust molecular signatures and mechanistic pathways affected by environmental toxins, namely mycotoxins. This work will significantly advance the field by providing stratification of ME/CFS by mycotoxin exposure and providing novel insights into environmentally driven immune reprogramming in ME/CFS. Outcomes include identification of candidate biomarkers, deeper understanding of the contribution of mycotoxins to immune exhaustion mechanisms, and a stratification framework for future diagnostics and therapeutic interventions. Our study represents a critical step toward developing precision medicine strategies for ME/CFS patients with environmentally exacerbated disease symptoms. Project Number: 1R01NS148845-01 | Fiscal Year: 2026 | NIH Institute/Center: National Institute of Neurological Disorders and Stroke (NINDS) | Principal Investigator: Lubov Nathanson | Institution: NOVA SOUTHEASTERN UNIVERSITY, Fort Lauderdale-Davie, FL | Award Amount: $614,458 | Activity Code: R01 | Study Section: Special Emphasis Panel[ZRG1 CFS-W (80)] View on NIH RePORTER: https://reporter.nih.gov/project-details/11347476