0033 Proteomic Profiling of Plasma Biomarkers in Men and Women with Diagnosed Insomnia

Abstract Introduction Chronic insomnia disorder affects approximately 10% of the U.S. population and is linked to various adverse health consequences. Despite advancements in pharmacological interventions, many individuals continue to experience recurrent symptoms. A comprehensive understanding of the molecular mechanisms underlying insomnia is crucial for developing innovative and effective treatments. This study aimed to profile plasma proteins using an unbiased high-throughput assay to identify potential biomarkers for insomnia, with a specific focus on discerning any differential responses between men and women. Methods This prospective, observational study (n=60; 40 insomnia, 20 control; 58.3% females) was part of a larger study completed at Joint Base San Antonio-Lackland. Participants completed an overnight in-lab diagnostic polysomnogram, blood collection and clinical evaluation. We applied an unbiased high multiplexed proteomic discovery technique using DNA aptamers to target 7,000 proteins in plasma samples. Chi-square test and Mann–Whitney U-test were performed to determine the group differences. Adjusted p-values were calculated by using Benjamini–Hochberg's false discovery rate (FDR). Dysregulated proteins were uploaded into the Ingenuity Pathway Analysis software (Qiagen IPA) to explore the mechanistic networks most significantly associated with the study outcome. Significance level was set at 0.05 in all tests. Results We identified 54 significantly dysregulated plasma proteins (29 upregulated, 25 downregulated) when comparing individuals with diagnosed insomnia, stratified by gender. The top three upregulated proteins were T cell surface antigen CD2, Leptin (LEP), and N-terminal pro-BNP (NPPB); the top three downregulated proteins were Kunitz-type protease inhibitor 3 (SPINT3), Beta-defensin 104 (DEFB104A), and Benign Prostate-specific Antigen (KLK3). Pathway analysis revealed associations with proinflammatory cascades and metabolic signaling pathways. Conclusion Our findings highlight the feasibility of multiplex proteomic profiling in identifying blood-based biomarkers for understanding the pathophysiological mechanisms of insomnia disorder. Further research is warranted to ascertain the clinical utility of these candidate proteins. Support (if any) This research was supported by grants from the American Academy of Sleep Medicine Foundation (supported by Eisai, Inc.), the Defense Health Agency, Defense Medical Research and Development Program, Clinical Research Intramural Initiative for Military Women’s Health (DM170708; Mysliwiec), and the US Air Force Air Force Materiel Command (AFMC), Wright-Patterson Air Force Base, Ohio (FA8650-18-2-6953; Peterson).