Bielowka, Adrianna M.Patel, DilipLi, DongyangBernabeu-Herrero, Maria E.Game, LaurenceAldred, Micheala A.Mollet, Inês G.Shovlin, Claire L.2026-03-092026-03-092025-09-011460-2725PURE: 156370598PURE UUID: 99ff150b-4c36-47bc-8fdf-824d4a723c0eScopus: 105023546309PubMed: 39658243WOS: 001404190400001ORCID: /0000-0002-1422-1336/work/207951048http://hdl.handle.net/10362/201124Publisher Copyright: © 2025 The Author(s) 2024. Published by Oxford University Press on behalf of the Association of Physicians.Background: Disease biomarkers are often identified long after initiating pathologies, hampering mechanistic understanding and the development of preventative strategies. We hypothesized that aberrant cellular responses to normally-encountered stresses may be relevant to later disease states. Aim: To model two under-explored acute cellular stresses for blood-exposed cells, and cross-reference to known biomarkers of disease. Methods: Normal primary human endothelial cells (ECs) were treated for 1–6h with cycloheximide (CHX) 100μg/ml to inhibit protein translation and nonsense-mediated decay (modelling the integrated stress response), or 10μmol/l ferric citrate (modelling diurnal variation in serum iron that can be augmented by treatments prescribed 8 million times/year in England). Directional whole transcriptome RNA-seq identified differentially expressed genes and micro(mi)RNAs. Customized novel scripts examined the expression of 517 225 exons to predict 1h CHX-stabilized exons. Validations were by cel-miR-39-spiked qRT-PCR and RNA-seq in other EC types, peripheral blood mononuclear cells (PBMCs) and plasma. Results: miRNAs fell transiently at 1h after 10μmol/l ferric citrate (P<0.01), specifically in let-7 family member pre-miRNAs (‘let-7’, P<0.05), where there was an accompanying differential 6h increased expression of 570 let-7-target mRNAs identified through TargetScan (P<0.0001). qRT-PCR and RNA-seq validations in other normal ECs, plasma and PBMCs confirmed up to 80% falls in pre-let-7b/let-7b-5p after 1h iron, and exon 3B of the SLC11A2 (NRAMP2/DMT1)-encoded iron/copper transporter as a novel exon most consistently stabilized following 1h treatment with CHX. Overlaps with disease biomarkers for cancer, growth retardation and multiple organ-specific diseases were identified. Conclusions: Biomarkers for normal, acute cellular responses overlap with disease-state biomarkers, warranting further study.101459608engGeneral MedicineSDG 3 - Good Health and Well-beingjournal article10.1093/qjmed/hcae235https://www.scopus.com/pages/publications/105023546309