{"doi":"10.1002/clt2.12057","title":"Lung miRNA profiles show a time‐of‐day response in house dust mite‐induced allergic asthma in mice","abstract":"Asthma is a chronic inflammatory lung disease that shows a time-of-day effect (increased airway inflammation and altered lung function) on the severity of the disease.1 Recent studies support the functional role of miRNAs in the molecular pathophysiology of asthma phenotypes/endotypes2, 3 through the post-transcriptional gene regulation of key signaling pathways and cellular processes in human airways, immune cells in asthmatics, and in mouse model of allergic asthma.6 However, whether a rhythmic change in expression of miRNAs is responsible for the time-of-day effects observed during asthma and its exacerbations are not known. In this study, we utilized high-throughput miRNA and mRNA profiling using NanoString to determine the time-of-day effects using the house dust mite (HDM)-induced allergic asthma model. We demonstrate that timed dosing of HDM at Zeitgeber time (ZT0: 6:00 a.m or ZT12: 6:00 p.m) shows differentially expressed (DE) miRNAs and their predicted mRNAs that are reflective of the asthma phenotype. DE analysis revealed a strong time-of-day difference in the miRNA expression at ZT0 versus ZT12 HDM exposed mice compared to PBS (control). We found 6 miRNAs (downregulated: miR-125b-5p, miR-125a-5p, miR-150, miR-23a, miR-23b and miR-15b) and 3 miRNAs (upregulated: miR-652, miR-200b and miR-200c) significantly in HDM versus PBS at ZT0 (Figure 1A,B), and 21 miRNAs downregulated and 17 miRNAs upregulated significantly in HDM versus PBS at ZT12 (Figure 1C,D). Hierarchical cluster analysis of DE miRNAs in HDM versus PBS groups at ZT0 and ZT12 pairwise comparison along with their fold change/ratio, p-value and the false discovery rate (FDR) adjusted p-value is provided (Figure 1A–D). miRNA expression signatures in PBS (control) and house dust mite (HDM) exposed mouse lung. (A) Heat map shows unsupervised hierarchical clustering of nine differentially expressed (DE) miRNAs in HDM versus PBS exposed mouse lung at ZT0 analyzed by Morpheus tool. (B) Pairwise comparison analysis performed by nSolver 4.0 software showing DE miRNAs in HDM versus PBS at ZT0. (C) Heat map shows unsupervised hierarchical clustering of 38 DE miRNAs in HDM versus PBS exposed mouse lung at ZT12 analyzed by Morpheus tool. (D) Pairwise comparison analysis performed by nSolver 4.0 software showing DE miRNAs in HDM versus PBS at ZT12. Data are shown as fold change/ratio, p-value (p < 0.05) and FDR adjusted p-value < 1 (n = 6/group/time point) When we compared the DE miRNAs in HDM versus PBS group, we found the same 6 miRNAs (downregulated) and 3 miRNAs (upregulated) at ZT0 pairwise comparisons were among the common miRNAs DE at both ZT0 and ZT12 (Figure S1A,B). Additionally, unsupervised clustering and Principal Component Analysis (PCA) of normalized counts from HDM versus PBS groups at ZT0 and ZT12 showed clusters separated based on the treatment groups (Figure S1C–D). Hierarchical cluster analysis of normalized counts from HDM versus PBS groups at ZT0 and ZT12 revealed close clustering of all DE miRNAs in PBS and HDM groups at ZT12 except for few HDM ZT0 samples that were clustered with either the PBS ZT0 or HDM ZT12 (Figure S1E). To assess the degree of DE miRNAs in the lung of HDM versus PBS at ZT0 and ZT12, we performed multiple comparison analyses. Several of the miRNAs were either significantly downregulated or upregulated in PBS and HDM inter-group comparisons (Figures 2A and S2A,–B). Eleven DE miRNAs (downregulated) and 8 DE miRNAs (upregulated) in the HDM-exposed group significantly compared to PBS at ZT12, and 3 DE miRNAs significantly upregulated in the HDM-exposed group compared to PBS at ZT0 (Figure S2A,B). However, several DE miRNA targets were either downregulated (2 miRNAs: miR-720 and miR-1944) or upregulated (9 miRNAs: miR-151-5p, miR-199a-3p, miR-30c, let-7d, miR-15a, miR-181a, miR-25, let-7e, and miR-126-3p) in PBS group at ZT0 versus ZT12 comparison (Figure S2A,B). The pairwise comparison data for HDM versus PBS at ZT0 and ZT12 revealed","journal":"Clinical and Translational Allergy","year":2021,"id":209095,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":5,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9532,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2021-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":435957,"name":"A. R. Srinivasan","orcid":"0000-0002-3272-9985","position":1,"is_corresponding":false},{"id":278343,"name":"Isaac K. Sundar","orcid":"0000-0001-6742-3460","position":0,"is_corresponding":true}],"reference_count":7,"raw_metadata":null,"created_at":"2026-07-18T23:52:01.526969Z","pmid":"34435755","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}