پردیس کشاورزی و منابع طبیعی دانشگاه تهران Iranian Journal of animal Science 2008-4773 56 3 2025 09 23 Integrative and comparative analysis of transcriptional profiles and competitive endogenous RNA regulatory networks to identify key genes associated with heat stress in broiler chickens Integrative and comparative analysis of transcriptional profiles and competitive endogenous RNA regulatory networks to identify key genes associated with heat stress in broiler chickens 665 688 101469 10.22059/ijas.2025.391124.654061 FA Faezeh Hesari Department of Animal Science, University College of Agriculture and Natural Resources, University of Tehran, Karaj, Alborz, Iran. Mostafa Sadeghi Department of Animal Science, University College of Agriculture and Natural Resources, University of Tehran, Karaj, Alborz, Iran. Seyed Reza Miraei-Ashtiani Department of Animal Science, University College of Agriculture and Natural Resources, University of Tehran, Karaj, Alborz, Iran. 0000-0003-3597-4385 Farzad Ghafouri Department of Animal Science, University College of Agriculture and Natural Resources, University of Tehran, Karaj, Alborz, Iran. Journal Article 2025 02 25 Broilers are particularly sensitive to high ambient temperatures, which may have a negative impact on their welfare and production efficiency. The aim of the study was to combine a literature review analysis of transcriptional profiling and a competitive endogenous regulatory network analysis (CERNA) to identify the key messenger and non-coding RNAs involved in the molecular regulation of the cells of the pituitary gland and the brain in heat stress broiler chickens. Overall, two long non-coding RNA, 11 microRNAs and 426 common mRNAs and genes were identified that showed significant differences in expression between the two tissues. From the analysis, 12 major genes (<em>HSP90A1, HSPA5, HSP90B1, HSPA4, HUSY1, AHSA2, ALB, DNAJA1, DNAJA4, HSPA4L, and HSPH1</em>) were found to be more extensively expressed in broiler chickens under heat stress than in control chickens. In addition, after reconstructing the competitive endogenous RNA regulatory network and the associated candidate modules, the main important metabolic signalling pathways identified were protein-folding chaperones, cellular component organisation, protein-binding to heat shock proteins, the endoplasmic reticulum chaperone complex, and protein-processing signalling pathways in the endoplasmic reticulum. Overall, considering the identified RNAs involved in the ceRNA regulatory network that underlie phenotypic differences in the severity of heat stress, this study may provide new insights into the molecular mechanisms underlying the control of heat stress severity and resistance in broiler chickens. Broilers are particularly sensitive to high ambient temperatures, which may have a negative impact on their welfare and production efficiency. The aim of the study was to combine a literature review analysis of transcriptional profiling and a competitive endogenous regulatory network analysis (CERNA) to identify the key messenger and non-coding RNAs involved in the molecular regulation of the cells of the pituitary gland and the brain in heat stress broiler chickens. Overall, two long non-coding RNA, 11 microRNAs and 426 common mRNAs and genes were identified that showed significant differences in expression between the two tissues. From the analysis, 12 major genes (<em>HSP90A1, HSPA5, HSP90B1, HSPA4, HUSY1, AHSA2, ALB, DNAJA1, DNAJA4, HSPA4L, and HSPH1</em>) were found to be more extensively expressed in broiler chickens under heat stress than in control chickens. In addition, after reconstructing the competitive endogenous RNA regulatory network and the associated candidate modules, the main important metabolic signalling pathways identified were protein-folding chaperones, cellular component organisation, protein-binding to heat shock proteins, the endoplasmic reticulum chaperone complex, and protein-processing signalling pathways in the endoplasmic reticulum. Overall, considering the identified RNAs involved in the ceRNA regulatory network that underlie phenotypic differences in the severity of heat stress, this study may provide new insights into the molecular mechanisms underlying the control of heat stress severity and resistance in broiler chickens. https://ijas.ut.ac.ir/article_101469_2ca8623ba6a3cf6da08d9488142531fe.pdf