Nuclear and chromatin rearrangement associate to epigenome and gene expression changes in a model of in vitro adipogenesis and hypertrophy |
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| Institution: | 1. Nanoscopy, Istituto Italiano Tecnologia, Via Enrico Melen 83, 16152, Genova, Italy;2. Department of Earth, Environment and Life Sciences (DISTAV), University of Genova, Corso Europa 26, 16132, Genova, Italy;3. Department of Physics (DIFILAB), University of Genoa, Via Dodecaneso 33, 16146, Genoa, Italy;4. DIMES, Department of Experimental Medicine, University of Genoa, Genoa, Italy;5. IRCCS Ospedale Policlinico San Martino, Genoa, Italy;6. Clinica Medica “A. Murri”, Department of Biomedical Sciences and Human Oncology, University of Bari, Medical School, Piazza Giulio Cesare 11, 70124, Bari, Italy;1. State Key Laboratory Incubation Base for Green Processing of Chemical Engineering, School of Chemistry and Chemical Engineering, Shihezi University, Shihezi 832003, China;2. College of Life Sciences, Zhejiang University, Hangzhou 310058, China;3. Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA;1. Centro de Endocrinología Experimental y Aplicada - CENEXA (UNLP-CONICET CCT LA PLATA CEAS CICPBA), Facultad de Ciencias Médicas, Universidad Nacional de La Plata, Street 60 and 120, La Plata 1900, Argentina;2. Facultad de Ciencias Médicas, Universidad Nacional de La Plata, Street 60 and 120, La Plata 1900, Argentina;3. Instituto de Ciencias de la Salud, UNAJ-CICPBA, Street Avenue Calchaqui 6200, Florencio Varela 1888, Argentina;1. Chromosome Dynamics Group, Molecular Oncology Programme, Spanish National Cancer Research Centre (CNIO), Madrid, Spain |
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| Abstract: | Hypertrophy of adipocytes represents the main cause of obesity. We investigated in vitro the changes associated with adipocyte differentiation and hypertrophy focusing on the nuclear morphometry and chromatin epigenetic remodelling. The 3 T3-L1 pre-adipocytes were firstly differentiated into mature adipocytes, then cultured with long-chain fatty acids to induce hypertrophy. Confocal and super-resolution stimulation emission depletion (STED) microscopy combined with ELISA assays allowed us to explore nuclear architecture, chromatin distribution and epigenetic modifications. In each condition, we quantified the triglyceride accumulation, the mRNA expression of adipogenesis and dysfunction markers, the release of five pro-inflammatory cytokines. Confocal microscopy revealed larger volume and less elongated shape of the nuclei in both mature and hypertrophic cells respect to pre-adipocytes, and a trend toward reduced chromatin compaction. Compared to mature adipocytes, the hypertrophic phenotype showed larger triglyceride content, increased PPARγ expression reduced IL-1a release, and up-regulation of a pool of genes markers for adipose tissue dysfunction. Moreover, a remodelling of both epigenome and chromatin organization was observed in hypertrophic adipocytes, with an increase in the average fluorescence of H3K9 acetylated domains in parallel with the increase in KAT2A expression, and a global hypomethylation of DNA. These findings making light on the nuclear changes during adipocyte differentiation and hypertrophy might help the strategies for treating obesity and metabolic complications. |
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