1. Deregulation of the imprinted DLK1-DIO3 locus ncRNAs is associated with replicative senescence of human adipose-derived stem cells
- Author
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Fátima Sánchez-Cabo, Susana Cañón, Silvia García-López, Alberto Martínez-Serrano, Carmen Albo-Castellanos, Mario F. Fraga, Rocío G. Urdinguio, Antonio Bernad, UAM. Departamento de Biología Molecular, Centro de Biología Molecular Severo Ochoa (CBM), European Commission, Ministerio de Economía, Industria y Competitividad (España), Instituto de Salud Carlos III, Comunidad de Madrid, Gobierno del Principado de Asturias (España), Asociación Española Contra el Cáncer, and Fundación Cajastur
- Subjects
0301 basic medicine ,Aging ,Physiology ,lcsh:Medicine ,Determinants of hADSC ,Biochemistry ,Pediatrics ,Epigenesis, Genetic ,Mice ,miRNA cluster (14q32.31) ,Medicine and Health Sciences ,European commission ,lcsh:Science ,Cells, Cultured ,Cellular Senescence ,Multidisciplinary ,DNA methylation ,Biología y Biomedicina / Biología ,Chromatin ,3. Good health ,Up-Regulation ,Nucleic acids ,Intercellular Signaling Peptides and Proteins ,Christian ministry ,Epigenetics ,RNA, Long Noncoding ,Biological Cultures ,DNA modification ,Chromatin modification ,Research Article ,Chromosome biology ,Cell biology ,Cells ,Research and Analysis Methods ,Senescence ,Iodide Peroxidase ,Deregulation ,03 medical and health sciences ,Genomic Imprinting ,Political science ,Genetics ,Animals ,Humans ,Non-coding RNA ,Cell Proliferation ,Chromosomes, Human, Pair 14 ,Natural antisense transcripts ,Biology and life sciences ,lcsh:R ,Calcium-Binding Proteins ,Membrane Proteins ,Mesenchymal Stem Cells ,DNA ,Cell Cultures ,Gene regulation ,MicroRNAs ,030104 developmental biology ,Gene Expression Regulation ,Genetic Loci ,Long non-coding RNAs ,RNA ,lcsh:Q ,Gene expression ,Physiological Processes ,Humanities ,Organism Development ,Developmental Biology - Abstract
Background Human adult adipose-derived stem cells (hADSCs) have become the most promising cell source for regenerative medicine. However the prolonged ex vivo expansion periods required to obtain the necessary therapeutic dose promotes progressive senescence, with the concomitant reduction of their therapeutic potential. Aim and scope A better understanding of the determinants of hADSC senescence is needed to improve biosafety while preserving therapeutic efficiency. Here, we investigated the association between deregulation of the imprinted DLK1-DIO3 region and replicative senescence in hADSC cultures. Methods We compared hADSC cultures at short (P) and prolonged (P) passages, both in standard and low [O] (21 and 3%, respectively), in relation to replicative senescence. hADSCs were evaluated for expression alterations in the DLK1-DIO3 region on chromosome 14q32, and particularly in its main miRNA cluster. Results Comparison of hADSCs cultured at P or P surprisingly showed a quite significant fraction (69%) of upregulated miRNAs in P cultures mapping to the imprinted 14q32 locus, the largest miRNA cluster described in the genome. In agreement, expression of the lncRNA MEG3 (Maternally Expressed 3; Meg3/Gtl2), cultured at 21 and 3% [O], was also significantly higher in P than in P passages. During hADSC replicative senescence the AcK16H4 activating mark was found to be significantly associated with the deregulation of the entire DLK1-DIO3 locus, with a secondary regulatory role for the methylation of DMR regions. Conclusion A direct relationship between DLK1-DIO3 deregulation and replicative senescence of hADSCs is reported, involving upregulation of a very significant fraction of its largest miRNA cluster (14q32.31), paralleled by the progressive overexpression of the lncRNA MEG3, which plays a central role in the regulation of Dlk1/Dio3 activation status in mice., Ministry of Economy, Industry (SAF2015-70882-R; AEI/FEDER, UE), Comunidad Autónoma de Madrid (S2010/BMD-2420), Instituto Salud Carlos III (RETICS TerCel, RD12/0019/0018) and the European Commission (FP7-HEALTH-2009/CARE-MI)
- Published
- 2018