Depletion of SUN1/2 induces heterochromatin accrual in mesenchymal stem cells during adipogenesis.

Critical to the mechano-regulation of mesenchymal stem cells (MSC), Linker of the Nucleoskeleton and Cytoskeleton (LINC) complex transduces cytoskeletal forces to the nuclei. The LINC complex contains outer nuclear membrane Nesprin proteins that associate with the cytoskeleton and their inner nuclear membrane couplers, SUN proteins. Here we tested the hypothesis that severing of the LINC complex-mediated cytoskeletal connections may have different effects on chromatin organization and MSC differentiation than those due to ablation of SUN proteins. In cells cultured under adipogenic conditions, interrupting LINC complex function through dominant-negative KASH domain expression (dnKASH) increased adipogesis while heterochromatin H3K27 and H3K9 methylation was unaltered. In contrast, SUN1/2 depletion inhibited adipogenic gene expression and fat droplet formation; as well the anti-adipogenic effect of SUN1/2 depletion was accompanied by increased H3K9me3, which was enriched on Adipoq, silencing this fat locus. We conclude that releasing the nucleus from cytoskeletal constraints via dnKASH accelerates adipogenesis while depletion of SUN1/2 increases heterochromatin accrual on adipogenic genes in a fashion independent of LINC complex function. Therefore, while these two approaches both disable LINC complex functions, their divergent effects on the epigenetic landscape indicate they cannot be used interchangeably to study mechanical regulation of cell differentiation.