345 LOSS OF Brm AND Baf170, COMPONENTS OF THE ATP-DEPENDENT CHROMATIN REMODELING COMPLEX, FACILITATES REPROGRAMMING OF SOMATIC CELL

Abstract

The SWI/SNF [SWItch/Sucrose NonFermentable; also known as BAF (Brg/Brahma-associated factors)] ATP-dependent chromatin remodelling complexes are epigenetic modifiers that change the structure of chromatin, and thereby modulate gene transcription. The BAF chromatin remodeling complexes undergo progressive changes in subunit composition during cellular differentiation. In embryonic stem cells (ESC), BAF complex, esBAF, contains Brg1 and Baf155, which are crucial for ESC self-renewal and also facilitate induced pluripotent stem cell (iPSC) reprogramming from somatic cells. Here we sought to determine the roles of somatic BAF components (Brm and Baf170, homologues of Brg1 and Baf155, respectively) in mouse iPSC reprogramming through shRNA-mediated knockdown studies. We found that the expression of Brm and Baf170 were inhibited in reprogramming by Janus kinase/signal transducer and activator of transcription 3 (Jak/Stat3) activity, which is essential for pluripotency establishment. We further found that knockdown of Brm and Baf170 in mouse somatic cells promotes reprogramming efficiency. Specifically, loss of Brm and Baf170 during early (Days 3 and 6 after initial iPSC induction) and later-stage (Day 9) reprogramming, respectively, improves the numbers of iPSC colonies formed. These also led to significant upregulation of pluripotent genes, including Sox2, Nanog, Fgf5, and Tbx3. Although the somatic Brm and Baf170 are believed to be absent in ESC, the expression levels of Nanog and Tbx3 were increased significantly by knockdown of either Brm or Baf170 in ESC. Finally, we showed that inhibition of these somatic BAF components also promotes complete reprogramming of partially reprogrammed cells (pre-iPSC). These data suggest that inhibiting somatic BAF complex improved complete reprogramming by facilitating the activation of the pluripotency circuitry. A reduced activity of the somatic BAF complex constitutes part of the Stat3 regulated epigenetic changes during pluripotency establishment.