Tread -8- EpiSci - We explicate how nanotopography-sensitive and mechanotransduction-dependent Nanog regulation is mediated by KDM3A These results show for the first time a molecular mechanism by which nanotopography-dependent stimulation specifically triggers a nuclear epigenetic mechanoresponse

Tread -7- EpiSci - 3. Nanotopography upregulates KDM3A,a specific Histone H3K9me2 demethylase. We discovered a new variant of KDM3A in mESCs. We developed a degron system which efficiently ablates all variants allowing to study the mechanistic role of the protein in an acute and reversible way.

Tread -6- EpiSci - RNA-seq analysis reveals that nanotopography induces a broad transcriptional response characterised by downregulation of integrin adhesion complexes, actin cytoskeleton organisation, and cell migration, alongside upregulation of core pluripotency factors such as Nanog.

Tread -5- Consistently with nuclear 3D changes, mESC/nanotopography interaction induces marked changes in chromatin organisation with reduced levels of H3K27me3 and H3K9me2, concomitant with increased H3K4me3. H3K9me3 levels remain unchanged, but is redistributed toward the nuclear periphery .

Tread -4- EpiSci - Our AFM measurements further reveal that nanotopography induces a progressive decrease in cellular tension and stiffness and importantly is accompanied by a change of the nuclear 3D shape.

Tread -3- EpiSci - We used nanofabrication technique Supersonic Cluster Beam Deposition (SCBD) to produce substrates with nanotopographical features of defined roughness that mimic ultrastructural ECM characteristics. Nanotopography strongly impact mESCs morphology.

Tread-2- EpiSci - The extracellular matrix (ECM) topography is increasingly recognised as a powerful regulator of cell behaviour, yet whether nanoscale topography can directly and deterministically instruct nuclear architecture and cell identity remains virtually unknown.