<i>Egr1</i> is a 3D matrix-specific mediator of mechanosensitive stem cell lineage commitment.

Abstract

While extracellular matrix (ECM) mechanics strongly regulate stem cell commitment, the field's mechanistic understanding of this phenomenon largely derives from simplified two-dimensional (2D) culture substrates. Here, we found a 3D matrix-specific mechanoresponsive mechanism for neural stem cell (NSC) differentiation. NSC lineage commitment in 3D is maximally stiffness sensitive in the range of 0.1 to 1.2 kPa, a narrower and more brain-mimetic range than we had previously identified in 2D (0.75 to 75 kPa). Transcriptomics revealed stiffness-dependent up-regulation of early growth response 1 (<i>Egr1</i>) in 3D but not in 2D. <i>Egr1</i> knockdown enhanced neurogenesis in stiff ECMs by driving β-catenin nuclear localization and activity in 3D, but not in 2D. Mechanical modeling and experimental studies under osmotic pressure indicate that stiff 3D ECMs are likely to stimulate <i>Egr1</i> via increases in confining stress during cell volumetric growth. To our knowledge, <i>Egr1</i> represents the first 3D-specific stem cell mechanoregulatory factor.