Poster #102, Cedars-Sinai Medical Center
Enhancing Maturation of Human iPSC-Derived Tenocytes Through PIEZO1 Activation in Tenoids
Mentors: Ahmet Engin Pazarceviren, PhD and Dmitriy Sheyn, PhD
Tendon injuries commonly heal through fibrotic remodeling rather than true regeneration, resultingin incomplete restoration of tendon function. Tendons are highly organized connective tissues that transmit forces from muscle to bone through a collagen-rich extracellular matrix (ECM) maintained by specialized cells called tenocytes. Following injury, the loss of organized ECM and replacement with scar tissue compromise tendon strength, elasticity, and limiting the tendon regenerative capacity. During embryonic development, tendon progenitor cells undergo tightly regulated biochemical and mechanical signaling to differentiate into mature tenocytes capable of producing and maintaining a highly organized ECM. We therefore hypothesize that mimicking developmental mechanotransduction will promote iTenocyte maturation. Our laboratory established a developmental differentiation platform capable of generating committed iTenocytes in a 2D environment from iPSCs through sequential WNT/BMP modulation followed by GDF5 (50ng/mL) and TGF-β3 (10ng/mL) treatment. Preliminary studies demonstrated efficient differentiation toward the tendon lineage; however, the resulting cells exhibit characteristics of an immature tenocyte phenotype. This study evaluates whether pharmacological activation of the mechanosensitive ion channel PIEZO1 using the small-molecule agonist Yoda1, in combination with a Type I collagen hydrogel that mimics the native tendon ECM, enhances iTenocyte maturation in a 3D environment. Activation of PIEZO1 induces calcium-dependent mechanotransduction signaling, providing a controllable approach to stimulate maturation without requiring complex mechanical loading systems. Maturation is evaluated by assessing the expression of tendon-associated proteins SCX, MKX, and TNMD while monitoring α-smooth muscle actin (α-SMA) as an indicator of myofibroblast differentiation. This study establishes a framework for incorporating developmental mechanotransduction into iPSC-derived tendon differentiation. The findings provided insight into whether PIEZO1-mediated mechanical signaling enhances iTenocyte maturation and may guide the development of more effective stem cell-based therapies for tendon regeneration.