title: A biomimetic gelatin-based platform elicits a pro-differentiation effect on podocytes through mechanotransduction.
abstract: Using a gelatin microbial transglutaminase (gelatin-mTG) cell culture platform tuned to exhibit stiffness spanning that of healthy and diseased glomeruli, we demonstrate that kidney podocytes show marked stiffness sensitivity. Podocyte-specific markers that are critical in the formation of the renal filtration barrier are found to be regulated in association with stiffness-mediated cellular behaviors. While podocytes typically de-differentiate in culture and show diminished physiological function in nephropathies characterized by altered tissue stiffness, we show that gelatin-mTG substrates with Young's modulus near that of healthy glomeruli elicit a pro-differentiation and maturation response in podocytes better than substrates either softer or stiffer. The pro-differentiation phenotype is characterized by upregulation of gene and protein expression associated with podocyte function, which is observed for podocytes cultured on gelatin-mTG gels of physiological stiffness independent of extracellular matrix coating type and density. Signaling pathways involved in stiffness-mediated podocyte behaviors are identified, revealing the interdependence of podocyte mechanotransduction and maintenance of their physiological function. This study also highlights the utility of the gelatin-mTG platform as an in vitro system with tunable stiffness over a range relevant for recapitulating mechanical properties of soft tissues, suggesting its potential impact on a wide range of research in cellular biophysics.
authors.affiliation: Department of Mechanical Engineering Columbia University, New York, NY 10027, USA.
authors.forename: Mufeng
authors.initials: M
authors.lastname: Hu
authors.affiliation: Department of Pharmacological Sciences and Systems Biology Center New York, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
authors.forename: Evren U
authors.initials: EU
authors.lastname: Azeloglu
authors.affiliation: Department of Mechanical Engineering Columbia University, New York, NY 10027, USA.
authors.forename: Amit
authors.initials: A
authors.lastname: Ron
authors.affiliation: Department of Chemistry Columbia University, New York, NY 10027, USA.
authors.forename: Khanh-Hoa
authors.initials: KH
authors.lastname: Tran-Ba
authors.affiliation: Department of Pharmacological Sciences and Systems Biology Center New York, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
authors.forename: Rhodora C
authors.initials: RC
authors.lastname: Calizo
authors.affiliation: Department of Pharmacological Sciences and Systems Biology Center New York, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
authors.forename: Iman
authors.initials: I
authors.lastname: Tavassoly
authors.affiliation: Department of Mechanical Engineering Columbia University, New York, NY 10027, USA.
authors.forename: Smiti
authors.initials: S
authors.lastname: Bhattacharya
authors.affiliation: Department of Pharmacological Sciences and Systems Biology Center New York, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
authors.forename: Gomathi
authors.initials: G
authors.lastname: Jayaraman
authors.affiliation: Department of Pharmacological Sciences and Systems Biology Center New York, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
authors.forename: Yibang
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of Pharmacological Sciences and Systems Biology Center New York, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
authors.forename: Vera
authors.initials: V
authors.lastname: Rabinovich
authors.affiliation: Department of Pharmacological Sciences and Systems Biology Center New York, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
authors.forename: Ravi
authors.initials: R
authors.lastname: Iyengar
authors.affiliation: Department of Mechanical Engineering Columbia University, New York, NY 10027, USA.
authors.forename: James C
authors.initials: JC
authors.lastname: Hone
authors.affiliation: Department of Pharmacological Sciences and Systems Biology Center New York, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.|Division of Nephrology, Department of Medicine, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
authors.forename: John C
authors.initials: JC
authors.lastname: He
authors.affiliation: Department of Chemistry Columbia University, New York, NY 10027, USA.
authors.forename: Laura J
authors.initials: LJ
authors.lastname: Kaufman
mesh_terms: D040761:Biomimetic Materials / Q000378:metabolism*; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D005780:Gelatin / Q000378:metabolism*; D006801:Humans; D040542:Mechanotransduction, Cellular; D050199:Podocytes / Q000187:drug effects* / Q000502:physiology*; D011503:Transglutaminases / Q000378:metabolism*
chemical_list: D005780:Gelatin; D011503:Transglutaminases