Wnt inhibition induces persistent increases in intrinsic stiffness of XXXX trabecular meshwork cells.

@entity1 @entity12393 @entity726 @entity3546 @entity1889 @entity71487
Wnt antagonism has been linked to @entity726 and intraocular pressure regulation, as has increased stiffness of @entity1 trabecular meshwork (HTM) tissue. We have shown culturing HTM cells on substrates that mimic the elevated stiffness of glaucomatous tissue leads to elevated expression of the Wnt antagonist @entity12393 ( @entity12393 ), suggesting a linkage between @entity12393 and HTM mechanobiology. In this study, we document biomechanical consequences of Wnt antagonism on HTM cells. Cells were treated with the Wnt antagonists ( @entity12393 , @entity71487 , and LGK-974) for 8 days and allowed to recover for 4 days. After recovery, intrinsic cell stiffness and activation of the Wnt pathway via @entity3546 staining and blotting were assayed. Basal cell stiffness values were 3.71      0.37, 4.33      3.07, and 3.07      kPa (median      S.D.) for cells derived from 3 donors. Cell stiffness increased after 0.25   g/mL (4.32      5.12, 8.86      8.51, 4.84      3.15  kPa) and 0.5   g/mL (16.75      5.59, 13.18      7.99, and 8.54      5.77  kPa) @entity12393 treatment. Stiffening was observed after 10   M @entity71487 (10.72      5.63 and 6.57      5.53  kPa) as well as LGK-974 (9.60      7.41 and 11.40      9.24  kPa) treatment compared with controls (3.79      1.01 and 5.16      2.14  kPa). Additionally, Wnt inhibition resulted in decreased @entity3546 staining and increased phosphorylation at @entity1889 41 after recovery. In conclusion, this work demonstrates a causal relationship between Wnt inhibition and cell stiffening. Additionally, these findings suggest transient Wnt inhibition resulted in durable modulation of the mechanical phenotype of HTM cells. When placed in context with previous results, these findings provide a causal link between Wnt antagonism and cell stiffness and suggest a feedback loop contributing to @entity726 progression.
