Transforming Growth Factor Beta 3 Modifies Mechanics and Composition of Extracellular Matrix Deposited by Human Trabecular Meshwork Cells

Raghunathan, Vijay Krishna; Morgan, Joshua T.; Chang, Yow Ren; Weber, Darren; Phinney, Brett S.; Murphy, Christopher J.; Russell, Paul

doi:10.1021/ab500060r

Cited by 26 publications

(37 citation statements)

References 69 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Despite the correlation of glaucoma with HTM stiffness, there is a paucity of data directly linking biological phenotypes of glaucoma with stiffening of the cells or matrix. Recently, we have shown that transforming growth factor β signaling, implicated in glaucoma, causes HTM cells to deposit a physically stiffer matrix with elevated SFRP1 secretion [ 92 ]. In a separate study, we demonstrated SFRP1, also associated with glaucoma, induces intrinsic stiffening of HTM cells [ 81 ].…”

Section: Discussionmentioning

confidence: 99%

The intrinsic stiffness of human trabecular meshwork cells increases with senescence

et al. 2015

Self Cite

View full text Add to dashboard Cite

Dysfunction of the human trabecular meshwork (HTM) plays a central role in the age-associated disease glaucoma, a leading cause of irreversible blindness. The etiology remains poorly understood but cellular senescence, increased stiffness of the tissue, and the expression of Wnt antagonists such as secreted frizzled related protein-1 (SFRP1) have been implicated. However, it is not known if senescence is causally linked to either stiffness or SFRP1 expression. In this study, we utilized in vitro HTM senescence to determine the effect on cellular stiffening and SFRP1 expression. Stiffness of cultured cells was measured using atomic force microscopy and the morphology of the cytoskeleton was determined using immunofluorescent analysis. SFRP1 expression was measured using qPCR and immunofluorescent analysis. Senescent cell stiffness increased 1.88±0.14 or 2.57±0.14 fold in the presence or absence of serum, respectively. This was accompanied by increased vimentin expression, stress fiber formation, and SFRP1 expression. In aggregate, these data demonstrate that senescence may be a causal factor in HTM stiffening and elevated SFRP1 expression, and contribute towards disease progression. These findings provide insight into the etiology of glaucoma and, more broadly, suggest a causal link between senescence and altered tissue biomechanics in aging-associated diseases.

show abstract

Section: Discussionmentioning

confidence: 99%

The intrinsic stiffness of human trabecular meshwork cells increases with senescence

et al. 2015

Self Cite

View full text Add to dashboard Cite

show abstract

“…Isolated tissue samples were immediately digested in RIPA buffer, homogenized, and stored at −80 °C until analysis. Samples were precipitated and analyzed as described previously 56 , 68 . Mass spectra were collected in a data-dependent mode.…”

Section: Methodsmentioning

confidence: 99%

Biomechanical, ultrastructural, and electrophysiological characterization of the non-human primate experimental glaucoma model

Raghunathan

Eaton

Christian

et al. 2017

Sci Rep

Self Cite

View full text Add to dashboard Cite

Laser-induced experimental glaucoma (ExGl) in non-human primates (NHPs) is a common animal model for ocular drug development. While many features of human hypertensive glaucoma are replicated in this model, structural and functional changes in the unlasered portions of trabecular meshwork (TM) of laser-treated primate eyes are understudied. We studied NHPs with ExGl of several years duration. As expected, ExGl eyes exhibited selective reductions of the retinal nerve fiber layer that correlate with electrophysiologic measures documenting a link between morphologic and elctrophysiologic endpoints. Softening of unlasered TM in ExGl eyes compared to untreated controls was observed. The degree of TM softening was consistent, regardless of pre-mortem clinical findings including severity of IOP elevation, retinal nerve fiber layer thinning, or electrodiagnostic findings. Importantly, this softening is contrary to TM stiffening reported in glaucomatous human eyes. Furthermore, microscopic analysis of unlasered TM from eyes with ExGl demonstrated TM thinning with collapse of Schlemm’s canal; and proteomic analysis confirmed downregulation of metabolic and structural proteins. These data demonstrate unexpected and compensatory changes involving the TM in the NHP model of ExGl. The data suggest that compensatory mechanisms exist in normal animals and respond to elevated IOP through softening of the meshwork to increase outflow.

show abstract

“…The OCT imaging probe was aimed at the nasal or temporal limbus and the image was centered and focused on the SC lumen. While collecting images, mouse eyes were subjected to a series of IOP steps (10,12,15,17, and 20 mm Hg) by adjusting the height of the fluid reservoir. At each IOP step, a sequence of repeated OCT B-scans (each with 1000 A-scans spanning 0.5 mm in lateral length) from spatially close positions was captured, registered, and averaged to create a high signalto-noise-ratio image from the iridocorneal angle region of each animal.…”

Section: Optical Coherence Tomographic Imagingmentioning

confidence: 99%

Integral role for lysyl oxidase‐like‐1 in conventional outflow tissue function and behavior

Schmitt

Johnson

et al. 2020

The FASEB Journal

View full text Add to dashboard Cite

Lysyl oxidase-like-1 (LOXL1), a vital crosslinking enzyme in elastin fiber maintenance, is essential for the stability and strength of elastic vessels and tissues. Variants in the LOXL1 locus associate with a dramatic increase in risk of exfoliation syndrome (XFS), a systemic fibrillopathy, which often presents with ocular hypertension and exfoliation glaucoma (XFG). We examined the role of LOXL1 in conventional outflow function, the prime regulator of intraocular pressure (IOP). Using Loxl1 −/− , Loxl1 +/− , and Loxl1 +/+ mice, we observed an inverse relationship between LOXL1 expression and IOP, which worsened with age. Elevated IOP in Loxl1 −/− mice was associated with a larger globe, decreased ocular compliance, increased outflow facility, extracellular matrix (ECM) abnormalities, and dilated intrascleral veins, yet, no dilation of arteries or capillaries. Interestingly, in living Loxl1 −/− mouse eyes, Schlemm's canal (SC) was less susceptible to collapse when challenged with acute elevations in IOP, suggesting elevated episcleral venous pressure (EVP). Thus, LOXL1 expression is required for normal IOP control, while ablation results in altered ECM repair/homeostasis and conventional outflow physiology. Dilation of SC and distal veins, but not arteries, is consistent with key structural and functional roles for elastin in low-pressure vessels subjected to cyclical mechanical stress.

show abstract

Transforming Growth Factor Beta 3 Modifies Mechanics and Composition of Extracellular Matrix Deposited by Human Trabecular Meshwork Cells

Cited by 26 publications

References 69 publications

The intrinsic stiffness of human trabecular meshwork cells increases with senescence

The intrinsic stiffness of human trabecular meshwork cells increases with senescence

Biomechanical, ultrastructural, and electrophysiological characterization of the non-human primate experimental glaucoma model

Integral role for lysyl oxidase‐like‐1 in conventional outflow tissue function and behavior

Contact Info

Product

Resources

About