Extracellular matrix stiffness and TGFβ2 regulate YAP/TAZ activity in human trabecular meshwork cells

Li, Haiyan; Raghunathan, Vijay Krishna; Stamer, W. Daniel; Ganapathy, Preethi S.; Herberg, Samuel

doi:10.1101/2021.09.29.462394

Cited by 6 publications

(35 citation statements)

References 84 publications

(131 reference statements)

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“…Recently, we demonstrated that TGFβ2 increases nuclear YAP and TAZ, a readout for active YAP/TAZ signaling, in HTM cells cultured atop or within ECM hydrogels 26 . To assess the effect of TGFβ2 on YAP/TAZ subcellular localization in HSC cells subjected to the different treatments, we evaluated both YAP and TAZ nuclear-to-cytoplasmic (N/C) ratios.…”

Section: Resultsmentioning

confidence: 99%

“…Therefore, we hypothesized that TGFβ2 may increase HSC cell contractility, which could be reduced by YAP/TAZ inhibition. To test this hypothesis, we encapsulated HSC cells in ECM hydrogels - a method developed in our laboratory to characterize HTM cell behavior 26, 30, 43 - and treated the constructs with TGFβ2, either alone or in combination with VP, to assess the level of hydrogel contraction at 5 d. TGFβ2-treated HSC hydrogels exhibited significantly greater contraction compared to controls (62.79% of controls; Fig. 5A,B ), consistent with our previous studies using HTM cells 26, 30, 43 .…”

Section: Resultsmentioning

confidence: 99%

“…To test this hypothesis, we encapsulated HSC cells in ECM hydrogels - a method developed in our laboratory to characterize HTM cell behavior 26, 30, 43 - and treated the constructs with TGFβ2, either alone or in combination with VP, to assess the level of hydrogel contraction at 5 d. TGFβ2-treated HSC hydrogels exhibited significantly greater contraction compared to controls (62.79% of controls; Fig. 5A,B ), consistent with our previous studies using HTM cells 26, 30, 43 . Co-treatment with VP significantly decreased pathologic HSC hydrogel contraction (81.56% of controls) compared to TGFβ2-treated samples ( Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Thirty microliters of the hydrogel solution were pipetted onto a Surfasil (Fisher Scientific)-coated 18 × 18-mm square glass coverslip followed by placing a regular 12-mm round glass coverslip onto the hydrogels. Constructs were crosslinked by exposure to UV light (OmniCure S1500 UV Spot Curing System; Excelitas Technologies, Mississauga, Ontario, Canada) at 320-500 nm, 2.2 W/cm 2 for 5 s, as previously described 26, 30, 43 . The hydrogel-adhered coverslips were removed with fine-tipped tweezers and placed in 24-well culture plates (Corning; Thermo Fisher Scientific).…”

Section: Methodsmentioning

confidence: 99%

“…YAP/TAZ can be activated by multiple stimuli, such as stiffened ECM, increased mechanical stress, and exposure to growth factors 41, 42 . Recently, we demonstrated that both ECM stiffening and TGFβ2 increase YAP/TAZ activity in human TM cells, which was linked to pathologic cell contractility and ECM remodeling that may exacerbate glaucoma pathology 26 .…”

Section: Introductionmentioning

confidence: 99%

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YAP/TAZ mediate TGFβ2-induced Schlemm’s canal cell dysfunction

Perkumas

Stamer

et al. 2022

Preprint

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Purpose: Elevated transforming growth factor beta2 (TGFβ2) levels in the aqueous humor have been linked to glaucomatous outflow tissue dysfunction. Potential mediators of dysfunction are the transcriptional co-activators, Yes-associated protein (YAP) and transcriptional coactivator with PDZ binding motif (TAZ). However, the molecular underpinnings of YAP/TAZ modulation in SC cells under glaucomatous conditions are not well understood. Here, we investigate how TGFβ2 regulates YAP/TAZ activity in human SC (HSC) cells using biomimetic extracellular matrix (ECM) hydrogels, and examine whether pharmacologic YAP/TAZ inhibition would attenuate TGFβ2-induced HSC cell dysfunction. Methods: Primary HSC cells were seeded atop photocrosslinked ECM hydrogels, made of collagen type I, elastin-like polypeptide and hyaluronic acid, or encapsulated within the hydrogels. Changes in actin cytoskeleton, YAP/TAZ activity, ECM production, phospho-myosin light chain levels, and hydrogel contraction were assessed. Results: TGFβ2 significantly increased YAP/TAZ nuclear localization in HSC cells, which was prevented by either filamentous (F)-actin relaxation or depolymerization. Pharmacologic YAP/TAZ inhibition using verteporfin decreased fibronectin expression and reduced actomyosin cytoskeletal rearrangement in HSC cells induced by TGFβ2. Similarly, verteporfin significantly attenuated TGFβ2-induced HSC cell-encapsulated hydrogel contraction. Conclusions: Our data provide evidence for a pathologic role of aberrant YAP/TAZ signaling in HSC cells under simulated glaucomatous conditions, and suggest that pharmacologic YAP/TAZ inhibition has promising potential to improve outflow tissue dysfunction.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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YAP/TAZ mediate TGFβ2-induced Schlemm’s canal cell dysfunction

Perkumas

Stamer

et al. 2022

Preprint

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Technological advances in ocular trabecular meshwork in vitro models for glaucoma research

Bikuna-Izagirre

Aldazábal

Extramiana

et al. 2022

Biotech & Bioengineering

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Glaucoma is the leading cause of irreversible blindness worldwide and is characterized by the progressive degeneration of the optic nerve. Intraocular pressure (IOP), which is considered to be the main risk factor for glaucoma development, builds up in response to the resistance (resistance to what?) provided by the trabecular meshwork (TM) to aqueous humor (AH) outflow. Although the TM and its relationship to AH outflow have remained at the forefront of scientific interest, researchers remain uncertain regarding which mechanisms drive the deterioration of the TM. Current tissue‐engineering fabrication techniques have come up with promising approaches to successfully recreate the TM. Nonetheless, more accurate models are needed to understand the factors that make glaucoma arise. In this review, we provide a chronological evaluation of the technological milestones that have taken place in the field of glaucoma research, and we conduct a comprehensive comparison of available TM fabrication technologies. Additionally, we also discuss AH perfusion platforms, since they are essential for the validation of these scaffolds, as well as pressure–outflow relationship studies and the discovery of new IOP‐reduction therapies.

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Role of protein phosphorylation in cell signaling, disease, and the intervention therapy

Pang

Wang

Qin

et al. 2022

MedComm

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Protein phosphorylation is an important post‐transcriptional modification involving an extremely wide range of intracellular signaling transduction pathways, making it an important therapeutic target for disease intervention. At present, numerous drugs targeting protein phosphorylation have been developed for the treatment of various diseases including malignant tumors, neurological diseases, infectious diseases, and immune diseases. In this review article, we analyzed 303 small‐molecule protein phosphorylation kinase inhibitors (PKIs) registered and participated in clinical research obtained in a database named Protein Kinase Inhibitor Database (PKIDB), including 68 drugs approved by the Food and Drug Administration of the United States. Based on previous classifications of kinases, we divided these human protein phosphorylation kinases into eight groups and nearly 50 families, and delineated their main regulatory pathways, upstream and downstream targets. These groups include: protein kinase A, G, and C (AGC) and receptor guanylate cyclase (RGC) group, calmodulin‐dependent protein kinase (CaMK) group, CMGC [Cyclin‐dependent kinases (CDKs), Mitogen‐activated protein kinases (MAPKs), Glycogen synthase kinases (GSKs), and C dc2‐like kinases (CLKs)] group, sterile (STE)‐MAPKs group, tyrosine kinases (TK) group, tyrosine kinase‐like (TKL) group, atypical group, and other groups. Different groups and families of inhibitors stimulate or inhibit others, forming an intricate molecular signaling regulatory network. This review takes newly developed new PKIs as breakthrough point, aiming to clarify the regulatory network and relationship of each pathway, as well as their roles in disease intervention, and provide a direction for future drug development.

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Extracellular matrix stiffness and TGFβ2 regulate YAP/TAZ activity in human trabecular meshwork cells

Cited by 6 publications

References 84 publications

YAP/TAZ mediate TGFβ2-induced Schlemm’s canal cell dysfunction

YAP/TAZ mediate TGFβ2-induced Schlemm’s canal cell dysfunction

Technological advances in ocular trabecular meshwork in vitro models for glaucoma research

Role of protein phosphorylation in cell signaling, disease, and the intervention therapy

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