Molecular Aspects of Medicine, Journal Year: 2023, Volume and Issue: 94, P. 101228 - 101228
Published: Nov. 27, 2023
Language: Английский
Scientific Reports, Journal Year: 2024, Volume and Issue: 14(1)
Published: March 23, 2024
Mutations in myocilin (MYOC) are the leading known genetic cause of primary open-angle glaucoma, responsible for about 4% all cases. MYOC a gain-of-function phenotype which mutant accumulates endoplasmic reticulum (ER) to ER stress and trabecular meshwork (TM) cell death. Therefore, knocking out at genome level is an ideal strategy permanently cure disease. We have previously utilized CRISPR/Cas9 editing successfully target using adenovirus 5 (Ad5). However, Ad5 not suitable vector clinical use. Here, we sought determine efficacy adeno-associated viruses (AAVs) lentiviruses (LVs) TM. First, examined TM tropism single-stranded (ss) self-complimentary (sc) AAV serotypes as well LV expressing GFP via intravitreal (IVT) intracameral (IC) injections. observed that LV_GFP expression was more specific injected IVT route. IC injections Trp-mutant scAAV2 showed prominent robust also ciliary body retina. next constructed lentiviral particles Cas9 guide RNA (gRNA) targeting (crMYOC) transduction cells stably with LV_crMYOC significantly reduced accumulation its associated chronic stress. A single injection Tg-MYOC
Language: Английский
Citations
10
Cell Death and Disease, Journal Year: 2023, Volume and Issue: 14(8)
Published: Aug. 24, 2023
Abstract Glaucoma is a group of diseases that leads to chronic degeneration retinal ganglion cell (RGC) axons and progressive loss RGCs, resulting in vision loss. While aging elevated intraocular pressure (IOP) have been identified as the main contributing factors glaucoma, molecular mechanisms signaling pathways triggering RGC death axonal are not fully understood. Previous studies our laboratory found overactivation autophagy DBA/2J::GFP-LC3 mice led optic nerve with glaucomatous IOP elevation. We similar findings GFP-LC3 subjected Here, we further investigated impact deficiency on autophagy-deficient DBA/2J- Atg4b ko +/− mice, generated via CRISPR/Cas9 technology; well experimental TGFβ2 ocular hypertensive model. Our data shows that, contrast DBA/2J littermates, do develop also protected against elevation Atg4 deletion did compromise or survival mice. Moreover, results indicate protective role ON atrophy Together, suggests pathogenic activation hypertension glaucoma.
Language: Английский
Citations
18
Investigative Ophthalmology & Visual Science, Journal Year: 2023, Volume and Issue: 64(10), P. 18 - 18
Published: July 17, 2023
Purpose: Transforming growth factor (TGF)-β2 has been widely implicated in human glaucoma pathology. The purpose of this study was to determine the source TGF-β2 aqueous humor (AH) and its relationship with intraocular pressure (IOP) an inherited large animal model glaucoma. Methods: Sixty-six glaucomatous cats homozygous for LTBP2 mutation, 42 normal were studied. IOP measured weekly by rebound tonometry. AH collected anterior chamber paracentesis from each eye under general anesthesia, serum samples venous blood concurrently. Concentrations total, active latent quantitative sandwich immunoassay. For comparisons between groups, unpaired t-test or Mann Whitney test used, P < 0.05 considered significant. relationships concentrations values examined Pearson's correlation coefficient generalized estimating equation. Results: significantly higher than cats. showed a significant, robust positive (r = 0.83, R2 0.70, 0.0001). Serum did not correlate different groups. mRNA protein expression increased local ocular tissues Conclusions: Enhanced, production association identified spontaneous feline model, providing foundation preclinical testing novel therapeutics limit disease-associated elevation signaling
Language: Английский
Citations
12
Published: Jan. 21, 2025
The risk for developing primary open-angle glaucoma (POAG) correlates with the magnitude of ocular hypertension (OHT) and concentration transforming growth factor-β2 (TGFβ2) in aqueous humor. Effective treatment POAG requires detailed understanding interaction between pressure sensing mechanisms trabecular meshwork (TM) biochemical factors. Here, we employed molecular, optical, electrophysiological tonometric strategies to establish role TGFβ2 transcription functional expression mechanosensitive channel isoforms alongside studies TM contractility biomimetic hydrogels, intraocular (IOP) regulation a mouse model -induced OHT. upregulated TRPV4 PIEZO1 transcripts time-dependently augmented activation. activation induced whereas pharmacological inhibition suppressed TGFβ2-induced hypercontractility abrogated OHT eyes overexpressing TGFβ2. Trpv4 -deficient mice resisted TGFβ2-driven increases IOP. Nocturnal was not additive TGFβ- evoked Our study establishes fundamental TGFβ as modulator mechanosensing nonexcitable cells, identifies final common mechanism circadian pathological offers insights future treatments that can lower IOP sizeable cohort hypertensive patients resist current treatments.
Language: Английский
Citations
0
Published: Jan. 21, 2025
The risk for developing primary open-angle glaucoma (POAG) correlates with the magnitude of ocular hypertension (OHT) and concentration transforming growth factor-β2 (TGFβ2) in aqueous humor. Effective treatment POAG requires detailed understanding interaction between pressure sensing mechanisms trabecular meshwork (TM) biochemical factors. Here, we employed molecular, optical, electrophysiological tonometric strategies to establish role TGFβ2 transcription functional expression mechanosensitive channel isoforms alongside studies TM contractility biomimetic hydrogels, intraocular (IOP) regulation a mouse model -induced OHT. upregulated TRPV4 PIEZO1 transcripts time-dependently augmented activation. activation induced whereas pharmacological inhibition suppressed TGFβ2-induced hypercontractility abrogated OHT eyes overexpressing TGFβ2. Trpv4 -deficient mice resisted TGFβ2-driven increases IOP. Nocturnal was not additive TGFβ- evoked Our study establishes fundamental TGFβ as modulator mechanosensing nonexcitable cells, identifies final common mechanism circadian pathological offers insights future treatments that can lower IOP sizeable cohort hypertensive patients resist current treatments.
Language: Английский
Citations
0
Experimental Eye Research, Journal Year: 2024, Volume and Issue: 244, P. 109939 - 109939
Published: May 22, 2024
Transforming growth factor-β2 (TGF-β2) induced fibrogenic changes in human trabecular meshwork (HTM) cells have been implicated (TM) damage and intraocular pressure (IOP) elevation primary open-angle glaucoma (POAG) patients. Silibinin (SIL) exhibited anti-fibrotic properties various organs tissues. This study aimed to assess the effects of SIL on TGF-β2-treated HTM elucidate underlying mechanisms. Our found that effectively inhibited cell proliferation, attenuated TGF-β2-induced migration, mitigated reorganization both actin vimentin filaments. Moreover, suppressed expressions fibronectin (FN), collagen type I alpha 1 chain (COL1A1), alpha-smooth muscle (α-SMA) cells. RNA sequencing indicated interfered with phosphoinositide 3-kinase (PI3K)/protein kinase B (PKB, also known as AKT) signaling pathway, extracellular matrix (ECM)-receptor interaction, focal adhesion Western blotting demonstrated activation Janus 2 (JAK2)/signal transducers activators transcription 3 (STAT3) downstream PI3K/AKT pathways by TGF-β2, potentially contributing its inhibitory ECM protein production ability inhibit could be a potential IOP-lowering agent reducing fibrotic TM tissue POAG patients, which warrants further investigation through additional animal clinical studies.
Language: Английский
Citations
3
Published: Feb. 17, 2024
Abstract Glaucoma, a major cause of blindness, is characterized by elevated intraocular pressure (IOP) due to improper drainage aqueous humor via the trabecular meshwork (TM) outflow pathway. Our recent work identified that loss clusterin resulted in IOP. This study delves deeper elucidate role IOP regulation. Employing an ex vivo human anterior segment perfusion model, we established constitutive expression and secretion as well exogenous addition can significantly lower Interestingly, lowered transforming growth factor β2 (TGFβ2)-induced elevation. effect was linked suppression extracellular matrix (ECM) deposition and, highlighting crucial maintaining ECM equilibrium. A comprehensive global proteomic approach revealed broad impact on TM cell structure function identifying alterations protein related cytoskeletal organization, processing, cellular mechanics, following induction. These findings underscore beneficial modulation functionality clusterin. Specifically, influences actin-cytoskeleton focal adhesion dynamics, which are instrumental contractility processes. Additionally, it suppresses activity proteins critical TGFβ2, G-protein, JAK-STAT signaling pathways, vital for regulation ocular pressure. By delineating these targeted effects within pathway, our pave way novel treatment strategies aimed at mitigating progression hypertension glaucoma through molecular interventions.
Language: Английский
Citations
2
Molecular Aspects of Medicine, Journal Year: 2023, Volume and Issue: 94, P. 101219 - 101219
Published: Oct. 13, 2023
Language: Английский
Citations
5
Frontiers in Molecular Biosciences, Journal Year: 2023, Volume and Issue: 10
Published: May 11, 2023
TGF-β2 is the predominant TGF-β isoform within eye. One function of to provide eye with immune protection against intraocular inflammation. The beneficial must be under tight control a network different factors. A disbalance can result in diseases. In Primary Open-Angle Glaucoma (POAG), one leading causes irreversible blindness worldwide, significantly elevated aqueous humor and antagonistic molecules like BMPs are reduced. changes provoke an altering quantity quality extracellular matrix actin cytoskeleton outflow tissues, increased resistance thereby pressure (IOP), major risk factor for primary open-angle glaucoma. pathologic effect glaucoma mainly meditated by CCN2/CTGF. CCN2/CTGF modulate BMP signaling direct binding. specific overexpression caused increase IOP led loss axons, hallmark appears play critical role homeostatic balance eye, so we investigated if pathways tissues. To this end, analyzed on both two transgenic mouse models moderate (βB1-CTGF1) high (βB1-CTGF6) immortalized human trabecular meshwork (HTM) cells. Additionally, investigate whether mediates effects via pathways. We observed developmental malformations ciliary body βB1-CTGF6 inhibition pathway. βB1-CTGF1, detected dysregulation pathways, reduced activity signaling. was shown HTM Finally, mediated its RhoA/ROCK ERK conclude that functions as modulator which shifted
Language: Английский
Citations
4
Biochemical Pharmacology, Journal Year: 2022, Volume and Issue: 206, P. 115340 - 115340
Published: Nov. 5, 2022
Language: Английский
Citations
7