Altering equine corneal fibroblast differentiation through Smad gene transfer

Marlo, Todd L.; Giuliano, Elizabeth A.; Tripathi, Ratnakar; Sharma, Ajay; Mohan, Rajiv R.

doi:10.1111/vop.12485

Cited by 11 publications

(5 citation statements)

References 37 publications

(48 reference statements)

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“…The phosphorylated pSMAD2/3 complex binds to SMAD4 to translocate into the nucleus. Inhibitory SMAD6 and SMAD7 prevent activation of SMAD signaling by binding to pSMAD2/3 prior to SMAD4 activity [48][49][50][51][52][53].…”

Section: Introductionmentioning

confidence: 99%

Mustard Gas Exposure Actuates SMAD2/3 Signaling to Promote Myofibroblast Generation in the Cornea

Sinha,

Tripathi,

Balne

et al. 2023

Cells

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Sulfur mustard gas (SM) is a vesicating and alkylating agent used as a chemical weapon in many mass-casualty incidents since World War I. Ocular injuries were reported in >90% of exposed victims. The mechanisms underlying SM-induced blindness remain elusive. This study tested the hypothesis that SM-induced corneal fibrosis occurs due to the generation of myofibroblasts from resident fibroblasts via the SMAD2/3 signaling pathway in rabbit eyes in vivo and primary human corneal fibroblasts (hCSFs) isolated from donor corneas in vitro. Fifty-four New Zealand White Rabbits were divided into three groups (Naïve, Vehicle, SM-Vapor treated). The SM-Vapor group was exposed to SM at 200 mg-min/m3 for 8 min at the MRI Global facility. Rabbit corneas were collected on day 3, day 7, and day 14 for immunohistochemistry, RNA, and protein lysates. SM caused a significant increase in SMAD2/3, pSMAD, and ɑSMA expression on day 3, day 7, and day 14 in rabbit corneas. For mechanistic studies, hCSFs were treated with nitrogen mustard (NM) or NM + SIS3 (SMAD3-specific inhibitor) and collected at 30 m, 8 h, 24 h, 48 h, and 72 h. NM significantly increased TGFβ, pSMAD3, and SMAD2/3 levels. On the contrary, inhibition of SMAD2/3 signaling by SIS3 treatment significantly reduced SMAD2/3, pSMAD3, and ɑSMA expression in hCSFs. We conclude that SMAD2/3 signaling appears to play a vital role in myofibroblast formation in the cornea following mustard gas exposure.

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Section: Introductionmentioning

confidence: 99%

Mustard Gas Exposure Actuates SMAD2/3 Signaling to Promote Myofibroblast Generation in the Cornea

Sinha,

Tripathi,

Balne

et al. 2023

Cells

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“…Other studies evaluated expression of transgenes coding for decorin (DCN), 36 bone morphogenic protein (BMP), 37,38 IGF‐1, 39–44 interleukin receptor antagonist (IL1‐RA), 44,45 VEGF, 14 and FGF), 14 interleukin 4 (IL‐4), 46 and interleukin 10 (IL‐10) 47,48 . Two studies investigated the effect of small interfering RNA (RNAi) to inhibit gene expression 49,50 . A variety of delivery systems were used including polyethyleneimine nanoparticles, plasmid DNA, and recombinant viral vectors derived from adenovirus (AdV), adeno‐associated virus (AAV), retrovirus, lentivirus, equine infectious anemia virus (EIAV), and human immunodeficiency virus (HIV).…”

Section: Introductionmentioning

confidence: 99%

Administration and detection of gene therapy in horses: A systematic review

Haughan

Ortved

Robinson

2022

Drug Testing and Analysis

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Gene therapy uses genetic modification of cells to produce a therapeutic effect.Defective or missing genes can be repaired or replaced, or gene expression can be modified using a variety of technologies. Repair of defective genes can be achieved using specialized gene editing tools. Gene addition promotes gene expression by introducing synthetic copies of genes of interest (transgenes) into cells where they are transcribed and translated into therapeutic proteins. Protein production can also be modified using therapies that regulate gene expression.Gene therapy is currently prohibited in both human and equine athletes because of the potential to induce production of performance-enhancing proteins in the athlete's body, also referred to as "gene doping." Detection of gene doping is challenging and necessitates development of creative, novel analytical methods for doping control. Methods for detection of gene doping must be specific to and will vary depending on the type of gene therapy. The purpose of this paper is to present the results of a systematic review of gene editing, gene therapy, and detection of gene doping in horses.Based on the published literature, gene therapy has been administered to horses in a large number of experimental studies and a smaller number of clinical cases. Detection of gene therapy is possible using a combination of PCR and sequencing technologies. This summary can provide a basis for discussion of appropriate and inappropriate uses for gene therapy in horses by the veterinary community and guide expansion of methods to detect inappropriate uses by the regulatory community.

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“…Corneal wound healing is a complex process involving numerous cytokines, activation of keratocytes, transdifferentiation of fibroblasts to myofibroblasts, angiogenesis, and increased extracellular matrix (ECM) deposition, increased secretion of matrix metallic proteinases (MMPs), and altered gene expression of genes such as the Smad family [3,[5][6][7]. The cytokine transforming growth factor beta (TGFβ) has been shown to play a major role in the formation of corneal fibrosis via activation of fibroblasts and differentiation of activated fibroblasts to…”

Section: Corneal Wound Healing and Major Playersmentioning

confidence: 99%

“…The effects of TGF-β1 are facilitated via various intracellular signaling pathways including the Smad-dependent intracellular signaling pathways [9]. The Smad protein family is classified based on function and is composed of three categories including receptorregulated Smads (R-Smads), common mediator Smads (co-Smads), and inhibitor Smads (I-Smads) [7,9,43].…”

Section: Chapter 2 Literature Reviewmentioning

confidence: 99%

Topical therapeutic strategies for the treatment of corneal fibrosis in veterinary species

Fuchs¹

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Purpose: To determine the safety and efficacy of a novel combination of TRAM-34 and ascorbic acid applied topically in vivo in native and alkali injured rabbit corneas. Methods: Twelve New Zealand rabbits were randomly assigned into 2 groups (6 rabbits/group). Native healthy eyes were treated OD BIDx5 days with either treatment (combination TRAM-34 25[mu]M (Tocris Biosciences, Bristol, UK) and ascorbic acid 10 percent (TVC)) or control (BSS). Rabbits underwent an axial corneal wound OS using an established model. Groups were treated OS BIDx5 days. Degree of corneal opacity, ocular health, safety, and efficacy were determined utilizing the Fantes grading scale and modified McDonald-Shadduck (mMS) scoring system. Immunohistochemical and microscopy techniques evaluated corneal fibrotic markers at study conclusion (day 28). Results: Combination therapy was well tolerated in all eyes, with no significant differences in mMS scores, IOP, or central corneal thickness (CCT) between treatment and control groups. Significant differences in mMS scores between groups were found at day 1 (p=0.0001), 2 (p=0.0001), 3 (p=0.0001), 4 (p=0.0285), 14 (p=0.0041) and 28 (p=0.0002). Significant differences in Fantes scores were detected between groups at day 7 (p=0.001), 14 (p=0.0027), and 28 (p=0.0001). Significant differences in CCT between groups were found at days 7 (p=0.036), 14 (p=0.0495), and 28 (p=0.0487). Laboratory testing of corneal tissues demonstrated decreased fibrosis in treatment versus control groups at day 28. Conclusions: Novel bi-modal TVC topical therapy was well tolerated and demonstrated improved corneal wound healing and reduction in fibrotic changes in TVC treated rabbits compared to controls.

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Altering equine corneal fibroblast differentiation through Smad gene transfer

Abstract: Using gene transfer technology involving profibrotic Smad silencing, antifibrotic Smad overexpression or its combination is a novel strategy to control TGF-β1-mediated fibrosis in equine fibroblasts. Combination gene therapy was not better than single gene therapy in this study.

Cited by 11 publications

References 37 publications

Mustard Gas Exposure Actuates SMAD2/3 Signaling to Promote Myofibroblast Generation in the Cornea

Mustard Gas Exposure Actuates SMAD2/3 Signaling to Promote Myofibroblast Generation in the Cornea

Administration and detection of gene therapy in horses: A systematic review

Topical therapeutic strategies for the treatment of corneal fibrosis in veterinary species

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