Non-Viral Gene Therapy in Trabecular Meshwork Cells to Prevent Fibrosis in Minimally Invasive Glaucoma Surgery

Kostas Kafetzis; ARISTIDES TAGALAKIS; Jinyuan Luo; Greymi Tan; Kai Xin Thong; Neeru Vallabh; Carl Sheridan; Yusuke Sato; Hideyoshi Harashima; Cynthia Yu-Wai-Man

doi:10.3390/pharmaceutics14112472

Non-Viral Gene Therapy in Trabecular Meshwork Cells to Prevent Fibrosis in Minimally Invasive Glaucoma Surgery

Kostas Kafetzis, ARISTIDES TAGALAKIS^*, Jinyuan Luo, Greymi Tan, Kai Xin Thong, Neeru Vallabh, Carl Sheridan, Yusuke Sato, Hideyoshi Harashima, Cynthia Yu-Wai-Man^*

^*Corresponding author for this work

Research output: Contribution to journal › Article (journal) › peer-review

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Abstract

: The primary cause of failure for minimally invasive glaucoma surgery (MIGS) is fibrosis in
the trabecular meshwork (TM) that regulates the outflow of aqueous humour, and no anti-fibrotic
drug is available for intraocular use in MIGS. The myocardin-related transcription factor/serum
response factor (MRTF/SRF) pathway is a promising anti-fibrotic target. This study aims to utilise
a novel lipid nanoparticle (LNP) to deliver MRTF-B siRNA into human TM cells and to compare
its effects with those observed in human conjunctival fibroblasts (FF). Two LNP formulations were
prepared with and without the targeting peptide cY, and with an siRNA concentration of 50 nM.
We examined the biophysical properties and encapsulation efficiencies of the LNPs, and evaluated
the effects of MRTF-B silencing on cell viability, key fibrotic genes expression and cell contractility.
Both LNP formulations efficiently silenced MRTF-B gene and were non-cytotoxic in TM and FF
cells. The presence of cY made the LNPs smaller and more cationic, but had no significant effect on
encapsulation efficiency. Both TM and FF cells also showed significantly reduced contractibility after
transfection with MRTF-B siRNA LNPs. In TM cells, LNPs with cY achieved a greater decrease in
contractility compared to LNPs without cY. In conclusion, we demonstrate that the novel CL4H6-
LNPs are able to safely and effectively deliver MRTF-B siRNA into human TM cells. LNPs can serve
as a promising non-viral gene therapy to prevent fibrosis in MIGS.

Original language	English
Article number	2472
Pages (from-to)	1-17
Journal	Pharmaceutics
Volume	14
Early online date	16 Nov 2022
DOIs	https://doi.org/10.3390/pharmaceutics14112472 https://doi.org/10.3390/pharmaceutics14112472
Publication status	Published - 16 Nov 2022

Keywords

nanoparticle
gene therapy
trabecular meshwork
fibrosis
MIGS

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Data Science STEM Research Centre

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This output contributes to the following UN Sustainable Development Goals (SDGs)

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pharmaceutics-14-02472Final published version, 4.61 MBLicence: CC BY

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Kafetzis, K., TAGALAKIS, ARISTIDES., Luo, J., Tan, G., Thong, K. X., Vallabh, N., Sheridan, C., Sato, Y., Harashima, H., & Yu-Wai-Man, C. (2022). Non-Viral Gene Therapy in Trabecular Meshwork Cells to Prevent Fibrosis in Minimally Invasive Glaucoma Surgery. Pharmaceutics, 14, 1-17. Article 2472. https://doi.org/10.3390/pharmaceutics14112472, https://doi.org/10.3390/pharmaceutics14112472

@article{0580682a590440ce9073bc0dd618cb3d,

title = "Non-Viral Gene Therapy in Trabecular Meshwork Cells to Prevent Fibrosis in Minimally Invasive Glaucoma Surgery",

abstract = ": The primary cause of failure for minimally invasive glaucoma surgery (MIGS) is fibrosis inthe trabecular meshwork (TM) that regulates the outflow of aqueous humour, and no anti-fibroticdrug is available for intraocular use in MIGS. The myocardin-related transcription factor/serumresponse factor (MRTF/SRF) pathway is a promising anti-fibrotic target. This study aims to utilisea novel lipid nanoparticle (LNP) to deliver MRTF-B siRNA into human TM cells and to compareits effects with those observed in human conjunctival fibroblasts (FF). Two LNP formulations wereprepared with and without the targeting peptide cY, and with an siRNA concentration of 50 nM.We examined the biophysical properties and encapsulation efficiencies of the LNPs, and evaluatedthe effects of MRTF-B silencing on cell viability, key fibrotic genes expression and cell contractility.Both LNP formulations efficiently silenced MRTF-B gene and were non-cytotoxic in TM and FFcells. The presence of cY made the LNPs smaller and more cationic, but had no significant effect onencapsulation efficiency. Both TM and FF cells also showed significantly reduced contractibility aftertransfection with MRTF-B siRNA LNPs. In TM cells, LNPs with cY achieved a greater decrease incontractility compared to LNPs without cY. In conclusion, we demonstrate that the novel CL4H6-LNPs are able to safely and effectively deliver MRTF-B siRNA into human TM cells. LNPs can serveas a promising non-viral gene therapy to prevent fibrosis in MIGS.",

keywords = "nanoparticle, gene therapy, trabecular meshwork, fibrosis, MIGS",

author = "Kostas Kafetzis and ARISTIDES TAGALAKIS and Jinyuan Luo and Greymi Tan and Thong, {Kai Xin} and Neeru Vallabh and Carl Sheridan and Yusuke Sato and Hideyoshi Harashima and Cynthia Yu-Wai-Man",

year = "2022",

month = nov,

day = "16",

doi = "10.3390/pharmaceutics14112472",

language = "English",

volume = "14",

pages = "1--17",

journal = "Pharmaceutics",

issn = "1999-4923",

publisher = "Multidisciplinary Digital Publishing Institute (MDPI)",

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Kafetzis, K, TAGALAKIS, ARISTIDES, Luo, J, Tan, G, Thong, KX, Vallabh, N, Sheridan, C, Sato, Y, Harashima, H & Yu-Wai-Man, C 2022, 'Non-Viral Gene Therapy in Trabecular Meshwork Cells to Prevent Fibrosis in Minimally Invasive Glaucoma Surgery', Pharmaceutics, vol. 14, 2472, pp. 1-17. https://doi.org/10.3390/pharmaceutics14112472, https://doi.org/10.3390/pharmaceutics14112472

TY - JOUR

T1 - Non-Viral Gene Therapy in Trabecular Meshwork Cells to Prevent Fibrosis in Minimally Invasive Glaucoma Surgery

AU - Kafetzis, Kostas

AU - TAGALAKIS, ARISTIDES

AU - Luo, Jinyuan

AU - Tan, Greymi

AU - Thong, Kai Xin

AU - Vallabh, Neeru

AU - Sheridan, Carl

AU - Sato, Yusuke

AU - Harashima, Hideyoshi

AU - Yu-Wai-Man, Cynthia

PY - 2022/11/16

Y1 - 2022/11/16

N2 - : The primary cause of failure for minimally invasive glaucoma surgery (MIGS) is fibrosis inthe trabecular meshwork (TM) that regulates the outflow of aqueous humour, and no anti-fibroticdrug is available for intraocular use in MIGS. The myocardin-related transcription factor/serumresponse factor (MRTF/SRF) pathway is a promising anti-fibrotic target. This study aims to utilisea novel lipid nanoparticle (LNP) to deliver MRTF-B siRNA into human TM cells and to compareits effects with those observed in human conjunctival fibroblasts (FF). Two LNP formulations wereprepared with and without the targeting peptide cY, and with an siRNA concentration of 50 nM.We examined the biophysical properties and encapsulation efficiencies of the LNPs, and evaluatedthe effects of MRTF-B silencing on cell viability, key fibrotic genes expression and cell contractility.Both LNP formulations efficiently silenced MRTF-B gene and were non-cytotoxic in TM and FFcells. The presence of cY made the LNPs smaller and more cationic, but had no significant effect onencapsulation efficiency. Both TM and FF cells also showed significantly reduced contractibility aftertransfection with MRTF-B siRNA LNPs. In TM cells, LNPs with cY achieved a greater decrease incontractility compared to LNPs without cY. In conclusion, we demonstrate that the novel CL4H6-LNPs are able to safely and effectively deliver MRTF-B siRNA into human TM cells. LNPs can serveas a promising non-viral gene therapy to prevent fibrosis in MIGS.

AB - : The primary cause of failure for minimally invasive glaucoma surgery (MIGS) is fibrosis inthe trabecular meshwork (TM) that regulates the outflow of aqueous humour, and no anti-fibroticdrug is available for intraocular use in MIGS. The myocardin-related transcription factor/serumresponse factor (MRTF/SRF) pathway is a promising anti-fibrotic target. This study aims to utilisea novel lipid nanoparticle (LNP) to deliver MRTF-B siRNA into human TM cells and to compareits effects with those observed in human conjunctival fibroblasts (FF). Two LNP formulations wereprepared with and without the targeting peptide cY, and with an siRNA concentration of 50 nM.We examined the biophysical properties and encapsulation efficiencies of the LNPs, and evaluatedthe effects of MRTF-B silencing on cell viability, key fibrotic genes expression and cell contractility.Both LNP formulations efficiently silenced MRTF-B gene and were non-cytotoxic in TM and FFcells. The presence of cY made the LNPs smaller and more cationic, but had no significant effect onencapsulation efficiency. Both TM and FF cells also showed significantly reduced contractibility aftertransfection with MRTF-B siRNA LNPs. In TM cells, LNPs with cY achieved a greater decrease incontractility compared to LNPs without cY. In conclusion, we demonstrate that the novel CL4H6-LNPs are able to safely and effectively deliver MRTF-B siRNA into human TM cells. LNPs can serveas a promising non-viral gene therapy to prevent fibrosis in MIGS.

KW - nanoparticle

KW - gene therapy

KW - trabecular meshwork

KW - fibrosis

KW - MIGS

U2 - 10.3390/pharmaceutics14112472

DO - 10.3390/pharmaceutics14112472

M3 - Article (journal)

SN - 1999-4923

VL - 14

SP - 1

EP - 17

JO - Pharmaceutics

JF - Pharmaceutics

M1 - 2472

ER -

Non-Viral Gene Therapy in Trabecular Meshwork Cells to Prevent Fibrosis in Minimally Invasive Glaucoma Surgery

Abstract

Keywords

Research Institutes

Research Centres

UN SDGs

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