Non-Viral Gene Therapy in Trabecular Meshwork Cells to Prevent Fibrosis in Minimally Invasive Glaucoma Surgery
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 (...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-11-01
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| Series: | Pharmaceutics |
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| Online Access: | https://www.mdpi.com/1999-4923/14/11/2472 |
| _version_ | 1797464261105025024 |
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| author | Jinyuan Luo Greymi Tan Kai Xin Thong Konstantinos N. Kafetzis Neeru Vallabh Carl M. Sheridan Yusuke Sato Hideyoshi Harashima Aristides D. Tagalakis Cynthia Yu-Wai-Man |
| author_facet | Jinyuan Luo Greymi Tan Kai Xin Thong Konstantinos N. Kafetzis Neeru Vallabh Carl M. Sheridan Yusuke Sato Hideyoshi Harashima Aristides D. Tagalakis Cynthia Yu-Wai-Man |
| author_sort | Jinyuan Luo |
| collection | DOAJ |
| description | 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 c<inline-formula><math xmlns="http://www.w3.org/1998/Math/MathML" display="inline"><semantics><mi mathvariant="sans-serif">Υ</mi></semantics></math></inline-formula>, and with an siRNA concentration of 50 nM. We examined the biophysical properties and encapsulation efficiencies of the LNPs, and evaluated the effects of <i>MRTF-B</i> silencing on cell viability, key fibrotic genes expression and cell contractility. Both LNP formulations efficiently silenced <i>MRTF-B</i> gene and were non-cytotoxic in TM and FF cells. The presence of c<inline-formula><math xmlns="http://www.w3.org/1998/Math/MathML" display="inline"><semantics><mi mathvariant="sans-serif">Υ</mi></semantics></math></inline-formula> 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 c<inline-formula><math xmlns="http://www.w3.org/1998/Math/MathML" display="inline"><semantics><mi mathvariant="sans-serif">Υ</mi></semantics></math></inline-formula> achieved a greater decrease in contractility compared to LNPs without c<inline-formula><math xmlns="http://www.w3.org/1998/Math/MathML" display="inline"><semantics><mi mathvariant="sans-serif">Υ</mi></semantics></math></inline-formula>. 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. |
| first_indexed | 2024-03-09T18:04:57Z |
| format | Article |
| id | doaj.art-f81134b72765434d8df5ccafcd8d7fdb |
| institution | Directory Open Access Journal |
| issn | 1999-4923 |
| language | English |
| last_indexed | 2024-03-09T18:04:57Z |
| publishDate | 2022-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pharmaceutics |
| spelling | doaj.art-f81134b72765434d8df5ccafcd8d7fdb2023-11-24T09:36:23ZengMDPI AGPharmaceutics1999-49232022-11-011411247210.3390/pharmaceutics14112472Non-Viral Gene Therapy in Trabecular Meshwork Cells to Prevent Fibrosis in Minimally Invasive Glaucoma SurgeryJinyuan Luo0Greymi Tan1Kai Xin Thong2Konstantinos N. Kafetzis3Neeru Vallabh4Carl M. Sheridan5Yusuke Sato6Hideyoshi Harashima7Aristides D. Tagalakis8Cynthia Yu-Wai-Man9Faculty of Life Sciences & Medicine, King’s College London, London SE1 7EH, UKFaculty of Life Sciences & Medicine, King’s College London, London SE1 7EH, UKFaculty of Life Sciences & Medicine, King’s College London, London SE1 7EH, UKDepartment of Biology, Edge Hill University, Ormskirk L39 4QP, UKDepartment of Eye and Vision Science, Institute of Life Course and Medical Sciences, University of Liverpool, Liverpool L69 3BX, UKDepartment of Eye and Vision Science, Institute of Life Course and Medical Sciences, University of Liverpool, Liverpool L69 3BX, UKFaculty of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-ku, Sapporo 060-0812, JapanFaculty of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-ku, Sapporo 060-0812, JapanDepartment of Biology, Edge Hill University, Ormskirk L39 4QP, UKFaculty of Life Sciences & Medicine, King’s College London, London SE1 7EH, UKThe 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 c<inline-formula><math xmlns="http://www.w3.org/1998/Math/MathML" display="inline"><semantics><mi mathvariant="sans-serif">Υ</mi></semantics></math></inline-formula>, and with an siRNA concentration of 50 nM. We examined the biophysical properties and encapsulation efficiencies of the LNPs, and evaluated the effects of <i>MRTF-B</i> silencing on cell viability, key fibrotic genes expression and cell contractility. Both LNP formulations efficiently silenced <i>MRTF-B</i> gene and were non-cytotoxic in TM and FF cells. The presence of c<inline-formula><math xmlns="http://www.w3.org/1998/Math/MathML" display="inline"><semantics><mi mathvariant="sans-serif">Υ</mi></semantics></math></inline-formula> 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 c<inline-formula><math xmlns="http://www.w3.org/1998/Math/MathML" display="inline"><semantics><mi mathvariant="sans-serif">Υ</mi></semantics></math></inline-formula> achieved a greater decrease in contractility compared to LNPs without c<inline-formula><math xmlns="http://www.w3.org/1998/Math/MathML" display="inline"><semantics><mi mathvariant="sans-serif">Υ</mi></semantics></math></inline-formula>. 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.https://www.mdpi.com/1999-4923/14/11/2472nanoparticlegene therapytrabecular meshworkfibrosisMIGS |
| spellingShingle | Jinyuan Luo Greymi Tan Kai Xin Thong Konstantinos N. Kafetzis Neeru Vallabh Carl M. Sheridan Yusuke Sato Hideyoshi Harashima Aristides D. Tagalakis Cynthia Yu-Wai-Man Non-Viral Gene Therapy in Trabecular Meshwork Cells to Prevent Fibrosis in Minimally Invasive Glaucoma Surgery Pharmaceutics nanoparticle gene therapy trabecular meshwork fibrosis MIGS |
| title | Non-Viral Gene Therapy in Trabecular Meshwork Cells to Prevent Fibrosis in Minimally Invasive Glaucoma Surgery |
| title_full | Non-Viral Gene Therapy in Trabecular Meshwork Cells to Prevent Fibrosis in Minimally Invasive Glaucoma Surgery |
| title_fullStr | Non-Viral Gene Therapy in Trabecular Meshwork Cells to Prevent Fibrosis in Minimally Invasive Glaucoma Surgery |
| title_full_unstemmed | Non-Viral Gene Therapy in Trabecular Meshwork Cells to Prevent Fibrosis in Minimally Invasive Glaucoma Surgery |
| title_short | Non-Viral Gene Therapy in Trabecular Meshwork Cells to Prevent Fibrosis in Minimally Invasive Glaucoma Surgery |
| title_sort | non viral gene therapy in trabecular meshwork cells to prevent fibrosis in minimally invasive glaucoma surgery |
| topic | nanoparticle gene therapy trabecular meshwork fibrosis MIGS |
| url | https://www.mdpi.com/1999-4923/14/11/2472 |
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