Cargando…

Blocking neuropilin-2 enhances corneal allograft survival by selectively inhibiting lymphangiogenesis on vascularized beds

PURPOSE: To investigate the potential inhibitory effects of RNA interference-mediated knockdown of neuropilin-2 (NP2) on inflammation-induced corneal hemangiogenesis and lymphangiogenesis, and whether selective inhibition of lymphangiogenesis on vascularized recipient beds before transplantation imp...

Descripción completa

Detalles Bibliográficos
Autores principales: Tang, Xian-ling, Sun, Jun-feng, Wang, Xi-ying, Du, Ling-ling, Liu, Ping
Formato: Texto
Lenguaje:English
Publicado: Molecular Vision 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2994732/
https://www.ncbi.nlm.nih.gov/pubmed/21139694
_version_ 1782192984748457984
author Tang, Xian-ling
Sun, Jun-feng
Wang, Xi-ying
Du, Ling-ling
Liu, Ping
author_facet Tang, Xian-ling
Sun, Jun-feng
Wang, Xi-ying
Du, Ling-ling
Liu, Ping
author_sort Tang, Xian-ling
collection PubMed
description PURPOSE: To investigate the potential inhibitory effects of RNA interference-mediated knockdown of neuropilin-2 (NP2) on inflammation-induced corneal hemangiogenesis and lymphangiogenesis, and whether selective inhibition of lymphangiogenesis on vascularized recipient beds before transplantation improves the graft survival. METHODS: Mouse lymphatic endothelial cells were transfected with the plasmid expressing artificial microRNA (amiRNA) against mouse NP2, and the down-regulation of VEGF-C-induced NP2 expression by NP2 amiRNA was evaluated by real-time PCR and western blot assays. Next, NP2 amiRNA or negative control amiRNA was injected intrastromally into BALB/c mouse model of suture-induced corneal neovascularization three days after surgery. Corneas were harvested 1 week after suture placement and the formation of lymphatic and blood vessels as well as the recruitment of macrophage was evaluated by immunohistochemical staining. The neovascularized graft beds treated by NP2 amiRNA or control then served as recipients of orthotopic corneal transplants, and age-matched C57BL/6 donors were used. Corneal allografts were examined twice a week for 8 weeks, and graft clarity was quantified by means of an opacity score. RESULTS: VEGF-C-induced NP2 expression at both mRNA and protein levels was significantly suppressed by NP2 amiRNA in mouse lymphatic endothelial cells. Intrastromal administration of NP2 amiRNA reduced corneal lymphangiogenesis by 45% versus control (p=0.015), but corneal hemangiogenesis (p=0.815) and the recruitment of CD11 antigen-like family member B (CD11b)-positive macrophage (p=0.589) were unchanged. Kaplan–Meier survival analysis revealed a better graft survival rate in the vascularized recipient beds pre-treated by NP2 amiRNA in comparison to controls (p=0.014). CONCLUSIONS: Knockdown of NP2 improves corneal graft survival by selectively inhibiting lymphangiogenesis in vascularized beds before transplantation. Thus our results open new treatment options for transplant rejection and other lymphatic disorders.
format Text
id pubmed-2994732
institution National Center for Biotechnology Information
language English
publishDate 2010
publisher Molecular Vision
record_format MEDLINE/PubMed
spelling pubmed-29947322010-12-06 Blocking neuropilin-2 enhances corneal allograft survival by selectively inhibiting lymphangiogenesis on vascularized beds Tang, Xian-ling Sun, Jun-feng Wang, Xi-ying Du, Ling-ling Liu, Ping Mol Vis Research Article PURPOSE: To investigate the potential inhibitory effects of RNA interference-mediated knockdown of neuropilin-2 (NP2) on inflammation-induced corneal hemangiogenesis and lymphangiogenesis, and whether selective inhibition of lymphangiogenesis on vascularized recipient beds before transplantation improves the graft survival. METHODS: Mouse lymphatic endothelial cells were transfected with the plasmid expressing artificial microRNA (amiRNA) against mouse NP2, and the down-regulation of VEGF-C-induced NP2 expression by NP2 amiRNA was evaluated by real-time PCR and western blot assays. Next, NP2 amiRNA or negative control amiRNA was injected intrastromally into BALB/c mouse model of suture-induced corneal neovascularization three days after surgery. Corneas were harvested 1 week after suture placement and the formation of lymphatic and blood vessels as well as the recruitment of macrophage was evaluated by immunohistochemical staining. The neovascularized graft beds treated by NP2 amiRNA or control then served as recipients of orthotopic corneal transplants, and age-matched C57BL/6 donors were used. Corneal allografts were examined twice a week for 8 weeks, and graft clarity was quantified by means of an opacity score. RESULTS: VEGF-C-induced NP2 expression at both mRNA and protein levels was significantly suppressed by NP2 amiRNA in mouse lymphatic endothelial cells. Intrastromal administration of NP2 amiRNA reduced corneal lymphangiogenesis by 45% versus control (p=0.015), but corneal hemangiogenesis (p=0.815) and the recruitment of CD11 antigen-like family member B (CD11b)-positive macrophage (p=0.589) were unchanged. Kaplan–Meier survival analysis revealed a better graft survival rate in the vascularized recipient beds pre-treated by NP2 amiRNA in comparison to controls (p=0.014). CONCLUSIONS: Knockdown of NP2 improves corneal graft survival by selectively inhibiting lymphangiogenesis in vascularized beds before transplantation. Thus our results open new treatment options for transplant rejection and other lymphatic disorders. Molecular Vision 2010-11-09 /pmc/articles/PMC2994732/ /pubmed/21139694 Text en Copyright © 2010 Molecular Vision. http://creativecommons.org/licenses/by/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Tang, Xian-ling
Sun, Jun-feng
Wang, Xi-ying
Du, Ling-ling
Liu, Ping
Blocking neuropilin-2 enhances corneal allograft survival by selectively inhibiting lymphangiogenesis on vascularized beds
title Blocking neuropilin-2 enhances corneal allograft survival by selectively inhibiting lymphangiogenesis on vascularized beds
title_full Blocking neuropilin-2 enhances corneal allograft survival by selectively inhibiting lymphangiogenesis on vascularized beds
title_fullStr Blocking neuropilin-2 enhances corneal allograft survival by selectively inhibiting lymphangiogenesis on vascularized beds
title_full_unstemmed Blocking neuropilin-2 enhances corneal allograft survival by selectively inhibiting lymphangiogenesis on vascularized beds
title_short Blocking neuropilin-2 enhances corneal allograft survival by selectively inhibiting lymphangiogenesis on vascularized beds
title_sort blocking neuropilin-2 enhances corneal allograft survival by selectively inhibiting lymphangiogenesis on vascularized beds
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2994732/
https://www.ncbi.nlm.nih.gov/pubmed/21139694
work_keys_str_mv AT tangxianling blockingneuropilin2enhancescornealallograftsurvivalbyselectivelyinhibitinglymphangiogenesisonvascularizedbeds
AT sunjunfeng blockingneuropilin2enhancescornealallograftsurvivalbyselectivelyinhibitinglymphangiogenesisonvascularizedbeds
AT wangxiying blockingneuropilin2enhancescornealallograftsurvivalbyselectivelyinhibitinglymphangiogenesisonvascularizedbeds
AT dulingling blockingneuropilin2enhancescornealallograftsurvivalbyselectivelyinhibitinglymphangiogenesisonvascularizedbeds
AT liuping blockingneuropilin2enhancescornealallograftsurvivalbyselectivelyinhibitinglymphangiogenesisonvascularizedbeds