title: Intraocular pressure reduction and neuroprotection conferred by bone marrow-derived mesenchymal stem cells in an animal model of glaucoma.
abstract: BACKGROUND
  Glaucoma is a sight-threatening retinal neuropathy associated with elevated intraocular pressure (IOP) due to degeneration and fibrosis of the trabecular meshwork (TM). Glaucoma medications aim to reduce IOP without targeting the specific TM pathology, Bone-marrow mesenchymal stem cells (MSCs) are used today in various clinical studies. Here, we investigated the potential of MSCs therapy in an glaucoma-like ocular hypertension (OHT) model and decipher in vitro the effects of MSCs on primary human trabecular meshwork cells.
  
  
  METHODS
  Ocular hypertension model was performed by cauterization of 3 episcleral veins (EVC) of Long-Evans male rat eyes. MSCs were isolated from rat bone marrow, amplified in vitro and tagged with quantum dot nanocrystals. Animals were distributed as 1) MSCs group receiving 5.10(5)cells/6μl Minimum Essential Medium and 2) MEM group receiving 6μl MEM (n = 10 each). Injections were performed into the anterior chamber of 20 days-hypertensive eyes and IOP was monitored twice a week for 4 weeks. At the end of experiment, cell distribution in the anterior segment was examined in confocal microscopy on flat mounted corneas. Moreover, we tested in vitro effects of MSCs conditioned medium (MSC-CM) on primary human trabecular meshwork cells (hTM cells) using Akt activation, myosin phosphorylation and TGF-β2-dependent profibrotic phenotype in hTM cells.
  
  
  RESULTS
  We demonstrated a rapid and long-lasting in vivo effect of MSCs transplantation that significantly reduced IOP in hypertensive eyes induced by EVC. MSCs were located to the ciliary processes and the TM. Enumeration of RGCs on whole flat-mounted retina highlighted a protective effect of MSCs on RGCs death. In vitro, MSC-CM promotes: (i) hTM cells survival by activating the antiapoptotic pathway, Akt, (ii) hTM cells relaxation as analyzed by the decrease in myosin phosphorylation and (iii) inhibition of TGF-β2-dependent profibrotic phenotype acquisition in hTM cells.
  
  
  CONCLUSIONS
  MSCs injection in the ocular anterior chamber in a rat model of OHT provides neuroprotective effect in the glaucoma pathophysiology via TM protection. These results demonstrate that MSCs constitute promising tool for treating ocular hypertension and retinal cell degeneration.
authors.affiliation: INSERM, U968, Paris, F-75012, France. chris.roubeix@gmail.com.|UPMC Université Paris 06, UMR_S 968, Institut de la Vision, Paris, F-75012, France. chris.roubeix@gmail.com.|CNRS, UMR_7210, Paris, F-75012, France. chris.roubeix@gmail.com.
authors.forename: Christophe
authors.initials: C
authors.lastname: Roubeix
authors.affiliation: INSERM, U968, Paris, F-75012, France. david.godefroy@inserm.fr.|UPMC Université Paris 06, UMR_S 968, Institut de la Vision, Paris, F-75012, France. david.godefroy@inserm.fr.|CNRS, UMR_7210, Paris, F-75012, France. david.godefroy@inserm.fr.
authors.forename: David
authors.initials: D
authors.lastname: Godefroy
authors.affiliation: Institut des Maladies Métaboliques et Cardiovasculaires, INSERM UMR 1048, 31432 Toulouse cedex 4, France, Toulouse, France. celine.mias@inserm.fr.
authors.forename: Céline
authors.initials: C
authors.lastname: Mias
authors.affiliation: INSERM, U968, Paris, F-75012, France. anais.sapienza@inserm.fr.|UPMC Université Paris 06, UMR_S 968, Institut de la Vision, Paris, F-75012, France. anais.sapienza@inserm.fr.|CNRS, UMR_7210, Paris, F-75012, France. anais.sapienza@inserm.fr.
authors.forename: Anaïs
authors.initials: A
authors.lastname: Sapienza
authors.affiliation: INSERM, U968, Paris, F-75012, France. luisa.riancho@inserm.fr.|UPMC Université Paris 06, UMR_S 968, Institut de la Vision, Paris, F-75012, France. luisa.riancho@inserm.fr.|CNRS, UMR_7210, Paris, F-75012, France. luisa.riancho@inserm.fr.
authors.forename: Luisa
authors.initials: L
authors.lastname: Riancho
authors.affiliation: INSERM, U968, Paris, F-75012, France. julie.degardin@inserm.fr.|UPMC Université Paris 06, UMR_S 968, Institut de la Vision, Paris, F-75012, France. julie.degardin@inserm.fr.|CNRS, UMR_7210, Paris, F-75012, France. julie.degardin@inserm.fr.
authors.forename: Julie
authors.initials: J
authors.lastname: Degardin
authors.affiliation: INSERM, U968, Paris, F-75012, France. valerie.fradot@inserm.fr.|UPMC Université Paris 06, UMR_S 968, Institut de la Vision, Paris, F-75012, France. valerie.fradot@inserm.fr.|CNRS, UMR_7210, Paris, F-75012, France. valerie.fradot@inserm.fr.
authors.forename: Valérie
authors.initials: V
authors.lastname: Fradot
authors.affiliation: INSERM, U968, Paris, F-75012, France. ivana.ivkovic@inserm.fr.|UPMC Université Paris 06, UMR_S 968, Institut de la Vision, Paris, F-75012, France. ivana.ivkovic@inserm.fr.|CNRS, UMR_7210, Paris, F-75012, France. ivana.ivkovic@inserm.fr.
authors.forename: Ivana
authors.initials: I
authors.lastname: Ivkovic
authors.affiliation: INSERM, U968, Paris, F-75012, France. serge.picaud@inserm.fr.|UPMC Université Paris 06, UMR_S 968, Institut de la Vision, Paris, F-75012, France. serge.picaud@inserm.fr.|CNRS, UMR_7210, Paris, F-75012, France. serge.picaud@inserm.fr.
authors.forename: Serge
authors.initials: S
authors.lastname: Picaud
authors.affiliation: INSERM, U968, Paris, F-75012, France. florian.sennlaub@inserm.fr.|UPMC Université Paris 06, UMR_S 968, Institut de la Vision, Paris, F-75012, France. florian.sennlaub@inserm.fr.|CNRS, UMR_7210, Paris, F-75012, France. florian.sennlaub@inserm.fr.
authors.forename: Florian
authors.initials: F
authors.lastname: Sennlaub
authors.affiliation: INSERM, U968, Paris, F-75012, France. alexandre.denoyer@gmail.com.|UPMC Université Paris 06, UMR_S 968, Institut de la Vision, Paris, F-75012, France. alexandre.denoyer@gmail.com.|CNRS, UMR_7210, Paris, F-75012, France. alexandre.denoyer@gmail.com.|Centre Hospitalier National d'Ophtalmologie des Quinze-Vingts, INSERM-DHOS CIC 503, Paris, F-75012, France. alexandre.denoyer@gmail.com.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Denoyer
authors.affiliation: INSERM, U968, Paris, F-75012, France. william.rostene@inserm.fr.|UPMC Université Paris 06, UMR_S 968, Institut de la Vision, Paris, F-75012, France. william.rostene@inserm.fr.|CNRS, UMR_7210, Paris, F-75012, France. william.rostene@inserm.fr.
authors.forename: William
authors.initials: W
authors.lastname: Rostene
authors.affiliation: INSERM, U968, Paris, F-75012, France. jose.sahel@institut-vision.org.|UPMC Université Paris 06, UMR_S 968, Institut de la Vision, Paris, F-75012, France. jose.sahel@institut-vision.org.|CNRS, UMR_7210, Paris, F-75012, France. jose.sahel@institut-vision.org.|Centre Hospitalier National d'Ophtalmologie des Quinze-Vingts, INSERM-DHOS CIC 503, Paris, F-75012, France. jose.sahel@institut-vision.org.
authors.forename: José Alain
authors.initials: JA
authors.lastname: Sahel
authors.affiliation: INSERM, U968, Paris, F-75012, France. stephane.melik-parsadaniantz@inserm.fr.|UPMC Université Paris 06, UMR_S 968, Institut de la Vision, Paris, F-75012, France. stephane.melik-parsadaniantz@inserm.fr.|CNRS, UMR_7210, Paris, F-75012, France. stephane.melik-parsadaniantz@inserm.fr.
authors.forename: Stéphane Melik
authors.initials: SM
authors.lastname: Parsadaniantz
authors.affiliation: INSERM, U968, Paris, F-75012, France. frbaudouin@aol.com.|UPMC Université Paris 06, UMR_S 968, Institut de la Vision, Paris, F-75012, France. frbaudouin@aol.com.|CNRS, UMR_7210, Paris, F-75012, France. frbaudouin@aol.com.|Centre Hospitalier National d'Ophtalmologie des Quinze-Vingts, INSERM-DHOS CIC 503, Paris, F-75012, France. frbaudouin@aol.com.|University Paris Descartes, Sorbonne Paris Cité, Paris, F-75006, France. frbaudouin@aol.com.|Faculté de Pharmacie de Paris, University Paris Descartes, Sorbonne Paris Cité, Paris, F-75006, France. frbaudouin@aol.com.
authors.forename: Françoise
authors.initials: F
authors.lastname: Brignole-Baudouin
authors.affiliation: INSERM, U968, Paris, F-75012, France. cbaudouin@quinze-vingts.fr.|UPMC Université Paris 06, UMR_S 968, Institut de la Vision, Paris, F-75012, France. cbaudouin@quinze-vingts.fr.|CNRS, UMR_7210, Paris, F-75012, France. cbaudouin@quinze-vingts.fr.|Centre Hospitalier National d'Ophtalmologie des Quinze-Vingts, INSERM-DHOS CIC 503, Paris, F-75012, France. cbaudouin@quinze-vingts.fr.|Department of Ophthalmology, Hôpital Ambroise Pare, AP HP, Boulogne, F-92100, France. cbaudouin@quinze-vingts.fr.|University Versailles St Quentin en Yvelines, Montigny-Le-Bretonneux, F-78180, France. cbaudouin@quinze-vingts.fr.
authors.forename: Christophe
authors.initials: C
authors.lastname: Baudouin
mesh_terms: D000818:Animals; D017209:Apoptosis; D002478:Cells, Cultured; D005901:Glaucoma / Q000628:therapy*; D007429:Intraocular Pressure; D008297:Male; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology; D051381:Rats; D020318:Rats, Long-Evans