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Theranostics 2021; 11(13):6154-6172. doi:10.7150/thno.55472 This issue Cite

Research Paper

Inner retinal injury in experimental glaucoma is prevented upon AAV mediated Shp2 silencing in a caveolin dependent manner

Mojdeh Abbasi^1✉*, Vivek K. Gupta^1✉*, Nitin Chitranshi^1✉*, Veer Gupta², Reza Ranjbaran³, Rashi Rajput¹, Kanishka Pushpitha¹, Devaraj KB¹, Yuyi You^1,4, Ghasem Hosseini Salekdeh^5,6, Robert G. Parton^7,8, Mehdi Mirzaei¹, Stuart L. Graham^1,4

1. Department of Clinical Medicine, Faculty of Medicine and Health Sciences, Macquarie University, North Ryde, Sydney, NSW 2109, Australia.
2. School of Medicine, Deakin University, Melbourne, VIC, Australia.
3. Diagnostic Laboratory Sciences and Technology Research Center, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran.
4. Save Sight Institute, Sydney University, Sydney NSW 2000, Australia.
5. Department of Molecular Systems Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, Tehran, Iran.
6. Department of Molecular Sciences, Macquarie University, North Ryde, Sydney, NSW 2109, Australia.
7. Institute for Molecular Bioscience and Centre for Microscopy and Microanalysis, The University of Queensland, Queensland 4072, Australia.
8. Institute for Molecular Bioscience, The University of Queensland, Brisbane, QLD, Australia.
* Equivalent first author.

✉ Corresponding authors: Vivek Gupta, Faculty of Medicine and Health Sciences, Macquarie University, Sydney, 2109, Australia, vivek.guptaedu.au; Mojdeh Abbasi, Faculty of Medicine and Health Sciences, Macquarie University, Sydney, 2109, Australia, mojdeh.abbasimq.edu.au; Nitin Chitranshi, Faculty of Medicine and Health Sciences, Macquarie University, Sydney, 2109, Australia, nitin.chitranshiedu.au. More

Abstract

SH2 domain containing tyrosine phosphatase 2 (Shp2; PTPN11) regulates several intracellular pathways downstream of multiple growth factor receptors. Our studies implicate that Shp2 interacts with Caveolin-1 (Cav-1) protein in retinal ganglion cells (RGCs) and negatively regulates BDNF/TrkB signaling. This study aimed to investigate the mechanisms underlying the protective effects of shp2 silencing in the RGCs in glaucomatous conditions.

Methods: Shp2 was silenced in the Cav-1 deficient mice and the age matched wildtype littermates using adeno-associated viral (AAV) constructs. Shp2 expression modulation was performed in an acute and a chronic mouse model of experimental glaucoma. AAV2 expressing Shp2 eGFP-shRNA under a strong synthetic CAG promoter was administered intravitreally in the animals' eyes. The contralateral eye received AAV-eGFP-scramble-shRNA as control. Animals with Shp2 downregulation were subjected to either microbead injections or acute ocular hypertension experimental paradigm. Changes in inner retinal function were evaluated by measuring positive scotopic threshold response (pSTR) while structural and biochemical alterations were evaluated through H&E staining, western blotting and immunohistochemical analysis of the retinal tissues.

Results: A greater loss of pSTR amplitudes was observed in the WT mice compared to Cav-1^-/- retinas in both the models. Silencing of Shp2 phosphatase imparted protection against inner retinal function loss in chronic glaucoma model in WT mice. The functional rescue also translated to structural preservation of ganglion cell layer in the chronic glaucoma condition in WT mice which was not evident in Cav-1^-/- mice retinas.

Conclusions: This study indicates that protective effects of Shp2 ablation under chronic experimental glaucoma conditions are dependent on Cav-1 in the retina, suggesting in vivo interactions between the two proteins.

Keywords: Glaucoma, Retinal Ganglion cells, Shp2 phosphatase, Caveolin, TrkB.

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