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

Theranostics. 2021 Apr 15;11(13):6154-6172. doi: 10.7150/thno.55472. eCollection 2021.

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

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alpha-Globulins / genetics
  • Animals
  • Apoptosis
  • Brain-Derived Neurotrophic Factor / physiology
  • Caveolin 1 / deficiency
  • Caveolin 1 / genetics
  • Caveolin 1 / physiology*
  • DNA, Complementary / genetics
  • Dependovirus / genetics
  • Focal Adhesion Kinase 1 / physiology
  • Gene Knockdown Techniques
  • Genes, Reporter
  • Genes, Synthetic
  • Genetic Therapy*
  • Genetic Vectors / therapeutic use*
  • Glaucoma / metabolism
  • Glaucoma / pathology
  • Glaucoma / therapy*
  • Integrin beta1 / physiology
  • Intraocular Pressure
  • Intravitreal Injections
  • Membrane Glycoproteins / physiology
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Promoter Regions, Genetic
  • Protein Tyrosine Phosphatase, Non-Receptor Type 11 / antagonists & inhibitors*
  • Protein Tyrosine Phosphatase, Non-Receptor Type 11 / biosynthesis
  • Protein Tyrosine Phosphatase, Non-Receptor Type 11 / genetics
  • Protein-Tyrosine Kinases / physiology
  • Retina / pathology*
  • Up-Regulation

Substances

  • Alpha-Globulins
  • Bdnf protein, mouse
  • Brain-Derived Neurotrophic Factor
  • Caveolin 1
  • DNA, Complementary
  • Integrin beta1
  • Membrane Glycoproteins
  • chorionic alpha(2)-microglobulin
  • Ntrk2 protein, mouse
  • Protein-Tyrosine Kinases
  • Focal Adhesion Kinase 1
  • Ptk2 protein, mouse
  • Protein Tyrosine Phosphatase, Non-Receptor Type 11
  • Ptpn11 protein, mouse