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M. L. Bajenaru, R. A. Santos, B. A. Obesso, R. G. Corredor, E. Hernandez, M. E. Fini, J. L. Goldberg; B1-Integrin Activating Antibody HUTS-21 Exhibits Retinal Ganglion Cell Neuroprotective Properties. Invest. Ophthalmol. Vis. Sci. 2010;51(13):3696.
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RGC survival and neurite outgrowth are promoted by neurotrophins, and extracellular matrix (ECM) molecules such as laminin. Degradation of laminin in the ECM of RGC in experimental animal models relevant to retinal disease contributes to RGC death. Integrins are the major ECM receptors. The purpose of this study is to evaluate HUTS-21, a β1 integrin activating antibody as neuroprotective agent to improve RGC regeneration after retinal ischemia reperfusion injury (RIRI).
Studies were performed in rat RGC cultures, and in a rat RIRI model. Purified rat RGC cultures were prepared by immunopanning. Expression of β1 integrin, FAK, and P-FAK was analyzed by immunofluorescence in RGC cultured on laminin, or poly-D-lysine (PDL). RGC in culture were treated with β1 integrin activating, and blocking antibodies, and FAK pharmacological inhibitors, genistein, and PP2. RGC survival, and neurites were examined and counted by live cell imaging. RIRI was induced in Sprague-Dawley rats by unilateral elevation of the intraocular pressure to 110 mm Hg for 60 min. HUTS-21 activating antibodies were administered intravitreally in the rat eye 30 min prior to RIRI. RGC loss was quantified in the retina after retrograde labeling with FG. Expression and activation of β1 integrin, and FAK was determined by western blotting in retinal extracts.
We have previously identified β1 integrin and FAK as key regulators of the integrin survival pathway, and neurite outgrowth in RGC. We showed this pathway is disrupted after RIRI. When cultured on laminin RGC exhibited increased survival, and neurite outgrowth. These properties correlated with an up-regulation of the integrin survival pathway, and increased focal contacts on laminin, versus PDL. Treatment with β1 activating antibodies HUTS-21 of RGC cultured on PDL mimics the neuroprotective effect of laminin, and enhances the RGC survival, and neurite outgrowth. HUTS-21 protective effect can be abolished by co-treatment with the FAK inhibitor PP2. Intravitreal admistration of HUTS-21 in the rat RIRI model partially rescues RGC death and prevents disruption of the β1 integrin survival pathway in RGC.
We have identified HUTS-21 as a potential neuroprotective agent that promotes RGC survival, and regeneration after RIRI.
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