Evaluation of the neuroprotective efficacy of the gramine derivative ITH12657 against NMDA-induced excitotoxicity in the rat retina

Johnny Di Pierdomenico; Alejandro Gallego-Ortega; María Norte-Muñoz; Beatriz Vidal-Villegas; Isaac Bravo; María Boluda-Ruiz; Jose Manuel Bernal-Garro; Iván Fernandez-Bueno; Jose Carlos Pastor-Jimeno; María Paz Villegas-Pérez; Marcelino Avilés-Trigueros; Cristobal de Los Ríos; Manuel Vidal-Sanz

doi:10.3389/fnana.2024.1335176

Evaluation of the neuroprotective efficacy of the gramine derivative ITH12657 against NMDA-induced excitotoxicity in the rat retina

Front Neuroanat. 2024 Feb 13:18:1335176. doi: 10.3389/fnana.2024.1335176. eCollection 2024.

Authors

Johnny Di Pierdomenico^#¹, Alejandro Gallego-Ortega^#¹, María Norte-Muñoz¹, Beatriz Vidal-Villegas¹, Isaac Bravo^{2

3}, María Boluda-Ruiz¹, Jose Manuel Bernal-Garro¹, Iván Fernandez-Bueno⁴, Jose Carlos Pastor-Jimeno⁴, María Paz Villegas-Pérez¹, Marcelino Avilés-Trigueros¹, Cristobal de Los Ríos^{2

3}, Manuel Vidal-Sanz¹

Affiliations

¹ Departamento de Oftalmología, Universidad de Murcia e IMIB-Arrixaca, Murcia, Spain.
² Instituto de Investigación Sanitaria, Hospital Universitario de la Princesa, Madrid, Spain.
³ Departamento de Ciencias Básicas de la Salud, Universidad Rey Juan Carlos, Alcorcón, Spain.
⁴ Instituto Universitario de Oftalmobiología Aplicada (IOBA), Retina Group, Universidad de Valladolid, Valladolid, Spain.

^# Contributed equally.

Abstract

Purpose: The aim of this study was to investigate, the neuroprotective effects of a new Gramine derivative named: ITH12657, in a model of retinal excitotoxicity induced by intravitreal injection of NMDA.

Methods: Adult Sprague Dawley rats received an intravitreal injection of 100 mM NMDA in their left eye and were treated daily with subcutaneous injections of ITH12657 or vehicle. The best dose-response, therapeutic window study, and optimal treatment duration of ITH12657 were studied. Based on the best survival of Brn3a + RGCs obtained from the above-mentioned studies, the protective effects of ITH12657 were studied in vivo (retinal thickness and full-field Electroretinography), and ex vivo by quantifying the surviving population of Brn3a + RGCs, αRGCs and their subtypes α-ONsRGCs, α-ONtRGCs, and α-OFFRGCs.

Results: Administration of 10 mg/kg ITH12657, starting 12 h before NMDA injection and dispensed for 3 days, resulted in the best significant protection of Brn3a + RGCs against NMDA-induced excitotoxicity. In vivo, ITH12657-treated rats showed significant preservation of retinal thickness and functional protection against NMDA-induced retinal excitotoxicity. Ex vivo results showed that ITH12657 afforded a significant protection against NMDA-induced excitotoxicity for the populations of Brn3a + RGC, αRGC, and αONs-RGC, but not for the population of αOFF-RGC, while the population of α-ONtRGC was fully resistant to NMDA-induced excitotoxicity.

Conclusion: Subcutaneous administration of ITH12657 at 10 mg/kg, initiated 12 h before NMDA-induced retinal injury and continued for 3 days, resulted in the best protection of Brn3a + RGCs, αRGC, and αONs-RGC against excitotoxicity-induced RGC death. The population of αOFF-RGCs was extremely sensitive while α-ONtRGCs were fully resistant to NMDA-induced excitotoxicity.

Keywords: Brn3a + RGCs; NMDA excitotoxicity; SD-OCT; glaucoma; neuroprotection; retina calcium blocker; retina ganglion cell; αRGCs.

Copyright © 2024 Di Pierdomenico, Gallego-Ortega, Norte-Muñoz, Vidal-Villegas, Bravo, Boluda-Ruiz, Bernal-Garro, Fernandez-Bueno, Pastor-Jimeno, Villegas-Pérez, Avilés-Trigueros, de los Ríos and Vidal-Sanz.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This research was funded by PID2019-106498GB-I00 funded by Ministerio de Ciencia, Innovación y Universidades (MCIN)/AEI/ 10.13039/501100011033 to MV-S and MA-T; (RetiBrain) RED2018-102499-T to MV-S; and by the Instituto de Salud Carlos III (PI19/01724 to CdlR) and co-funded with the European Regional Development Fund (ERDF) within the “Plan Estatal de Investigación Científica y Técnica y de Innovación 2017–2020” (FIS/PI 18–00754) to MV-P.