The role of IP3R1 in neuroinflammation triggered by retinitis pigmentosa

• Grant number: 21/11969-9
• Support Opportunities: Scholarships in Brazil - Master
• Start date: July 01, 2022
• End date: June 30, 2024
• Field of knowledge: Biological Sciences - Genetics - Animal Genetics
• Principal Investigator: Alexandre Hiroaki Kihara
• Grantee: Théo Henrique de Lima Vasconcellos
• Host Institution: Centro de Matemática, Computação e Cognição (CMCC). Universidade Federal do ABC (UFABC). Ministério da Educação (Brasil). Santo André , SP, Brazil
• Associated scholarship(s): 23/11449-0 - Cloning of cone-specific inducible expression using CRISPR Cre-Lox system, BE.EP.MS

Abstract

The retina is a photosensitive neuroepithelium found in the inner region of the eye considered an integral component of the central nervous system (CNS). An imbalance in the physiological conditions or changes in the number of functional photoreceptors can lead to important visual deficits. One such disease that virtually leads to vision impairment is retinitis pigmentosa (RP). Among the causes of autosomal recessive RP are mutations in the PDE6B gene, culminating in the initial cell death of rod-type photoreceptors and subsequent degradation of cones. However, the processes underlying the primary death of rods are not fully known, although studies point to mechanisms involved in the inflammatory response, oxidative and metabolic stress, epigenetic factors, and calcium dysregulation. Among the latter, the type 1 inositol-1,4,5-triphosphate receptor (IP3R1), responsible for intracellular calcium homeostasis, could participate in the progression of this pathology. IP3R1 is associated with several important cellular events, such as cell death and proliferation. In animal models for autosomal recessive RP, several alterations related to calcium dysregulation have been described, like mitochondrial and endoplasmic reticulum overload, affecting the photoreceptors' metabolism. In this scenario, this project aims to analyze the effects of the pharmacological blocking of IP3R1 in an animal model of autosomal recessive RP and in vitro cell cultures, focusing on analyses of morphometric and inflammatory aspects. The pharmacological blockade will be performed using the drug 2-APB by subretinal injection. To analyze the effects caused by the intervention, immunofluorescence techniques will be used for protein localization analysis, accompanied by the analysis of protein levels by western blot. Also in vitro experiments will be used to evaluate neuroinflammation and the role of IP3R1 glial response. The multiplex bead assay will be used to determine the inflammatory response combined with specific cellular markers. Finally, we will analyze cell death by apoptosis, using the TUNEL assay. With this study, we intend to evaluate the role of the IP3R1 protein in the progression of RP and its role in neuroinflammatory processes, which may open perspectives for therapies focused on calcium dysregulation.

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

MOVIO, MARILIA INES; DE LIMA-VASCONCELLOS, THEO HENRIQUE; DOS SANTOS, GABRIELI BOVI; ECHEVERRY, MARCELA BERMUDEZ; COLOMBO, ELISABETTA; MATTOS, LEONARDO S.; RESENDE, RODRIGO RIBEIRO; KIHARA, ALEXANDRE HIROAKI. Retinal organoids from human-induced pluripotent stem cells: From studying retinal dystrophies to early diagnosis of Alzheimer?s and Parkinson?s disease. SEMINARS IN CELL & DEVELOPMENTAL BIOLOGY, v. 144, p. 10-pg., 2023-03-17. (21/14227-3, 21/03711-1, 19/17892-8, 20/02035-0, 20/11667-0, 21/11969-9, 17/26388-6)

DA SILVA, VICTOR A.; BOBOTIS, BIANCA C.; CORREIA, FELIPE F.; LIMA-VASCONCELLOS, THEO H.; CHIARANTIN, GABRIELLY M. D.; DE LA VEGA, LAURA; LOMBELLO, CHRISTIANE B.; WILLERTH, STEPHANIE M.; MALMONGE, SONIA M.; PASCHON, VERA; et al. The Impact of Biomaterial Surface Properties on Engineering Neural Tissue for Spinal Cord Regeneration. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, v. 24, n. 17, p. 25-pg., 2023-09-01. (17/12112-9, 18/06316-3, 18/19300-8, 19/17892-8, 19/18921-1, 20/02035-0, 20/11667-0, 21/11969-9, 19/24406-2)

MARINHO, LUCIANA SIMOES RAFAGNIN; CHIARANTIN, GABRIELLY MARIA DENADAI; IKEBARA, JULIANE MIDORI; CARDOSO, DEBORA STERZECK; DE LIMA-VASCONCELLOS, THEO HENRIQUE; HIGA, GUILHERME SHIGUETO VILAR; FERRAZ, MARIANA SACRINI AYRES; DE PASQUALE, ROBERTO; TAKADA, SILVIA HONDA; PAPES, FABIO; et al. The impact of antidepressants on human neurodevelopment: Brain organoids as experimental tools. SEMINARS IN CELL & DEVELOPMENTAL BIOLOGY, v. 144, p. 10-pg., 2023-03-17. (19/17892-8, 20/16268-6, 20/11451-7, 20/02035-0, 21/11969-9, 20/11667-0, 19/15024-9, 16/17329-3)