Rescue of Defective Electroretinographic Responses in Dp71-Null Mice With AAV-Mediated Reexpression of Dp71

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Salgueiro Barboni, Mirella Telles ^{[1, 2]} ; Vaillend, Cyrille ^[3] ; Joachimsthaler, Anneka ^{[4, 5]} ; Passos Liber, Andre Mauricio ^[1] ; Khabou, Hanen ^[6] ; Roux, Michel J. ^[7] ; Vacca, Ophelie ^{[6, 3]} ; Vignaud, Lucile ^[6] ; Dalkara, Deniz ^[6] ; Guillonneau, Xavier ^[6] ; Ventura, Dora Fix ^[1] ; Rendon, Alvaro ^[6] ; Kremers, Jan ^{[4, 5]}

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Dept Expt Psychol, Sao Paulo - Brazil

^[2] Semmelweis Univ, Dept Ophthalmol, Budapest - Hungary

^[3] Univ Paris Saclay, Univ Paris Sud, Neurosci Paris Saclay Inst Neuro PSI, CNRS, UMR 9197, Orsay - France

^[4] Univ Hosp Erlangen, Dept Ophthalmol, Erlangen - Germany

^[5] FAU Erlangen Nurnberg, Dept Biol, Anim Physiol, Erlangen - Germany

^[6] Sorbonne Univ, Inst Vis, Dept Therapeut, Paris - France

^[7] Univ Strasbourg, Dept Translat Med & Neurogenet, IGBMC ICS Phenomin, Illkirch Graffenstaden - France

• Tipo de documento: Artigo Científico
• Fonte: INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE; v. 61, n. 2 FEB 2020.
• Citações Web of Science: 0

Resumo

PURPOSE. To study the potential effect of a gene therapy, designed to rescue the expression of dystrophin Dp71 in the retinas of Dp71-null mice, on retinal physiology. METHODS. We recorded electroretinograms (ERGs) in Dp71-null and wild-type littermate mice. In dark-adapted eyes, responses to flashes of several strengths were measured. In addition, flash responses on a 25-candela/square meters background were measured. On- and Off-mediated responses to sawtooth stimuli and responses to photopic sine-wave modulation (3-30 Hz) were also recorded. After establishing the ERG phenotype, the ShH10-GFP adeno-associated virus (AAV), which has been previously shown to target specifically Muller glial cells (MGCs), was delivered intravitreously with or without (sham therapy) the Dp71 coding sequence under control of a CBA promoter. ERG recordings were repeated three months after treatment. Real-time quantitative PCR and Western blotting analyses were performed in order to quantify Dp71 expression in the retinas. RESULTS. Dp71-null mice displayed reduced b-waves in dark- and light-adapted flash ERGs and smaller response amplitudes to photopic rapid-on sawtooth modulation and to sine-wave stimuli. Three months after intravitreal injections of the ShH10-GFP-2A-Dp71 AAV vector, ERG responses were completely recovered in treated eyes of Dp71-null mice. The functional rescue was associated with an overexpression of Dp71 in treated retinas. CONCLUSIONS. The present results show successful functional recovery accompanying the reexpression of Dp71. In addition, this experimental model sheds light on MGCs influencing ERG components, since previous reports showed that aquaporin 4 and Kir4.1 channels were mislocated in MGCs of Dp71-null mice, while their distribution could be normalized following intravitreal delivery of the same ShH10-GFP-2A-Dp71 vector. (AU)

• Processo FAPESP: 14/26818-2 - Desenvolvimento e implantação de métodos de avaliação visual: aplicações clínicas e em modelos animais
• Beneficiário: Dora Selma Fix Ventura
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 16/22007-5 - Optimização da capacidade funcional visual de pacientes com maculopatias através de protocolos de treinamento do sistema visual
• Beneficiário: Mirella Telles Salgueiro Barboni
• Modalidade de apoio: Bolsas no Exterior - Pesquisa
• Processo FAPESP: 19/00777-1 - Relação entre hábitos circadianos e funções visuais em serpentes diurnas e noturnas (serpentes, Colubridae)
• Beneficiário: André Maurício Passos Liber
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 16/04538-3 - Distrofinas e o sistema visual humano: estudo dos diferentes fenótipos visuais de pacientes com distrofia muscular de Duchenne
• Beneficiário: Dora Selma Fix Ventura
• Modalidade de apoio: Auxílio à Pesquisa - Regular