Factors Impacting Efficacy of AAV-Mediated CRISPR-Based Genome Editing for Treatment of Choroidal Neovascularization
| Autor: | Eric Tieu, Sook Hyun Chung, Jared H. Fong, Uyen Thao Nguyen, Glenn Yiu, Emerald Lovece Nguyen, Amy Nguyen, Ratheesh Kumar Meleppat, Natalie Stucka, Robert J. Zawadzki, Pengfei Zhang, Suman K. Manna, Iris Natalie Mollhoff |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Aging
macular degeneration lcsh:QH426-470 genetic structures Angiogenesis CNV AMD Neurodegenerative Biology Article angiogenesis 03 medical and health sciences Transduction (genetics) 0302 clinical medicine Genome editing Genetics medicine genome editing CRISPR Guide RNA lcsh:QH573-671 Cas9 age-related macular degeneration Eye Disease and Disorders of Vision Molecular Biology 030304 developmental biology VEGFa 0303 health sciences Gene knockdown 5.2 Cellular and gene therapies lcsh:Cytology Gene Therapy VEGF eye diseases 3. Good health lcsh:Genetics Choroidal neovascularization 030221 ophthalmology & optometry Cancer research Molecular Medicine sense organs Development of treatments and therapeutic interventions medicine.symptom Biotechnology |
| Zdroj: | Molecular Therapy: Methods & Clinical Development, Vol 17, Iss, Pp 409-417 (2020) Molecular Therapy. Methods & Clinical Development |
| ISSN: | 2329-0501 |
| DOI: | 10.1016/j.omtm.2020.01.006 |
| Popis: | Frequent injections of anti-vascular endothelial growth factor (anti-VEGF) agents are a clinical burden for patients with neovascular age-related macular degeneration (AMD). Genomic disruption of VEGF-A using adeno-associated viral (AAV) delivery of clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 has the potential to permanently suppress aberrant angiogenesis, but the factors that determine the optimal efficacy are unknown. Here, we investigate two widely used Cas9 endonucleases, SpCas9 and SaCas9, and evaluate the relative contribution of AAV-delivery efficiency and genome-editing rates invivo to determine the mechanisms that drive successful CRISPR-based suppression of VEGF-A, using a mouse model of laser-induced choroidal neovascularization (CNV). We found that SpCas9 demonstrated higher genome-editing rates, greater VEGF reduction, and more effective CNV suppression than SaCas9, despite similar AAV transduction efficiency between a dual-vector approach for SpCas9 and single-vector system for SaCas9 to deliver the Cas9 orthologs and single guide RNAs (gRNAs). Our results suggest that successful VEGF knockdown using AAV-mediated CRISPR systems may be determined more by the efficiency of genome editing rather than viral transduction and that SpCas9 may be more effective than SaCas9 as a potential therapeutic strategy for CRISPR-based treatment of CNV in neovascular AMD. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|