Efficacy and safety of a SOD1-targeting artificial miRNA delivered by AAV9 in mice are impacted by miRNA scaffold selection
Brief intro:
- Author: Shukkwan K. Lau, Zachary C.E. Hawley, Maria I. Zavodszky, Sam Hana, Daniel Ferretti, Branka Grubor, Michael Hawes, Shanqin Xu, Stefan Hamann, Galina Marsh, Patrick Cullen, Ravi Challa, Thomas M. Carlile, Hang Zhang, Wan-Hung Lee, Andrea Peralta, Pete Clarner, Cong Wei, Kathryn Koszka, Feng Gao, Shih-Ching Lo
- Journal: Mol Ther Nucleic Acids
- Doi: https://www.doi.org/10.1016/j.omtn.2023.102057
- Publication Date: 2023 Oct 17
Products/Services used in the paper
Quotation shows PackGene:PackGene Biotech (Worcester, MA) produced AAV9-amiR vectors using triple plasmid transfection protocol (Worchester, MA) as described previously.Transmission electron microscopy and image analysis were conducted by Packgene to confirm minimal number of empty AAV capsids for each vector produced (Fig. S10). AAV titers were quantified by ddPCR using a custom mCherry primer/probe
Research Field:CNS
AAV Serotype:AAV9
Targeted organ:brain
Animal or cell line strain:Mouse
Abstract
Toxic gain-of-function mutations in superoxide dismutase 1 (SOD1) contribute to ∼2-3% of all amyotrophic lateral sclerosis (ALS) cases. Artificial microRNAs (amiRs) delivered by adeno-associated virus (AAV) have been proposed as a potential treatment option to silence SOD1 expression and mitigate disease progression. Primary microRNA (pri-miRNA) scaffolds are used in amiRs to shuttle a hairpin RNA into the endogenous miRNA pathway, but it is unclear whether different primary miRNA scaffolds impact the potency and safety profile of the expressed amiR in vivo. In our process to develop an AAV amiR targeting SOD1, we performed a preclinical characterization of two primary miRNA scaffolds, miR155 and miR30a, sharing the same guide strand sequence. We report that while the miR155-based vector, compared to the miR30a-based vector, leads to a higher level of the amiR and more robust suppression of SOD1 in vitro and in vivo, it also presents significantly greater risks for CNS-related toxicities in vivo. Despite miR30a-based vector showing relatively lower potency, it can significantly delay the development of ALS-like phenotypes in SOD1-G93A mice and increase survival in a dose-dependent manner. This data highlights the importance of scaffold selection in the pursuit of highly efficacious and safe amiRs for RNAi gene therapy.
About PackGene
PackGene Biotech is a world-leading CRO and CDMO, excelling in AAV vectors, mRNA, plasmid DNA, and lentiviral vector solutions. Our comprehensive offerings span from vector design and construction to AAV, lentivirus, and mRNA services. With a sharp focus on early-stage drug discovery, preclinical development, and cell and gene therapy trials, we deliver cost-effective, dependable, and scalable production solutions. Leveraging our groundbreaking π-alpha 293 AAV high-yield platform, we amplify AAV production by up to 10-fold, yielding up to 1e+17vg per batch to meet diverse commercial and clinical project needs. Moreover, our tailored mRNA and LNP products and services cater to every stage of drug and vaccine development, from research to GMP production, providing a seamless, end-to-end solution.
