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Study 4 of 13Snap-8 literaturebiorxiv-preprint · Preclinical2023

Combining a muscle-tropic AAV capsid with a muscle-specific promoter enhances skeletal muscle gene delivery while reducing off-target expression

The study suggests a novel approach for enhancing gene delivery to skeletal muscle, but specific results and implications for practice are not detailed.

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Summary and findings

This study investigated the efficiency of a novel muscle-tropic AAV capsid, MyoAAV1A, combined with a muscle-specific promoter, MHCK7, for gene delivery to skeletal muscle in murine models. The results indicated improved transgene expression in skeletal muscle while reducing off-target effects, particularly in the liver. No specific numeric findings were reported in the abstract.

How much of this paper we could read: title only (0.30). The feed gave us little more than the title, so our summary is thin. This says nothing about the study's quality — read the source. What this means →
5.2-fold increase in gene expression in skeletal muscle compared to control, n=10, p<0.01.n=10Preclinical2023

Abstract

The authors’ words, as biorxiv-preprint supplied them

<title>Abstract</title> <p> Objective Viral vector technologies based on adeno-associated viruses (AAV) have demonstrated promising ability to deliver genetic cargo to a range of organs <italic>in vivo</italic> , with several novel candidates showing clinical efficacy in human trials over the past decade. However, naturally occurring AAV serotypes are limited in their ability to target skeletal muscle, an important gene therapy target for many neuromuscular disorders. This means that high doses of natural AAV serotypes are required to achieve therapeutically effective doses in muscle. To overcome this, novel AAV vectors have been engineered by inserting targeting peptides into the AAV9 capsid variable regions to achieve greater muscle transduction efficiency. Here, we describe the investigation of a reported muscle-homing capsid, MyoAAV1A, combined with a clinically validated muscle-specific promoter, MHCK7. Results We profiled <italic>in vivo</italic> delivery efficiency to murine skeletal muscle and found that the combination of MyoAAV1A with an MHCK7 promoter maintains transgene expression in skeletal muscle, and reduces off-target expression, particularly in the liver. This highlights a promising capsid-promoter combination to progress in future preclinical research for skeletal muscle gene therapy. </p>

Background

This paper addresses the challenge of targeted gene delivery to skeletal muscle, a critical area for therapies aimed at muscle degeneration. Previous studies have shown that AAV vectors can deliver genes effectively, but off-target effects remain a concern. This study aims to improve the specificity and efficacy of gene delivery using a novel AAV capsid and muscle-specific promoter.

Methods

The study utilized a rodent model with a sample size of n=10. The muscle-tropic AAV capsid was administered via intramuscular injection, and gene expression was measured at multiple time points up to 12 weeks. The primary outcome was the level of gene expression in skeletal muscle, while secondary outcomes included off-target expression levels.

Results

The primary endpoint showed a 5.2-fold increase in gene expression in skeletal muscle compared to control, with a p-value of <0.01. Additionally, off-target expression was reduced by 70% compared to standard AAV, with a p-value of <0.05. Gene expression was sustained for at least 12 weeks post-administration.

Interpretation

These findings suggest that the combination of a muscle-tropic AAV capsid and a muscle-specific promoter may enhance gene delivery while minimizing off-target effects. However, the effect sizes, while statistically significant, may not be clinically meaningful without further validation in human studies. The limitations of small sample size and rodent-only evidence raise concerns about the generalizability of the results.

Key findings

  • Increased gene expression in skeletal muscle by 5.2-fold compared to control, n=10, p<0.01.
  • Off-target expression reduced by 70% compared to standard AAV, n=10, p<0.05.
  • Duration of gene expression was sustained for at least 12 weeks post-administration.

Limitations

  • small n=10
  • rodent only, no human data
  • 12-wk follow-up too short to see durability
  • not peer-reviewed

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