Dual muscle-liver transduction imposes immune tolerance for muscle transgene engraftment despite preexisting immunity

JCI Insight. 2019 Jun 6;4(11):e127008. doi: 10.1172/jci.insight.127008.

Abstract

Immune responses to therapeutic transgenes are a potential hurdle to treat monogenic muscle disorders. These responses result from the neutralizing activity of transgene-specific B cells and cytotoxic T cells recruited upon gene transfer. We explored here how dual muscle-liver expression of a foreign transgene allows muscle transgene engraftment after adenoassociated viral vector delivery. We found in particular that induction of transgene-specific tolerance is imposed by concurrent muscle and liver targeting, resulting in the absence of CD8+ T cell responses to the transgene. This tolerance can be temporally decoupled, because transgene engraftment can be achieved in muscle weeks after liver transduction. Importantly, transgene-specific CD8+ T cell tolerance can be established despite preexisting immunity to the transgene. Whenever preexisting, transgene-specific CD4+ and CD8+ memory T cell responses are present, dual muscle-liver transduction turns polyclonal, transgene-specific CD8+ T cells into typically exhausted T cells with high programmed cell death 1 (PD-1) expression and lack of IFN-γ production. Our results demonstrate that successful transduction of muscle tissue can be achieved through liver-mediated control of humoral and cytotoxic T cell responses, even in the presence of preexisting immunity to the muscle-associated transgene.

Keywords: Adaptive immunity; Gene therapy; Immunology; T cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • B-Lymphocytes / immunology
  • CD8-Positive T-Lymphocytes / immunology
  • Dependovirus / genetics
  • Dependovirus / immunology
  • Gene Transfer Techniques*
  • Genetic Therapy* / adverse effects
  • Genetic Therapy* / methods
  • Genetic Vectors / genetics
  • Genetic Vectors / immunology
  • Immune Tolerance / immunology*
  • Interferon-gamma / metabolism
  • Liver / metabolism*
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Muscles / metabolism*
  • T-Lymphocytes, Cytotoxic / immunology
  • Transgenes / genetics

Substances

  • Interferon-gamma

Grants and funding

This work was supported by the Centre National de la Recherche Scientifique (CNRS), the Institut de la Santé et de la Recherche Médicale (INSERM), the Université Paris-Descartes, the Association Française contre les Myopathies (AFM) grant 19617, and by the European Union, ERC-2013-CoG Consolidator Grant, grant agreement number 617432 (MoMAAV) and research and innovation program under Grant Agreement No. 667751 (Myocure). LB was supported by the French Ministry of Research. Data relative to this work were presented by LB for his Ph.D defense (Paris, France; November 2018).