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Expression of therapeutic proteins after delivery of chemically modified mRNA in mice

Nature Biotechnology volume 29, pages 154–157 (2011)Cite this article

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Abstract

Current viral vectors for gene therapy1,2,3 are associated with serious safety concerns, including leukemogenesis4, and nonviral vectors are limited by low gene transfer efficiency5. Here we investigate the therapeutic utility of chemically modified mRNA as an alternative to DNA-based gene therapy. A combination of nucleotide modifications abrogates mRNA interaction with Toll-like receptor (TLR)3, TLR7, TLR8 and retinoid-inducible gene I (RIG-I), resulting in low immunogenicity and higher stability in mice. A single intramuscular injection of modified murine erythropoietin mRNA raises the average hematocrit in mice from 51.5% to 64.2% after 28 days. In a mouse model of a lethal congenital lung disease caused by a lack of surfactant protein B (SP-B), twice weekly local application of an aerosol of modified SP-B mRNA to the lung restored 71% of the wild-type SP-B expression, and treated mice survived until the predetermined end of the study after 28 days.

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Figure 1: Modification of mRNA enhances transgene expression and decreases immune responses by reducing immunoreceptor binding.
Figure 2: Dual-modification of mEpo mRNA reduces binding to innate immune receptors, prevents immune responses and prolongs transgene expression in vivo.
Figure 3: Rescue of SP-B deficient mice with repeated doses of modified SP-B mRNA.

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Acknowledgements

We are grateful to J. Whitsett (Department of Pediatrics, Division of Pulmonary Biology, University of Cincinnati College of Medicine) for providing the conditional SP-B knockout mice. We thank R. Imker for technical assistance and A. Roscher for careful proofreading of the manuscript. This work was supported by the German Federal Ministry of Education and Research, grants 01GU0616, 13N9182, BioFuture (0311898) and LMUexcellent (Investitionsfonds). A T7 promoter containing pST1-2β-globin UTR-A(120) construct was kindly provided by U. Sahin (Department of Internal Medicine III, Division of Experimental and Translational Oncology, Johannes-Gutenberg University). Polyclonal rabbit antiserum directed against SP-B and anti-human/anti-mouse SP-B antibody were gifts of W. Steinhilber (Altana AG, Wesel, Germany).

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Authors and Affiliations

  1. Department of Pediatrics, Ludwig Maximilian's University Munich, Munich, Germany

    Michael S D Kormann, Günther Hasenpusch, Manish K Aneja, Gabriela Nica, Marta Illenyi, Andrea Schams, Matthias Griese, Joseph Rosenecker & Carsten Rudolph

  2. Division of Neonatology, Children's Hospital and Perinatal Center, Grosshadern, Ludwig Maximilian's University Munich, Munich, Germany

    Andreas W Flemmer, Susanne Herber-Jonat & Marceline Huppmann

  3. Children's Hospital and Interdisciplinary Center for Infectious Diseases, University of Tübingen, Tübingen, Germany

    Michael S D Kormann, Lauren E Mays, Rupert Handgretinger & Dominik Hartl

  4. Institute for Pathology, Diakonien Hospital Rotenburg (Wümme) LTD, Rotenburg (Wümme), Germany.,

    Iris Bittmann

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  1. Michael S D Kormann
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Contributions

M.S.D.K. designed research, conceived and performed experiments, and wrote the manuscript. G.H., A.W.F. and S.H.-J. conceived and performed animal experiments. M.K.A. performed animal experiments and cloning. M.H. contributed to animal experiments. G.N. performed cloning. M.I. and A.S. performed in vitro experiments. L.E.M., D.H., M.G. and R.H. contributed materials and support in drafting the manuscript. I.B. performed histology. C.R. and J.R. designed and supervised the research and composed and wrote the manuscript.

Corresponding authors

Correspondence to Michael S D Kormann or Carsten Rudolph.

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Competing interests

C.R. recently founded a company, ethris GmbH, dedicated to the clinical translation of the technology described in this paper. The other authors declare that they have no competing financial interests.

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Kormann, M., Hasenpusch, G., Aneja, M. et al. Expression of therapeutic proteins after delivery of chemically modified mRNA in mice. Nat Biotechnol 29, 154–157 (2011). https://doi.org/10.1038/nbt.1733

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