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Differential effects of toll-like receptor stimulation on mRNA-driven myogenic conversion of human and mouse fibroblasts.

Publication ,  Journal Article
Lee, J; Xu, L; Gibson, TM; Gersbach, CA; Sullenger, BA
Published in: Biochem Biophys Res Commun
September 23, 2016

Transfection with in vitro transcribed mRNAs is a safe and effective tool to convert somatic cells to any cell type of interest. One caveat of mRNA transfection is that mRNAs are recognized by multiple RNA-sensing toll like receptors (TLRs). These TLRs can both promote and inhibit cellular reprogramming. We demonstrated that mRNA transfection stimulated TLR3 and TLR7 and induced cytotoxicity and IFN-β expression in human and mouse fibroblasts. Furthermore, mRNA transfection induced paracrine inhibition of repeated mRNA transfection through type I IFNs. Modified mRNAs (mmRNAs) containing pseudouridine and 5-methycytosine reduced TLR stimulation, cytotoxicity and IFN-β expression in fibroblasts. Repeated liposomal transfection with MyoD mmRNAs significantly enhanced myogenic conversion of human and mouse fibroblasts compared with repeated transfection with MyoD mRNAs. Interestingly, electroporation of mRNAs and mmRNAs completely abrogated cytotoxicity and IFN-β expression and also abolished myogenic conversion of fibroblasts. At a low concentration, TLR7/8 agonist R848 enhanced MyoD mmRNA-driven conversion of human fibroblasts into skeletal muscle cells, whereas high concentrations of R848 inhibited myogenic conversion of fibroblasts. Our study suggests that deliberate control of TLR signaling is a key factor in the success of mRNA-driven cellular reprogramming.

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Published In

Biochem Biophys Res Commun

DOI

EISSN

1090-2104

Publication Date

September 23, 2016

Volume

478

Issue

3

Start / End Page

1484 / 1490

Location

United States

Related Subject Headings

  • Transfection
  • Transcription, Genetic
  • Toll-Like Receptor 7
  • Toll-Like Receptor 3
  • Signal Transduction
  • RNA, Messenger
  • Paracrine Communication
  • MyoD Protein
  • Muscle Development
  • Mice
 

Citation

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Lee, J., Xu, L., Gibson, T. M., Gersbach, C. A., & Sullenger, B. A. (2016). Differential effects of toll-like receptor stimulation on mRNA-driven myogenic conversion of human and mouse fibroblasts. Biochem Biophys Res Commun, 478(3), 1484–1490. https://doi.org/10.1016/j.bbrc.2016.08.159
Lee, Jaewoo, Li Xu, Tyler M. Gibson, Charles A. Gersbach, and Bruce A. Sullenger. “Differential effects of toll-like receptor stimulation on mRNA-driven myogenic conversion of human and mouse fibroblasts.Biochem Biophys Res Commun 478, no. 3 (September 23, 2016): 1484–90. https://doi.org/10.1016/j.bbrc.2016.08.159.
Lee J, Xu L, Gibson TM, Gersbach CA, Sullenger BA. Differential effects of toll-like receptor stimulation on mRNA-driven myogenic conversion of human and mouse fibroblasts. Biochem Biophys Res Commun. 2016 Sep 23;478(3):1484–90.
Lee, Jaewoo, et al. “Differential effects of toll-like receptor stimulation on mRNA-driven myogenic conversion of human and mouse fibroblasts.Biochem Biophys Res Commun, vol. 478, no. 3, Sept. 2016, pp. 1484–90. Pubmed, doi:10.1016/j.bbrc.2016.08.159.
Lee J, Xu L, Gibson TM, Gersbach CA, Sullenger BA. Differential effects of toll-like receptor stimulation on mRNA-driven myogenic conversion of human and mouse fibroblasts. Biochem Biophys Res Commun. 2016 Sep 23;478(3):1484–1490.
Journal cover image

Published In

Biochem Biophys Res Commun

DOI

EISSN

1090-2104

Publication Date

September 23, 2016

Volume

478

Issue

3

Start / End Page

1484 / 1490

Location

United States

Related Subject Headings

  • Transfection
  • Transcription, Genetic
  • Toll-Like Receptor 7
  • Toll-Like Receptor 3
  • Signal Transduction
  • RNA, Messenger
  • Paracrine Communication
  • MyoD Protein
  • Muscle Development
  • Mice