miRNA-mediated control of exogenous OCT4 during mesenchymal-epithelial transition increases measles vector reprogramming efficiency

Ramya Rallabandi; Brenna Sharp; Conrad Cruz; Qi Wang; Alexis Locsin; Christopher B Driscoll; Ella Lee; Tim Nelson; Patricia Devaux

doi:10.1016/j.omtm.2021.11.012

miRNA-mediated control of exogenous OCT4 during mesenchymal-epithelial transition increases measles vector reprogramming efficiency

Mol Ther Methods Clin Dev. 2021 Nov 29:24:48-61. doi: 10.1016/j.omtm.2021.11.012. eCollection 2022 Mar 10.

Authors

Ramya Rallabandi^{1

2}, Brenna Sharp³, Conrad Cruz³, Qi Wang³, Alexis Locsin³, Christopher B Driscoll¹, Ella Lee³, Tim Nelson⁴, Patricia Devaux^{3

1

2}

Affiliations

¹ Virology and Gene Therapy Graduate Track, Mayo Clinic Graduate School of Biomedical Sciences, Mayo Clinic, Rochester, MN 55905, USA.
² Regenerative Sciences PhD Program, Mayo Clinic Graduate School of Biomedical Sciences, Mayo Clinic, Rochester, MN 55905, USA.
³ Department of Molecular Medicine, Mayo Clinic, Rochester, MN 55905, USA.
⁴ Department of Molecular Pharmacology and Experimental Therapeutics, Mayo Clinic College of Medicine, Rochester MN 55905, USA.

Abstract

OCT4 is a key mediator of induced pluripotent stem cell (iPSC) reprogramming, but the mechanistic insights into the role of exogenous OCT4 and timelines that initiate pluripotency remain to be resolved. Here, using measles reprogramming vectors, we present microRNA (miRNA) targeting of exogenous OCT4 to shut down its expression during the mesenchymal to the epithelial transition phase of reprogramming. We showed that exogenous OCT4 is required only for the initiation of reprogramming and is dispensable for the maturation stage. However, the continuous expression of SOX2, KLF4, and c-MYC is necessary for the maturation stage of the iPSC. Additionally, we demonstrate a novel application of miRNA targeting in a viral vector to contextually control the vector/transgene, ultimately leading to an improved reprogramming efficiency. This novel approach could be applied to other systems for improving the efficiency of vector-induced processes.

Keywords: OCT4; iPSC; measles virus; mesenchymal-epithelial transition; microRNA targeting; reprogramming; viral vectors.

Abstract

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