Conditional deletion of miR-204 and miR-211 in murine retinal pigment epithelium results in retinal degeneration

Samuel W Du; Ravikiran Komirisetty; Dominik Lewandowski; Elliot H Choi; Damian Panas; Susie Suh; Marcin Tabaka; Roxana A Radu; Krzysztof Palczewski

doi:10.1016/j.jbc.2024.107344

Conditional deletion of miR-204 and miR-211 in murine retinal pigment epithelium results in retinal degeneration

J Biol Chem. 2024 May 4;300(6):107344. doi: 10.1016/j.jbc.2024.107344. Online ahead of print.

Authors

Samuel W Du¹, Ravikiran Komirisetty², Dominik Lewandowski³, Elliot H Choi³, Damian Panas⁴, Susie Suh³, Marcin Tabaka⁴, Roxana A Radu², Krzysztof Palczewski⁵

Affiliations

¹ Gavin Herbert Eye Institute-Center for Translational Vision Research, Department of Ophthalmology, University of California, Irvine, California, USA; Department of Physiology and Biophysics, University of California, Irvine, California, USA. Electronic address: swdu@uci.edu.
² Department of Ophthalmology and UCLA Stein Eye Institute, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, California, USA.
³ Gavin Herbert Eye Institute-Center for Translational Vision Research, Department of Ophthalmology, University of California, Irvine, California, USA.
⁴ International Centre for Translational Eye Research, Warsaw, Poland; Institute of Physical Chemistry, Polish Academy of Sciences, Warsaw, Poland.
⁵ Gavin Herbert Eye Institute-Center for Translational Vision Research, Department of Ophthalmology, University of California, Irvine, California, USA; Department of Physiology and Biophysics, University of California, Irvine, California, USA; Department of Chemistry, University of California, Irvine, Irvine, California, USA; Department of Molecular Biology and Biochemistry, University of California, Irvine, Irvine, California, USA. Electronic address: kpalczew@uci.edu.

Abstract

MicroRNAs (miRs) are short, evolutionarily conserved noncoding RNAs that canonically downregulate expression of target genes. The miR family composed of miR-204 and miR-211 is among the most highly expressed miRs in the retinal pigment epithelium (RPE) in both mouse and human and also retains high sequence identity. To assess the role of this miR family in the developed mouse eye, we generated two floxed conditional KO mouse lines crossed to the RPE65-ERT2-Cre driver mouse line to perform an RPE-specific conditional KO of this miR family in adult mice. After Cre-mediated deletion, we observed retinal structural changes by optical coherence tomography; dysfunction and loss of photoreceptors by retinal imaging; and retinal inflammation marked by subretinal infiltration of immune cells by imaging and immunostaining. Single-cell RNA sequencing of diseased RPE and retinas showed potential miR-regulated target genes, as well as changes in noncoding RNAs in the RPE, rod photoreceptors, and Müller glia. This work thus highlights the role of miR-204 and miR-211 in maintaining RPE function and how the loss of miRs in the RPE exerts effects on the neural retina, leading to inflammation and retinal degeneration.

Keywords: RPE; electrophysiology; gene KO; inflammation; microRNA (miRNA); retinal degeneration.

Grants and funding

P30 EY000331/EY/NEI NIH HHS/United States