Cargando…

Protecting Pax6 3′ UTR from MicroRNA-7 Partially Restores PAX6 in Islets from an Aniridia Mouse Model

Aniridia is a rare congenital syndrome that is associated with reduced visual acuity and progressive loss of vision. Aniridia patients may also develop systemic health issues associated with defects in the pancreas, digestive, and central nervous systems. The spectrum of symptoms associated with ani...

Descripción completa

Detalles Bibliográficos
Autores principales: Yongblah, Kevin, Alford, Spencer C., Ryan, Bridget C., Chow, Robert L., Howard, Perry L.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society of Gene & Cell Therapy 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6171161/
https://www.ncbi.nlm.nih.gov/pubmed/30290306
http://dx.doi.org/10.1016/j.omtn.2018.08.018
_version_ 1783360739413590016
author Yongblah, Kevin
Alford, Spencer C.
Ryan, Bridget C.
Chow, Robert L.
Howard, Perry L.
author_facet Yongblah, Kevin
Alford, Spencer C.
Ryan, Bridget C.
Chow, Robert L.
Howard, Perry L.
author_sort Yongblah, Kevin
collection PubMed
description Aniridia is a rare congenital syndrome that is associated with reduced visual acuity and progressive loss of vision. Aniridia patients may also develop systemic health issues associated with defects in the pancreas, digestive, and central nervous systems. The spectrum of symptoms associated with aniridia is due to haploinsufficiency of the paired box 6 gene (PAX6) and its role in the development and maintenance of the affected tissues. Here, we isolated pancreatic islets from mice heterozygous for Pax6 to test whether a Pax6-specific miRNA suppression (target protector) strategy can restore PAX6 protein levels. We show that miR-7 and miR-375 target specific sites within the Pax6 3′ UTR in a mouse pancreatic β-insulinoma cell line. Tough decoys (Tuds) against miR-7 and miR-375 increase expression of a mouse Pax6 3′ UTR luciferase reporter and increase PAX6 protein levels in these cells. Finally, we demonstrate that the shielding of the miR-7 binding site with a target protector restores PAX6 protein levels in the Pax6 heterozygous islets. The data presented here represent a proof of concept for RNA-based therapy for the progressive defects associated with aniridia and suggest the target protector approach may be a useful therapeutic strategy for other haploinsufficiency diseases.
format Online
Article
Text
id pubmed-6171161
institution National Center for Biotechnology Information
language English
publishDate 2018
publisher American Society of Gene & Cell Therapy
record_format MEDLINE/PubMed
spelling pubmed-61711612018-10-05 Protecting Pax6 3′ UTR from MicroRNA-7 Partially Restores PAX6 in Islets from an Aniridia Mouse Model Yongblah, Kevin Alford, Spencer C. Ryan, Bridget C. Chow, Robert L. Howard, Perry L. Mol Ther Nucleic Acids Article Aniridia is a rare congenital syndrome that is associated with reduced visual acuity and progressive loss of vision. Aniridia patients may also develop systemic health issues associated with defects in the pancreas, digestive, and central nervous systems. The spectrum of symptoms associated with aniridia is due to haploinsufficiency of the paired box 6 gene (PAX6) and its role in the development and maintenance of the affected tissues. Here, we isolated pancreatic islets from mice heterozygous for Pax6 to test whether a Pax6-specific miRNA suppression (target protector) strategy can restore PAX6 protein levels. We show that miR-7 and miR-375 target specific sites within the Pax6 3′ UTR in a mouse pancreatic β-insulinoma cell line. Tough decoys (Tuds) against miR-7 and miR-375 increase expression of a mouse Pax6 3′ UTR luciferase reporter and increase PAX6 protein levels in these cells. Finally, we demonstrate that the shielding of the miR-7 binding site with a target protector restores PAX6 protein levels in the Pax6 heterozygous islets. The data presented here represent a proof of concept for RNA-based therapy for the progressive defects associated with aniridia and suggest the target protector approach may be a useful therapeutic strategy for other haploinsufficiency diseases. American Society of Gene & Cell Therapy 2018-09-01 /pmc/articles/PMC6171161/ /pubmed/30290306 http://dx.doi.org/10.1016/j.omtn.2018.08.018 Text en © 2018 The Author(s) http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Yongblah, Kevin
Alford, Spencer C.
Ryan, Bridget C.
Chow, Robert L.
Howard, Perry L.
Protecting Pax6 3′ UTR from MicroRNA-7 Partially Restores PAX6 in Islets from an Aniridia Mouse Model
title Protecting Pax6 3′ UTR from MicroRNA-7 Partially Restores PAX6 in Islets from an Aniridia Mouse Model
title_full Protecting Pax6 3′ UTR from MicroRNA-7 Partially Restores PAX6 in Islets from an Aniridia Mouse Model
title_fullStr Protecting Pax6 3′ UTR from MicroRNA-7 Partially Restores PAX6 in Islets from an Aniridia Mouse Model
title_full_unstemmed Protecting Pax6 3′ UTR from MicroRNA-7 Partially Restores PAX6 in Islets from an Aniridia Mouse Model
title_short Protecting Pax6 3′ UTR from MicroRNA-7 Partially Restores PAX6 in Islets from an Aniridia Mouse Model
title_sort protecting pax6 3′ utr from microrna-7 partially restores pax6 in islets from an aniridia mouse model
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6171161/
https://www.ncbi.nlm.nih.gov/pubmed/30290306
http://dx.doi.org/10.1016/j.omtn.2018.08.018
work_keys_str_mv AT yongblahkevin protectingpax63utrfrommicrorna7partiallyrestorespax6inisletsfromananiridiamousemodel
AT alfordspencerc protectingpax63utrfrommicrorna7partiallyrestorespax6inisletsfromananiridiamousemodel
AT ryanbridgetc protectingpax63utrfrommicrorna7partiallyrestorespax6inisletsfromananiridiamousemodel
AT chowrobertl protectingpax63utrfrommicrorna7partiallyrestorespax6inisletsfromananiridiamousemodel
AT howardperryl protectingpax63utrfrommicrorna7partiallyrestorespax6inisletsfromananiridiamousemodel