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MicroRNA inhibition using antimiRs in acute human brain tissue sections

Antisense inhibition of microRNAs is an emerging preclinical approach to pharmacoresistant epilepsy. A leading candidate is an "antimiR" targeting microRNA‐134 (ant‐134), but testing to date has used rodent models. Here, we develop an antimiR testing platform in human brain tissue sections...

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Autores principales: Morris, Gareth, Langa, Elena, Fearon, Conor, Conboy, Karen, Lau E‐How, Kelvin, Sanz‐Rodriguez, Amaya, O'Brien, Donncha F., Sweeney, Kieron, Lacey, Austin, Delanty, Norman, Beausang, Alan, Brett, Francesca M., Cryan, Jane B., Cunningham, Mark O., Henshall, David C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9546319/
https://www.ncbi.nlm.nih.gov/pubmed/35656590
http://dx.doi.org/10.1111/epi.17317
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author Morris, Gareth
Langa, Elena
Fearon, Conor
Conboy, Karen
Lau E‐How, Kelvin
Sanz‐Rodriguez, Amaya
O'Brien, Donncha F.
Sweeney, Kieron
Lacey, Austin
Delanty, Norman
Beausang, Alan
Brett, Francesca M.
Cryan, Jane B.
Cunningham, Mark O.
Henshall, David C.
author_facet Morris, Gareth
Langa, Elena
Fearon, Conor
Conboy, Karen
Lau E‐How, Kelvin
Sanz‐Rodriguez, Amaya
O'Brien, Donncha F.
Sweeney, Kieron
Lacey, Austin
Delanty, Norman
Beausang, Alan
Brett, Francesca M.
Cryan, Jane B.
Cunningham, Mark O.
Henshall, David C.
author_sort Morris, Gareth
collection PubMed
description Antisense inhibition of microRNAs is an emerging preclinical approach to pharmacoresistant epilepsy. A leading candidate is an "antimiR" targeting microRNA‐134 (ant‐134), but testing to date has used rodent models. Here, we develop an antimiR testing platform in human brain tissue sections. Brain specimens were obtained from patients undergoing resective surgery to treat pharmacoresistant epilepsy. Neocortical specimens were submerged in modified artificial cerebrospinal fluid (ACSF) and dissected for clinical neuropathological examination, and unused material was transferred for sectioning. Individual sections were incubated in oxygenated ACSF, containing either ant‐134 or a nontargeting control antimiR, for 24 h at room temperature. RNA integrity was assessed using BioAnalyzer processing, and individual miRNA levels were measured using quantitative reverse transcriptase polymerase chain reaction. Specimens transported in ACSF could be used for neuropathological diagnosis and had good RNA integrity. Ant‐134 mediated a dose‐dependent knockdown of miR‐134, with approximately 75% reduction of miR‐134 at 1 μmol L(−1) and 90% reduction at 3 μmol L(−1). These doses did not have off‐target effects on expression of a selection of three other miRNAs. This is the first demonstration of ant‐134 effects in live human brain tissues. The findings lend further support to the preclinical development of a therapy that targets miR‐134 and offer a flexible platform for the preclinical testing of antimiRs, and other antisense oligonucleotide therapeutics, in human brain.
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spelling pubmed-95463192022-10-14 MicroRNA inhibition using antimiRs in acute human brain tissue sections Morris, Gareth Langa, Elena Fearon, Conor Conboy, Karen Lau E‐How, Kelvin Sanz‐Rodriguez, Amaya O'Brien, Donncha F. Sweeney, Kieron Lacey, Austin Delanty, Norman Beausang, Alan Brett, Francesca M. Cryan, Jane B. Cunningham, Mark O. Henshall, David C. Epilepsia Brief Communication Antisense inhibition of microRNAs is an emerging preclinical approach to pharmacoresistant epilepsy. A leading candidate is an "antimiR" targeting microRNA‐134 (ant‐134), but testing to date has used rodent models. Here, we develop an antimiR testing platform in human brain tissue sections. Brain specimens were obtained from patients undergoing resective surgery to treat pharmacoresistant epilepsy. Neocortical specimens were submerged in modified artificial cerebrospinal fluid (ACSF) and dissected for clinical neuropathological examination, and unused material was transferred for sectioning. Individual sections were incubated in oxygenated ACSF, containing either ant‐134 or a nontargeting control antimiR, for 24 h at room temperature. RNA integrity was assessed using BioAnalyzer processing, and individual miRNA levels were measured using quantitative reverse transcriptase polymerase chain reaction. Specimens transported in ACSF could be used for neuropathological diagnosis and had good RNA integrity. Ant‐134 mediated a dose‐dependent knockdown of miR‐134, with approximately 75% reduction of miR‐134 at 1 μmol L(−1) and 90% reduction at 3 μmol L(−1). These doses did not have off‐target effects on expression of a selection of three other miRNAs. This is the first demonstration of ant‐134 effects in live human brain tissues. The findings lend further support to the preclinical development of a therapy that targets miR‐134 and offer a flexible platform for the preclinical testing of antimiRs, and other antisense oligonucleotide therapeutics, in human brain. John Wiley and Sons Inc. 2022-06-20 2022-08 /pmc/articles/PMC9546319/ /pubmed/35656590 http://dx.doi.org/10.1111/epi.17317 Text en © 2022 The Authors. Epilepsia published by Wiley Periodicals LLC on behalf of International League Against Epilepsy. https://creativecommons.org/licenses/by-nc/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc/4.0/ (https://creativecommons.org/licenses/by-nc/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.
spellingShingle Brief Communication
Morris, Gareth
Langa, Elena
Fearon, Conor
Conboy, Karen
Lau E‐How, Kelvin
Sanz‐Rodriguez, Amaya
O'Brien, Donncha F.
Sweeney, Kieron
Lacey, Austin
Delanty, Norman
Beausang, Alan
Brett, Francesca M.
Cryan, Jane B.
Cunningham, Mark O.
Henshall, David C.
MicroRNA inhibition using antimiRs in acute human brain tissue sections
title MicroRNA inhibition using antimiRs in acute human brain tissue sections
title_full MicroRNA inhibition using antimiRs in acute human brain tissue sections
title_fullStr MicroRNA inhibition using antimiRs in acute human brain tissue sections
title_full_unstemmed MicroRNA inhibition using antimiRs in acute human brain tissue sections
title_short MicroRNA inhibition using antimiRs in acute human brain tissue sections
title_sort microrna inhibition using antimirs in acute human brain tissue sections
topic Brief Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9546319/
https://www.ncbi.nlm.nih.gov/pubmed/35656590
http://dx.doi.org/10.1111/epi.17317
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