MicroRNA inhibition using antimiRs in acute human brain tissue sections
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Published on: July 23, 2012
Gareth Morris1,2,3, Elena Langa2,3, Conor Fearon4,5
1Neuroscience, Physiology, and Pharmacology, University College London, London, UK.
Abstract:
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.
Insights
Antisense inhibition targeting microRNA-134 (ant-134) effectively reduces miR-134 levels in human brain tissue. This study establishes a platform for testing antimiRs in human tissue for epilepsy treatment.
Area of Science:
- Neuroscience
- Molecular Biology
- Pharmacology
Background:
- Pharmacoresistant epilepsy presents a significant clinical challenge.
- MicroRNA (miRNA) inhibition is a promising preclinical therapeutic strategy.
- Current antimiR testing relies on animal models, limiting direct human relevance.
Purpose of the Study:
- To establish a novel platform for testing antimiR efficacy in human brain tissue.
- To evaluate the effectiveness of an antimiR targeting microRNA-134 (ant-134) in human neocortical sections.
- To assess the safety and specificity of ant-134 in a human ex vivo model.
Main Methods:
- Human neocortical specimens from epilepsy surgery were used.
- Tissue sections were incubated with ant-134 or control antimiR in artificial cerebrospinal fluid (ACSF).
- RNA integrity was assessed, and miR-134 levels were quantified using RT-qPCR.
Main Results:
- Human brain tissue maintained good RNA integrity after transport in ACSF.
- Ant-134 demonstrated dose-dependent knockdown of miR-134, achieving up to 90% reduction.
- No off-target effects were observed on other selected miRNAs at tested doses.
Conclusions:
- This study provides the first evidence of ant-134 activity in live human brain tissue.
- The developed platform enables preclinical testing of antimiRs and other antisense oligonucleotides in human tissue.
- Findings support the further preclinical development of miR-134 targeted therapies for pharmacoresistant epilepsy.
