血浆微小RNA作为实验性创伤性脑损伤后严重癫痫发生的预后生物标志物——EpiBioS4Rx项目1研究

Plasma microRNAs as prognostic biomarkers for development of severe epilepsy after experimental traumatic brain injury-EpiBioS4Rx Project 1 study.

作者信息

Heiskanen Mette, Ndode-Ekane Xavier Ekolle, Ali Idrish, Santana-Gomez Cesar, Puhakka Noora, Gupta Shalini Das, Andrade Pedro, Immonen Riikka, Casillas-Espinosa Pablo, Manninen Eppu, Smith Gregory, Brady Rhys D, Silva Juliana, Braine Emma, Hudson Matt, Yamakawa Glen R, Jones Nigel C, Shultz Sandy R, Harris Neil G, Wright David K, Gröhn Olli, Staba Richard J, O'Brien Terence J, Pitkänen Asla

机构信息

A. I. Virtanen Institute for Molecular Sciences, University of Eastern Finland, Kuopio, Finland.

Department of Neuroscience, Monash University, Melbourne, Victoria, Australia.

出版信息

Epilepsia. 2025 Mar;66(3):870-885. doi: 10.1111/epi.18219. Epub 2024 Dec 11.

DOI:10.1111/epi.18219

PMID:39661396

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11908664/

Abstract

OBJECTIVE

To test a hypothesis that acutely regulated plasma microRNAs (miRNAs) can serve as prognostic biomarkers for the development of post-traumatic epilepsy (PTE).

METHODS

Adult male Sprague-Dawley rats (n = 245) were randomized to lateral fluid-percussion-induced traumatic brain injury (TBI) or sham operation at three study sites (Finland, Australia, United States). Video-electroencephalography (vEEG) was performed on the seventh post-injury month to detect spontaneous seizures. Tail vein plasma collected 48 h after TBI for miRNA analysis was available from 209 vEEG monitored animals (45 sham, 164 TBI [32 with epilepsy]). Based on small RNA sequencing and previous data, the seven most promising brain enriched miRNAs (miR-183-5p, miR-323-3p, miR-434-3p, miR-9a-3p, miR-124-3p, miR-132-3p, and miR-212-3p) were validated by droplet digital polymerase chain reaction (ddPCR).

RESULTS

All seven plasma miRNAs differentiated between TBI and sham-operated rats. None of the seven miRNAs differentiated TBI rats that did and did not develop epilepsy (p > .05), or rats with ≥3 vs <3 seizures in a month (p > .05). However, miR-212-3p differentiated rats that developed epilepsy with seizure clusters (i.e., ≥3 seizures within 24 h) from those without seizure clusters (.34 ± .14 vs .60 ± .34, adj. p < .05) with an area under the curve (AUC) of .81 (95% confidence interval [CI] .65-.97, p < .01, 64% sensitivity, 95% specificity). Lack of elevation in miR-212-3p also differentiated rats that developed epilepsy with seizure clusters from all other TBI rats (n = 146, .34 ± .14 vs .55 ± .31, p < .01) with an AUC of .74 (95% CI .61-.87, p < .01, 82% sensitivity, 62% specificity). Glmnet analysis identified a combination of miR-212-3p and miR-132-3p as an optimal set to differentiate TBI rats with vs without seizure clusters (cross-validated AUC .75, 95% CI .47-.92, p < .05).

SIGNIFICANCE

miR-212-3p alone or in combination with miR-132-3p shows promise as a translational prognostic biomarker for the development of severe PTE with seizure clusters.

摘要

目的

验证急性调控的血浆微小RNA（miRNA）能否作为创伤后癫痫（PTE）发生的预后生物标志物这一假设。

方法

成年雄性Sprague-Dawley大鼠（n = 245）在三个研究地点（芬兰、澳大利亚、美国）被随机分为侧方流体冲击诱导的创伤性脑损伤（TBI）组或假手术组。在伤后第7个月进行视频脑电图（vEEG）检查以检测自发癫痫发作。209只接受vEEG监测的动物（45只假手术组，164只TBI组[32只发生癫痫]）在TBI后48小时采集尾静脉血浆用于miRNA分析。基于小RNA测序和先前数据，通过液滴数字聚合酶链反应（ddPCR）验证了7种最有前景的脑富集miRNA（miR-183-5p、miR-323-3p、miR-434-3p、miR-9a-3p、miR-124-3p、miR-132-3p和miR-212-3p）。

结果

所有7种血浆miRNA均可区分TBI大鼠和假手术大鼠。这7种miRNA均不能区分发生和未发生癫痫的TBI大鼠（p > 0.05），也不能区分一个月内发作≥3次和<3次的大鼠（p > 0.05）。然而，miR-212-3p可区分有癫痫发作簇（即24小时内发作≥3次）的癫痫大鼠和无发作簇的大鼠（0.34±0.14 vs 0.60±0.34，校正p < 0.05），曲线下面积（AUC）为0.81（95%置信区间[CI] 0.65 - 0.97，p < 0.01，敏感性64%，特异性95%）。miR-212-3p缺乏升高也可区分有癫痫发作簇的癫痫大鼠和所有其他TBI大鼠（n = 146，0.34±0.14 vs 0.55±0.31，p < 0.01），AUC为0.74（95% CI 0.61 - 0.87，p < 0.01，敏感性82%，特异性62%）。Glmnet分析确定miR-212-3p和miR-132-3p的组合是区分有和无发作簇的TBI大鼠的最佳组合（交叉验证AUC 0.75，95% CI 0.47 - 0.92，p < 0.05）。