Exploring the Diagnostic Potential of miRNA Signatures in the Fabry Disease Serum: A Comparative Study of Automated and Manual Sample Isolations

Josephine Yen Fang, Saravanan Ayyadurai, Alyssa F. Pybus, Hiroshi Sugimoto, Mark G. Qian
{"title":"Exploring the Diagnostic Potential of miRNA Signatures in the Fabry Disease Serum: A Comparative Study of Automated and Manual Sample Isolations","authors":"Josephine Yen Fang, Saravanan Ayyadurai, Alyssa F. Pybus, Hiroshi Sugimoto, Mark G. Qian","doi":"10.1101/2024.03.25.24304836","DOIUrl":null,"url":null,"abstract":"Fabry disease, an X-linked lysosomal storage disorder caused by galactosidase alpha (GLA) gene mutations, exhibits diverse clinical manifestations, and poses significant diagnostic challenges. Early diagnosis and treatment are crucial for improved patient outcomes, pressing the need for reliable biomarkers. In this study, we aimed to identify miRNA candidates as potential biomarkers for Fabry disease using the KingFisher™ automated isolation method and NanoString nCounter® miRNA detection assay. Clinical serum samples were collected from both healthy subjects and Fabry disease patients. RNA extraction from the samples was performed using the KingFisher™ automated isolation method with the MagMAX mirVanaTM kit or manually using the Qiagen miRNeasy kit. The subsequent NanoString nCounter® miRNA detection assay showed consistent performance and no correlation between RNA input concentration and raw count, ensuring reliable and reproducible results. Interestingly, the detection range and highly differential miRNA between the control and disease groups were found to be distinct depending on the isolation method employed. Nevertheless, enrichment analysis of miRNA-targeting genes consistently revealed significant associations with angiogenesis pathways in both isolation methods. Additionally, our investigation into the impact of enzyme replacement therapy on miRNA expression indicated that some differential miRNAs may be sensitive to treatment. Our study provides valuable insights to identify miRNA biomarkers for Fabry disease. While different isolation methods yielded various detection ranges and highly differential miRNAs, the consistent association with angiogenesis pathways suggests their significance in disease progression. These findings lay the groundwork for further investigations and validation studies, ultimately leading to the development of non-invasive and reliable biomarkers to aid in early diagnosis and treatment monitoring for Fabry disease.","PeriodicalId":501212,"journal":{"name":"medRxiv - Rheumatology","volume":"99 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2024-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"medRxiv - Rheumatology","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1101/2024.03.25.24304836","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0

Abstract

Fabry disease, an X-linked lysosomal storage disorder caused by galactosidase alpha (GLA) gene mutations, exhibits diverse clinical manifestations, and poses significant diagnostic challenges. Early diagnosis and treatment are crucial for improved patient outcomes, pressing the need for reliable biomarkers. In this study, we aimed to identify miRNA candidates as potential biomarkers for Fabry disease using the KingFisher™ automated isolation method and NanoString nCounter® miRNA detection assay. Clinical serum samples were collected from both healthy subjects and Fabry disease patients. RNA extraction from the samples was performed using the KingFisher™ automated isolation method with the MagMAX mirVanaTM kit or manually using the Qiagen miRNeasy kit. The subsequent NanoString nCounter® miRNA detection assay showed consistent performance and no correlation between RNA input concentration and raw count, ensuring reliable and reproducible results. Interestingly, the detection range and highly differential miRNA between the control and disease groups were found to be distinct depending on the isolation method employed. Nevertheless, enrichment analysis of miRNA-targeting genes consistently revealed significant associations with angiogenesis pathways in both isolation methods. Additionally, our investigation into the impact of enzyme replacement therapy on miRNA expression indicated that some differential miRNAs may be sensitive to treatment. Our study provides valuable insights to identify miRNA biomarkers for Fabry disease. While different isolation methods yielded various detection ranges and highly differential miRNAs, the consistent association with angiogenesis pathways suggests their significance in disease progression. These findings lay the groundwork for further investigations and validation studies, ultimately leading to the development of non-invasive and reliable biomarkers to aid in early diagnosis and treatment monitoring for Fabry disease.
查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
探索法布里病血清中 miRNA 标志的诊断潜力:自动样本分离与人工样本分离的比较研究
法布里病是一种由半乳糖苷酶α(GLA)基因突变引起的X连锁溶酶体贮积症,临床表现多种多样,给诊断带来了巨大挑战。早期诊断和治疗对改善患者预后至关重要,因此迫切需要可靠的生物标志物。在这项研究中,我们采用 KingFisher™ 自动分离方法和 NanoString nCounter® miRNA 检测分析法,旨在鉴定作为法布里病潜在生物标志物的候选 miRNA。临床血清样本采集自健康受试者和法布里病患者。样本的 RNA 提取采用 KingFisher™ 自动分离法和 MagMAX mirVanaTM 试剂盒,或使用 Qiagen miRNeasy 试剂盒手工提取。随后进行的 NanoString nCounter® miRNA 检测分析表明,该方法性能稳定,RNA 输入浓度与原始计数之间无相关性,确保了结果的可靠性和可重复性。有趣的是,根据采用的分离方法,对照组和疾病组之间的 miRNA 检测范围和高度差异被发现是不同的。然而,对 miRNA 靶向基因的富集分析一致显示,两种分离方法都与血管生成通路有显著关联。此外,我们对酶替代疗法对 miRNA 表达影响的调查表明,一些不同的 miRNA 可能对治疗敏感。我们的研究为确定法布里病的 miRNA 生物标志物提供了宝贵的见解。虽然不同的分离方法会产生不同的检测范围和高度差异的 miRNA,但它们与血管生成途径的一致关联表明,它们在疾病进展中具有重要意义。这些发现为进一步的调查和验证研究奠定了基础,最终将开发出无创、可靠的生物标记物,帮助法布里病的早期诊断和治疗监测。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 去求助
来源期刊
自引率
0.00%
发文量
0
期刊最新文献
Mosaic loss of chromosome Y characterizes late-onset rheumatoid arthritis and contrasting associations of polygenic risk score based on age at onset. Use of Metagenomic Microbial Plasma Cell-Free DNA Next-Generation Sequencing Assay in Outpatient Rheumatology Practice Proteomic profiling of the large vessel vasculitis spectrum identifies shared signatures of innate immune activation and stromal remodelling Pre-trained convolutional neural network with transfer learning by artificial illustrated images classify power Doppler ultrasound images of rheumatoid arthritis joints Associations between exposure to OPEs and rheumatoid arthritis risk among adults in NHANES, 2011-2018
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1