Effect of a 1-week mind-body intervention retreat on exosome-derived miRNA for 21 healthy human participants

We employed exosome-specific small RNA transcriptomics to analyze extracellular RNA (exRNA) profiles in human plasma-derived exosomes isolated via differential ultracentrifugation. Total exRNAs were extracted, quantified, and used to generate small RNA libraries, targeting transcripts between 16-120 nucleotides for sequencing on an Illumina NovaSeq X Plus platform. Sequencing reads were processed using Cutadapt, STAR aligner, and MultiQC, and differential expression analysis was conducted using EdgeR, applying TMM normalization and a paired negative binomial generalized linear model to assess intervention effects. Functional annotation of differentially expressed exRNAs was performed using ClusterProfiler and Reactome Pathways. The analysis accounted for participant-specific factors such as experience level, sex, and age. This is the first study to analyze human exosomal RNA changes following an intensive mind-body (meditation, reconceptualization, open label placebo healing ritual) intervention. Overall design: Exosomes were isolated from whole plasma from 21 human participants (15 females; mean age=46 yrs.) before ("pre") and after ("post") a 6-day intensive mind-body retreat, and miRNAseq was performed on an Illumina NovaSeq X Plus platform.

本研究采用外泌体特异性小RNA转录组学技术，分析通过差速超速离心法分离的人血浆来源外泌体中的细胞外RNA（extracellular RNA，简称exRNA）谱特征。我们对总exRNA进行提取、定量，并构建靶向覆盖16~120个核苷酸转录本的小RNA文库，随后在Illumina NovaSeq X Plus测序平台上完成测序。测序reads经Cutadapt、STAR比对软件及MultiQC进行预处理与质控，随后采用EdgeR开展差异表达分析，通过TMM标准化及配对负二项广义线性模型评估干预效应。差异表达exRNA的功能注释采用ClusterProfiler软件及Reactome通路数据库完成。本分析纳入了受试者个体特异性因素，包括经验水平、性别与年龄。本研究为首个分析高强度心身干预（冥想、认知重构、开放标签安慰剂疗愈仪式）后人外泌体RNA变化的研究。整体实验设计：从21名受试者（15名女性，平均年龄46岁）的全血浆中分离外泌体，分别采集为期6天的高强度心身静修营干预前（"pre"）及干预后（"post"）的样本，并在Illumina NovaSeq X Plus平台上开展miRNAseq测序。