PO.PR01.04 · 预防研究
用于早期检测结直肠癌及癌前病变的粪便微生物组特征
Fecal microbiome signatures for early detection of colorectal cancer and precursor lesions
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景：结直肠癌（CRC）仍是全球重大健康负担，开发用于早期检测的非侵入性生物标志物是一项关键的未满足需求。越来越多的证据表明，肠道微生物组失调参与腺瘤-癌序列，并可能支持 CRC 风险分层。
目的：研究来自粪便免疫化学检测（FIT）样本残余物的肠道微生物组分析是否可作为 CRC 筛查的生物标志物。
方法：将来自巴西 FIT 阳性个体的残余粪便样本（n=133）根据结肠镜检查分类为无病变（N=30）、非进展期腺瘤（NAA=46）、进展期腺瘤（AA=13）或结直肠癌（CA=44）。在三名个体中，在三种采集管（两种 FIT 管和 Omnigut）上比较了微生物组谱。来自 FIT 样本的细菌 DNA 在 Oxford Nanopore MinION（R10.4.1 流动池，持续 48 小时）上进行 16S rRNA 测序。使用 Dorado 处理读段，用 Emu 与精选的全长 16S 数据库进行比对，并在 QIIME2 中使用 Python 进行分析。评估了 Alpha 多样性（25 种不同指标）、Beta 多样性（Jaccard、Bray-Curtis）和差异丰度（ANCOMBC）。
结果：CA 样本的 Alpha 多样性显著高于 N 和 NAA 患者（Chao 1，p < 0.001 和 p=0.006；观察到的特征，p < 0.001 和 p=0.007），与 AA 相比无差异。Beta 多样性（PERMANOVA，两种指标 p = 0.001）显示 CA 与 AA（Jaccard，p=0.001，Bray Curtis，p=0.001）、NAA（Jaccard，p=0.001，Bray Curtis，p=0.001）和 N（Jaccard，p=0.001，Bray Curtis，p=0.003）之间存在显著的组成分离。差异丰度表明随着病变进展菌群失调加剧。与 N 样本相比，NAA 表现出 42 个差异丰度物种（9 个减少，33 个增加），AA 显示 65 个物种（15 个减少，50 个增加），CRC 则显示出明显的菌群失调，有 179 个物种（28 个减少，151 个增加）。初步检查凸显了 CA 样本中 Fusobacterium nucleatum 的富集。分析分类学谱显示，所分析的三种采集管之间物种组成无差异。
结论：使用常规残余 FIT 样本进行微生物组分析是可行的，并揭示了沿腺瘤-癌谱的显著多样性变化。这些发现支持 FIT 来源的微生物组特征作为巴西筛查人群中 CRC 早期检测非侵入性生物标志物的潜力。
查看英文原文 English abstract
Background: Colorectal cancer (CRC) remains a major global health burden, and the development of non-invasive biomarkers for early detection is a critical unmet need. Growing evidence indicates that intestinal microbiome dysbiosis contributes to the adenoma-carcinoma sequence and may support CRC risk stratification.
Aim: To investigate whether intestinal microbiome profiling from residual material of Fecal Immunochemical Test (FIT) samples can serve as a biomarker for CRC screening.
Methods: Residual stool samples from Brazilian FIT-positive individuals (n=133) were classified according to colonoscopy as no lesions (N=30), non-advanced adenoma (NAA=46), advanced adenoma (AA=13), or colorectal cancer (CA=44). In three individuals, the microbiome profile was compared on three collection tubes (two FIT tubes and Omnigut). Bacterial DNA from FIT samples underwent 16S rRNA sequencing on Oxford Nanopore MinION (R10.4.1 flow cells, for 48 hours). Reads were processed using Dorado, aligned with Emu to a curated full-length 16S database, and analyzed in QIIME2 using Python. Alpha diversity (25 different metrics), beta diversity (Jaccard, Bray-Curtis), and differential abundance (ANCOMBC) were assessed.
Results: Alpha diversity was significantly higher in CA samples compared with N and NAA patients (Chao 1, p < 0.001 and p=0.006; Observed features, p < 0.001 and p=0.007 ), with no difference compared to AA. Beta diversity (PERMANOVA, p = 0.001 for both metrics) showed significant compositional separation among CA and AA (Jaccard, p=0.001 , Bray Curtis, p=0.001 ), NAA (Jaccard, p=0.001 , Bray Curtis, p=0.001 ), and N (Jaccard, p=0.001 , Bray Curtis, p=0.003 ). Differential abundance indicated increasing dysbiosis as lesions progressed. Compared with N samples, NAA exhibited 42 differentially abundant species (9 decreased, 33 increased), AA showed 65 species (15 decreased, 50 increased), and CRC showed pronounced dysbiosis with 179 species (28 decreased, 151 increased). Preliminary inspection highlights enrichment of Fusobacterium nucleatum in CA samples. Analysis of the taxonomic profiles revealed no differences in species composition among the three collection tubes analyzed.
Conclusion: Microbiome profiling using routine residual FIT samples is feasible and reveals a marked diversity shift along the adenoma-carcinoma spectrum. These findings
support the potential of FIT-derived microbiome signatures as non-invasive biomarkers for CRC early detection in Brazilian screening populations.
利益披露 Disclosure
M. Bisarro Dos Reis, None..
M. T. Reis, None..
A. Peres, None..
C. L. Hashimoto, None..
D. P. Guimarães, None..
R. M. Reis, None.