16S rRNA Gene Pyrosequencing Reveals Shift in Patient Faecal Microbiota During High-Dose Chemotherapy as Conditioning Regimen for Bone Marrow Transplantation
16S rRNA Gene Pyrosequencing Reveals Shift in Patient Faecal Microbiota During High-Dose Chemotherapy as Conditioning Regimen for Bone Marrow Transplantation
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DOI：
10.1007/s00248-013-0355-4
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发表时间：
2014-04-01
影响因子：
3.6
通讯作者：
de La Cochetiere, Marie France
中科院分区：
文献类型：
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作者：
Montassier, Emmanuel;Batard, Eric;de La Cochetiere, Marie France
由于骨髓移植（BMT）相关的强化细胞毒性治疗，胃肠道紊乱是血液学恶性肿瘤的常见副作用。然而，化疗期间肠道微生物群的变化仍然缺乏描述，这可能是由于以前的研究使用了基于培养和低分辨率的分子方法。我们研究的目的是应用下一代DNA测序技术来分析化疗引起的粪便微生物群的变化。我们纳入了8例非霍奇金淋巴瘤患者，他们接受了一个疗程的BMT调理化疗。我们在化疗开始的前一天收集了化疗前的粪便样本，在化疗开始后1周收集了化疗后的样本。从粪便样本中提取总DNA，利用变性高效液相色谱法扩增16S核糖体RNA （rRNA）基因V6至V8区域，利用PCR引物靶向16S rRNA基因V5和V6高变区，对16S rRNA基因进行454焦磷酸测序。使用QIIME管道对原始序列数据进行筛选、修剪和过滤。我们观察到α多样性的急剧减少和肠道微生物群组成在化疗反应中的显着差异。化疗与粪杆菌的急剧下降有关，并伴有埃希氏菌的增加。化疗引起的肠道微生物群的变化可能会对免疫功能低下的癌症患者产生严重的副作用。我们的研究是确定有胃肠道紊乱风险的患者的第一步，并促进预防肠道微生物群急剧变化的策略。
Gastrointestinal disturbances are a side-effect frequently associated with haematological malignancies due to the intensive cytotoxic treatment given in connection with bone marrow transplantation (BMT). However, intestinal microbiota changes during chemotherapy remain poorly described, probably due to the use of culture-based and low-resolution molecular methods in previous studies. The objective of our study was to apply a next generation DNA sequencing technology to analyse chemotherapy-induced changes in faecal microbiota. We included eight patients with non-Hodgkin's lymphoma undergoing one course of BMT conditioning chemotherapy. We collected a prechemotherapy faecal sample, the day before chemotherapy was initiated, and a postchemotherapy sample, collected 1 week after the initiation of chemotherapy. Total DNA was extracted from faecal samples, denaturing high-performance liquid chromatography based on amplification of the V6 to V8 region of the 16S ribosomal RNA (rRNA) gene, and 454-pyrosequencing of the 16 S rRNA gene, using PCR primers targeting the V5 and V6 hypervariable 16S rRNA gene regions were performed. Raw sequence data were screened, trimmed, and filtered using the QIIME pipeline. We observed a steep reduction in alpha diversity and significant differences in the composition of the intestinal microbiota in response to chemotherapy. Chemotherapy was associated with a drastic drop in Faecalibacterium and accompanied by an increase of Escherichia. The chemotherapy-induced shift in the intestinal microbiota could induce severe side effects in immunocompromised cancer patients. Our study is a first step in identifying patients at risk for gastrointestinal disturbances and to promote strategies to prevent this drastic shift in intestinal microbiota.