DataSheet_1_The oral microbiome of patients with ischemic stroke predicts their severity and prognosis.pdf

Background and objectivesStroke is a common group of cerebrovascular diseases that can lead to brain damage or death. Several studies have shown a close link between oral health and stroke. However, the oral microbiome profiling of ischemic stroke (IS) and its potential clinical implication are unclear. This study aimed to describe the oral microbiota composition of IS, the high risk of IS, and healthy individuals and to profile the relationship between microbiota and IS prognosis.
MethodsThis observational study recruited three groups: IS, high-risk IS (HRIS), and healthy control (HC) individuals. Clinical data and saliva were collected from participants. The modified Rankin scale score after 90 days was used to assess the prognosis of stroke. Extracted DNA from saliva and performed 16S ribosomal ribonucleic acid (rRNA) gene amplicon sequencing. Sequence data were analyzed using QIIME2 and R packages to evaluate the association between the oral microbiome and stroke.
ResultsA total of 146 subjects were enrolled in this study according to the inclusion criteria. Compared with HC, HRIS and IS demonstrated a progressive increase trend in Chao1, observed species richness, and Shannon and Simpson diversity index. On the basis of permutational multivariate analysis of variance, the data indicate a great variation in the saliva microbiota composition between HC and HRIS (F = 2.40, P < 0.001), HC and IS (F = 5.07, P < 0.001), and HRIS and IS (F = 2.79, P < 0.001). The relative abundance of g_Streptococcus, g_Prevotella, g_Veillonella, g_Fusobacterium, and g_Treponema was higher in HRIS and IS compared with that in HC. Furthermore, we constructed the predictive model by differential genera to effectively distinguish patients with IS with poor 90-day prognoses from those with good (area under the curve = 79.7%; 95% CI, 64.41%–94.97%; p < 0.01).
DiscussionIn summary, the oral salivary microbiome of HRIS and IS subjects have a higher diversity, and the differential bacteria have some predictive value for the severity and prognosis of IS. Oral microbiota may be used as potential biomarkers in patients with IS.

研究背景与目的
脑卒中是一类可导致脑损伤甚至死亡的常见脑血管疾病。多项研究已证实口腔健康与脑卒中存在密切关联，但目前针对缺血性脑卒中（ischemic stroke, IS）的口腔微生物组谱特征及其潜在临床应用价值仍不明确。本研究旨在明确缺血性脑卒中患者、缺血性脑卒中高危人群及健康对照者的口腔菌群组成，并解析菌群与缺血性脑卒中预后之间的关联。

研究方法
本项观察性研究共纳入三组研究对象：缺血性脑卒中患者、缺血性脑卒中高危人群（high-risk IS, HRIS）及健康对照（healthy control, HC）。收集所有研究对象的临床资料与唾液样本，于受试者发病后90天采用改良Rankin量表评分评估脑卒中预后情况。提取唾液样本中的DNA，并开展16S核糖体核糖核酸（16S ribosomal ribonucleic acid, rRNA）基因扩增子测序。采用QIIME2与R软件包对测序数据进行分析，以评估口腔微生物组与脑卒中之间的关联。

研究结果
本研究共按照纳入标准纳入146名研究对象。与健康对照相比，缺血性脑卒中高危人群及缺血性脑卒中患者的Chao1指数、观测物种丰富度、Shannon指数及Simpson指数均呈逐步升高趋势。基于置换多元方差分析结果，健康对照与缺血性脑卒中高危人群、健康对照与缺血性脑卒中患者、缺血性脑卒中高危人群与缺血性脑卒中患者的唾液菌群组成均存在显著差异（F=2.40，P<0.001；F=5.07，P<0.001；F=2.79，P<0.001）。与健康对照相比，缺血性脑卒中高危人群及缺血性脑卒中患者的链球菌属（g_Streptococcus）、普雷沃菌属（g_Prevotella）、韦荣球菌属（g_Veillonella）、梭杆菌属（g_Fusobacterium）以及密螺旋体属（g_Treponema）的相对丰度均显著升高。此外，本研究基于差异菌属构建了预测模型，可有效区分90天预后不良与预后良好的缺血性脑卒中患者（曲线下面积=79.7%；95%置信区间：64.41%~94.97%；P<0.01）。

讨论
综上，缺血性脑卒中高危人群及缺血性脑卒中患者的唾液口腔微生物组具有更高的多样性，差异菌属对缺血性脑卒中的病情严重程度及预后具有一定预测价值。口腔微生物组有望成为缺血性脑卒中患者的潜在生物标志物。