口腔微生物组与头颈鳞状细胞癌的后续风险

口腔微生物群可能参与头颈鳞状细胞癌（HNSCC）的发生发展，但目前的证据主要局限于细菌16S扩增子测序或小规模回顾性病例对照研究。为检测口腔细菌和真菌微生物组是否与后续HNSCC发生风险相关，在3个流行病学队列中进行前瞻性嵌套病例对照研究，包括美国癌症学会癌症预防研究二期营养队列、前列腺、肺、结直肠和卵巢癌筛查试验以及南方社区队列研究。在随访平均（标准差）5.1（3.6）年期间，共识别出236例前瞻性发展为HNSCC的患者。选择未发生HNSCC的对照参与者，按队列、年龄、性别、种族与族裔以及采样时间的2:1频率匹配。数据分析于2023年进行。利用全基因组鸟枪法测序对口腔细菌微生物组进行表征，利用内部转录间隔（ITS）测序分析口腔真菌微生物组。通过偏差校正的微生物群落组成分析评估细菌和真菌分类单元与HNSCC的关联。采用逻辑回归检验红色和橙色口腔致病菌复合体的关联。基于与HNSCC风险相关的微生物，计算微生物风险评分。主要结局指标为HNSCC的发生率。研究包括236例HNSCC病例，平均（SD）年龄60.9（9.5）岁，女性占24.6%，随访平均（SD）5.1（3.6）年，配对对照485例。整体微生物组多样性在基线时与后续HNSCC风险无显著相关，但发现13种口腔细菌种类与HNSCC的发生具有差异性相关性。这些细菌包括新鉴定的Prevotella salivae、Streptococcus sanguinis和Leptotrichia属，以及几种β和γ变形菌门的菌种。红色/橙色牙周病致病菌复合体与HNSCC风险呈中等相关（每1个标准差，OR=1.06；95% CI，1.00-1.12）。基于22种细菌构建的微生物风险评分每升高1个标准差，HNSCC风险增加50%（多变量OR=1.50；95% CI，1.21-1.85）。未发现与HNSCC风险显著相关的真菌分类单元。本研究提供了有力证据支持口腔细菌是HNSCC发生的风险因素。鉴定出的细菌及其复合体与其他风险因素结合，有望用于识别高风险人群，进行个性化预防。

The oral microbiota may be involved in development of head and neck squamous cell cancer (HNSCC), yet current evidence is largely limited to bacterial 16S amplicon sequencing or small retrospective case-control studies.To test whether oral bacterial and fungal microbiomes are associated with subsequent risk of HNSCC development.Prospective nested case-control study among participants providing oral samples in 3 epidemiological cohorts, the American Cancer Society Cancer Prevention Study II Nutrition Cohort, the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial, and the Southern Community Cohort Study. Two hundred thirty-six patients who prospectively developed HNSCC were identified during a mean (SD) of 5.1 (3.6) years of follow-up. Control participants who remained HNSCC free were selected by 2:1 frequency matching on cohort, age, sex, race and ethnicity, and time since oral sample collection. Data analysis was conducted in 2023.Characterization of the oral bacterial microbiome using whole-genome shotgun sequencing and the oral fungal microbiome using internal transcribed spacer sequencing. Association of bacterial and fungal taxa with HNSCC was assessed by analysis of compositions of microbiomes with bias correction. Association with red and orange oral pathogen complexes was tested by logistic regression. A microbial risk score for HNSCC risk was calculated from risk-associated microbiota.The primary outcome was HNSCC incidence.The study included 236 HNSCC case participants with a mean (SD) age of 60.9 (9.5) years and 24.6% women during a mean of 5.1 (3.6) years of follow-up, and 485 matched control participants. Overall microbiome diversity at baseline was not related to subsequent HNSCC risk; however 13 oral bacterial species were found to be differentially associated with development of HNSCC. The species included the newly identified Prevotella salivae, Streptococcus sanguinis, and Leptotrichia species, as well as several species belonging to beta and gamma Proteobacteria. The red/orange periodontal pathogen complex was moderately associated with HNSCC risk (odds ratio, 1.06 per 1 SD; 95% CI, 1.00-1.12). A 1-SD increase in microbial risk score (created based on 22 bacteria) was associated with a 50% increase in HNSCC risk (multivariate odds ratio, 1.50; 95% CI, 1.21-1.85). No fungal taxa associated with HNSCC risk were identified.This case-control study yielded compelling evidence that oral bacteria are a risk factor for HNSCC development. The identified bacteria and bacterial complexes hold promise, along with other risk factors, to identify high-risk individuals for personalized prevention of HNSCC.