Objective: Head and neck cancer (HNC) is a common cancer represented by nearly 80% oral cavity (OC) and oropharyngeal cancers (OPCs). Seventy percent of OPCs are associated with the Human Papilloma Virus (HPV). Immunotherapy holds the promise of future improvements in treating HNC patients. The study objective was to determine whether durvalumab immunotherapy alone, prior to curative surgery, would significantly impact the oral salivary microbiome in a pilot cohort of HPV negative and positive OC and OPC patients. Methods: Early stage OPC patients with squamous cell carcinoma were recruited: 5 HPV+ and 12 HPV−, and treated with two or three administrations of durvalumab given every two weeks, prior to surgery. Unstimulated saliva was collected and processed for bacterial DNA Isolation and V1–V3 16S rRNA gene next generation sequencing, taxa identification, and determination of relative abundance at four time points: baseline prior to surgery (A) and weekly durvalumab treatment timepoints (B, C, and D). Alpha- and beta-diversity differences for the time series were determined in Primerv7. MaAsLin2 in R was used to identify potential associations with the time series and/or HPV status. Linear decomposition model (LDM) R-package was used to investigate the relationship of salivary microbiome with HPV status. ROC curves were plotted for significant species in common between MaAsLin2 analysis and FDR-corrected Mann-Whitney U-test using XLSTAT. Results: Longitudinal microbiome data across four timepoints (A, B, C, D) were obtained (HPV+: n = 18 samples; HPV−: n = 46 samples). A total of 416 taxa were detected across all time points, ranging from 336 to 373 per group. There were no differences inα- andβ-diversities for all longitudinal comparisons (C vs. BCD, AB vs. CD, or A vs. B, C, or D). However, comparison A vs. D showed a significant increase inPrevotella melaninogenicarelative abundance, a potentially pathogenic species able to evade the immune system, after three weeks treatment. Moreover, differences inbeta-diversity based on HPV status were found. LDM analysis identifiedVeillonella atypica, overrepresented in HPV+ group, as the top species accounting for HPV status. Conclusions: The results are consistent with findings from previous studies investigating HNC patients treated with chemoradiotherapy. More research is needed to understand possible impact of immunotherapy on opportunistic bacterial species, although negligible impact from durvalumab treatment on salivary microbiome was observed.
目的：头颈癌（HNC）是一种常见癌症，其中近80%为口腔癌（OC）和口咽癌（OPC）。70%的口咽癌与人乳头瘤病毒（HPV）相关。免疫疗法有望在未来改善头颈癌患者的治疗效果。本研究旨在探讨，在HPV阴性和阳性的口腔癌及口咽癌患者组成的试点队列中，于根治性手术前单独使用度伐利尤单抗免疫治疗是否会显著影响口腔唾液微生物组。方法：招募早期鳞状细胞癌口咽癌患者：5例HPV阳性，12例HPV阴性，在手术前每两周接受两到三次度伐利尤单抗治疗。收集非刺激性唾液样本，进行细菌DNA提取、V1–V3区16S rRNA基因二代测序、分类单元鉴定，并在四个时间点测定相对丰度：手术前基线（A）及每周度伐利尤单抗治疗时间点（B、C、D）。使用Primer v7分析时间序列的α和β多样性差异。使用R语言中的MaAsLin2识别与时间序列和/或HPV状态的可能关联。使用R包线性分解模型（LDM）研究唾液微生物组与HPV状态的关系。利用XLSTAT对MaAsLin2分析与经FDR校正的Mann-Whitney U检验中共同出现的显著物种绘制ROC曲线。结果：获得了四个时间点（A、B、C、D）的纵向微生物组数据（HPV+：n=18个样本；HPV−：n=46个样本）。所有时间点共检测到416个分类单元，每组样本数量在336至373之间。所有纵向比较（C vs. BCD、AB vs. CD或A vs. B、C、D）的α和β多样性均无差异。然而，A与D的比较显示，经过三周治疗后，能够逃避免疫系统的潜在致病菌——产黑色素普雷沃菌的相对丰度显著增加。此外，基于HPV状态的β多样性存在差异。LDM分析发现，在HPV阳性组中过度代表的非典型韦荣球菌是解释HPV状态的首要物种。结论：这些结果与先前研究放化疗头颈癌患者的发现一致。尽管观察到度伐利尤单抗治疗对唾液微生物组的影响微乎其微，但仍需更多研究来理解免疫疗法对机会性细菌物种的可能影响。
肿瘤(癌症)患者之家