人乳头瘤病毒（HPV）相关的癌症通过促进抑制性肿瘤微环境继续逃避免疫系统的攻击。因此，免疫治疗似乎是一种有前途的方法来针对HPV相关的肿瘤。我们假设作为一个表观遗传剂的丙戊酸（VA）与阿维鲁马布的联合可以增强HPV相关实体肿瘤的抗肿瘤免疫。我们在七个非响应者（NRs）和四个响应者（Rs）的全外周血单个核细胞（PBMCs）上进行了总体RNA测序（RNA-seq）。共分析了39个样本（如术前、VA后、阿维鲁马布后和末期）。此外，我们对血浆分析物进行了量化，并进行了流式细胞术。我们观察到，在NRs与Rs中，VA和/或avelumab治疗后，免疫反应的差异模式。与NRs中髓源性抑制细胞特征相关的主要中性粒细胞外陷阱（NETs）形成和中性粒细胞脱颗粒途径相关联的转录本的显著上调。我们注意到IL-8 / IL-18细胞因子的升高和NRs基线和末期的明显转录组特征。通过使用接受者操作特征，我们确定了血浆IL-8 / IL-18的临界值，用以区分NRs和Rs。我们发现VA和avelumab在NRs与Rs中的治疗效果有差异。因此，我们的结果暗示测量血浆IL-8 / IL-18和PBMCs的总体RNA测序可能作为有价值的生物标志物，以预测免疫治疗的结果。本文受版权保护。保留所有权利。

Human papillomavirus (HPV)-associated cancer continues to evade the immune system by promoting a suppressive tumor-microenvironment. Therefore, immunotherapy appears to be a promising approach for targeting HPV-associated tumors. We hypothesized that valproic acid (VA) as an epigenetic agent combined with avelumab may enhance the anti-tumor immunity in HPV-associated solid tumors. We performed bulk RNA-sequencing (RNA-seq) on total peripheral blood mononuclear cells (PBMCs) of seven non-responders (NRs) and four responders (Rs). A total of 39 samples (e.g. pre-treatment, post-VA, post-avelumab, and endpoint) were analyzed. Also, we quantified plasma analytes and performed flow cytometry. We observed a differential pattern in immune response following treatment with VA and/or avelumab in NRs vs Rs. A significant upregulation of transcripts associated with NETosis [the formation of neutrophil extracellular traps (NETs)] and neutrophil degranulation pathways was linked to the presence of a myeloid-derived suppressor cell signature in NRs. We noted the elevation of IL-8/IL-18 cytokines and a distinct transcriptome signature at the baseline and endpoint in NRs. By using the receiver operator characteristics, we identified a cut-off value for the plasma IL-8/IL-18 to discriminate NRs form Rs. We found differential therapeutic effects for VA and avelumab in NRs vs Rs. Thus, our results imply that measuring the plasma IL-8/IL-18 and bulk RNA-seq of PBMCs may serve as valuable biomarkers to predict immunotherapy outcomes.This article is protected by copyright. All rights reserved.