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Abstract：
BACKGROUND: Oropharyngeal squamous cell carcinoma (OPSCC) induced by human papillomavirus (HPV-positive) is associated with better clinical outcomes than HPV-negative OPSCC. However, the clinical benefits of immunotherapy in patients with HPV-positive OPSCC remain unclear.
METHODS: To identify the cellular and molecular factors that limited the benefits associated with HPV in OPSCC immunotherapy, we performed single-cell RNA (n=20) and T-cell receptor sequencing (n=10) analyses of tonsil or base of tongue tumor biopsies prior to immunotherapy. Primary findings from our single-cell analysis were confirmed through immunofluorescence experiments, and secondary validation analysis were performed via publicly available transcriptomics data sets.
RESULTS: We found significantly higher transcriptional diversity of malignant cells among non-responders to immunotherapy, regardless of HPV infection status. We also observed a significantly larger proportion of CD4+ follicular helper T cells (Tfh) in HPV-positive tumors, potentially due to enhanced Tfh differentiation. Most importantly, CD8+ resident memory T cells (Trm) with elevated KLRB1 (encoding CD161) expression showed an association with dampened antitumor activity in patients with HPV-positive OPSCC, which may explain their heterogeneous clinical outcomes. Notably, all HPV-positive patients, whose Trm presented elevated KLRB1 levels, showed low expression of CLEC2D (encoding the CD161 ligand) in B cells, which may reduce tertiary lymphoid structure activity. Immunofluorescence of HPV-positive tumors treated with immune checkpoint blockade showed an inverse correlation between the density of CD161+ Trm and changes in tumor size.
CONCLUSIONS: We found that CD161+ Trm counteracts clinical benefits associated with HPV in OPSCC immunotherapy. This suggests that targeted inhibition of CD161 in Trm could enhance the efficacy of immunotherapy in HPV-positive oropharyngeal cancers.
BACKGROUND: NCT03737968.
METHODS: To identify the cellular and molecular factors that limited the benefits associated with HPV in OPSCC immunotherapy, we performed single-cell RNA (n=20) and T-cell receptor sequencing (n=10) analyses of tonsil or base of tongue tumor biopsies prior to immunotherapy. Primary findings from our single-cell analysis were confirmed through immunofluorescence experiments, and secondary validation analysis were performed via publicly available transcriptomics data sets.
RESULTS: We found significantly higher transcriptional diversity of malignant cells among non-responders to immunotherapy, regardless of HPV infection status. We also observed a significantly larger proportion of CD4+ follicular helper T cells (Tfh) in HPV-positive tumors, potentially due to enhanced Tfh differentiation. Most importantly, CD8+ resident memory T cells (Trm) with elevated KLRB1 (encoding CD161) expression showed an association with dampened antitumor activity in patients with HPV-positive OPSCC, which may explain their heterogeneous clinical outcomes. Notably, all HPV-positive patients, whose Trm presented elevated KLRB1 levels, showed low expression of CLEC2D (encoding the CD161 ligand) in B cells, which may reduce tertiary lymphoid structure activity. Immunofluorescence of HPV-positive tumors treated with immune checkpoint blockade showed an inverse correlation between the density of CD161+ Trm and changes in tumor size.
CONCLUSIONS: We found that CD161+ Trm counteracts clinical benefits associated with HPV in OPSCC immunotherapy. This suggests that targeted inhibition of CD161 in Trm could enhance the efficacy of immunotherapy in HPV-positive oropharyngeal cancers.
BACKGROUND: NCT03737968.
摘要：
背景：人乳头瘤病毒（HPV阳性）诱导的口咽鳞状细胞癌（OPSCC）比HPV阴性的OPSCC具有更好的临床结局。然而,免疫治疗在HPV阳性OPSCC患者中的临床获益尚不清楚.
方法：为了确定在OPSCC免疫治疗中限制HPV相关益处的细胞和分子因素，我们在免疫治疗前对扁桃体或舌根肿瘤活检进行了单细胞RNA(n=20)和T细胞受体测序(n=10)分析.我们的单细胞分析的主要发现通过免疫荧光实验得到证实，和二次验证分析通过公开可用的转录组学数据集进行.
结果：我们发现免疫疗法无反应者中恶性细胞的转录多样性明显更高，无论HPV感染状况如何。我们还观察到在HPV阳性肿瘤中CD4+滤泡辅助性T细胞（Tfh）的比例明显更高，可能是由于Tfh分化增强。最重要的是,KLRB1（编码CD161）表达升高的CD8常驻记忆T细胞（Trm）与HPV阳性OPSCC患者的抗肿瘤活性减弱有关，这可以解释他们的异质性临床结果。值得注意的是,所有HPV阳性患者，Trm的KLRB1水平升高，显示CLEC2D（编码CD161配体）在B细胞中的低表达，这可能会降低三级淋巴结构的活性。用免疫检查点阻断治疗的HPV阳性肿瘤的免疫荧光显示CD161+Trm的密度与肿瘤大小的变化之间的负相关。
结论：我们发现CD161+Trm抵消了OPSCC免疫治疗中与HPV相关的临床益处。这表明在Trm中靶向抑制CD161可以增强HPV阳性口咽癌中免疫疗法的功效。
背景：NCT03737968。
方法：为了确定在OPSCC免疫治疗中限制HPV相关益处的细胞和分子因素，我们在免疫治疗前对扁桃体或舌根肿瘤活检进行了单细胞RNA(n=20)和T细胞受体测序(n=10)分析.我们的单细胞分析的主要发现通过免疫荧光实验得到证实，和二次验证分析通过公开可用的转录组学数据集进行.
结果：我们发现免疫疗法无反应者中恶性细胞的转录多样性明显更高，无论HPV感染状况如何。我们还观察到在HPV阳性肿瘤中CD4+滤泡辅助性T细胞（Tfh）的比例明显更高，可能是由于Tfh分化增强。最重要的是,KLRB1（编码CD161）表达升高的CD8常驻记忆T细胞（Trm）与HPV阳性OPSCC患者的抗肿瘤活性减弱有关，这可以解释他们的异质性临床结果。值得注意的是,所有HPV阳性患者，Trm的KLRB1水平升高，显示CLEC2D（编码CD161配体）在B细胞中的低表达，这可能会降低三级淋巴结构的活性。用免疫检查点阻断治疗的HPV阳性肿瘤的免疫荧光显示CD161+Trm的密度与肿瘤大小的变化之间的负相关。
结论：我们发现CD161+Trm抵消了OPSCC免疫治疗中与HPV相关的临床益处。这表明在Trm中靶向抑制CD161可以增强HPV阳性口咽癌中免疫疗法的功效。
背景：NCT03737968。
