围手术期免疫检查点抑制剂（ICI）用于中度高危清除型肾细胞癌（ccRCC）的临床试验未能一致证明患者预后的改善。这些未成功的ICI试验表明，肿瘤浸润免疫表型，这里称之为免疫细胞类型、状态及其在肿瘤微环境（TME）中的空间位置不利于ICI治疗。定义肿瘤浸润免疫细胞可能有助于在TME中确定适合ICI治疗的预测性免疫表型。为了界定ccRCC TME中的免疫表型，我们对来自六名在三级转诊医院就诊的ccRCC患者的新鲜邻近肿瘤（pTME，n = 2），低级（LG，n = 4，G1-G2）和高级（HG，n = 4，G3-G4）组织样本进行了空间转录组测序（ST-seq）。在生成的ST-seq数据集中，我们使用多个公共可用的单细胞RNA和T细胞受体测序数据集作为参考，鉴定出了免疫细胞类型和状态，这里称之为精疲力竭/促肿瘤状态或非精疲力竭/抗肿瘤状态。HG TME表现出丰富的精疲力竭/促肿瘤免疫细胞，但未能一致增加PD-1、PD-L1和CTLA4检查点及血管生成基因的表达。HG TME的其他表型特征包括：与组织密切相关的促肿瘤单核细胞，在HAVCR2和LAG3检查点的表达持续增加；一种表达干细胞样祖细胞基因的精疲力竭CD8 + T细胞亚群；以及在复发TME中表达TREM2的促肿瘤肿瘤相关巨噬细胞和单核细胞。尽管样本数量较小，但本研究是迄今为止关于人体ccRCC的最大ST-seq数据集。我们的研究揭示了高风险ccRCC TME受精疲力竭/促肿瘤免疫表型的浸润，缺乏特定检查点基因的表达，证实了HG ccRCC TME具有免疫原性但不利于ICI治疗。©2023. Nature Publishing Group UK.

Perioperative immune checkpoint inhibitor (ICI) trials for intermediate high-risk clear cell renal cell carcinoma (ccRCC) have failed to consistently demonstrate improved patient outcomes. These unsuccessful ICI trials suggest that the tumour infiltrating immunophenotypes, termed here as the immune cell types, states and their spatial location within the tumour microenvironment (TME), were unfavourable for ICI treatment. Defining the tumour infiltrating immune cells may assist with the identification of predictive immunophenotypes within the TME that are favourable for ICI treatment. To define the immunophenotypes within the ccRCC TME, fresh para-tumour (pTME, n = 2), low-grade (LG, n = 4, G1-G2) and high-grade (HG, n = 4, G3-G4) tissue samples from six patients with ccRCC presenting at a tertiary referral hospital underwent spatial transcriptomics sequencing (ST-seq). Within the generated ST-seq datasets, immune cell types and states, termed here as exhausted/pro-tumour state or non-exhausted/anti-tumour state, were identified using multiple publicly available single-cell RNA and T-cell receptor sequencing datasets as references. HG TMEs revealed abundant exhausted/pro-tumour immune cells with no consistent increase in expression of PD-1, PD-L1 and CTLA4 checkpoints and angiogenic genes. Additional HG TME immunophenotype characteristics included: pro-tumour tissue-resident monocytes with consistently increased expression of HAVCR2 and LAG3 checkpoints; an exhausted CD8+ T cells sub-population with stem-like progenitor gene expression; and pro-tumour tumour-associated macrophages and monocytes within the recurrent TME with the expression of TREM2. Whilst limited by a modest sample size, this study represents the largest ST-seq dataset on human ccRCC. Our study reveals that high-risk ccRCC TMEs are infiltrated by exhausted/pro-tumour immunophenotypes lacking specific checkpoint gene expression confirming that HG ccRCC TME are immunogenic but not ICI favourable.© 2023. Nature Publishing Group UK.