Department of Pathology, Yale University School of Medicine, New Haven, Connecticut, USA.
Department of Pathology, Yale School of Medicine, New Haven, Connecticut, USA.
J Immunother Cancer. 2024 Jun 10;12(6):e009039. doi: 10.1136/jitc-2024-009039.
Despite the impressive outcomes with immune checkpoint inhibitor (ICI) in non-small cell lung cancer (NSCLC), only a minority of the patients show long-term benefits from ICI. In this study, we used retrospective cohorts of ICI treated patients with NSCLC to discover and validate spatially resolved protein markers associated with resistance to programmed cell death protein-1 (PD-1) axis inhibition.
Pretreatment samples from 56 patients with NSCLC treated with ICI were collected and analyzed in a tissue microarray (TMA) format in including four different tumor regions per patient using the GeoMx platform for spatially informed transcriptomics. 34 patients had assessable tissue with tumor compartment in all 4 TMA spots, 22 with leukocyte compartment and 12 with CD68 compartment. The patients' tissue that was not assessable in fourfold redundancy in each compartment was designated as the validation cohort; cytokeratin (CK) (N=22), leukocytes CD45 (N=31), macrophages, CD68 (N=43). The human whole transcriptome, represented by~18,000 individual genes assessed by oligonucleotide-tagged in situ hybridization, was sequenced on the NovaSeq platform to quantify the RNAs present in each region of interest.
54,000 gene variables were generated per case, from them 25,740 were analyzed after removing targets with expression lower than a prespecified frequency. Cox proportional-hazards model analysis was performed for overall and progression-free survival (OS, PFS, respectively). After identifying genes significantly associated with limited survival benefit (HR>1)/progression per spot per patient, we used the intersection of them across the four TMA spots per patient. This resulted in a list of 12 genes in the tumor-cell compartment ( in tumor-cell compartment were also significantly associated with OS and PFS, respectively, in the validation cohort (CK: HR, 2.48; p=0.02 and HR, 5.33; p=0.04). In CD45 compartment, secreted frizzled-related protein 2, was associated with OS in the discovery cohort but not in the validation cohort. Similarly, in the CD68 compartment and PNN interacting serine and arginine rich protein were significantly associated with PFS and OS, respectively, in the majority but not all four spots per patient.
This work highlights and as potential indicative biomarkers of resistance to PD-1 axis blockade that might help to improve precision immunotherapy strategies for lung cancer.
尽管免疫检查点抑制剂（ICI）在非小细胞肺癌（NSCLC）中取得了令人瞩目的疗效，但只有少数患者能从中长期获益。在这项研究中，我们使用接受 ICI 治疗的 NSCLC 患者的回顾性队列，发现并验证了与程序性死亡蛋白-1（PD-1）轴抑制耐药相关的空间分辨蛋白标志物。
收集了 56 名接受 ICI 治疗的 NSCLC 患者的预处理样本，并在组织微阵列（TMA）格式中进行分析，每个患者包括四个不同的肿瘤区域，使用 GeoMx 平台进行空间转录组学分析。34 名患者的 TMA 四个重复区域均有可评估的肿瘤组织，22 名患者有可评估的白细胞组织，12 名患者有可评估的 CD68 组织。每个区域均有不可评估的组织被指定为验证队列；细胞角蛋白（CK）（N=22）、白细胞 CD45（N=31）、巨噬细胞、CD68（N=43）。通过寡核苷酸标记原位杂交评估了约 18000 个个体基因的人类全转录组，并在 NovaSeq 平台上对其进行测序，以定量分析每个感兴趣区域的 RNA。
每个病例生成了 54000 个基因变量，其中 25740 个在去除表达频率低于预设值的靶标后进行了分析。对总生存期（OS）和无进展生存期（PFS）进行了 Cox 比例风险模型分析。在确定了与生存获益受限（HR>1）/每个患者每个 TMA 点进展相关的基因后，我们在每个患者的四个 TMA 点之间使用它们的交集。这导致了肿瘤细胞区室中 12 个基因的列表（在肿瘤细胞区室中也与 OS 和 PFS 显著相关，在验证队列中也是如此（CK：HR，2.48；p=0.02 和 HR，5.33；p=0.04）。在 CD45 区室中，分泌卷曲相关蛋白 2 与发现队列中的 OS 相关，但与验证队列无关。同样，在 CD68 区室中，PNN 相互作用丝氨酸和精氨酸丰富蛋白与 PFS 和 OS 显著相关，分别在大多数患者中，而不是所有四个患者的每个 TMA 点。
这项工作突出了 和 作为 PD-1 轴阻断耐药的潜在指示性生物标志物，可能有助于改善肺癌的精准免疫治疗策略。
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