近年来，随着免疫检查点抑制剂（ICIs）的出现，肾透明细胞癌（ccRCC）的管理发生了快速变化。然而，只有有限数量的患者可以持续对免疫检查点抑制剂产生积极反应，许多患者会对治疗产生耐药性，这为改善全身治疗的疗效提出了额外需求。为了选择最有前景的植物化学物质（PTCs），我们使用在线数据库进行结合概率和靶基因预测，并在ccRCC细胞株中使用侵袭、迁移和凋亡实验，以及抑制癌症干细胞（CSCs）标记物。我们使用混合淋巴细胞肿瘤细胞培养（MLTC）系统和流式细胞术来确认潜在的组合策略。通过NanoString分析，我们确定了潜在的免疫治疗靶点和新型CSC标记物。通过生物信息分析，我们分析了癌组织中VCAM1、CXCL1和IL8的mRNA和蛋白质表达情况，以及在ccRCC中的免疫特征和生存特征。由于其对CSCs的强抑制作用、对患者外周血单个核细胞（PBMCs）中FoxP3+调节性T细胞的显著减少以及对肿瘤特异性抗原的识别导致内源性效应细胞CD3+CD8+和CD3+CD4+ T细胞的活化，shikonin被选为对于ccRCC患者ipilimumab最有前景的有益组合伙伴。根据NanoString分析，VCAM1、CXCL1和IL8被探索为ccRCC潜在的免疫治疗靶点和新型CSC标记物。与正常组织相比，ccRCC肿瘤组织中VCAM1的mRNA和蛋白质水平较高，并且与ccRCC患者的免疫特征和生存特征显著正相关。我们认为，shikonin和ipilimumab的组合可能是一种有前景的治疗策略，而VCAM1则是ccRCC治疗中的一种新型免疫治疗靶点。版权所有 © 2023 Lyu、Stadlbauer、Wang、Buchner和Pohla。

Management of clear cell renal cell carcinoma (ccRCC) has changed rapidly in recent years with the advent of immune checkpoint inhibitors (ICIs). However, only a limited number of patients can sustainably respond to immune checkpoint inhibitors and many patients develop resistance to therapy, creating an additional need for therapeutic strategies to improve the efficacy of systemic therapies.Binding probability and target genes prediction using online databases, invasion, migration, and apoptosis assays as well as the inhibition of cancer stem cells (CSCs) markers in ccRCC cell lines were used to select the most promising phytochemicals (PTCs). Mixed lymphocyte tumor cell culture (MLTC) system and flow cytometry were performed to confirm the potential combination strategy. The potential immunotherapeutic targets and novel CSC markers were identified via the NanoString analysis. The mRNA and protein expression, immune signatures as well as survival characteristics of the marker in ccRCC were analyzed via bioinformation analysis.Shikonin was selected as the most promising beneficial combination partner among 11 PTCs for ipilimumab for the treatment of ccRCC patients due to its strong inhibitory effect on CSCs, the significant reduction of FoxP3+ Treg cells in peripheral blood mononuclear cells (PBMCs) of patients and activation of the endogenous effector CD3+CD8+ and CD3+CD4+ T cells in response to the recognition of tumor specific antigens. Based on NanoString analysis VCAM1, CXCL1 and IL8 were explored as potential immunotherapeutic targets and novel CSC markers in ccRCC. The expression of VCAM1 was higher in the tumor tissue both at mRNA and protein levels in ccRCC compared with normal tissue, and was significantly positively correlated with immune signatures and survival characteristics in ccRCC patients.We propose that a combination of shikonin and ipilimumab could be a promising treatment strategy and VCAM1 a novel immunotherapeutic target for the treatment of ccRCC.Copyright © 2023 Lyu, Stadlbauer, Wang, Buchner and Pohla.