Feilen Tobias, Rogg Manuel, Andreev Grigor, Pinter Niko, Wess Maximilian, Kössinger Anna L, Diel Nastasja, Neumann-Haefelin Elke, Ganner Athina, Grabbert Markus, Schell Christoph, Schilling Oliver
Institute of Surgical Pathology, Medical Center, Faculty of Medicine, University of Freiburg, Freiburg, Germany; Spemann Graduate School of Biology and Medicine (SGBM), University of Freiburg, Freiburg, Germany; Faculty of Biology, University of Freiburg, Freiburg, Germany.
Institute of Surgical Pathology, Medical Center, Faculty of Medicine, University of Freiburg, Freiburg, Germany.
Neoplasia. 2025 Oct;68:101214. doi: 10.1016/j.neo.2025.101214. Epub 2025 Aug 5.
Von Hippel-Lindau (VHL) disease describes a hereditary tumor predisposition syndrome, caused by germline mutations in the VHL tumor suppressor gene, resulting in the functional loss of the VHL protein (pVHL). pVHL loss translates into a pseudo-hypoxic state that drives clear cell renal cell carcinoma (ccRCC) development. ccRCC tumors frequently form a pseudocapsule (PC) at the tumor boundary. This study describes the first comprehensive proteomic analysis of the PC in ccRCC patients with hereditary VHL inactivation, revealing a distinctive matrisomal signature. We conducted a deep, mass spectrometry-based proteomic analysis of 130 formalin-fixed paraffin-embedded (FFPE) ccRCC samples, comprising 54 tumor, 45 PC, and 31 non-malignant adjacent tissue (NAT) specimens from 34 patients. The PC exhibited unique matrisomal features, with pronounced enrichment of structural extracellular matrix (ECM) components, ECM processing enzymes, and secreted signaling proteins such as TGFβ2. Its proteome composition, including proteins involved in immune response, varied with tumor size and semi-tryptic peptide analysis indicated selective ECM processing in the PC and elevated levels of proteolysis within the tumor. Further, tumor proteomes reflected canonical VHL-driven metabolic reprogramming, including upregulated glycolysis and hypoxia markers, suppressed aerobic metabolism, and dysregulated fatty acid metabolism. Enriched immunoproteasome, MHC-I, and inflammasome proteins indicated an active immune response. Pro-angiogenic factors enriched in the tumor partially extended into the PC. Comparison of primary vs metachronous ccRCC cases uncovered proteomic tumor plasticity in VHL disease. Together, our study delineates the PC as an active, signaling-rich compartment at the ccRCC boundary with potential implications for tumor progression and clinical relevance beyond a mere structural scaffold.
冯·希佩尔-林道（VHL）病是一种遗传性肿瘤易感性综合征，由VHL肿瘤抑制基因的种系突变引起，导致VHL蛋白（pVHL）功能丧失。pVHL缺失转化为一种假性缺氧状态，从而驱动透明细胞肾细胞癌（ccRCC）的发展。ccRCC肿瘤在肿瘤边界处常形成假包膜（PC）。本研究描述了对遗传性VHL失活的ccRCC患者的PC进行的首次全面蛋白质组学分析，揭示了一种独特的基质特征。我们对130个福尔马林固定石蜡包埋（FFPE）的ccRCC样本进行了深度的基于质谱的蛋白质组学分析，这些样本包括来自34名患者的54个肿瘤、45个PC和31个非恶性邻近组织（NAT）标本。PC表现出独特的基质特征，结构细胞外基质（ECM）成分、ECM加工酶和分泌信号蛋白如TGFβ2显著富集。其蛋白质组组成，包括参与免疫反应的蛋白质，随肿瘤大小而变化，半胰蛋白酶肽分析表明PC中存在选择性ECM加工，且肿瘤内蛋白水解水平升高。此外，肿瘤蛋白质组反映了典型的VHL驱动的代谢重编程，包括糖酵解和缺氧标志物上调、有氧代谢受抑制以及脂肪酸代谢失调。富集的免疫蛋白酶体、MHC-I和炎性小体蛋白表明存在活跃的免疫反应。肿瘤中富集的促血管生成因子部分延伸至PC。原发性与异时性ccRCC病例的比较揭示了VHL病中蛋白质组学肿瘤可塑性。总之，我们的研究将PC描绘为ccRCC边界处一个活跃的、富含信号的区域，对肿瘤进展具有潜在影响，且具有超出单纯结构支架的临床相关性。
