上尿路尿路上皮癌（UTUC）是一种罕见的尿路上皮癌，复发率高，生存率低。几乎三分之二的 UTUC 在诊断时是侵入性的；因此，改进诊断方法是提高生存率的关键。 UTUC 的组织病理学分析对于诊断至关重要，通常需要内窥镜活检、组织切片和标记。然而，内窥镜活检很微小，并且切割成薄片进行常规组织病理学分析具有挑战性；这使诊断变得复杂。在这里，我们使用体积 3 维 (3D) 成像来探索临床 UTUC 活检的内部景观，无需切片，表明磷酸化核糖体蛋白 S6 (pS6) 的 3D 分析可以更准确地预测肿瘤分级和预后。通过观察肿瘤脉管系统，我们发现 pS6 细胞位于血管附近，在高级别肿瘤中的水平显着高于低级别肿瘤中。此外，pS6 细胞的聚集与较短的无复发生存期相关。我们的结果表明，对 UTUC 样本深处 pS6 细胞的结构壁龛进行 3D 体积成像可以提高诊断率、分级和预后预测。

Upper tract urothelial carcinoma (UTUC) is a rare form of urothelial cancer with a high incidence of recurrence and a low survival rate. Almost two-thirds of UTUCs are invasive at the time of diagnosis; therefore, improving diagnostic methods is key to increasing survival rates. Histopathological analysis of UTUC is essential for diagnosis and typically requires endoscopy biopsy, tissue sectioning, and labeling. However, endoscopy biopsies are minute, and it is challenging to cut into thin sections for conventional histopathology; this complicates diagnosis. Here, we used volumetric 3-dimensional (3D) imaging to explore the inner landscape of clinical UTUC biopsies, without sectioning, revealing that 3D analysis of phosphorylated ribosomal protein S6 (pS6) could predict tumor grade and prognosis with improved accuracy. By visualizing the tumor vasculature, we discovered that pS6+ cells were localized near blood vessels at significantly higher levels in high-grade tumors than in low-grade tumors. Furthermore, the clustering of pS6+ cells was associated with shorter relapse-free survival. Our results demonstrate that 3D volume imaging of the structural niches of pS6 cells deep inside the UTUC samples improved diagnostic yield, grading, and prognosis prediction.