Integrated Transcriptomics and Machine Learning Reveal REN as a Dual Regulator of Tumor Stemness and NK Cell Evasion in Wilms Tumor Progression

Qingfei Cao¹Junyi Li^1*Yunfei Zou²Changwen Xu¹Huihui Tang¹Meixue Chen^3*

• ¹Department of Urology, First Affiliated Hospital of Jinzhou Medical University, Jinzhou, China
• ²Department of Cardiology, First Affiliated Hospital of Jinzhou Medical University, Jinzhou, China
• ³Department of Pediatric, First Affiliated Hospital of Jinzhou Medical University, Jinzhou, China

Wilms tumor (WT), the most prevalent pediatric kidney cancer, poses a clinical challenge in highrisk cases due to chemotherapy resistance and immunosuppressive tumor microenvironments (TME).Through an integrative analysis combining single-cell RNA sequencing (scRNA-seq), spatial transcriptomics, bulk RNA-seq, and advanced machine learning techniques, we identify the renin gene (REN) as a central regulator of tumor stemness and natural killer (NK) cell evasion. Our novel machine learning-derived Cancer Stemness Prognostic Index (CSPI) stratifies WT patients into distinct risk groups and accurately distinguishes histological subtypes of WT. High-CSPI tumors exhibit enhanced tumor stemness, metabolic reprogramming (ROS/oxidative phosphorylation), and suppressed immune activity. Spatial transcriptomics validated the spatial distribution of key stemness-related genes and revealed physical proximity between tumor cells and NK cells among different WT histological subtypes. Moreover, at spatial and single-cell resolution, our data show that REN-expressing tumor cells drive NK cell exhaustion-rather than T cell dysfunction-via PTN-NCL and COL4A1-CD44 ligand-receptor interactions. Molecular docking further identified estrogen-based compounds as potential REN inhibitors. Functional assays revealed that REN knockdown significantly hampers tumor proliferation, migration, and survival in vitro. These findings establish REN as a promising dual therapeutic target for inhibiting tumor stemness and restoring NK-mediated immune surveillance, thus offering transformative opportunities for precision oncology in WT.