A Lactate-Related tSNE Signature Defines Prognostic Subtypes of Bladder Cancer and Reveals LINC01094-Mediated VIM Stabilization in Metastasis and Drug Resistance Author List and Affiliations

Pu Zhang¹Hegan Zhang²Wanli Yu³Dage Fan⁴Yao Pan^5*Zhuang Wei^6*Fangzhen Cai^6*Qingliu He^7*

• ¹Department of Urology, Sichuan Academy of Medical Sciences and Sichuan Provincial People's Hospital, Chengdu, Sichuan Province, China
• ²Department of Gynecology, Quanzhou Women and Children’s Hospital, Quanzhou, Fujian Province, China
• ³Department of Vascular Surgery, First Affiliated Hospital of Chongqing Medical University, Chongqing, Chongqing Municipality, China
• ⁴Department of pathology, The Second Affiliated Hospital of Fujian Medical University, Quanzhou, Fujian Province, China
• ⁵Department of Obstetrics and Gynecology, The Second Affiliated Hospital of Fujian Medical University, Quanzhou, Fujian Province, China
• ⁶Department of Urology, The Second Affiliated Hospital of Fujian Medical University, Quanzhou, Fujian Province, China
• ⁷The Second Affiliated Hospital of Fujian Medical University, Quanzhou, China

Background: Bladder cancer (BLCA) is prone to metastasis and often shows poor responses to chemotherapy and immunotherapy. Investigating the underlying mechanisms of metastasis and drug resistance may therefore offer new therapeutic strategies for BLCA.Methods: Publicly available datasets were analyzed using consensus clustering and t-distributed stochastic neighbor embedding (tSNE) to characterize a lactate-related gene signature in BLCA. Gene set variation analysis (GSVA) was employed to assess signaling pathway activity, while immune cell infiltration in the tumor microenvironment (TME) was evaluated using single-sample gene set enrichment analysis (ssGSEA), the Estimation of Stromal and Immune cells in Malignant Tumors using Expression data (ESTIMATE), and CIBERSORT. RNA pull-down and RNAbinding protein immunoprecipitation (RIP) assays were then performed to confirm molecular interactions.Results: Two distinct BLCA subtypes were identified based on lactate-related gene expression, and a lactate-based tSNE score was constructed. This score demonstrated prognostic value and was integrated into a nomogram confirmed by a calibration curve.Functionally, higher tSNE scores correlated with immune-and inflammation-related pathways, as well as with immunotherapy efficacy in BLCA. Among candidate regulators identified, LINC01094 emerged as a key factor in BLCA metastasis and drug resistance. LINC01094 was predominantly localized in the cytoplasm and was upregulated in tumor tissues compared with adjacent normal tissues, acting as an unfavorable prognostic factor. In vitro, LINC01094 promoted metastasis and chemotherapy resistance, potentially by stabilizing VIM protein levels and inhibiting its ubiquitination.This comprehensive analysis of lactate-related genes reveals how this gene signature may shape the tumor microenvironment and affect BLCA patient prognosis. Additionally, our data suggest that targeting LINC01094 with antisense oligonucleotides (ASOs) could reduce BLCA cell metastasis and enhance their sensitivity to chemotherapy.