ALDH1L1過度表現促進尿路上皮癌腫瘤進展與血管新生，並為不良預後的預測指標

葉碧雯^1,2,8、李威明^1,2,3、李經家^1,2,3、韋又菁⁴、葉信志^1,2、李香瑩^1,2、林崇裕^1,2、孔美蘭⁵、

李學德⁶、李健逢^7*、吳文正^1,2,8,9*

¹高雄醫學大學 泌尿學科，²高雄醫學大學附設醫院 泌尿科，³高雄醫學大學岡山醫院 泌尿科，⁴高高雄醫學大學岡山醫院 病理科， ⁵高雄榮民總醫院 醫學教育與研究科，⁶國立陽明交通大學 解剖與細胞生物學研究所，⁷奇美醫院 醫學研究部，⁸高雄醫學大學 再生醫學與細胞治療中心，⁹國立中山大學 醫學科學技術研究所

ALDH1L1 Overexpression Promotes Tumor Progression and Angiogenesis and Serves as a Predictor of Poor Prognosis in Urothelial Carcinoma

Bi-Wen Yeh^1,2,8、Wei-Ming Li^1,2,3、Ching-Chia Li^1,2,3、Yu-Ching Wei⁴、Hsin-Chih Yeh^1,2、Hsiang-Ying Lee^1,2、Chung Yu Lin^1,2、Mei-Lang Kung⁵、Hsueh-Te Lee⁶、 Chien-Feng Li⁷*、 and Wen-Jeng Wu^1,2,8,9*

¹Department of Urology, School of Medicine, College of Medicine, Kaohsiung Medical University (KMU), Kaohsiung; ²Department of Urology, KMU Hospital, Kaohsiung; ³Department of Urology, Kaohsiung Medical University Gangshan Hospital, Kaohsiung; ⁴Department of Pathology, Kaohsiung Medical University Gangshan Hospital, Kaohsiung; ⁵Department of Medical Education and Research, Kaohsiung Veterans General Hospital, Kaohsiung; ⁶Institute of Anatomy and Cell Biology, National Yang Ming Chiao Tung University, Taipei; ⁷Department of Medical Research, Chi-Mei Medical Center, Tainan; ⁸Regenerative Medicine and Cell Therapy Research Center, KMU, Kaohsiung; ⁹Institute of Medical Science and Technology, National Sun Yat-sen University, Kaohsiung, Taiwan

Purposes: Cancer progression is driven by a complex interplay of molecular events. In this study, we investigated the pivotal role of ALDH1L1 in urothelial carcinoma (UC), with particular emphasis on its impact on angiogenesis and tumor aggressiveness. By manipulating ALDH1L1 expression in UC cells, we observed significant changes in angiogenesis, cell proliferation, migration, and invasion, highlighting its multifaceted role in UC progression. Among these, angiogenesis-a hallmark of cancer-emerged as the most prominent feature associated with ALDH1L1. This study further aimed to elucidate the roles of ALDH1L1 in tumor progression, inflammation, angiogenesis, and its potential as a therapeutic target in UC.

Materials and Methods: We established a well-characterized cohort comprising 295 patients with urinary bladder urothelial carcinoma (UBUC) and 340 patients with upper tract urothelial carcinoma (UTUC) to evaluate the clinical significance of ALDH1L1. ALDH1L1 expression levels were assessed and correlated with various clinicopathological parameters.

Results: In UTUC, high ALDH1L1 expression was significantly associated with advanced pathological stage (P<0.001), high histological grade (P<0.001), vascular invasion (VI; P<0.001), increased mitotic activity (P<0.001), lymph node metastasis (P=0.049), and showed a borderline association with perineural invasion (PNI; P = 0.098). Similar findings were observed in UBUC, where elevated ALDH1L1 expression correlated with advanced stage (P<0.001), high grade (P<0.001), vascular invasion (P<0.001), lymph node metastasis (P=0.004), and perineural invasion (P=0.001). Overall, high ALDH1L1 expressions was associated with advanced tumor stage, nodal metastasis, high histological grade, vascular and perineural invasion, and increased mitotic activity. Moreover, patients with high ALDH1L1 expression had significantly worse disease-specific survival (DSS) and metastasis-free survival (MeFS) compared with those with low expression levels. In multivariate analysis, ALDH1L1 expression remained an independent predictor of DSS and MeFS, suggesting its potential as a novel prognostic biomarker and therapeutic target. To further investigate its functional significance, we evaluated the association between ALDH1L1 expression and tumor microvascular density (MVD). Quantification of CD31-positive vessels using ImageJ revealed that patients with high ALDH1L1 expression exhibited significantly higher MVD in both UTUC and UBUC cohorts, supporting a pro-angiogenic role of ALDH1L1 in UC progression.

Conclusions: Our findings demonstrate that ALDH1L1 plays a critical role in the pathogenesis and progression of UC. Targeting ALDH1L1, either as monotherapy or in combination with the inhibition of related signaling pathways, may offer therapeutic benefits for patients with ALDH1L1-overexpressing UC. Ongoing studies are focused on elucidating the underlying molecular mechanisms and developing effective targeted therapeutic strategies, including potential combination approaches with immune checkpoint inhibitors (ICIs).