Background: While the genetic characteristics of muscle-invasive bladder cancer have been studied in detail, there are few reports using the same genetic analysis platform to investigate differences and similarities among urothelial carcinoma (UC) derived from different sites and with differing degrees of invasion. Methods: We used targeted exome sequencing with Oncopanel to study 82 low-grade (LG) non-muscle invasive bladder cancer tumors (LG-NMIBC), 127 high-grade (HG) NMIBC, 199 muscle-invasive bladder cancer tumors (MIBC), and 55 HG invasive upper tract urothelial carcinoma (UTUC) tissue samples. Oncopanel assesses 275-447 cancer genes for somatic mutations and copy number alterations without a paired normal. All single nucleotide variants (SNVs) that were considered likely germline were excluded. Clinico-pathological characteristics, mutation frequencies for 238, and tumor mutation burden (TMB) were determined for the 4 types of UC, and were compared using the χ² test and Kruskall Wallis Test. Nominal p values were obtained, and the Benjamini-Hochberg fdr correction was employed to determine statistical significance (q < 0.1). Results: Key results are shown in the Table. Conclusions: In this series of uniformly analyzed UC samples of different kinds, we found that: 1) FGFR3, STAG2 and KDM6A mutations are highly enriched in LG-NMIBC; 2) HG-NMIBC, MIBC, and UTUC all have similar genetic alterations; and 3) TP53 and RB1 mutations are associated with all of the latter 3 types of UC in contrast to LG-NMIBC.