Objective　To investigate the epidemiology and genomic characterizations of carbapenem-resistant Escherichia coli (CREC) from a tertiary hospital in Guangzhou, Guangdong province, China from January 2021 to December 2023. Methods　The non-repetitive CREC isolates identified at the hospital from January 2021 to December 2023 were included and their clinical characteristics were analyzed through the clinical record system. A total of 42 CREC isolates were randomly selected for species identification, antimicrobial susceptibility testing, and multilocus sequence typing. Whole-genome sequencing and bioinformatics analysis were conducted to investigate the antimicrobial resistance genes, virulence-related genes, replicons, and plasmid types carrying the carbapenem resistance genes in CREC. Results　A total of 2 887 Escherichia coli (EC) isolates were collected from the hospital between January 2021 to December 2023, including 105 CREC strains. The prevalence rates of CREC over the three years were 1.76%, 2.80%, and 5.69%, respectively, showing a significant upward trend (P<0.001). All 42 CREC isolates showed resistance to multiple antibiotics, with resistance rates for amoxicillin-clavulanate, ceftazidime, and cefepime all at 100.00%, ertapenem and imipenem at 97.62%, levofloxacin at 83.33%, and amikacin at 45.24%. All CREC isolates were susceptible to tigecycline. The 42 CREC strains were classified into 14 ST types, with the main types being ST167 (28.57%), ST410 (26.19%), and ST131 (9.52%). ST410 CREC isolates carried significantly more fim, algA, icsP/sopA, and high-pathogenicity island (HPI) related virulence genes than ST167 isolates (P<0.05), whereas aslA and hcp-2 were significantly lower than ST167 (P<0.05). The carbapenem resistance genes carried by CREC were primarily blaNDM-5 (n=34), followed by blaNDM-1 (n=6), with only one isolate carrying blaKPC-2. These carbapenem resistance genes were mainly located on IncF (n=27) and IncX3 (n=11) type of plasmids. Conclusions　The prevalence of CREC significantly increased from 2021 to 2023 in this hospital, with ST167 and ST410 as the dominant types. ST410 CREC is a newly prevalent high-risk clone carrying adhesion, biofilm formation, and HPI-related virulence genes, and there is an urgent need to monitor its epidemiological status and transmission characteristics to provide a basis for the formulation of control measures.