Background: Ventilator-associated pneumonia (VAP) is an important cause of healthcare-associated infections, resulting in prolonged hospitalization with increased morbidity and mortality. Knowledge of predominant local pathogens and their antimicrobial susceptibility patterns helps in selection of appropriate initial antibiotic therapy in these critical cases. Aim and objective: The aim and objective of this study is to characterize the microbiology and antimicrobial susceptibility patterns of VAP isolates in a tertiary cancer center. Materials and methods: This is a 4-year qualitative observational study carried out at a tertiary care cancer hospital in Mumbai. All nondirect bronchoalveolar lavage specimens from patients with a clinical suspicion of VAP sent from the critical care unit to the department of microbiology were processed as per standard laboratory procedures. All isolates were identified to species level and an antimicrobial susceptibility testing was performed by the Kirby–Bauer disk diffusion method and/or the VITEK 2 automated identification and susceptibility system, according to Clinical and Laboratory Standards Institute guidelines. Results: The study comprised 1,074 patients: 710 (66.10%) men and 364 (33.90%) women. A total of 827 bacterial isolates were obtained with 780 (94.32%) gram-negative organisms and 47 (5.68%) gram-positive organisms; of which Acinetobacter baumannii (38.7%), Pseudomonas aeruginosa (17.5%), and Klebsiella pneumoniae (16.6%) were the commonest. Of gram-negative bacilli, multidrug-resistant organisms constituted 87.50% and were susceptible to colistin. Conclusions: VAP is associated with pathogens, such as A. baumannii, P. aeruginosa, and K. pneumoniae in our setting. High rates of resistance to aminoglycosides, β-lactam-β-lactamase inhibitor combinations, and carbapenems were noted.