Retinoblastoma is a pediatric eye cancer affecting children usually less than five years of age. Inactivation of both the copies of the RB1 gene in a child's retina initiates cancer formation. While a lot of literature exists regarding pathogenesis and genetic changes in retinoblastoma, there is still a lack of revelation of specific progression markers and correlation of expression markers with disease mechanism. In this study, we aim to determine transcriptomic, metabolomic and proteomic profile of this cancer from different sample types viz. tissue, aqueous humor, vitreous humor and tear from enucleated eyes of 9 subjects and 2 deceased controls, whose cause of death is not due to any eye related disease. In the first stage of the study, we performed pathway analysis of mRNA and miRNA microarray data looking for gene enrichment that would enable functional characterization of tumors. We were able to identify many of the key pathways that are known to be involved in the progression of retinoblastoma including cell cycle pathway. The study also revealed 18 novel miRNAs which has not been previously implicated in the disease. Combined pathway analysis of metabolomics and transcriptomics data reveal a significant overlap in many of the underlying pathways. The overlapping pathways form the basis of targeted proteomics analysis in tears, that will enable to identify a non-invasive marker to categorize the stage of the disease. We are also studying metabolic dysfunction in the Rb cell line samples using Seahorse analyzer. This will enable measurement of extracellular flux (oxygen consumption rate and extracellular acidification rate) to provide insight into mitochondrial function and glycolytic activity. Overall, the study shows the utility of applying an integrative approach to study molecular mechanisms in retinoblastoma by co-analyzing multi-omics data that will also help identify signatures that are unique to aggressive retinoblastoma.