This study aimed to identify genes that cooperate with a heterozygous Pten mutation to promote prostate tumor formation. Through in vivo mutagenesis screening, researchers pinpointed two genes, Bzw2 and Eif5a2, linked to protein translation. Further validation using prostate organoid models revealed that either downregulation of Bzw2 or overexpression of EIF5A2 increased organoid size and prostate growth in vivo. Both genes impacted the PI3K pathway, leading to sustained elevation of phospho-AKT levels and reduced PTEN protein expression. Mechanistic investigations unveiled EIF5A2’s direct involvement in PTEN protein translation. Analysis of patient datasets identified EIF5A2 amplifications in various cancers, including prostate, correlating with upregulation of PI3K pathway genes. Organoids with high EIF5A2 levels exhibited sensitivity to AKT inhibitors. These findings suggest a potential treatment strategy for patients with genetic aberrations in these genes, especially EIF5A2-amplified tumors, emphasizing the clinical relevance of targeting the PI3K pathway in prostate cancer.