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TLS are organized ectopic lymphoid aggregates that facilitate local antigen presentation, T and B cell activation, and memory immune responses within the tumor microenvironment. Their presence and maturation status correlate with favorable clinical outcomes and enhanced responsiveness to immune checkpoint inhibitors. Despite these insights, several obstacles hinder clinical translation. Current approaches remain limited in dynamically monitoring TLS evolution during treatment, and the underlying immunoregulatory mechanisms\u2014particularly interactions among immune subsets, stromal components, and tumor cells\u2014are not fully elucidated. Additionally, translation faces challenges including inter\u2010institutional variability, lack of standardized assessment criteria, and the need for multi\u2010center prospective validation. Integrating multimodal technologies\u2014high\u2010plex imaging, spatial transcriptomics, and digital pathology\u2014with AI\u2010based platforms holds promise for refined TLS characterization and real\u2010time monitoring. Near\u2010term priorities include optimizing dynamic monitoring techniques and establishing standardized scoring systems. Medium\u2010to\u2010long\u2010term priorities encompass multi\u2010center trials to validate TLS utility across diverse populations, identify novel drivers of TLS formation, and develop optimized combinatorial immunotherapy regimens that leverage TLS as both predictive biomarkers and therapeutic targets. 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