Immunotherapy has reshaped the oncology landscape by enabling the immune system to recognize and eliminate malignant cells. Although immune checkpoint inhibitors (ICIs) targeting cytotoxic T-lymphocyte-associated protein 4 (CTLA-4), programmed cell death-1 (PD-1), and programmed death-ligand 1 (PD-L1) have achieved durable responses in several cancers, their therapeutic benefit remains limited to a subset of patients, largely due to immune evasion, tumor heterogeneity, and immunosuppressive features of the tumor microenvironment (TME). This review comprehensively examines the expanding landscape of next-generation immune checkpoints, encompassing both co-inhibitory (lymphocyte activation gene-3 [LAG-3], T cell immunoglobulin and mucin-domain containing-3 [TIM-3], TIGIT, VISTA, and IGSF8) and co-stimulatory (ICOS, OX40, GITR, 4-1BB, CD40, and CD27) pathways that collectively regulate the balance between immune activation and tolerance. We discuss their molecular mechanisms, translational rationale, and emerging clinical evidence, highlighting their potential to reinvigorate antitumor immunity, particularly in ICI-refractory settings. Beyond checkpoint modulation, we explore complementary strategies aimed at remodeling the TME and enhancing immune responsiveness, including targeting immunometabolic pathways (IDO1 and CD73), innate immune sensing (toll-like receptors [TLRs]), cytokine signaling (IL-2), micronutrient immunoregulators (vitamin D), and the gut microbiota. The integration of these approaches into rational combination regimens, guided by predictive features such as T cell infiltration, tumor mutational burden (TMB), and microbiome composition, holds promise for extending the clinical impact of immunotherapy across malignancies. We further advocate for a precision immuno-oncology framework that leverages multiomic profiling, systems biology, and artificial intelligence (AI) to optimize therapeutic selection and mitigate immune-related toxicities. Emerging advances in nanomedicine, synthetic biology, and chronotherapy offer additional opportunities to enhance therapeutic specificity and durability, collectively charting a path from mechanistic insight to clinical translation toward realizing the full curative potential of cancer immunotherapy. Trial Registration: ClinicalTrials.gov identifier: LAG-3 (NCT03470922, NCT04082364, NCT05064059, NCT05352672, NCT02614833, NCT03625323, NCT01968109), TIM-3 (NCT03307785, NCT03680508, NCT02608268, NCT02817633), TIGIT (NCT03563716), VISTA (NCT02671955, NCT02812875), IGSF8 (NCT05669430), CD73 (NCT02503774), B7-H3 (NCT02475213, NCT01391143, NCT02628535, NCT03406949, NCT00089245, NCT01099644, NCT01502917), OX40 (NCT01862900, NCT02315066, NCT02410512, NCT02221960, NCT02528357, NCT02923349, NCT02705482), CD27 (NCT02335918, NCT02924038, NCT02302339, NCT02386111, NCT02543645), 4-1BB (NCT01307267, NCT02444793, NCT01471210, NCT02253992, NCT02554812), CD40 (NCT02588443, NCT03329950), ICOS (NCT02904226, NCT02723955, NCT03251924), GITR (NCT02583165, NCT02132754, NCT02697591, NCT03126110, NCT02740270, NCT02598960, NCT01239134, NCT02628574), IDO1 (NCT02752074, NCT02658890, NCT02077881, NCT01560923, NCT02073123, NCT02327078, NCT02178722), TLRs (NCT02556463, NCT02042781), and IL-2-based therapies (NCT02869295, NCT02983045).
