This study investigates the use of mitochondria-targeted nano-adjuvants to enhance the effectiveness of radio-immunotherapy (RT) in treating tumors. The researchers found that collagen acts as a physical barrier, inhibiting oxygen perfusion and immune cell infiltration in solid tumors. Additionally, RT can induce DNA damage repair and T cell exclusion, reducing its efficacy. To address these issues, the team developed IR-LND@Lip nano-adjuvants, which combine the mitochondria-targeting molecule IR-68 with the oxidative phosphorylation inhibitor Lonidamine (LND) and are encapsulated in liposomes. This combination effectively sensitizes RT by generating more DNA damage and converting cold tumors into hot ones through the co-inhibition of PD-L1, collagen, and TGF-β. In vitro and in vivo experiments demonstrated that IR-LND@Lip significantly enhances the efficacy of RT by improving oxygen perfusion, reducing tumor hypoxia, and activating T cells. The combination therapy of IR-LND@Lip and RT almost completely suppressed the growth of bladder and breast tumors, highlighting its potential for clinical application in tumor therapy.This study investigates the use of mitochondria-targeted nano-adjuvants to enhance the effectiveness of radio-immunotherapy (RT) in treating tumors. The researchers found that collagen acts as a physical barrier, inhibiting oxygen perfusion and immune cell infiltration in solid tumors. Additionally, RT can induce DNA damage repair and T cell exclusion, reducing its efficacy. To address these issues, the team developed IR-LND@Lip nano-adjuvants, which combine the mitochondria-targeting molecule IR-68 with the oxidative phosphorylation inhibitor Lonidamine (LND) and are encapsulated in liposomes. This combination effectively sensitizes RT by generating more DNA damage and converting cold tumors into hot ones through the co-inhibition of PD-L1, collagen, and TGF-β. In vitro and in vivo experiments demonstrated that IR-LND@Lip significantly enhances the efficacy of RT by improving oxygen perfusion, reducing tumor hypoxia, and activating T cells. The combination therapy of IR-LND@Lip and RT almost completely suppressed the growth of bladder and breast tumors, highlighting its potential for clinical application in tumor therapy.