Canagliflozin as a Potential Preclinical Therapy for Tuberous Sclerosis Complex: Inhibition of Tsc2 -/- Cell Proliferation via Cell Cycle Arrest and Mitochondrial Dysfunction.

Purpose: Canagliflozin (Ca), a sodium-glucose cotransporter 2 (SGLT2) inhibitor traditionally used for type 2 diabetes, has shown potential in the treatment of lymphangiomatosis (the pulmonary lesion phenotype of TSC). However, its effects on Tsc2 -/- cells, a key feature of tuberous sclerosis complex (TSC), have not been previously explored. This preclinical study aimed to investigate Ca's inhibitory mechanisms on Tsc2 -/- cell proliferation and its therapeutic potential in TSC-related lesions. Methods: The effects of Ca on Tsc2 -/- cells were evaluated using in vitro cellular assays, including proliferation, cell cycle, and mitochondrial function analyses, as well as proteomics. In vivo, a mouse xenograft model was employed to assess tumor growth inhibition and safety profile. Comparative studies with other SGLT2 inhibitors were conducted to identify compound-specific mechanisms. Results: Ca significantly inhibited Tsc2 -/- cell proliferation in a dose-dependent manner, inducing G1 phase cell cycle arrest and impairing mitochondrial function, as evidenced by reduced membrane potential and ATP production. Proteomic analysis revealed mitochondrial protein alterations, and Ca-induced ROS accumulation promoted apoptosis. In vivo, Ca (100 mg/kg/day) effectively suppressed tumor growth without significant adverse effects. Notably, Ca's effects were unique compared to other SGLT2 inhibitors, indicating mechanisms independent of SGLT2 inhibition. Conclusion: Ca inhibits Tsc2 -/- cell proliferation through dual mechanisms of cell cycle arrest and mitochondrial impairment, demonstrating significant therapeutic potential for TSC-related lesions. These findings highlight Ca as a promising alternative to current mTOR inhibitors, warranting further investigation into its molecular targets and clinical applications.