Through genetic and cellular studies, we demonstrated that nutrition repletion inactivates AMPK-TBC1D1 signaling, augments the level of GTP-bound Rab2A, and then increases the protein stability of PPARγ and the expression of PPARγ target genes, and ultimately contributes to the development of NAFLD. Here, TBC1D1 is linked to metabolic dysfunction-associated steatotic liver disease.