Acute lysosomal membrane damage reduces the cellular population of functional lysosomes. However, these damaged lysosomes have a remarkable recovery potential independent of lysosomal biogenesis and remain unaffected in cells depleted in TFEB and TFE3. We combined proximity-labelling-based proteomics, biochemistry and high-resolution microscopy to unravel a lysosomal membrane regeneration pathway that depends on ATG8, the lysosomal membrane protein LIMP2, the RAB7 GTPase-activating protein TBC1D15 and proteins required for autophagic lysosomal reformation, including dynamin-2, kinesin-5B and clathrin. Following lysosomal damage, LIMP2 acts as a lysophagy receptor to bind ATG8, which in turn recruits TBC1D15 to damaged membranes. TBC1D15 interacts with ATG8 proteins on damaged lysosomes and provides a scaffold to assemble and stabilize the autophagic lysosomal reformation machinery. This potentiates the formation of lysosomal tubules and subsequent dynamin-2-dependent scission. TBC1D15-mediated lysosome regeneration was also observed in a cell culture model of oxalate nephropathy.

急性溶酶体膜损伤会减少功能性溶酶体的细胞数量。然而，这些受损的溶酶体具有独立于溶酶体生物发生的显着恢复潜力，并且在 TFEB 和 TFE3 耗尽的细胞中不受影响。我们结合了基于邻近标记的蛋白质组学、生物化学和高分辨率显微镜，揭示了依赖于 ATG8、溶酶体膜蛋白 LIMP2、RAB7 GTPase 激活蛋白 TBC1D15 和自噬溶酶体重组所需的蛋白质（包括动力蛋白）的溶酶体膜再生途径。 -2、驱动蛋白-5B和网格蛋白。溶酶体损伤后，LIMP2 作为溶酶体受体结合 ATG8，ATG8 又将 TBC1D15 募集到受损的膜上。TBC1D15 与受损溶酶体上的 ATG8 蛋白相互作用，并提供组装和稳定自噬溶酶体重组机制的支架。这增强了溶酶体小管的形成和随后的 dynamin-2 依赖性分裂。在草酸盐肾病的细胞培养模型中也观察到 TBC1D15 介导的溶酶体再生。