Zhang Xiaotian, Jeong Heykyeong, Niu Jingwen, Holland Sabrina M, Rotanz Brittany N, Gordon John, Einarson Margret B, Childers Wayne E, Thomas Gareth M
Center for Neural Development and Repair, Lewis Katz School of Medicine at Temple University, Philadelphia, PA, 19140, USA.
Moulder Center for Drug Discovery, School of Pharmacy, Temple University, Philadelphia, PA, USA.
Nat Commun. 2025 Apr 3;16(1):3031. doi: 10.1038/s41467-025-58036-6.
Inhibiting dual leucine-zipper kinase (DLK) could potentially ameliorate diverse neuropathological conditions, but a direct inhibitor of DLK's kinase domain caused unintended side effects in human patients, indicative of neuronal cytoskeletal disruption. We sought a more precise intervention and show here that axon-to-soma pro-degenerative signaling requires acute, axonal palmitoylation of DLK. To identify potential modulators of this modification, we screened >28,000 compounds using a high-content imaging readout of DLK's palmitoylation-dependent subcellular localization. Several hits alter DLK localization in non-neuronal cells, reduce DLK retrograde signaling and protect cultured dorsal root ganglion neurons from neurodegeneration. Mechanistically, the two most neuroprotective compounds selectively prevent DLK's stimulus-dependent palmitoylation and subsequent recruitment to axonal vesicles, but do not affect palmitoylation of other axonal proteins assessed and avoid the cytoskeletal disruption associated with direct DLK inhibition. Our hit compounds also reduce pro-degenerative retrograde signaling in vivo, revealing a previously unrecognized neuroprotective strategy.
抑制双亮氨酸拉链激酶（DLK）可能会改善多种神经病理状况，但DLK激酶结构域的直接抑制剂在人类患者中会产生意外的副作用，这表明神经元细胞骨架受到破坏。我们寻求一种更精确的干预措施，并在此表明轴突到胞体的促变性信号传导需要DLK的急性轴突棕榈酰化。为了确定这种修饰的潜在调节剂，我们使用DLK棕榈酰化依赖性亚细胞定位的高内涵成像读数筛选了超过28000种化合物。有几种化合物改变了非神经元细胞中DLK的定位，减少了DLK的逆行信号传导，并保护培养的背根神经节神经元免于神经变性。从机制上讲，两种最具神经保护作用的化合物选择性地阻止了DLK依赖刺激的棕榈酰化以及随后向轴突囊泡的募集，但不影响所评估的其他轴突蛋白的棕榈酰化，并且避免了与直接抑制DLK相关的细胞骨架破坏。我们筛选出的化合物还减少了体内促变性逆行信号传导，揭示了一种以前未被认识的神经保护策略。
