Gómez-Deza Jorge, Nebiyou Matthew, El Touny Lara H, Alkaslasi Mor R, Zuo Zhenyu, Wlaschin Josette J, Nadal-Nicolas Francisco M, Slavutsky Anastasia L, Lloyd Eliza Y H, Hayashi Peter M, Ashby Nathan, Sohn Mira, Dale Ryan, Li Wei, Cheng Ken Chih-Chien, Rocha Pedro P, Le Pichon Claire E
Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, USA.
current affiliation: Department of Cancer and Cellular Biology, Lewis Katz School of Medicine, Temple University, Philadelphia, PA, USA; Fox Chase Cancer Center, Cancer Signaling and Microenvironment Program, Philadelphia, PA, USA.
bioRxiv. 2025 May 8:2023.09.27.559856. doi: 10.1101/2023.09.27.559856.
Apoptotic neuron death is a key feature of neurodegenerative disease. Considerable efforts have been made to target this pathway but the molecular mechanisms remain incompletely understood. Here, we conducted an unbiased whole genome CRISPR inhibition screen in human neurons to discover genes required for their death and identified known targets including the kinase MAP3K12 (DLK) and the transcription factor JUN. In addition, this screen revealed a potential role for the transcription factor ATF2. We demonstrate that ATF2 phosphorylation by MAP3 kinases is the core driver of the pro-apoptotic transcriptional response. Surprisingly, JUN phosphorylation is not required for apoptosis. However, the phosphorylation of ATF2 and upregulation of JUN expression are crucial. ATF2 therefore converts the kinase signal into a transcriptional response. Inhibiting ATF2 in cultured human neurons prevents cell death. Notably we show that ATF2 knockdown is neuroprotective in injury models . Thus, ATF2 provides a promising new target for a wide range of neurodegenerative disorders.
凋亡性神经元死亡是神经退行性疾病的一个关键特征。人们已经付出了相当大的努力来针对这一途径，但分子机制仍未完全清楚。在这里，我们在人类神经元中进行了无偏差的全基因组CRISPR抑制筛选，以发现其死亡所需的基因，并确定了已知的靶点，包括激酶MAP3K12（DLK）和转录因子JUN。此外，该筛选揭示了转录因子ATF2的潜在作用。我们证明，MAP3激酶对ATF2的磷酸化是促凋亡转录反应的核心驱动因素。令人惊讶的是，凋亡并不需要JUN的磷酸化。然而，ATF2的磷酸化和JUN表达的上调至关重要。因此，ATF2将激酶信号转化为转录反应。在培养的人类神经元中抑制ATF2可防止细胞死亡。值得注意的是，我们表明在损伤模型中敲低ATF2具有神经保护作用。因此，ATF2为广泛的神经退行性疾病提供了一个有前景的新靶点。
