UK Dementia Research Institute, Department of Clinical Neurosciences, Cambridge Biomedical Campus, University of Cambridge, Cambridge, CB2 0AH, UK.
Open Targets, Wellcome Genome Campus, Hinxton, Cambridge, CB10 1SA, UK.
Sci Rep. 2023 Mar 9;13(1):3934. doi: 10.1038/s41598-023-31141-6.
Accumulation of aggregated and misfolded proteins, leading to endoplasmic reticulum stress and activation of the unfolded protein response, is a hallmark of several neurodegenerative disorders, including Alzheimer's and Parkinson's disease. Genetic screens are powerful tools that are proving invaluable in identifying novel modulators of disease associated processes. Here, we performed a loss-of-function genetic screen using a human druggable genome library, followed by an arrayed-screen validation, in human iPSC-derived cortical neurons. We identified and genetically validated 13 genes, whose knockout was neuroprotective against Tunicamycin, a glycoprotein synthesis inhibitor widely used to induce endoplasmic reticulum stress. We also demonstrated that pharmacological inhibition of KAT2B, a lysine acetyltransferase identified by our genetic screens, by L-Moses, attenuates Tunicamycin-mediated neuronal cell death and activation of CHOP, a key pro-apoptotic member of the unfolded protein response in both cortical and dopaminergic neurons. Follow-up transcriptional analysis suggested that L-Moses provided neuroprotection by partly reversing the transcriptional changes caused by Tunicamycin. Finally, L-Moses treatment attenuated total protein levels affected by Tunicamycin, without affecting their acetylation profile. In summary, using an unbiased approach, we identified KAT2B and its inhibitor, L-Moses, as potential therapeutic targets for neurodegenerative diseases.
蛋白质聚集和错误折叠的积累导致内质网应激和未折叠蛋白反应的激活,是包括阿尔茨海默病和帕金森病在内的几种神经退行性疾病的标志。遗传筛选是一种强大的工具,它在鉴定新型疾病相关过程的调节剂方面证明是非常宝贵的。在这里,我们使用人类可用药基因组文库进行了功能丧失性遗传筛选,然后在人诱导多能干细胞衍生的皮质神经元中进行了阵列筛选验证。我们鉴定并遗传验证了 13 个基因,其敲除对衣霉素(一种广泛用于诱导内质网应激的糖蛋白合成抑制剂)具有神经保护作用。我们还证明,通过我们的遗传筛选鉴定的赖氨酸乙酰转移酶 KAT2B 的药理学抑制剂 L-Moses,可减轻衣霉素介导的皮质神经元和多巴胺能神经元中的细胞死亡和 CHOP(未折叠蛋白反应的关键促凋亡成员)的激活。后续的转录分析表明,L-Moses 通过部分逆转衣霉素引起的转录变化提供了神经保护作用。最后,L-Moses 处理减轻了衣霉素影响的总蛋白水平,而不影响它们的乙酰化谱。总之,我们使用一种无偏倚的方法,鉴定了 KAT2B 及其抑制剂 L-Moses,作为神经退行性疾病的潜在治疗靶点。