Ging Kathi, Frick Lukas, Schlachetzki Johannes, Armani Andrea, Zhu Yanping, Gilormini Pierre-André, Dhingra Ashutosh, Böck Desirée, Marques Ana, Deen Matthew, Chen Xi, Serdiuk Tetiana, Trevisan Chiara, Sellitto Stefano, Pisano Claudio, Glass Christopher K, Heutink Peter, Yin Jiang-An, Vocadlo David J, Aguzzi Adriano
Institute of Neuropathology, University of Zurich, Zurich, Switzerland.
Department of Cellular and Molecular Medicine, University of California San Diego, La Jolla, CA, USA.
NPJ Parkinsons Dis. 2024 Oct 22;10(1):192. doi: 10.1038/s41531-024-00819-7.
Mutations in GBA1 encoding the lysosomal enzyme β-glucocerebrosidase (GCase) are among the most prevalent genetic susceptibility factors for Parkinson's disease (PD), with 10-30% of carriers developing the disease. To identify genetic modifiers contributing to the incomplete penetrance, we examined the effect of 1634 human transcription factors (TFs) on GCase activity in lysates of an engineered human glioblastoma line homozygous for the pathogenic GBA1 L444P variant. Using an arrayed CRISPR activation library, we uncovered 11 TFs as regulators of GCase activity. Among these, activation of MITF and TFEC increased lysosomal GCase activity in live cells, while activation of ONECUT2 and USF2 decreased it. While MITF, TFEC, and USF2 affected GBA1 transcription, ONECUT2 might control GCase trafficking. The effects of MITF, TFEC, and USF2 on lysosomal GCase activity were reproducible in iPSC-derived neurons from PD patients. Our study provides a systematic approach to identifying modulators of GCase activity and deepens our understanding of the mechanisms regulating GCase.
编码溶酶体酶β-葡萄糖脑苷脂酶（GCase）的GBA1基因突变是帕金森病（PD）最常见的遗传易感性因素之一，10%至30%的携带者会患上这种疾病。为了确定导致不完全显性的基因修饰因子，我们研究了1634种人类转录因子（TFs）对工程化人类胶质母细胞瘤细胞系裂解物中GCase活性的影响，该细胞系对致病性GBA1 L444P变体纯合。使用阵列CRISPR激活文库，我们发现了11种TFs作为GCase活性的调节因子。其中，MITF和TFEC的激活增加了活细胞中溶酶体GCase的活性，而ONECUT2和USF2的激活则降低了它。虽然MITF、TFEC和USF2影响GBA1转录，但ONECUT2可能控制GCase的运输。MITF、TFEC和USF2对溶酶体GCase活性的影响在PD患者诱导多能干细胞衍生的神经元中是可重复的。我们的研究提供了一种系统的方法来识别GCase活性的调节因子，并加深了我们对调节GCase机制的理解。
