Ryan Veronica H, Lawton Sydney, Reyes Joel F, Hawrot James, Frankenfield Ashley M, Seddighi Sahba, Ramos Daniel M, Faghri Faraz, Johnson Nicholas L, Zou Jizhong, Kampmann Martin, Replogle John, Yuan Hebao, Johnson Kory R, Maric Dragan, Hao Ling, Nalls Mike A, Ward Michael E
National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD, USA.
Center for Alzheimer's and Related Dementias, National Institute on Aging, National Institutes of Health, Bethesda, MD, USA.
bioRxiv. 2024 Oct 1:2024.09.30.615241. doi: 10.1101/2024.09.30.615241.
TDP-43 mislocalization and pathology occurs across a range of neurodegenerative diseases, but the pathways that modulate TDP-43 in neurons are not well understood. We generated a Halo-TDP-43 knock-in iPSC line and performed a genome-wide CRISPR interference FACS-based screen to identify modifiers of TDP-43 levels in neurons. A meta-analysis of our screen and publicly available screens identified both specific hits and pathways present across multiple screens, the latter likely responsible for generic protein level maintenance. We identified BORC, a complex required for anterograde lysosome transport, as a specific modifier of TDP-43 protein, but not mRNA, levels in neurons. BORC loss led to longer half-life of TDP-43 and other proteins, suggesting lysosome location is required for proper protein turnover. As such, lysosome location and function are crucial for maintaining TDP-43 protein levels in neurons.
TDP-43的错误定位和病理变化在一系列神经退行性疾病中都会出现，但调节神经元中TDP-43的途径尚未得到充分了解。我们构建了一个Halo-TDP-43基因敲入的诱导多能干细胞系，并进行了基于全基因组CRISPR干扰荧光激活细胞分选的筛选，以确定神经元中TDP-43水平的调节因子。对我们的筛选结果和公开筛选数据进行的荟萃分析确定了多个筛选中都存在的特定命中靶点和途径，后者可能负责维持一般蛋白质水平。我们发现BORC（一种顺行性溶酶体运输所需的复合物）是神经元中TDP-43蛋白而非mRNA水平的特定调节因子。BORC缺失导致TDP-43和其他蛋白质的半衰期延长，这表明溶酶体定位对于正确的蛋白质周转是必需的。因此，溶酶体定位和功能对于维持神经元中TDP-43蛋白水平至关重要。
