Identification of Epigenetic Regulators of Vascular Calcification with a CRISPR-Based Screen

作者：Yaxin Lian, Chen Xie, Minjia Feng, Huijin Zhu, Xin Chen, Xiaolin Liu, Yun Kong, Dajiang He, Jianshuai Ma, Yuxi Chen, H. Zhang, Aoran Huang, Yanlian Chen, Hui Huang · 发表于：Journal of the American Society of Nephrology · 年份：2025 · DOI：10.1681/asn.0000000793 · 被引用次数：1 · 研究领域：CRISPR and Genetic Engineering

Key Points Magnetic-activated cell sorting–based clustered regularly interspaced short palindromic repeats (CRISPR) screen was used for the first time to identify critical genes and pathways involved in vascular calcification. Epigenetic-focused CRISPR screen identified novel vascular calcification regulators, providing potential targets when integrated with transcriptomics. Background Vascular calcification, mainly driven by osteogenic transdifferentiation of vascular smooth muscle cells (VSMCs), is a common pathologic condition in patients with CKD. However, the roles of other epigenetic regulators in this process remain largely unexplored. Clustered regularly interspaced short palindromic repeats (CRISPR) screen is a highly efficient strategy widely used in identifying genes related to various biologic processes. However, the lack of suitable cell sorting strategies combined with CRISPR screen meant this technology had not been applied to gene screening in vascular calcification. Methods We performed an epigenetic-focused CRISPR screen in primary human VSMCs and identified key genes and pathways underlying osteogenic transdifferentiation, based on small guide RNA enrichment in receptor activator of NF-kappa B ligand-positive (calcified) and receptor activator of NF-kappa B ligand-negative (noncalcified) VSMCs isolated by magnetic-activated cell sorting. To validate the screen results, potential genes with different rankings were validated by small interfering RNA intervent...