Heparan sulfate (HS) proteoglycans bind extracellular proteins that participate in cell signaling, attachment and endocytosis. These interactions depend on the arrangement of sulfated sugars in the HS chains generated by well-characterized biosynthetic enzymes; however, the regulation of these enzymes is largely unknown. We conducted genome-wide CRISPR–Cas9 screens with a small-molecule ligand that binds to HS. Screening of A375 melanoma cells uncovered additional genes and pathways impacting HS formation. The top hit was the epigenetic factor KDM2B, a histone demethylase. KDM2B inactivation suppressed multiple HS sulfotransferases and upregulated the sulfatase SULF1. These changes differentially affected the interaction of HS-binding proteins. KDM2B-deficient cells displayed decreased growth rates, which was rescued by SULF1 inactivation. In addition, KDM2B deficiency altered the expression of many extracellular matrix genes. Thus, KDM2B controls proliferation of A375 cells through the regulation of HS structure and serves as a master regulator of the extracellular matrix.

硫酸乙酰肝素 (HS) 蛋白聚糖结合参与细胞信号转导、附着和内吞作用的胞外蛋白。这些相互作用取决于由充分表征的生物合成酶产生的 HS 链中硫酸化糖的排列；然而，这些酶的调节在很大程度上是未知的。我们使用与 HS 结合的小分子配体进行了全基因组 CRISPR–Cas9 筛选。筛选 A375 黑色素瘤细胞发现了影响 HS 形成的其他基因和途径。最热门的是表观遗传因子KDM2B，一种组蛋白去甲基化酶。KDM2B失活抑制了多种 HS 磺基转移酶并上调了硫酸酯酶 SULF1。这些变化对 HS 结合蛋白的相互作用有不同的影响。KDM2B-缺陷细胞显示出降低的生长速率，这通过SULF1失活得以挽救。此外，KDM2B缺陷改变了许多细胞外基质基因的表达。因此，KDM2B通过调节 HS 结构来控制 A375 细胞的增殖，并作为细胞外基质的主要调节剂。