TFCP2 is a transcriptional regulator of heparan sulfate assembly and melanoma cell growth.
TFCP2 is a transcriptional regulator of heparan sulfate assembly and melanoma cell growth.
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DOI：
10.1016/j.jbc.2023.104713
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发表时间：
2023-06
影响因子：
4.8
通讯作者：
Weiss, Ryan J
中科院分区：
文献类型：
--
作者：
Basu, Amrita;Champagne, Rachel N;Patel, Neil G;Nicholson, Elijah D;Weiss, Ryan J
硫酸乙酰肝素（HS）是一种长的线性多糖，其在所有动物细胞中普遍表达，并且在许多细胞过程（包括细胞信号传导和发育）中起关键作用。HS组装失调与病理生理学条件有关，如肿瘤发生和罕见遗传疾病。HS生物合成通过一大群生物合成酶的活性以非模板驱动的方式在内质网和高尔基体中发生。虽然对它的生物合成了解很多，但对HS组装在不同组织类型和疾病状态中的调节知之甚少。为了解决这一知识缺口，我们最近进行了全基因组CRISPR/Cas9筛选，以确定HS生物合成的新调控因子。从这些筛选中，我们确定了α珠蛋白转录因子TFCP 2作为热门。为了研究TFCP 2在HS组装中的作用，我们使用CRISPR/Cas9系统靶向人黑素瘤细胞中的TFCP 2表达。与野生型细胞相比，TFCP 2敲除细胞表现出与细胞表面HS结合的成纤维细胞生长因子减少、HS组成改变和细胞生长减慢。此外，RNA测序显示TFCP 2调节参与HS组装的多种酶的表达，包括分泌的硫酸酯酶SULF 1。TFCP 2活性的药理学靶向类似地减少生长因子结合并增加SULF 1表达，并且TFCP 2突变细胞中SULF 1表达的敲低恢复了黑素瘤细胞生长。总体而言，这些研究确定TFCP 2作为HS的一种新型转录调节因子，并强调HS-蛋白相互作用作为减缓黑色素瘤生长的可能靶点。
Heparan sulfate (HS) is a long, linear polysaccharide that is ubiquitously expressed in all animal cells and plays a key role in many cellular processes, including cell signaling and development. Dysregulation of HS assembly has been implicated in pathophysiological conditions, such as tumorigenesis and rare genetic disorders. HS biosynthesis occurs in a non-template-driven manner in the endoplasmic reticulum and Golgi through the activity of a large group of biosynthetic enzymes. While much is known about its biosynthesis, little is understood about the regulation of HS assembly across diverse tissue types and disease states. To address this gap in knowledge, we recently performed genome-wide CRISPR/Cas9 screens to identify novel regulatory factors of HS biosynthesis. From these screens, we identified the alpha globin transcription factor, TFCP2, as a top hit. To investigate the role of TFCP2 in HS assembly, we targeted TFCP2 expression in human melanoma cells using the CRISPR/Cas9 system. TFCP2 knockout cells exhibited decreased fibroblast growth factor binding to cell surface HS, alterations in HS composition, and slowed cell growth compared to wild-type cells. Additionally, RNA sequencing revealed that TFCP2 regulates the expression of multiple enzymes involved in HS assembly, including the secreted endosulfatase, SULF1. Pharmacological targeting of TFCP2 activity similarly reduced growth factor binding and increased SULF1 expression, and the knockdown of SULF1 expression in TFCP2 mutant cells restored melanoma cell growth. Overall, these studies identify TFCP2 as a novel transcriptional regulator of HS and highlight HS–protein interactions as a possible target to slow melanoma growth.