The function of TWIST1 acetylation in cell fate and tissue development

摘要 颅面疾病是第二常见的先天性出生缺陷，而儿科癌症是第二常见的先天性出生缺陷。 儿童死亡的主要原因。这些异常是由调节神经管（NT）的中断引起的， 颅神经嵴细胞（CNCC）的发育。尽管流行率很高，但我们的国家在这方面仍存在重大差距。 了解导致这些疾病的遗传风险因素和潜在机制。我们有 最近发现了Twist 1在神经管和CNCC命运转换中的新功能。我们最近的发现 显示Twist 1参与NT关闭期间的细胞形状变化，并在NT关闭期间抑制上皮基因。 迁移前CNCC向迁移间充质细胞转化（EMT） 细胞此外，我们的支持性数据表明，TWIST 1在癌细胞中的K73、K76处被乙酰化，而在癌细胞中的K73、K76处被乙酰化。 二乙酰化的K73 Ac; K76 Ac使TWIST 1能够与组蛋白乙酰转移酶因子，BRD 8，p400， EPC 1和TIP 60，它们是NuA 4复合物的成员，在癌细胞中。为跟进调查结果， 贝勒医学院的Xu实验室将与UTHealth的Fakhouri实验室建立新的合作关系 牙科学院确定TWIST 1乙酰化在发展和CNCC中的新意义- 衍生肿瘤形成。对于这个合作项目，我们假设TWIST 1乙酰化是必要的， 决定了与染色质修饰因子的相互作用，以控制颅面过程中的细胞命运决定 发育和CNCC衍生的组织分化。为了检验这个高风险、高回报的假设，我们将 产生乙酰化-不合格（K73 A; K76 A）、乙酰化-模拟物（K73 Q; K76 Q）和脱乙酰化- 模拟/无能（K73 R; K76 R）小鼠系和具有相似点的遗传修饰的09 -1 CNCC系， 突变。小鼠品系的表型、组织学和细胞分析将评估以下功能： TWIST 1乙酰化在组织发育和潜在CNCC衍生肿瘤形成中的作用我们将调查 09 -1稳定系中TWIST 1乙酰化和CNCC分化能力。本项目旨在：（1）发现 TWIST 1乙酰化在颅颌面发育和肿瘤形成中的作用;（2）探讨TWIST 1乙酰化在颅颌面发育和肿瘤形成中的作用。 TWIST 1和NuA 4染色质修饰复合物相互作用及其在CNCC中的作用 分化能力。我们将确定Twist 1乙酰化不胜任（K73 A/R; K76 A/R）和 乙酰化模拟物（K73 A; K76 A）小鼠系对NT闭合、迁移前CNCC的命运转变和CNCC- 衍生肿瘤形成。我们将在O 9 -1 CNCC线路中检查与 NuA 4染色质修饰复合物和O 9 -1 CNCC迁移和分化能力。该项目将 提供了一个巨大的机会，建立跨学科的合作，将利用我们的 在小鼠遗传学、颅面发育和癌症生物学方面优势互补。

Abstract Craniofacial disorders are the second most common congenital birth defects, while pediatric cancers are the leading cause of death in children. These anomalies arise from a disruption in regulating neural tube (NT) and cranial neural crest cell (CNCC) development. Despite the high prevalence, significant gaps remain in our knowledge of the genetic risk factors and the underlying mechanisms that lead to these disorders. We have recently identified a novel function of Twist1 in the neural tube and CNCC fate transition. Our recent findings showed that Twist1 is involved in cell shape changes during NT closure and suppresses epithelial genes during the epithelial-to-mesenchymal transition (EMT) of pre-migratory CNCCs to become migratory mesenchymal cells. In addition, our supporting data indicates that TWIST1 is acetylated at K73; K76 in cancer cells, and the di-acetylated K73Ac;K76Ac enables TWIST1 to interact with histone acetyl-transferase factors, BRD8, p400, EPC1, and TIP60, which are members of the NuA4 complex, in cancer cells. To follow up on our findings, the Xu lab at Baylor College of Medicine (BCM) will establish a new collaboration with Fakhouri lab at UTHealth School of Dentistry to determine the novel significance of TWIST1 acetylation in the development and CNCC- derived tumor formation. For this collaborative project, we hypothesize that TWIST1 acetylation is necessary to dictate the interaction with chromatin modifying factors to control cell fate determination during craniofacial development and CNCC-derived tissue differentiation. To test this high-risk, high-reward hypothesis, we will generate acetylated-incompetent (K73A; K76A), acetylated-mimic (K73Q; K76Q), and deacetylated- mimic/incompetent (K73R; K76R) mouse lines and genetically-modified O9-1 CNCC lines with similar point- mutations. The phenotypical, histological, and cellular analyses of the mouse lines will assess the function of TWIST1 acetylation in tissue development and potentially CNCC-derived tumor formation. We will investigate TWIST1 acetylation and CNCC differentiation capacity in O9-1 stable lines. This project aims to: (1) Uncover the function of TWIST1 acetylation in craniofacial development and tumor formation, (2) Investigate the interaction between TWIST1 and NuA4 chromatin modifying complex and their involvement in CNCC differentiation capacity. We will determine the impact of Twist1 acetylated-incompetent (K73A/R; K76A/R) and acetylated-mimic (K73A; K76A) mouse lines on NT closure, fate transition in pre-migratory CNCCs, and CNCC- derived tumor formation. We will examine in O9-1 CNCC line the significance of interaction with the members of NuA4 chromatin-modifying complex, and O9-1 CNCC migratory and differentiation capacity. This project will provide a tremendous opportunity to establish a cross-disciplinary collaboration that will leverage our complementary strengths in mouse genetics, craniofacial development, and cancer biology.