Linking nucleotide and amino acid metabolism to cholesterol synthesis by MYCN

MYCN 将核苷酸和氨基酸代谢与胆固醇合成联系起来

基本信息

批准号：
10589091
负责人：
HAN-FEI DING
金额：
$ 33.29万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-02-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10589091
关键词：
Acetyl Coenzyme A Amino Acids Animal Model Automobile Driving Biochemical Cancer Cell Growth Carbon Cell Line Cell Proliferation Cells Child Cholesterol Cholesterol Homeostasis Clinical Code Combined Modality Therapy Data Development Down-Regulation Enzyme Inhibitor Drugs Enzymes Feedback Gene Expression Profiling Genes Genetic Transcription Genomics Glutamine Glycine Goals Growth Human Knowledge Link MYCN gene Malignant Childhood Neoplasm Metabolic Metabolic Pathway Metabolism Modeling Molecular Mus Neuroblastoma Nucleotides Oncogenes Oncogenic Output Pathway interactions Population Prediction of Response to Therapy Primary Neoplasm Process Production Proliferating Publishing Repression Research Role Serine Source Survival Rate Testing Therapeutic Therapeutic Intervention Tracer Transcriptional Activation Tumor Suppressor Proteins Tumorigenicity Up-Regulation Xenograft procedure alpha ketoglutarate amino acid metabolism anticancer research cancer cell carboxylate carboxylation enzyme pathway high risk in vivo mevalonate mouse model neoplastic cell neuroblastoma cell novel strategies novel therapeutic intervention nucleotide metabolism patient derived xenograft model predictive marker programs self-renewal small hairpin RNA stable isotope transcription factor tumor tumor progression

项目摘要

How oncogenes integrate metabolic pathways to meet the biosynthetic demands of cancer cell growth and proliferation is a central question of cancer research with broad clinical implications. In high-risk neuroblastoma, one of the deadliest childhood cancers, the mevalonate pathway is transcriptionally activated. This metabolic pathway uses acetyl-CoA carbon to produce cholesterol and other metabolites essential to sustain the proliferation of neuroblastoma cell lines in culture and the growth of neuroblastoma tumors in mouse models. We recently obtained evidence that the oncogenic transcription factor MYCN is required for the transcriptional activation of mevalonate pathway enzymes and the increased production of cholesterol in cell lines derived from high-risk neuroblastoma tumors with genomic amplification of MYCN. The proposed research will address two major questions concerning the molecular basis of MYCN action in reprogramming of mevalonate metabolism and cholesterol synthesis: 1) how MYCN activates the mevalonate pathway to increase its output; and 2) how MYCN coordinates other metabolic pathways to increase the supply of acetyl- CoA. In Aim 1, we will use a combination of cellular and molecular approaches to test the hypothesis that MYCN disrupts end-product feedback inhibition of the mevalonate pathway by transcriptional activation of SCAP, a positive regulator of mevalonate metabolism and repression of INSIG2, a negative regulator, leading to constitutive activation of the mevalonate pathway. We will further investigate the molecular basis for MYCN repression of INSIG2 expression. In Aim 2, we will use stable isotope tracers in combination with shRNA silencing and enzyme inhibitors to delineate the metabolic pathways that supply the substrate acetyl-CoA. We will test the hypothesis that MYCN increases the flux of glutamine carbon into the mevalonate pathway for cholesterol synthesis via transcriptional activation of nucleotide and serine-glycine synthesis pathways. In Aim 3, we will use stable isotope tracers in combination with enzyme inhibitors to provide in vivo evidence for glutamine as a major source of carbon for cholesterol synthesis via nucleotide and serine-glycine synthesis pathways in patient-derived xenografts and the TH-MYCN mouse model of high-risk neuroblastoma. Successful completion of this project will define a molecular mechanism for MYCN to integrate nucleotide, amino acid, and cholesterol metabolism in driving and sustaining high-risk neuroblastoma, which may suggest new avenues of therapeutic intervention.

致癌基因如何整合代谢途径以满足癌细胞生长的生物合成需求和扩散是具有广泛临床意义的癌症研究的核心问题。在高风险中神经母细胞瘤是最致命的童年癌症之一，甲谷酸途径已被转录激活。这种代谢途径使用乙酰辅酶A碳生产胆固醇和其他代谢物必不可少的代谢物维持神经母细胞瘤细胞系在培养和神经母细胞瘤肿瘤中的增殖鼠标模型。我们最近获得了证据，表明致癌转录因子MYCN是必需的大甲酸酯途径酶的转录激活和细胞中胆固醇的产生增加源自具有MYCN基因组扩增的高危神经母细胞瘤肿瘤的线。提议研究将解决有关MYCN作用的分子基础的两个主要问题大甲酸盐代谢和胆固醇的合成：1）MyCN如何激活Mevalonate途径增加其产出； 2）MyCN如何协调其他代谢途径以增加乙酰基的供应 COA。在AIM 1中，我们将使用细胞和分子方法的组合来检验以下假设。 MYCN通过转录激活的转录激活来破坏对甲龙酸盐途径的最终产品反馈抑制 SCAP是甲戊酸代谢的积极调节剂和对Indig2的抑制，负面的调节剂，领先者用于构成型甲硅酸盐途径的激活。我们将进一步研究MYCN的分子基础抑制Insig2表达。在AIM 2中，我们将使用稳定的同位素示踪剂与shRNA结合沉默和酶抑制剂描绘供应底物乙酰辅酶A的代谢途径。我们将测试MYCN将谷氨酰胺碳的通量增加到甲谷酸盐途径的假设胆固醇通过核苷酸和丝氨酸 - 甘氨酸合成途径的转录激活而合成。目标 3，我们将使用稳定的同位素示踪剂与酶抑制剂联合使用体内证据谷氨酰胺是通过核苷酸和丝氨酸合成的胆固醇合成碳的主要来源患者衍生异种移植物和高危神经母细胞瘤的Th-Mycn小鼠模型的途径。该项目的成功完成将定义MYCN整合核苷酸的分子机制，氨基酸和胆固醇代谢在驾驶和维持高危神经细胞瘤方面可能表明治疗干预的新途径。