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Metabolic Drivers of Melanoma Initiation, Progression and Therapy Response

黑色素瘤发生、进展和治疗反应的代谢驱动因素

基本信息

批准号：
8985670
负责人：
Narendra Wajapeyee
金额：
$ 18.11万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-12-15 至 2016-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8985670
关键词：
Accounting BRAF gene Biochemical Biochemical Genetics Cell Culture Techniques Cell Line Ceramidase Ceramides Cessation of life Clinical Disease Farber&apos s lipogranulomatosis Gene Expression Profiling Genes Genomic approach Growth Health Human Hydrolase Link Lipids MAP Kinase Signaling Pathways Mediating Mediator of activation protein Melanoma Cell Metabolic Metabolic Pathway Mitogen-Activated Protein Kinases Modeling Molecular Mutation NRAS gene Neoplasm Metastasis Oncogenic Pathogenesis Pathway interactions Patients Play RNA Interference RNA interference screen Resistance Role Sampling Signal Transduction Site Skin Cancer Staging Structure Testing Therapeutic United States Up-Regulation base cancer cell cancer type clinically significant effective therapy functional genomics glycosylation improved in vivo inhibitor/antagonist knock-down melanocyte melanoma mouse model mutant new therapeutic target novel research study resistance mechanism response small hairpin RNA targeted treatment tumor tumor initiation tumor progression

项目摘要

DESCRIPTION (provided by applicant): Melanoma account for <10,000 deaths annually in the United States alone and current therapies for advanced-stage melanoma do not provide durable clinical benefit. Therefore, improved molecular understanding of melanoma initiation and progression is essential for developing effective therapies against melanoma. Metabolic deregulation due to the alterations in metabolic pathways is one of the key hallmarks of cancer cells. However, the role of metabolic alterations in melanoma has neither been comprehensively analyzed nor fully understood. The central hypothesis is that metabolic alterations are crucial regulators of melanoma initiation, progression and therapy response. Therefore, to identify metabolic drivers of melanoma, we developed an integrative genomics approach by combining patient-derived melanoma sample gene expression profiling with the functional genomics approach of an in vivo RNAi screening and identified N-Acylsphingosine amidohydrolase (ASAH1) as a gene necessary for melanoma growth and metastasis. The overall objective is to determine the role of ASAH1 in melanoma initiation and progression, ascertain its mechanisms-of-action and determine its role in oncogenic BRAF inhibitor vemurafenib resistance. Specifically, in Aim 1, we will determine the role of ASAH1 in melanoma initiation and progression and ascertain whether ASAH1 is necessary for oncogenic BRAF- and NRAS-induced transformation and tumor progression. To do so, we will use cell culture-based models of immortalized human melanocytes, established and patient-derived short-term melanoma cultures as well as mouse models of melanomagenesis and melanoma metastasis. In Aim 2, we will determine the mechanism(s) of ASAH1 action. First, we will perform structure-function analyses to determine whether the lipid hydrolase activity of ASAH1 is necessary for its ability to promote melanoma initiation and progression. Additionally, guided by our preliminary results, we will test whether ASAH1 promotes melanoma initiation and progression by reducing cellular ceramide level and/or by activating MAP kinase pathway. The experiments will include biochemical, genetic, cell culture and in vivo mouse model-based approaches. In Aim 3, we will determine the role of ASAH1 in oncogenic BRAF inhibitor vemurafenib resistance. We will test whether inhibition of ASAH1 can restore vemurafenib sensitivity in BRAF mutant melanoma cells and determine if ASAH1 induced vemurafenib resistance depend upon its ability to regulate cellular ceramide level and/or MAP kinase pathway. Finally, using melanoma patients-derived short-term melanoma culture and primary sample of pre- and post-vemurafenib treatment we will establish the clinical significance of our results. Collectively, the results of ur experiments will uncover novel driver genes and pathways of melanoma initiation and progression, improve the molecular understanding of melanoma and reveal new therapeutic targets and mechanisms of resistance to melanoma therapies.

描述（由申请人提供）：仅在美国，黑色素瘤每年就造成<10，000例死亡，目前用于晚期黑色素瘤的治疗方法不能提供持久的临床益处。因此，提高对黑色素瘤发生和进展的分子理解对于开发针对黑色素瘤的有效疗法至关重要。由于代谢途径的改变而导致的代谢失调是癌细胞的关键标志之一。然而，代谢改变在黑色素瘤中的作用既没有得到全面分析，也没有得到充分理解。核心假设是代谢改变是黑色素瘤发生、进展和治疗反应的关键调节因子。因此，为了识别黑色素瘤的代谢驱动因素，我们通过将患者来源的黑色素瘤样品基因表达谱与体内RNAi筛选的功能基因组学方法相结合，开发了一种整合基因组学方法，并将N-酰基鞘氨醇酰胺水解酶（ASAH 1）鉴定为黑色素瘤生长和转移所必需的基因。总体目标是确定ASAH 1在黑色素瘤发生和进展中的作用，确定其作用机制，并确定其在致癌BRAF抑制剂维罗非尼耐药性中的作用。具体而言，在目标1中，我们将确定ASAH 1在黑色素瘤发生和进展中的作用，并确定ASAH 1是否是致癌BRAF和NRAS诱导的转化和肿瘤进展所必需的。为此，我们将使用基于细胞培养的永生化人黑素细胞模型、已建立的和患者来源的短期黑色素瘤培养物以及黑色素瘤发生和黑色素瘤转移的小鼠模型。在目标2中，我们将确定ASAH 1的作用机制。首先，我们将进行结构-功能分析，以确定ASAH 1的脂质水解酶活性是否是其能力所必需的，促进黑色素瘤的发生和发展。此外，根据我们的初步结果，我们将测试ASAH 1是否通过降低细胞神经酰胺水平和/或激活MAP激酶途径促进黑色素瘤的发生和进展。这些实验将包括生物化学、遗传学、细胞培养和基于体内小鼠模型的方法。在目标3中，我们将确定ASAH 1在致癌BRAF抑制剂维罗非尼耐药中的作用。我们将测试ASAH 1的抑制是否可以恢复BRAF突变黑素瘤细胞中的维罗非尼敏感性，并确定ASAH 1诱导的维罗非尼抗性是否取决于其调节细胞神经酰胺水平和/或MAP激酶途径的能力。最后，使用黑色素瘤患者来源的短期黑色素瘤培养物和vemurafenib治疗前后的原始样本，我们将确定我们结果的临床意义。总的来说，我们的实验结果将揭示新的驱动基因和黑色素瘤发生和发展的途径，提高黑色素瘤的分子认识，并揭示新的治疗靶点和黑色素瘤耐药机制。

项目成果

期刊论文数量（7）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Paraoxonase 2 Facilitates Pancreatic Cancer Growth and Metastasis by Stimulating GLUT1-Mediated Glucose Transport.

二氧氧蛋白酶2通过刺激GLUT1介导的葡萄糖转运来促进胰腺癌的生长和转移。

DOI：
10.1016/j.molcel.2017.07.014
发表时间：
2017-08-17
期刊：
Molecular cell
影响因子：
16
作者：
Nagarajan A;Dogra SK;Sun L;Gandotra N;Ho T;Cai G;Cline G;Kumar P;Cowles RA;Wajapeyee N
通讯作者：
Wajapeyee N

Serine hydroxymethyl transferase 1 stimulates pro-oncogenic cytokine expression through sialic acid to promote ovarian cancer tumor growth and progression.