Role of AMPK in melanoma brain metastasis

项目摘要： AMP激活的蛋白激酶（AMPK）是一种重要的进化保守的能量传感器， 通过监测细胞内AMP或ADP与ATP比率的变化来维持能量稳态。越来越多的证据 支持AMPK通路是代谢和癌症界面的主要信号传导参与者之一。 然而，AMPK在肿瘤进展和转移中的作用仍然不清楚。我们最近对 黑色素瘤癌症基因组学数据显示，编码α-羟色胺的PRKAA 2基因突变， AMPK的2催化亚基，发生在8-10%的皮肤黑色素瘤中，并且倾向于与NF 1突变共同发生。 在我们的初步研究中，我们发现AMPKα2的敲除促进了 在软琼脂测定中的突变NF 1-黑色素瘤细胞及其作为裸鼠中异种移植肿瘤的生长。 重要的是，我们发现AMPKα2在人黑色素瘤脑中的表达显著下调， 转移样品与患者匹配的颅外转移样品相比。此外，敲除 AMPKα2促进NF 1突变Mewo黑色素瘤细胞裸鼠脑转移基于这些 初步结果，我们假设AMPKα2在黑色素瘤中的失活促进肿瘤的发生， 进展为黑色素瘤脑转移在目的1中，我们将确定AMPKα2在黑色素瘤脑中的作用， 使用小鼠模型和人样品进行转移。我们计划描述AMPKα2缺失对 同基因和基因工程小鼠模型。人类黑色素瘤脑转移样本将 用于通过空间单细胞RNA-seq分析探索AMPKα2损失的贡献。在目标2中，我们将 表征参与黑色素瘤脑转移代谢调节的AMPK下游靶点。 我们将研究AMPKα2的一种新的假定底物在从头脂肪生成中的作用， 在我们的初步研究中发现，黑色素瘤脑转移，并探索其他新的AMPKα2靶点， 通过对来自小鼠模型的肿瘤样本进行全面的代谢组学和RPPA分析。在目标3中，我们 将评估靶向AMPK通路的代谢药物对增强 使用人黑色素瘤异种移植物的黑色素瘤脑转移的靶向和免疫治疗，同基因 和基因工程小鼠模型。为了实现这些目标，我们组建了一个调查小组， 拥有各种专业知识，包括癌细胞代谢，黑色素瘤脑转移生物学，黑色素瘤 肿瘤学家/外科医生和计算生物学。我们的研究不仅将为人们提供关于 AMPK代谢信号在黑色素瘤脑转移中解决了该领域的一个重大知识缺口， 还确定了新的治疗方法，以靶向黑色素瘤脑转移。

Project Summary: AMP activated protein kinase (AMPK) is a critical evolutionarily conserved energy sensor that regulates energy homeostasis by monitoring changes in the intracellular AMP or ADP to ATP ratios. Mounting evidence supports that the AMPK pathway is one of the major signaling players at the interface of metabolism and cancer. However, the role of AMPK in tumor progression and metastasis remains obscure. Our recent analysis on melanoma cancer genomics data has revealed that the mutations in the PRKAA2 gene, which encodes the alpha 2 catalytic subunit of AMPK, occur in 8-10% of cutaneous melanomas and tend to co-occur with NF1 mutations. In our preliminary studies, we found that knockout of AMPKα2 promoted anchorage-independent growth of mutant NF1-melanoma cells in soft agar assays and their growth as xenografted tumors in nude mice. Importantly, we found that expression of AMPKα2 is significantly downregulated in human melanoma brain metastasis samples compared to patient-matched extracranial metastasis samples. Furthermore, knockout of AMPKα2 promoted brain metastasis of NF1-mutant Mewo melanoma cells in nude mice. Based on these preliminary results, we hypothesize that inactivation of AMPKα2 in melanoma promotes tumor progression to melanoma brain metastasis. In Aim 1, we will establish the role of AMPKα2 in melanoma brain metastasis using mouse models and human samples. We plan to characterize the effects of AMPKα2 loss in both syngeneic and genetically engineered mouse models. Human melanoma brain metastasis samples will be used to explore the contribution of AMPKα2 loss through spatial single cell RNA-seq analyses. In aim 2, we will characterize downstream targets of AMPK involved in the metabolic regulation of melanoma brain metastasis. We will investigate the contribution of a novel putative substrate of AMPKα2 involved in de novo lipogenesis, identified in our preliminary studies, to melanoma brain metastasis, and explore additional new AMPKα2 targets through comprehensive metabolomics and RPPA analyses of tumor samples from mouse models. In aim 3, we will assess the therapeutic effects of metabolic drugs targeting the AMPK pathway on enhancing the efficacies of targeted- and immune- therapies in melanoma brain metastasis using human melanoma xenografts, syngeneic and genetically engineered mouse models. To achieve these aims, we have assembled a team of investigators with various expertise including cancer cell metabolism, melanoma brain metastasis biology, melanoma oncologists/surgeon and computational biology. Our study will not only provide critical insights into the role of AMPK metabolic signaling in melanoma brain metastasis addressing a significant knowledge gap in the field, but also identify novel therapeutic approaches to target melanoma brain metastasis.