Elucidating the therapeutic utility of targeting metabolic dependencies in osteosarcoma

阐明针对骨肉瘤代谢依赖性的治疗效用

基本信息

批准号：
9975390
负责人：
Heather Lynn Gardner
金额：
$ 12.63万
依托单位：
TUFTS UNIVERSITY BOSTON
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：
2020-03-01 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/9975390
关键词：
Address Adolescent Affect Amputation Animal Disease Models Animal Model Binding Bioinformatics Canis familiaris Cell Proliferation Cell membrane Cells ChIP-seq Chemicals Chromatin Conformation Capture and Sequencing Chromatin Interaction Analysis by Paired-End Tag Sequencing Clinical Clinical Trials Comparative Genomic Analysis Complement Data Dependence Development Disease Disease Outcome Down-Regulation Doxorubicin Drug Combinations Ensure Environment Exhibits FOXM1 gene Future Gene Expression Profile Genetic Transcription Genome Genomics Glutaminase Glycolysis Growth Human Immune checkpoint inhibitor Immune response Immunosuppression Immunotherapeutic agent Immunotherapy Impairment In Vitro Lactic acid Link Long Term Survivorship Malignant Bone Neoplasm Mediating Mediator of activation protein Medical Genetics Mentors Metabolic Metabolic Pathway Metabolism Metastatic Osteosarcoma Metastatic to Microscopic Modeling Molecular Mus Mutation Neoplasm Metastasis Nutrient Oncology Operative Surgical Procedures Pathway interactions Phenotype Phosphotransferases Positioning Attribute Primary Neoplasm Recording of previous events Refractory Regulation Research Resistance Resources STAT3 gene Scientist Somatic Mutation Structure Therapeutic Training Transcriptional Regulation Tumor Immunity Universities Variant Work Xenograft procedure Zebrafish aerobic glycolysis bisulfite sequencing cancer cell cancer clinical trial cancer genome cell growth chemotherapy clinical development clinical translation comparative design flexibility genetic manipulation immune checkpoint blockade in vivo inhibitor/antagonist innovation kinase inhibitor knockout gene loss of function metabolic profile mouse model neoplastic cell osteosarcoma programs skills small molecule inhibitor targeted agent therapeutic target tumor tumor growth tumor metabolism tumor microenvironment

项目摘要

PROJECT SUMMARY/ABSTRACT Osteosarcoma (OS), the most common malignant bone tumor in humans and dogs, shares several features in both species including clinical presentation and molecular alterations. Despite numerous efforts there have been no improvements in disease outcome for either species in the past three decades: 30% of people and 90% of dogs still die of metastasis. While some approaches have shown promise in the setting of microscopic disease, in both species, macroscopic metastasis exhibits inherent resistance to multiple agents (kinase inhibitors, chemotherapy, immunotherapy, among others). I recently characterized the canine OS genome and found that as with human OS, the somatic mutational load is low, copy number aberrations/structural variants predominate, and no clear molecular drivers are evident. These data, along with a history of failed clinical trial efforts suggest that contemporary approaches to therapeutic advancement such as kinase inhibitors and immune checkpoint blockade will likely have limited clinical impact, necessitating the development of innovative therapeutic strategies. A distinguishing feature of cancer cells is their ability to undergo aerobic glycolysis, allowing them to thrive in a variety of microenvironments. Monocarboxylate transporters (MCTs) are key facilitators of this, moving lactic acid across the plasma membrane, and are critical for growth and metastasis of glycolytic tumors, such as OS. In previous work, I found that loss of MCT1 or MCT4 function in OS cells decreases basal and compensatory glycolysis, cellular proliferation and invasive capacity. I also showed that MCT4 is a direct transcriptional target of STAT3 and FOXM1, both of which exhibit constitutive activation in OS, supporting a link between MCT4/STAT3/FOXM1 and aerobic glycolysis. Building on these data, I will by leverage a comparative cross- species approach to first define regulatory circuits that support aerobic glycolysis mediated by MCT1/MCT4 in OS and then identify and validate therapeutic vulnerabilities related to MCT control of cellular metabolism. I hypothesize that in OS cells, sustained MCT1/MCT4 expression is driven by constitutive STAT3/FOXM1 activation and increased MYC copy number, thereby promoting aerobic glycolysis. I further predict that loss of MCT1/MCT4 through genetic manipulation or targeted inhibitors will impair tumor growth in vivo, and that this can be maximized through rational drug combination. To accomplish this, I will first characterize the transcriptional regulation of MCT1/MCT4 in OS using a combination of ChIP-sequencing, bisulfite sequencing and 4C/ChiA-PET analysis. Xenograft studies in mice will complement in vitro analyses to assess how loss of MCT1/4 function affects OS metabolic profile, tumor phenotype and tumor growth. Lastly, I will use a zebrafish model to screen select agents for synthetic lethality with MCT blockade, then validate findings in mice. The rich research environment afforded by Tufts University and its partners ensures access to resources and expertise necessary for completion of the proposed work. My training in veterinary oncology, clinical trials and genetics along with guidance from my mentoring team with expertise in genomics, bioinformatics, animal models and translational oncology make me well positioned for successful transition to independence.

项目概要/摘要骨肉瘤 (OS) 是人类和狗中最常见的恶性骨肿瘤，具有以下几个共同特征：这两个物种包括临床表现和分子改变。尽管做出了许多努力过去三十年里，这两个物种的疾病结果都没有改善：30% 的人和 90% 的人狗仍然死于转移。虽然一些方法在微观疾病的治疗中显示出了希望，在这两个物种中，宏观转移表现出对多种药物（激酶抑制剂、化疗、免疫疗法等）。我最近对犬类操作系统基因组进行了表征，发现与人类操作系统一样，体细胞突变负荷较低，拷贝数畸变/结构变异占主导地位，并且没有明显的分子驱动因素。这些数据以及临床试验失败的历史表明当代治疗进展的方法，例如激酶抑制剂和免疫检查点封锁可能只会产生有限的临床影响，因此需要开发创新的治疗方法策略。癌细胞的一个显着特征是它们能够进行有氧糖酵解，从而使它们能够在各种微环境中茁壮成长。单羧酸转运蛋白 (MCT) 是这一过程的关键促进者，乳酸穿过质膜，对于糖酵解肿瘤的生长和转移至关重要，例如操作系统。在之前的工作中，我发现 OS 细胞中 MCT1 或 MCT4 功能的丧失会降低基础和代偿性糖酵解、细胞增殖和侵袭能力。我还表明 MCT4 是直接转录靶标 STAT3 和 FOXM1，两者都在 OS 中表现出组成型激活，支持之间的联系 MCT4/STAT3/FOXM1 和有氧糖酵解。基于这些数据，我将利用比较交叉物种方法首先定义支持 MCT1/MCT4 介导的有氧糖酵解的调节回路 OS，然后识别并验证与 MCT 控制细胞代谢相关的治疗漏洞。我假设在 OS 细胞中，持续的 MCT1/MCT4 表达是由组成型 STAT3/FOXM1 驱动的激活并增加 MYC 拷贝数，从而促进有氧糖酵解。我进一步预测通过基因操作或靶向抑制剂损失 MCT1/MCT4 将损害体内肿瘤生长，并且可以通过合理的药物组合来最大化这一点。为了实现这个目标，我首先要使用 ChIP 测序组合表征 OS 中 MCT1/MCT4 的转录调控，亚硫酸氢盐测序和 4C/ChiA-PET 分析。小鼠异种移植研究将补充体外分析评估 MCT1/4 功能丧失如何影响 OS 代谢特征、肿瘤表型和肿瘤生长。最后，我将使用斑马鱼模型筛选具有 MCT 阻断作用的合成致死剂，然后验证结果在小鼠中。塔夫茨大学及其合作伙伴提供的丰富的研究环境确保了资源的获取以及完成拟议工作所需的专业知识。我在兽医肿瘤学、临床试验方面的培训和遗传学以及我的指导团队的指导，他们具有基因组学、生物信息学、动物学方面的专业知识模型和转化肿瘤学使我能够成功过渡到独立。