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

• 批准号: 10578687
• 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
• 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/10578687
• 关键词: 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; Credentialing; 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; Invaded; Lactic acid; Link; Long Term Survivorship; Malignant Bone Neoplasm; Mediating; Mediator; Medical Genetics; Mentors; Metabolic; Metabolic Pathway; Metabolism; Metastatic Osteosarcoma; Microscopic; Modeling; Molecular; Mus; Mutation; Neoplasm Metastasis; Nutrient; Oncology; Operative Surgical Procedures; Pathway interactions; Persons; Phenotype; Phosphotransferases; Positioning Attribute; Primary Neoplasm; Proliferating; Recording of previous events; Refractory; Regulation; Research; Resistance; Resources; STAT3 gene; Scientist; Somatic Mutation; Therapeutic; Training; Transcriptional Regulation; Tumor Immunity; Universities; Variant; Work; Xenograft procedure; Zebrafish; aerobic glycolysis; arginase; bisulfite sequencing; cancer cell; cancer genome; cell growth; chemotherapy; clinical development; clinical translation; comparative; design; flexibility; genetic manipulation; immune checkpoint blockade; improved; in vivo; inhibitor; innovation; kinase inhibitor; knockout gene; loss of function; metabolic profile; mouse model; neoplastic cell; osteosarcoma; programs; skills; small molecule inhibitor; targeted agent; therapeutic target; translational oncology; tumor; tumor growth; tumor metabolism; tumor microenvironment

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.

项目总结/摘要 骨肉瘤（OS）是人类和犬中最常见的恶性骨肿瘤， 包括临床表现和分子改变。尽管做出了许多努力， 在过去的三十年里，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背景下显示出了希望， 在这两个物种中，肉眼可见的转移表现出对多种药物（激酶抑制剂， 化疗、免疫疗法等）。我最近描述了狗的OS基因组，发现 与人OS一样，体细胞突变负荷低，拷贝数畸变/结构变异占主导地位， 没有明显的分子驱动。这些数据，沿着临床试验失败的历史表明 当代的治疗方法，如激酶抑制剂和免疫检查点， 阻断的临床效果可能有限，因此需要开发创新的治疗药物。 战略布局癌细胞的一个显著特征是它们能够进行有氧糖酵解，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，并且对于糖酵解肿瘤的生长和转移至关重要，例如 OS.在以前的工作中，我发现OS细胞中MCT 1或MCT 4功能的丧失降低了基础和代偿性功能， 糖酵解、细胞增殖和侵袭能力。我还证明了MCT 4是一个直接的转录靶点， STAT 3和FOXM 1，这两者都表现出组成性激活OS，支持之间的联系， MCT 4/STAT 3/FOXM 1和有氧糖酵解。在这些数据的基础上，我将利用一个比较交叉- 种方法，首先定义支持由MCT 1/MCT 4介导的有氧糖酵解的调节回路， OS，然后识别和验证与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MCT 1/MCT 4表达由组成型STAT 3/FOXM 1驱动 激活和增加MYC拷贝数，从而促进有氧糖酵解。我进一步预测， 通过遗传操作或靶向抑制剂而导致的MCT 1/MCT 4的损失将损害体内肿瘤生长， 通过合理的药物组合可以最大限度地发挥这一作用。为了做到这一点，我将首先 使用ChIP测序的组合表征OS中MCT 1/MCT 4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MCT 1/4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 将使用斑马鱼模型来筛选具有MCT阻断的合成致死性的选择剂，然后验证发现 对小鼠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 和完成拟议工作所需的专业知识。我在兽医肿瘤学，临床试验 和遗传学沿着与指导我的导师团队的专业知识，在基因组学，生物信息学，动物 模型和转化肿瘤学使我能够成功地过渡到独立。