Comparative Oncology Program Laboratory

COP实验室开展的实验室活动和项目的具体目标是，通过在各种体外、体外/体内(肺转移试验或彪马)模型系统中对小鼠、犬和人类细胞系进行平行研究，提高对抗癌药物对转移生物学比较方面的影响的了解。以这种方式产生的数据提高了对自然发生的犬骨肉瘤(OS)模型的理解，并可通过COTC临床试验机制用于回答有关药物抗转移潜力的独特体内问题。在过去的两年里，我们通过与Peter Choyke/MIP博士的合作，将COP实验室的工作重点放在OS代谢组学和代谢成像上，因为COP现在是CCR分支的一部分。我们拥有的数据表明，细胞能量学在克隆相关的一对低转移和高转移OS细胞系与关键代谢基因的药理和基因组靶向研究之间存在混乱，包括但不限于TCA循环、糖酵解和谷氨酰胺分解途径。我们正在使用各种技术，如海马细胞外流量分析和13C-丙酮酸超极化磁共振成像，分别用于体外和体内研究，定量询问这些差异。此外，我们已经证明，将线粒体复合体1抑制剂(二甲双胍)和谷氨酰胺1抑制剂(CB-839)相结合可以有效地治疗骨肉瘤转移小鼠模型中的转移进展，其潜在机制主要与抑制天冬氨酸生物合成和通过细胞内脂质积累诱导难以处理的氧化还原应激有关。我们还开展了一个比较代谢成像项目，用MIP扫描仪(PET/CT和MRI)对患有自然发生的犬OS的狗进行成像，以建立和研究犬OS代谢表型的翻译相关性。成像研究完成后，狗在其家庭兽医办公室接受截肢手术，作为治疗这种疾病的常规标准的一部分。在其中一个病例中，几个地理上不同的骨肉瘤病变区域已经通过WGS进行了分析，我们正在继续招募狗，我们可以在手术时收集新鲜组织样本，以支持全球代谢谱(13C-葡萄糖核磁共振流量分析、代谢谱分析、RNAseq)。我们的目标是更充分地了解狗作为新陈代谢靶向药物的预测模型的相关性和适用性，这些药物可以转化为人类儿科骨肉瘤临床。

Laboratory activities and projects carried out by the COP laboratory have the specific goal of improving the understanding of the impact of anti-cancer agents on comparative aspects of metastasis biology by virtue of parallel study of murine, canine and human cell lines in a variety of in vitro, ex vivo/in vivo (Pulmonary Metastasis Assay or PuMA) model systems. Data generated in this manner improves understanding of naturally-occurring canine osteosarcoma (OS) models and could be employed to answer unique in vivo questions regarding the anti-metastatic potential of agents, via the COTC clinical trial mechanism. Over the last 2 years, we have focused the work of the COP laboratory on OS metabolomics and metabolic imaging via collaboration with Dr. Peter Choyke/MIP, as the COP now is a part of this CCR branch. We possess data to suggest that cellular energetics are deranged between our clonally-related pair of low and high metastasizing OS cell lines and studies of pharmacologic and genomic targeting of key metabolism genes, including but not limited to TCA cycle, glycolysis and glutaminolysis pathways. We are interrogating these differences quantitatively using a variety of techniques such as Seahorse extracellular flux analysis and 13C-pyruvate hyperpolarized MRI, for in vitro and in vivo investigations, respectively. Further, we have shown that combining a mitochondrial complex 1 inhibitor (metformin) with a glutamine 1 inhibitor (CB-839) can effectively treat metastatic progression within mouse models of osteosarcoma metastasis, and that the underlying mechanism is primarily related to suppression of aspartate biosynthesis and induction of unmanageable redox stress through cellular accumulation of lipids. We have also embarked upon a comparative metabolic imaging project wherein dog with naturally-occurring canine OS are imaged with MIP scanners (PET/CT and MRI) to both establish and investigate the translational relevance of the metabolic phenotype of canine OS. After imaging studies are complete, dogs undergo limb amputation at their family veterinary office as a part of routine standard of care for this disease. From one of these cases, several geographically distinct areas of the osteosarcoma lesion have been analyzed with WGS, and we are continuing to enroll dogs wherein we can collect fresh tissue specimens at the time of surgery in support of global metabolic profiling (13C-glucose NMR flux analysis, Metabolon profiling, RNAseq). Our goal is to more fully understand the relevance and applicability of the dog as a predictive model of metabolically-targeted agents that can be translated into the human pediatric osteosarcoma clinic.