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、离体/体内（肺转移试验或PuMA）模型系统中对鼠、犬和人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影响的理解。以这种方式生成的数据提高了对自然发生的犬骨肉瘤（OS）模型的理解，并可用于通过COTC临床试验机制回答关于药物抗转移潜力的独特体内问题。在过去的2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，在我们的克隆相关的一对低和高转移OS细胞系与关键代谢基因的药理学和基因组靶向研究之间，细胞能量学紊乱，包括但不限于TCA循环、糖酵解和β-氨基葡萄糖分解途径。我们正在询问这些差异定量使用各种技术，如海马细胞外通量分析和13 C-丙酮酸超极化MRI，在体外和体内的调查，分别。此外，我们已经表明，线粒体复合物1抑制剂（二甲双胍）与谷氨酰胺1抑制剂（CB-839）的组合可以有效地治疗骨肉瘤转移的小鼠模型中的转移进展，并且潜在的机制主要与抑制天冬氨酸生物合成和通过脂质的细胞积累诱导不可控制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，其中狗与自然发生的犬OS成像MIP扫描仪（PET/CT和MRI），以建立和研究犬OS的代谢表型的翻译相关性。成像研究完成后，狗接受截肢在他们的家庭兽医办公室作为这种疾病的常规护理标准的一部分。从其中一个病例中，已经使用WGS分析了骨肉瘤病变的几个地理上不同的区域，并且我们继续招募犬，其中我们可以在手术时收集新鲜组织标本，以支持全局代谢谱分析（13 C-葡萄糖NMR通量分析，代谢子谱分析，RNAseq）。我们的目标是更充分地了解狗作为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.