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理解。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，并且可以通过COTC临床试验机制来回答有关药物抗转移潜力的独特体内问题。在过去的两年中，我们通过与Peter Choyke博士/MIP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，细胞能量学在我们克隆相关的低转移性和高转移性OS细胞系和关键代谢基因的药理学和基因组靶向研究之间是紊乱的，包括但不限于TCA循环、糖酵解和谷氨酰胺解途径。我们正在使用各种技术，如海马细胞外通量分析和13c -丙酮酸超极化MRI，分别进行体外和体内研究，定量地询问这些差异。此外，我们已经证明，将线粒体复合物1抑制剂（二甲双胍）与谷氨酰胺1抑制剂（CB-839）联合使用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脂质积累诱导难以控制的氧化还原应激有关。我们还开展了一项比较代谢成像项目，其中使用MIP扫描仪（PET/CT和MRI）对患有自然发生的犬类OS的狗进行成像，以建立和研究犬类OS代谢表型的翻译相关性。在影像学检查完成后，狗在其家庭兽医办公室接受肢体截肢，作为该疾病常规标准护理的一部分。从这些病例中，我们用WGS分析了骨肉瘤病变的几个地理位置不同的区域，我们继续招募狗，在手术时收集新鲜的组织标本，以支持全球代谢谱（13c -葡萄糖核磁共振通量分析，代谢谱，rna - seq）。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，这些预测模型可以转化为人类儿童骨肉瘤临床。

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.