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.
| Ren, Ling; Hong, Ellen S; Mendoza, Arnulfo et al. (2017) Metabolomics uncovers a link between inositol metabolism and osteosarcoma metastasis. Oncotarget 8:38541-38553 |
