Immunohistochemical Characterisation of GLUT1, MMP3 and NRF2 in Osteosarcoma

Abstract

Osteosarcoma (OSA) is an aggressive bone malignancy. Unlike many other malignancies, OSA outcomes have not improved in recent decades. One challenge to the development of better diagnostic and therapeutic methods for OSA has been the lack of well characterized experimental model systems. Spontaneous OSA in dogs provides a good model for the disease seen in people and also remains an important veterinary clinical challenge. We recently used RNA sequencing and qRT-PCR to provide a detailed molecular characterization of OSA relative to non-malignant bone in dogs. We identified differential mRNA expression of the solute carrier family 2 member 1 (SLC2A1/GLUT1), matrix metallopeptidase 3 (MMP3) and nuclear factor erythroid 2–related factor 2 (NFE2L2/NRF2) genes in canine OSA tissue in comparison to paired non-tumor tissue. Our present work characterizes protein expression of GLUT1, MMP3 and NRF2 using immunohistochemistry. As these proteins affect key processes such as Wnt activation, heme biosynthesis, glucose transport, understanding their expression and the enriched pathways and gene ontologies enables us to further understand the potential molecular pathways and mechanisms involved in OSA. This study further supports spontaneous OSA in dogs as a model system to inform the development of new methods to diagnose and treat OSA in both dogs and people.

Deletion of Trp53 and Rb1 in Ctsk‐expressing cells drives osteosarcoma progression by activating glucose metabolism and YAP signaling

“… 16 , 19 , 40 , 41 Overexpression of Glut1 has been commonly observed in many cancers, such as osteosarcoma, lung cancer, and lymphoma. 41 , 42 In addition, Glut1 is considered to be the main isoform expressed in human osteosarcoma cell lines (SaOS2, MG‐63, and U2‐OS cell lines). 43 Previous studies have also shown that targeting glucose metabolism has the potential to provide an effective approach for cancer therapy.…”

“…Glucose is a major source of energy in mammalian cells because it upregulates the expression of glycolytic enzymes such as Glut1, HK2, Pfkfb3/4, and LDHA to stimulate tumorigenesis 16,19,40,41 . Overexpression of Glut1 has been commonly observed in many cancers, such as osteosarcoma, lung cancer, and lymphoma 41,42 . In addition, Glut1 is considered to be the main isoform expressed in human osteosarcoma cell lines (SaOS2, MG‐63, and U2‐OS cell lines) 43 .…”

Canine osteosarcoma in comparative oncology: Molecular mechanisms through to treatment discovery

“…None of these loci are consistently associated across breeds, suggesting there may be a difference between breeds regarding genetic predisposition to developing OSA (89,90). In addition to the 34 genetic loci identified, genes have been identified as differentially expressed in canine OSA compared to non-tumor tissue, many of which have implications for growth and metastasis, and are potential drug targets (55, [91][92][93][94][95][96]. These genes have been identified utilizing canine OSA tumor tissue, and/or canine OSA cell lines.…”

“…Interestingly, increased expression of SLC2A1 (GLUT1) within tumors also resulted in poorer prognosis and a shorter disease free interval in people (101). SLC2A1/GLUT1 (see Figure 1) levels were also significantly increased in naturally occurring canine OSA tissue compared to normal bone tissue (55, 95). Inhibition of SLC2A1 and cellular glucose transport has been achieved by a number of pharmaceuticals, however, as with the MMP3 inhibitors, these have yet to be utilized in OSA trials in either people or dogs (102)(103)(104)(105).…”

“…These genes have been identified utilizing canine OSA tumor tissue, and/or canine OSA cell lines. Some proteins of interest have also had histological work undertaken to start identifying their presence and relevance in OSA ( 55 , 95 ). There has also been variation in the expression of genes within tumors associated with survival time in canine OSA ( 97 – 100 ).…”