glycogen and Osteosarcoma

Glycogen dysregulation is a hallmark of aging, and aberrant glycogen drives metabolic reprogramming and pathogenesis in multiple diseases. However, glycogen heterogeneity in healthy and diseased tissues remains largely unknown. Herein, we describe a method to define spatial glycogen architecture in mouse and human tissues using matrix-assisted laser desorption/ionization mass spectrometry imaging. This assay provides robust and sensitive spatial glycogen quantification and architecture characterization in the brain, liver, kidney, testis, lung, bladder, and even the bone. Armed with this tool, we interrogated glycogen spatial distribution and architecture in different types of human cancers. We demonstrate that glycogen stores and architecture are heterogeneous among diseases. Additionally, we observe unique hyperphosphorylated glycogen accumulation in Ewing sarcoma, a pediatric bone cancer. Using preclinical models, we correct glycogen hyperphosphorylation in Ewing sarcoma through genetic and pharmacological interventions that ablate in vivo tumor growth, demonstrating the clinical therapeutic potential of targeting glycogen in Ewing sarcoma.

Topics: Animals; Bone Neoplasms; Child; Glycogen; Humans; Male; Mice; Osteosarcoma; Sarcoma, Ewing; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Osteosarcoma (OS) is the most common malignant bone tumor in children and adolescents. The survival rate of patients with metastatic disease remains very dismal. Nevertheless, metastasis is a complex process and a single-level analysis is not likely to identify its key biological determinants. In this study, we used a systems biology approach to identify common metastatic pathways that are jointly supported by both mRNA and protein expression data in two distinct human metastatic OS models.. mRNA expression microarray and N-linked glycoproteomic analyses were performed on two commonly used isogenic pairs of human metastatic OS cell lines, namely HOS/143B and SaOS-2/LM7. Pathway analysis of the differentially regulated genes and glycoproteins separately revealed pathways associated to metastasis including cell cycle regulation, immune response, and epithelial-to-mesenchymal-transition. However, no common significant pathway was found at both genomic and proteomic levels between the two metastatic models, suggesting a very different biological nature of the cell lines. To address this issue, we used a topological significance analysis based on a "shortest-path" algorithm to identify topological nodes, which uncovered additional biological information with respect to the genomic and glycoproteomic profiles but remained hidden from the direct analyses. Pathway analysis of the significant topological nodes revealed a striking concordance between the models and identified significant common pathways, including "Cytoskeleton remodeling/TGF/WNT", "Cytoskeleton remodeling/Cytoskeleton remodeling", and "Cell adhesion/Chemokines and adhesion". Of these, the "Cytoskeleton remodeling/TGF/WNT" was the top ranked common pathway from the topological analysis of the genomic and proteomic profiles in the two metastatic models. The up-regulation of proteins in the "Cytoskeleton remodeling/TGF/WNT" pathway in the SaOS-2/LM7 and HOS/143B models was further validated using an orthogonal Reverse Phase Protein Array platform.. In this study, we used a systems biology approach by integrating genomic and proteomic data to identify key and common metastatic mechanisms in OS. The use of the topological analysis revealed hidden biological pathways that are known to play critical roles in metastasis. Wnt signaling has been previously implicated in OS and other tumors, and inhibitors of Wnt signaling pathways are available for clinical testing. Further characterization of this common pathway and other topological pathways identified from this study may lead to a novel therapeutic strategy for the treatment of metastatic OS.

Topics: Animals; Cell Line, Tumor; Cytoskeleton; Gene Expression Regulation, Neoplastic; Glycogen; Glycoproteins; Humans; Lung Neoplasms; Mice; Neoplasm Metastasis; Osteosarcoma; Proteomics; RNA, Messenger; Signal Transduction; Systems Biology; Transcriptome; Tumor Necrosis Factor-alpha; Wheat Germ Agglutinins; Wnt Proteins

The insulin-like growth factor type 1 receptor (IGF 1R) mediates the acute metabolic effects of IGF I as well as IGF I-stimulated cell proliferation and protection from apoptosis. IGF binding proteins (IGFBPs) can modulate these responses. We, therefore, investigated whether intrinsic IGFBPs interfere with IGF I-induced regulation of IGF 1R expression and with the biological response to IGF I in two human tumor cell lines, the non-small-cell lung cancer cell line A549 and the osteoblastic osteosarcoma cell line Saos-2/B-10. We compared the growth rates, IGFBP production, IGF I binding characteristics, IGF 1R protein and mRNA levels, and the acute IGF I response (stimulation of glycogen synthesis) after pretreatment of the cells in serum-free medium with or without added IGF I or medium supplemented with 5% fetal calf serum (FCS). In contrast to A549 cells, which produce IGF I and significant amounts of IGFBPs, survival and proliferation of Saos-2/B-10 cells, which do not produce IGF I or significant amounts of IGFBPs, depended on the addition of exogenous IGF I. IGF I increased the concentration of IGFBP-2 and -3 and decreased the concentration of IGFBP-4 in the medium of A549 cells. As compared to FCS, IGF I pretreatment in both cell lines decreased the number of specific IGF I binding sites, down-regulated total and membrane IGF 1R protein, and largely reduced or abolished the acute IGF I response without affecting IGF 1R mRNA levels. The data suggest that the IGF 1R protein of the two cell lines is translationally and/or posttranslationally down-regulated by its ligand in the presence and in the absence of locally produced IGFBPs and that the cell lines have retained this negative feedback to counteract IGF I stimulation.

Topics: Apoptosis; Binding Sites; Binding, Competitive; Blood Proteins; Bone Neoplasms; Carcinoma, Non-Small-Cell Lung; Cell Division; Culture Media, Conditioned; Down-Regulation; Feedback; Gene Expression Regulation, Neoplastic; Glucose; Glycogen; Humans; Insulin-Like Growth Factor Binding Proteins; Insulin-Like Growth Factor I; Iodine Radioisotopes; Lung Neoplasms; Osteosarcoma; Protein Biosynthesis; Radioligand Assay; Receptor, IGF Type 1; RNA, Messenger; Tumor Cells, Cultured

IGFBP-3 is the predominant IGFBP in serum and the major IGFBP secreted by osteoblasts. Native and recombinant IGFBP-3 and a truncated form lacking the carboxyterminal third were tested for their effects on 2 osteoblastic cell lines. Intact but not truncated IGFBP-3 blocked IGF I-stimulated DNA and glycogen synthesis. Inhibition was dose-dependent and found whenever the concentration of intact IGFBP-3 exceeded the concentration of IGF I. Truncated IGFBP-3 appears to result from proteolytic cleavage and does occur in vivo. The loss of inhibition by IGFBP-3 may be regulated at the site of IGF target cells and thus be essential for IGF I-induced osteoblast growth.

Topics: Animals; Carrier Proteins; Cell Line; DNA Replication; Glucose; Glycogen; Humans; Insulin; Insulin-Like Growth Factor Binding Proteins; Insulin-Like Growth Factor I; Osteoblasts; Osteosarcoma; Rats; Recombinant Proteins; Signal Transduction

The reports concerns the light microscopical and ultrastructural findings obtained in three conventional osteosarcomas with an unusually high admixture of clear cells, whose presence appeared to be responsible for the marked change in the histological pattern of these tumours. In the tumours with a prevailing fibroblastic component the clear cells were either irregularly scattered throughout the tumour in the form of small groups, or they formed large groups sharply demarcated against the fusicellular areas of the tumours. In two cases it was shown that their cytoplasm contained exaggerated glycogen deposits accompanied by the formation of glycogen-containing phagolysosomes and occasional empty vacuoles. In the third case the clear cells showed vacuolar degeneration with numerous single-membrane-bound, empty vacuoles. In contrast to the clear-cell chondrosarcoma we did not find S-100 protein in clear cells of our osteosarcomas. Such findings could be particularly significant in the differential diagnosis of bone tumours.

Topics: Adolescent; Cytoplasmic Granules; Female; Glycogen; Humans; Male; Microscopy, Electron; Middle Aged; Osteosarcoma

The ultrastructure of three cases of osteoblastoma is described. The osteoblasts resemble normal osteoblasts with a few exceptions: irregular, indented nuclei, and occasional mitochondria with curved cristae and electron-lucent areas. The osteocytes and osteoclasts basically resemble their normal counterparts. There are also osteoprogenitor cells in different stages of maturation, some containing glycogen. The ultrastructure of an additional case histologically classified as aggressive osteoblastoma is described. It is essentially similar to the previous three cases of typical osteoblastoma. The only exception is the presence of osteoblasts with electron-lucent nuclei and less prominent organelles than the typical osteoblastoma cells have markedly indented and multilobed nuclei, dilated rough endoplasmic reticulum and lipid droplets. However, these differences from osteoblastoma cells are not pathognomonic. The final diagnosis of osteoblastic tumors rests at the light microscopy level.

Topics: Adolescent; Adult; Bone Neoplasms; Cell Differentiation; Female; Glycogen; Humans; Humerus; Lumbar Vertebrae; Male; Metatarsus; Mitochondria; Osteoblasts; Osteoclasts; Osteocytes; Osteoma, Osteoid; Osteosarcoma; Spinal Neoplasms; Thoracic Vertebrae

Cytologic and cytochemical examination of eighteen cases of round-cell sarcoma of bone allowed classification of these tumors into four cytologic groups. Additional cytochemical examinations based on the PAS and D-PAS reactions, and the demonstration of the activity of peroxidase, naphtol-ASD-Chloracetate esterase, alpha-naphthylacetate esterase, naphthol-AS-acetate esterase with and without sodium fluoride inhibition, acid and alkaline phosphatases yielded no evidence of uniform behavior among the individual groups or within any single group. The studies showed that a positive glycogen reaction cannot be used as a basic criterion for the classification of such tumors as Ewing's sarcoma and for regarding them as a uniform tumor group. It is possible that a pool of tumors is involved, including tumors of monocytic and probably of lymphocytic origin, reticulum-cell sarcoma, tumors of myelocytic and erythroplastic origin, stem-cell tumors, and endothelial-cell tumors. Histologic examination alone is not sufficient for the classification of round-cell sarcomas of bone, and it should be supplemented by cytologic and cytochemical or histochemical methods. Osteosarcomas (23 cases) and chondrosarcomas (8 cases) display cells which are characteristic for these tumors and which could be correlated with their benign counterparts, osteoblasts and chondroid cells. The histologically recognizable degree of malignancy of chondrosarcoma can be evaluated better with the cytologic than with the histologic technic. Indications of the possibilities of differential diagnosis based on the cytologic pictures of benign and malignant osteoplastic and chondroplastic tumors, giant-cell tumors and chordoma are discussed.

Topics: Adolescent; Bone Neoplasms; Cell Nucleolus; Cell Nucleus; Child; Chondroblastoma; Chondroma; Chondrosarcoma; Chordoma; Cytoplasm; Female; Giant Cell Tumors; Glycogen; Humans; Infant; Male; Naphthol AS D Esterase; Osteosarcoma; Periodic Acid-Schiff Reaction; Phosphoric Monoester Hydrolases; Sarcoma; Sarcoma, Ewing

Topics: Aneuploidy; Biopsy; Cell Line; Cells, Cultured; Chromosome Aberrations; Culture Techniques; Female; Glycogen; Histocytochemistry; Humans; Karyotyping; Microscopy, Electron; Middle Aged; Neoplasm Recurrence, Local; Osteosarcoma; Skull Neoplasms

Topics: Animals; Beryllium; Bone Neoplasms; Calcium; Esterases; Glycogen; Glycosaminoglycans; Histocytochemistry; Magnesium; Osteosarcoma; Oxidoreductases; Phosphoric Monoester Hydrolases; Phosphorus; Rabbits; Sarcoma, Experimental

Topics: Alkaline Phosphatase; Biopsy; Bone Cysts; Bone Diseases; Bone Neoplasms; Chondroma; Chondrosarcoma; Clinical Enzyme Tests; Diagnosis, Differential; Femoral Neoplasms; Fibrosarcoma; Giant Cell Tumors; Glycogen; Hemangioma; Hemangiosarcoma; Histocytochemistry; Humans; Ilium; Lymphoma, Large B-Cell, Diffuse; Multiple Myeloma; Neoplasm Metastasis; Osteoma; Osteoma, Osteoid; Osteomyelitis; Osteosarcoma; Ribs; Sarcoma, Ewing; Tibia; Trephining; Tuberculosis, Osteoarticular

Topics: Adolescent; Calcification, Physiologic; Calcinosis; Cell Membrane; Cell Nucleus; Endoplasmic Reticulum; Female; Femoral Neoplasms; Glycogen; Golgi Apparatus; Humans; Hydroxyapatites; Lipid Metabolism; Lysosomes; Microscopy, Electron; Mitochondria; Osteoblasts; Osteosarcoma; Ribosomes

Topics: Bone Diseases; Bone Neoplasms; Chondroma; Chondrosarcoma; Glycogen; Humans; Osteoma; Osteosarcoma