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IR/IGF1R signaling as potential target for treatment of high-grade osteosarcoma.

Kuijjer ML, Peterse EF, van den Akker BE, Briaire-de Bruijn IH, Serra M, Meza-Zepeda LA, Myklebost O, Hassan AB, Hogendoorn PC, Cleton-Jansen AM - BMC Cancer (2013)

Bottom Line: This pathway plays a key role in the growth and development of bone.Inhibitory effects of this drug were measured by Western blotting and cell proliferation assays.OSI-906 had a strong inhibitory effect on proliferation of 3 of 4 osteosarcoma cell lines, with IC₅₀s below 100 nM at 72 hrs of treatment.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pathology, Leiden University Medical Center, Albinusdreef 2, Leiden 2300RC, the Netherlands.

ABSTRACT

Background: High-grade osteosarcoma is an aggressive tumor most often developing in the long bones of adolescents, with a second peak in the 5th decade of life. Better knowledge on cellular signaling in this tumor may identify new possibilities for targeted treatment.

Methods: We performed gene set analysis on previously published genome-wide gene expression data of osteosarcoma cell lines (n=19) and pretreatment biopsies (n=84). We characterized overexpression of the insulin-like growth factor receptor (IGF1R) signaling pathways in human osteosarcoma as compared with osteoblasts and with the hypothesized progenitor cells of osteosarcoma - mesenchymal stem cells. This pathway plays a key role in the growth and development of bone. Since most profound differences in mRNA expression were found at and upstream of the receptor of this pathway, we set out to inhibit IR/IGF1R using OSI-906, a dual inhibitor for IR/IGF1R, on four osteosarcoma cell lines. Inhibitory effects of this drug were measured by Western blotting and cell proliferation assays.

Results: OSI-906 had a strong inhibitory effect on proliferation of 3 of 4 osteosarcoma cell lines, with IC₅₀s below 100 nM at 72 hrs of treatment. Phosphorylation of IRS-1, a direct downstream target of IGF1R signaling, was inhibited in the responsive osteosarcoma cell lines.

Conclusions: This study provides an in vitro rationale for using IR/IGF1R inhibitors in preclinical studies of osteosarcoma.

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Related in: MedlinePlus

Ingenuity pathways analysis canonical pathway IGF1 signaling This figure shows the IGF1 signaling pathway, with significantly upregulated genes in red, downregulated genes in green, and genes that did not meet our criteria for significance in gray. The left part of the symbols shows the analysis of osteosarcoma cell lines as compared with mesenchymal stem cells, the right part as compared with osteoblasts. Most consensus in gene expression is found upstream IGF1R signaling, in the expression of the IGF binding proteins.
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Figure 2: Ingenuity pathways analysis canonical pathway IGF1 signaling This figure shows the IGF1 signaling pathway, with significantly upregulated genes in red, downregulated genes in green, and genes that did not meet our criteria for significance in gray. The left part of the symbols shows the analysis of osteosarcoma cell lines as compared with mesenchymal stem cells, the right part as compared with osteoblasts. Most consensus in gene expression is found upstream IGF1R signaling, in the expression of the IGF binding proteins.

Mentions: To determine which genes have the most specific up- or downregulation in osteosarcoma, we combined lists of significantly differentially expressed genes of osteosarcoma cell lines (n=19) and a previously published set of osteosarcoma pretreatment biopsies (n=84, GEO accession GSE33382) in comparison with mesenchymal stem cells (n=12) and osteoblasts (n=3) by four-way Venn analysis of all significantly affected probes with the same direction of fold change (upregulated or downregulated in all four analyses) (Additional files 1 and 2). We identified IGFBP4 and GAS6 as the most downregulated genes in osteosarcoma (average log fold changes of -4.43 and -4.29, respectively). IGFBP2 was also present in the top 20 results from this four-way analysis (see Additional file 1). In addition, IGFBP3 and −7 were significantly downregulated, and IGF2BP3 was significantly upregulated in three out of the four analyses. Both IGFBP4 and GAS6 show high variability in expression in osteosarcoma cell lines and biopsies (Figure 1A). Patients of whom biopsies had very low expression of these genes had poor event-free survival profiles (log-rank test for trend, P = 0.01 for IGFBP4 and P = 0.04 for GAS6, Figure 1B). To visualize mRNA expression of the IGF1R signaling pathway members, we used Ingenuity Pathways Analysis on LIMMA toptables from osteosarcoma cells as compared with mesenchymal stem cells and from osteosarcoma cells as compared with osteoblasts (Figure 2). As can be seen in this figure, overlap of differentially expressed genes between these analyses was detected upstream of IGF1R.


IR/IGF1R signaling as potential target for treatment of high-grade osteosarcoma.

Kuijjer ML, Peterse EF, van den Akker BE, Briaire-de Bruijn IH, Serra M, Meza-Zepeda LA, Myklebost O, Hassan AB, Hogendoorn PC, Cleton-Jansen AM - BMC Cancer (2013)

Ingenuity pathways analysis canonical pathway IGF1 signaling This figure shows the IGF1 signaling pathway, with significantly upregulated genes in red, downregulated genes in green, and genes that did not meet our criteria for significance in gray. The left part of the symbols shows the analysis of osteosarcoma cell lines as compared with mesenchymal stem cells, the right part as compared with osteoblasts. Most consensus in gene expression is found upstream IGF1R signaling, in the expression of the IGF binding proteins.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3672007&req=5

Figure 2: Ingenuity pathways analysis canonical pathway IGF1 signaling This figure shows the IGF1 signaling pathway, with significantly upregulated genes in red, downregulated genes in green, and genes that did not meet our criteria for significance in gray. The left part of the symbols shows the analysis of osteosarcoma cell lines as compared with mesenchymal stem cells, the right part as compared with osteoblasts. Most consensus in gene expression is found upstream IGF1R signaling, in the expression of the IGF binding proteins.
Mentions: To determine which genes have the most specific up- or downregulation in osteosarcoma, we combined lists of significantly differentially expressed genes of osteosarcoma cell lines (n=19) and a previously published set of osteosarcoma pretreatment biopsies (n=84, GEO accession GSE33382) in comparison with mesenchymal stem cells (n=12) and osteoblasts (n=3) by four-way Venn analysis of all significantly affected probes with the same direction of fold change (upregulated or downregulated in all four analyses) (Additional files 1 and 2). We identified IGFBP4 and GAS6 as the most downregulated genes in osteosarcoma (average log fold changes of -4.43 and -4.29, respectively). IGFBP2 was also present in the top 20 results from this four-way analysis (see Additional file 1). In addition, IGFBP3 and −7 were significantly downregulated, and IGF2BP3 was significantly upregulated in three out of the four analyses. Both IGFBP4 and GAS6 show high variability in expression in osteosarcoma cell lines and biopsies (Figure 1A). Patients of whom biopsies had very low expression of these genes had poor event-free survival profiles (log-rank test for trend, P = 0.01 for IGFBP4 and P = 0.04 for GAS6, Figure 1B). To visualize mRNA expression of the IGF1R signaling pathway members, we used Ingenuity Pathways Analysis on LIMMA toptables from osteosarcoma cells as compared with mesenchymal stem cells and from osteosarcoma cells as compared with osteoblasts (Figure 2). As can be seen in this figure, overlap of differentially expressed genes between these analyses was detected upstream of IGF1R.

Bottom Line: This pathway plays a key role in the growth and development of bone.Inhibitory effects of this drug were measured by Western blotting and cell proliferation assays.OSI-906 had a strong inhibitory effect on proliferation of 3 of 4 osteosarcoma cell lines, with IC₅₀s below 100 nM at 72 hrs of treatment.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pathology, Leiden University Medical Center, Albinusdreef 2, Leiden 2300RC, the Netherlands.

ABSTRACT

Background: High-grade osteosarcoma is an aggressive tumor most often developing in the long bones of adolescents, with a second peak in the 5th decade of life. Better knowledge on cellular signaling in this tumor may identify new possibilities for targeted treatment.

Methods: We performed gene set analysis on previously published genome-wide gene expression data of osteosarcoma cell lines (n=19) and pretreatment biopsies (n=84). We characterized overexpression of the insulin-like growth factor receptor (IGF1R) signaling pathways in human osteosarcoma as compared with osteoblasts and with the hypothesized progenitor cells of osteosarcoma - mesenchymal stem cells. This pathway plays a key role in the growth and development of bone. Since most profound differences in mRNA expression were found at and upstream of the receptor of this pathway, we set out to inhibit IR/IGF1R using OSI-906, a dual inhibitor for IR/IGF1R, on four osteosarcoma cell lines. Inhibitory effects of this drug were measured by Western blotting and cell proliferation assays.

Results: OSI-906 had a strong inhibitory effect on proliferation of 3 of 4 osteosarcoma cell lines, with IC₅₀s below 100 nM at 72 hrs of treatment. Phosphorylation of IRS-1, a direct downstream target of IGF1R signaling, was inhibited in the responsive osteosarcoma cell lines.

Conclusions: This study provides an in vitro rationale for using IR/IGF1R inhibitors in preclinical studies of osteosarcoma.

Show MeSH
Related in: MedlinePlus