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High neuropeptide Y release associates with Ewing sarcoma bone dissemination - in vivo model of site-specific metastases.

Hong SH, Tilan JU, Galli S, Izycka-Swieszewska E, Polk T, Horton M, Mahajan A, Christian D, Jenkins S, Acree R, Connors K, Ledo P, Lu C, Lee YC, Rodriguez O, Toretsky JA, Albanese C, Kitlinska J - Oncotarget (2015)

Bottom Line: Hypoxia up-regulates NPY and activates its pro-metastatic functions.To test the impact of NPY on ES metastatic pattern, ES cell lines, SK-ES1 and TC71, with high and low peptide release, respectively, were used in an orthotopic xenograft model.SK-ES1 xenografts resulted in thoracic extra-osseous metastases (67%) and dissemination to bone (50%) and brain (25%), while TC71 tumors metastasized to the lungs (70%).

View Article: PubMed Central - PubMed

Affiliation: Department of Oncology, Lombardi Comprehensive Cancer Center, Georgetown University Medical Center, Georgetown University, Washington DC, USA.

ABSTRACT
Ewing sarcoma (ES) develops in bones or soft tissues of children and adolescents. The presence of bone metastases is one of the most adverse prognostic factors, yet the mechanisms governing their formation remain unclear. As a transcriptional target of EWS-FLI1, the fusion protein driving ES transformation, neuropeptide Y (NPY) is highly expressed and released from ES tumors. Hypoxia up-regulates NPY and activates its pro-metastatic functions. To test the impact of NPY on ES metastatic pattern, ES cell lines, SK-ES1 and TC71, with high and low peptide release, respectively, were used in an orthotopic xenograft model. ES cells were injected into gastrocnemius muscles of SCID/beige mice, the primary tumors excised, and mice monitored for the presence of metastases. SK-ES1 xenografts resulted in thoracic extra-osseous metastases (67%) and dissemination to bone (50%) and brain (25%), while TC71 tumors metastasized to the lungs (70%). Bone dissemination in SK-ES1 xenografts associated with increased NPY expression in bone metastases and its accumulation in bone invasion areas. The genetic silencing of NPY in SK-ES1 cells reduced bone degradation. Our study supports the role for NPY in ES bone invasion and provides new models for identifying pathways driving ES metastases to specific niches and testing anti-metastatic therapeutics.

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TC71 xenografts give rise to lung metastases and local relapses, while SK-ES1 xenografts metastasize to soft tissues of the thoracic region, bones and brainA. TC71 pulmonary metastases detected by magnetic resonance imaging (MRI) and macroscopic observation. The presence of metastases was confirmed by histological examination and positive staining for ES marker, CD99. B. MRI of recurrent TC71 tumor (white outline) adjacent to the amputation site (arrows). C. Soft tissue metastasis in thoracic region of mouse bearing an SK-ES1 xenograft. The tumor below lungs is identified by MRI (white outline), histopathology and positive staining for the ES marker, CD99. D. Distant metastasis to the contralateral tibia detected by MRI. Histopathology analysis confirms massive bone degradation within tumor. The presence of tumor cells as confirmed by CD99 staining. E. MRI of SK-ES1 metastasis to the brain (white outline) confirmed by histopathology and CD99 immunohistochemistry. F. NPY mRNA in SK-ES1 primary tumors and corresponding distant bone metastases measured by real-time RT-PCR. R – right; L – left; T – tumor; H&E – hematoxylin and eosin staining; * - p<0.05.
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Figure 2: TC71 xenografts give rise to lung metastases and local relapses, while SK-ES1 xenografts metastasize to soft tissues of the thoracic region, bones and brainA. TC71 pulmonary metastases detected by magnetic resonance imaging (MRI) and macroscopic observation. The presence of metastases was confirmed by histological examination and positive staining for ES marker, CD99. B. MRI of recurrent TC71 tumor (white outline) adjacent to the amputation site (arrows). C. Soft tissue metastasis in thoracic region of mouse bearing an SK-ES1 xenograft. The tumor below lungs is identified by MRI (white outline), histopathology and positive staining for the ES marker, CD99. D. Distant metastasis to the contralateral tibia detected by MRI. Histopathology analysis confirms massive bone degradation within tumor. The presence of tumor cells as confirmed by CD99 staining. E. MRI of SK-ES1 metastasis to the brain (white outline) confirmed by histopathology and CD99 immunohistochemistry. F. NPY mRNA in SK-ES1 primary tumors and corresponding distant bone metastases measured by real-time RT-PCR. R – right; L – left; T – tumor; H&E – hematoxylin and eosin staining; * - p<0.05.

Mentions: Consistent with the rapid growth of primary tumors, progression of the disease to metastasis in mice bearing TC71 xenografts was observed within 2-3 weeks (average of 18 days post-amputation to the first metastasis detection). The tumors metastasized exclusively to the lungs (70% of cases) and no other distant metastases were detected (Fig. 2A, Table 1). However, 90% of mice bearing TC71 xenografts developed local relapses in areas adjacent to the amputation sites, which gave rise to abdominal tumors (Fig. 2B), suggesting a high local invasiveness of TC71 cells.


High neuropeptide Y release associates with Ewing sarcoma bone dissemination - in vivo model of site-specific metastases.

Hong SH, Tilan JU, Galli S, Izycka-Swieszewska E, Polk T, Horton M, Mahajan A, Christian D, Jenkins S, Acree R, Connors K, Ledo P, Lu C, Lee YC, Rodriguez O, Toretsky JA, Albanese C, Kitlinska J - Oncotarget (2015)

TC71 xenografts give rise to lung metastases and local relapses, while SK-ES1 xenografts metastasize to soft tissues of the thoracic region, bones and brainA. TC71 pulmonary metastases detected by magnetic resonance imaging (MRI) and macroscopic observation. The presence of metastases was confirmed by histological examination and positive staining for ES marker, CD99. B. MRI of recurrent TC71 tumor (white outline) adjacent to the amputation site (arrows). C. Soft tissue metastasis in thoracic region of mouse bearing an SK-ES1 xenograft. The tumor below lungs is identified by MRI (white outline), histopathology and positive staining for the ES marker, CD99. D. Distant metastasis to the contralateral tibia detected by MRI. Histopathology analysis confirms massive bone degradation within tumor. The presence of tumor cells as confirmed by CD99 staining. E. MRI of SK-ES1 metastasis to the brain (white outline) confirmed by histopathology and CD99 immunohistochemistry. F. NPY mRNA in SK-ES1 primary tumors and corresponding distant bone metastases measured by real-time RT-PCR. R – right; L – left; T – tumor; H&E – hematoxylin and eosin staining; * - p<0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4466675&req=5

Figure 2: TC71 xenografts give rise to lung metastases and local relapses, while SK-ES1 xenografts metastasize to soft tissues of the thoracic region, bones and brainA. TC71 pulmonary metastases detected by magnetic resonance imaging (MRI) and macroscopic observation. The presence of metastases was confirmed by histological examination and positive staining for ES marker, CD99. B. MRI of recurrent TC71 tumor (white outline) adjacent to the amputation site (arrows). C. Soft tissue metastasis in thoracic region of mouse bearing an SK-ES1 xenograft. The tumor below lungs is identified by MRI (white outline), histopathology and positive staining for the ES marker, CD99. D. Distant metastasis to the contralateral tibia detected by MRI. Histopathology analysis confirms massive bone degradation within tumor. The presence of tumor cells as confirmed by CD99 staining. E. MRI of SK-ES1 metastasis to the brain (white outline) confirmed by histopathology and CD99 immunohistochemistry. F. NPY mRNA in SK-ES1 primary tumors and corresponding distant bone metastases measured by real-time RT-PCR. R – right; L – left; T – tumor; H&E – hematoxylin and eosin staining; * - p<0.05.
Mentions: Consistent with the rapid growth of primary tumors, progression of the disease to metastasis in mice bearing TC71 xenografts was observed within 2-3 weeks (average of 18 days post-amputation to the first metastasis detection). The tumors metastasized exclusively to the lungs (70% of cases) and no other distant metastases were detected (Fig. 2A, Table 1). However, 90% of mice bearing TC71 xenografts developed local relapses in areas adjacent to the amputation sites, which gave rise to abdominal tumors (Fig. 2B), suggesting a high local invasiveness of TC71 cells.

Bottom Line: Hypoxia up-regulates NPY and activates its pro-metastatic functions.To test the impact of NPY on ES metastatic pattern, ES cell lines, SK-ES1 and TC71, with high and low peptide release, respectively, were used in an orthotopic xenograft model.SK-ES1 xenografts resulted in thoracic extra-osseous metastases (67%) and dissemination to bone (50%) and brain (25%), while TC71 tumors metastasized to the lungs (70%).

View Article: PubMed Central - PubMed

Affiliation: Department of Oncology, Lombardi Comprehensive Cancer Center, Georgetown University Medical Center, Georgetown University, Washington DC, USA.

ABSTRACT
Ewing sarcoma (ES) develops in bones or soft tissues of children and adolescents. The presence of bone metastases is one of the most adverse prognostic factors, yet the mechanisms governing their formation remain unclear. As a transcriptional target of EWS-FLI1, the fusion protein driving ES transformation, neuropeptide Y (NPY) is highly expressed and released from ES tumors. Hypoxia up-regulates NPY and activates its pro-metastatic functions. To test the impact of NPY on ES metastatic pattern, ES cell lines, SK-ES1 and TC71, with high and low peptide release, respectively, were used in an orthotopic xenograft model. ES cells were injected into gastrocnemius muscles of SCID/beige mice, the primary tumors excised, and mice monitored for the presence of metastases. SK-ES1 xenografts resulted in thoracic extra-osseous metastases (67%) and dissemination to bone (50%) and brain (25%), while TC71 tumors metastasized to the lungs (70%). Bone dissemination in SK-ES1 xenografts associated with increased NPY expression in bone metastases and its accumulation in bone invasion areas. The genetic silencing of NPY in SK-ES1 cells reduced bone degradation. Our study supports the role for NPY in ES bone invasion and provides new models for identifying pathways driving ES metastases to specific niches and testing anti-metastatic therapeutics.

Show MeSH
Related in: MedlinePlus