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Expression of MYCN in Multipotent Sympathoadrenal Progenitors Induces Proliferation and Neural Differentiation, but Is Not Sufficient for Tumorigenesis.

Mobley BC, Kwon M, Kraemer BR, Hickman FE, Qiao J, Chung DH, Carter BD - PLoS ONE (2015)

Bottom Line: MYCN overexpression in spheres promoted commitment to the neural lineage, evidenced by an increased frequency of neuron-containing colonies.MYCN promoted proliferation of both sympathoadrenal progenitor spheres and differentiated neurons derived from these spheres, but there was also an increase in apoptosis.We find, however, that MYCN overexpression is not sufficient for these cells to form tumors in nude mice, suggesting that additional transforming mutations are necessary for tumorigenesis.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Microbiology, and Immunology, Division of Neuropathology, Vanderbilt University Medical Center, Nashville, Tennessee, the United States of America.

ABSTRACT
Neuroblastoma is a pediatric malignancy of the sympathetic ganglia and adrenal glands, hypothesized to originate from progenitors of the developing sympathetic nervous system. Amplification of the MYCN oncogene is a genetic marker of risk in this disease. Understanding the impact of oncogene expression on sympathoadrenal progenitor development may improve our knowledge of neuroblastoma initiation and progression. We isolated sympathoadrenal progenitor cells from the postnatal murine adrenal gland by sphere culture and found them to be multipotent, generating differentiated colonies of neurons, Schwann cells, and myofibroblasts. MYCN overexpression in spheres promoted commitment to the neural lineage, evidenced by an increased frequency of neuron-containing colonies. MYCN promoted proliferation of both sympathoadrenal progenitor spheres and differentiated neurons derived from these spheres, but there was also an increase in apoptosis. The proliferation, apoptosis, and neural lineage commitment induced by MYCN are tumor-like characteristics and thereby support the hypothesis that multipotent adrenal medullary progenitor cells are cells of origin for neuroblastoma. We find, however, that MYCN overexpression is not sufficient for these cells to form tumors in nude mice, suggesting that additional transforming mutations are necessary for tumorigenesis.

No MeSH data available.


Related in: MedlinePlus

MYCN Expressing Progenitors Do Not Form Tumors.(A) Tumors derived from subcutaneously injected BE (2)-C cells showed neuroblastoma-like histologic features including dense cellularity, high nuclear-to-cytoplasmic ratios, high mitotic activity, and frequent apoptotic bodies. Scale bar equals 100 microns. (B) BE (2)-C derived tumors showed diffuse nuclear expression of PHOX2B and (C) diffuse cytoplasmic TH expression. Scale bars equal 100 microns. (D) While human neuroblastoma cells formed tumors in 6/7 mice, MYCN overexpressing adrenal progenitor cells did not form tumors (0/7). Average tumor volumes are shown. 5 x 104 BE (2)-C human neuroblastoma cells or cells dissociated from Mycn-infected primary spheres were resuspended in HBSS with 30% Matrigel and delivered subcutaneously to nude mice.
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pone.0133897.g005: MYCN Expressing Progenitors Do Not Form Tumors.(A) Tumors derived from subcutaneously injected BE (2)-C cells showed neuroblastoma-like histologic features including dense cellularity, high nuclear-to-cytoplasmic ratios, high mitotic activity, and frequent apoptotic bodies. Scale bar equals 100 microns. (B) BE (2)-C derived tumors showed diffuse nuclear expression of PHOX2B and (C) diffuse cytoplasmic TH expression. Scale bars equal 100 microns. (D) While human neuroblastoma cells formed tumors in 6/7 mice, MYCN overexpressing adrenal progenitor cells did not form tumors (0/7). Average tumor volumes are shown. 5 x 104 BE (2)-C human neuroblastoma cells or cells dissociated from Mycn-infected primary spheres were resuspended in HBSS with 30% Matrigel and delivered subcutaneously to nude mice.

Mentions: Given that MYCN overexpression imparts tumor-like features including enhanced proliferation, neural lineage commitment, and colony forming potential, we assessed the ability of MYCN overexpressing adrenal progenitor cells to form tumors in vivo. Fifty thousand cells dissociated from primary spheres infected with MYCN expressing lentivirus were resuspended in HBSS with 30% matrigel and injected subcutaneously into the flanks of male nude mice. As a positive control, the same number of cells from the MYCN-amplified human neuroblastoma cell line BE (2)-C were delivered in an identical manner. While tumors with neuroblastoma-like histology and expression of the neuroblastoma SAP lineage markers Phox2B and TH developed in 6 out of 7 mice receiving BE (2)-C cells, no tumors formed following injection of the MYCN-overexpressing progenitor cells (Fig 5), indicating that overexpression of this oncogene in multipotent SAPs is not sufficient for tumorigenesis.


Expression of MYCN in Multipotent Sympathoadrenal Progenitors Induces Proliferation and Neural Differentiation, but Is Not Sufficient for Tumorigenesis.

Mobley BC, Kwon M, Kraemer BR, Hickman FE, Qiao J, Chung DH, Carter BD - PLoS ONE (2015)

MYCN Expressing Progenitors Do Not Form Tumors.(A) Tumors derived from subcutaneously injected BE (2)-C cells showed neuroblastoma-like histologic features including dense cellularity, high nuclear-to-cytoplasmic ratios, high mitotic activity, and frequent apoptotic bodies. Scale bar equals 100 microns. (B) BE (2)-C derived tumors showed diffuse nuclear expression of PHOX2B and (C) diffuse cytoplasmic TH expression. Scale bars equal 100 microns. (D) While human neuroblastoma cells formed tumors in 6/7 mice, MYCN overexpressing adrenal progenitor cells did not form tumors (0/7). Average tumor volumes are shown. 5 x 104 BE (2)-C human neuroblastoma cells or cells dissociated from Mycn-infected primary spheres were resuspended in HBSS with 30% Matrigel and delivered subcutaneously to nude mice.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4519318&req=5

pone.0133897.g005: MYCN Expressing Progenitors Do Not Form Tumors.(A) Tumors derived from subcutaneously injected BE (2)-C cells showed neuroblastoma-like histologic features including dense cellularity, high nuclear-to-cytoplasmic ratios, high mitotic activity, and frequent apoptotic bodies. Scale bar equals 100 microns. (B) BE (2)-C derived tumors showed diffuse nuclear expression of PHOX2B and (C) diffuse cytoplasmic TH expression. Scale bars equal 100 microns. (D) While human neuroblastoma cells formed tumors in 6/7 mice, MYCN overexpressing adrenal progenitor cells did not form tumors (0/7). Average tumor volumes are shown. 5 x 104 BE (2)-C human neuroblastoma cells or cells dissociated from Mycn-infected primary spheres were resuspended in HBSS with 30% Matrigel and delivered subcutaneously to nude mice.
Mentions: Given that MYCN overexpression imparts tumor-like features including enhanced proliferation, neural lineage commitment, and colony forming potential, we assessed the ability of MYCN overexpressing adrenal progenitor cells to form tumors in vivo. Fifty thousand cells dissociated from primary spheres infected with MYCN expressing lentivirus were resuspended in HBSS with 30% matrigel and injected subcutaneously into the flanks of male nude mice. As a positive control, the same number of cells from the MYCN-amplified human neuroblastoma cell line BE (2)-C were delivered in an identical manner. While tumors with neuroblastoma-like histology and expression of the neuroblastoma SAP lineage markers Phox2B and TH developed in 6 out of 7 mice receiving BE (2)-C cells, no tumors formed following injection of the MYCN-overexpressing progenitor cells (Fig 5), indicating that overexpression of this oncogene in multipotent SAPs is not sufficient for tumorigenesis.

Bottom Line: MYCN overexpression in spheres promoted commitment to the neural lineage, evidenced by an increased frequency of neuron-containing colonies.MYCN promoted proliferation of both sympathoadrenal progenitor spheres and differentiated neurons derived from these spheres, but there was also an increase in apoptosis.We find, however, that MYCN overexpression is not sufficient for these cells to form tumors in nude mice, suggesting that additional transforming mutations are necessary for tumorigenesis.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Microbiology, and Immunology, Division of Neuropathology, Vanderbilt University Medical Center, Nashville, Tennessee, the United States of America.

ABSTRACT
Neuroblastoma is a pediatric malignancy of the sympathetic ganglia and adrenal glands, hypothesized to originate from progenitors of the developing sympathetic nervous system. Amplification of the MYCN oncogene is a genetic marker of risk in this disease. Understanding the impact of oncogene expression on sympathoadrenal progenitor development may improve our knowledge of neuroblastoma initiation and progression. We isolated sympathoadrenal progenitor cells from the postnatal murine adrenal gland by sphere culture and found them to be multipotent, generating differentiated colonies of neurons, Schwann cells, and myofibroblasts. MYCN overexpression in spheres promoted commitment to the neural lineage, evidenced by an increased frequency of neuron-containing colonies. MYCN promoted proliferation of both sympathoadrenal progenitor spheres and differentiated neurons derived from these spheres, but there was also an increase in apoptosis. The proliferation, apoptosis, and neural lineage commitment induced by MYCN are tumor-like characteristics and thereby support the hypothesis that multipotent adrenal medullary progenitor cells are cells of origin for neuroblastoma. We find, however, that MYCN overexpression is not sufficient for these cells to form tumors in nude mice, suggesting that additional transforming mutations are necessary for tumorigenesis.

No MeSH data available.


Related in: MedlinePlus