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The bone marrow microenvironment in neuroblastoma progression
Abstracts
S35
A characteristic feature of classic PXE, an autosomal recessive disorder caused by mutations in the ABCC6 gene, is aberrant mineralization of connective tissues. Here, we report a family with PXE-like cutaneous features in association with multiple coagulation factor deficiency, an autosomal recessive disorder associated with GGCX mutations. The proband and her sister, both with severe skin findings including mineralization, were compound heterozygotes for missense mutations in GGCX, which were shown to result in reduced g-glutamyl carboxylase activity and in under-carboxylation of matrix gla protein. The proband's mother and aunt, also manifesting with PXE-like skin changes, were heterozygous carriers of a missense mutation (p.V255M) in GGCX and a null mutation (p.R1141X) in the ABCC6 gene, suggesting digenic nature of their skin findings. Thus, reduced gglutamyl carboxylase activity in individuals either compound heterozygous for a missense mutation in GGCX or with haploinsufficiency in GGCX in combination with heterozygosity for ABCC6 gene expression results in aberrant mineralization of skin leading to PXE-like phenotype. These findings expand the molecular basis of PXE-like phenotypes, and suggest a role for multiple genetic factors.
abilities and tumour growth/metastasis formation capacities in vivo. To this end we have developed a unique system for isolating native matrices from cultured cells; a system for the accurate monitoring and quantification of tumour cellendothelial interactions (under flow); and a novel system for examining the tumour cells' interactions with normal human tissues in vitro. Misexpression of glypican-4, -5 and -6 on diverse constitutive PG backgrounds modulates the in vitro and in vivo behaviour of the cells. To further define how given PG surface profiles may modify the behavioural traits of the cells, we combined the ectopic/overexpression of the above glypicans with RNAi knockdown of endogenous glypican-1, syndecan-1 and syndecan-4. The outcome of these investigations provide the first evidence that defined surface PGs pattern differentially control tumorigenesis.
doi:10.1016/j.matbio.2008.09.319
107 A central role for decorin during vertebrate convergent extension
105 The bone marrow microenvironment in neuroblastoma progression Yves A. De Clerck The Saban Research Institute of Childrens Hospital Los Angeles, USC Keck School of Medicine, United States It has become increasingly evident that the microenvironment contributes to tumor progression. In this aspect, the bone marrow and the bone are particularly fertile environments. In our laboratory, we have studied the contribution of the bone marrow to the progression of human neuroblastoma, the second most common solid tumor in children and a cancer that frequently metastasizes to the bone and the bone marrow. In the presence of bone marrow stromal cells (BMSC), human neuroblastoma cells stimulate the production of interleukin-6 (IL-6) by these cells. This stimulatory effect does not require cell–cell contact but is mediated by soluble factors like prostaglandin-E2 and galectin-3 binding protein, a glycosylated protein that binds to galectin-3, present at the surface of BMSC. The production of IL-6 in the bone marrow microenvironment has multiple effects that promote tumor progression. It stimulates osteoclast maturation and activity and the formation of osteolytic lesions. It also has a paracrine effect on neuroblastoma cells that express a functional IL-receptor (IL-6R). As a result, neuroblastoma cells proliferate more rapidly and become resistant to druginduced apoptosis. Consistent with IL-6 being a positive contributor to neuroblastoma progression, we found elevated levels of IL-6 in the serum and the bone marrow of patients with neuroblastoma bone metastasis and an increased expression of IL-6 in BMSC obtained from these patients. Thus IL-6 is a critical cytokine produced by BMSC in the bone marrow microenvironment that promotes the proliferation and survival of neuroblastoma cells. doi:10.1016/j.matbio.2008.09.320
106 The proteoglycan metastatic signature of a cancer cell Silvia Rossia, Katia Lacrimaa, Pier Andrea Nicolosib, Sabrina Cattaruzzab, Elena Garusia, Jorge Filmusc, Guido Davidd, Roberto Perrisa,b a Department of Genetics, University of Parma, Italy b National Cancer Institute Aviano, Italy c Sunnybrook and Women's College of Health Sciences, Toronto, Canada d Deparment of Mol. and Devel. Genetics, University of Leuven, Belgium Cell surface proteoglycans PGs are known key regulators of tumor progression and several of them are now emerging as clinically relevant targets. The tumorigenic function of the 11 primary PGs, including syndecans-1-4, glypicans-1-6 and NG2, is only in part resolved for some of these PGs individually, but not cumulatively. We have approached this problem by creating sarcoma cell lines with diverse PGs surface profiles and have comparatively assayed these engineered cells in vitro and for their migratory
doi:10.1016/j.matbio.2008.09.321
Jason J. Zoellera, Wittaya Pimtonga, Helen Corbya, Angela McQuillana, Silvia Goldonia, Alex E. Iozzoa, Rick T. Owensb, David J. McQuillanb, Shiu-Ying Hoa, Renato V. Iozzoa a Thomas Jefferson University, Philadelphia, PA 19107, United States b LifeCell Corporation, Branchburg, NJ 08876, United States Decorin, an archetypal member of the small leucine-rich proteoglycan gene family, regulates collagen fibrillogenesis and cell growth. To further explore its biological function, we examined the role of decorin during zebrafish development. Zebrafish decorin is a chondroitin sulfate proteoglycan which exhibits a high degree of conservation with its mammalian counterparts and displays a unique spatiotemporal expression pattern. Morpholino-mediated knockdown of zebrafish decorin identifies a developmental role for decorin during medial-lateral convergence and anteriorposterior extension of the body plan, as well as in craniofacial cartilage formation. Decorin morphants display a pronounced shortening of the head to tail axis as well as compression, flattening and extension of the jaw cartilages. The morphant phenotype could be rescued by zebrafish decorin mRNA. Unexpectedly, microinjection of excess zebrafish decorin mRNA or proteoglycan/protein core into one-cell stage embryos caused a defect in convergent extension associated with cyclopia. Overall, the results of our investigation indicate a central function for decorin during key early events of body plan development and cartilage formation, and further suggest that decorin may play a role in the Wnt and/or Hedgehog signaling pathways. doi:10.1016/j.matbio.2008.09.322
108 Prolyl 3-hydroxylase 1 null mice have abnormal bones and tendons Janice A. Vrankaa, Elena N. Pokidyshevaa, Kerry Maddoxa, Lauren K. Hayashia, Sara F. Tufaa, Doug R. Keenea, Emily Larsonc, Robert Kleinc, Hans Peter Bächingera,b a Research Department, Shriners Hospitals for Children, Portland, OR 97239, United States b Department of Biochem and Molec Biology, Oregon Health and Sci Uni., Portland, OR 97239, United States c Bone and Mineral Unit, Oregon Health and Science Uni., Portland, OR 97239, United States Osteogenesis imperfecta (OI) is a skeletal disorder primarily caused by mutations in collagen I genes. However, recent investigations have revealed that mutations in cartilage associated protein (CRTAP) or prolyl 3-hydroxylase 1 (P3H1) can cause a severe, recessive form of OI. These reports show minimal 3-hydroxylation of key residues in types I and II collagens as a result of CRTAP or P3H1 deficiency and demonstrate the importance of P3H1 and CRTAP to bone structure and development. P3H1 and CRTAP have previously been shown to form a stable complex with cyclophilin B (CYPB), and P3H1 was shown to catalyze the 3-hydroxylation of specific proline residues in
S35
A characteristic feature of classic PXE, an autosomal recessive disorder caused by mutations in the ABCC6 gene, is aberrant mineralization of connective tissues. Here, we report a family with PXE-like cutaneous features in association with multiple coagulation factor deficiency, an autosomal recessive disorder associated with GGCX mutations. The proband and her sister, both with severe skin findings including mineralization, were compound heterozygotes for missense mutations in GGCX, which were shown to result in reduced g-glutamyl carboxylase activity and in under-carboxylation of matrix gla protein. The proband's mother and aunt, also manifesting with PXE-like skin changes, were heterozygous carriers of a missense mutation (p.V255M) in GGCX and a null mutation (p.R1141X) in the ABCC6 gene, suggesting digenic nature of their skin findings. Thus, reduced gglutamyl carboxylase activity in individuals either compound heterozygous for a missense mutation in GGCX or with haploinsufficiency in GGCX in combination with heterozygosity for ABCC6 gene expression results in aberrant mineralization of skin leading to PXE-like phenotype. These findings expand the molecular basis of PXE-like phenotypes, and suggest a role for multiple genetic factors.
abilities and tumour growth/metastasis formation capacities in vivo. To this end we have developed a unique system for isolating native matrices from cultured cells; a system for the accurate monitoring and quantification of tumour cellendothelial interactions (under flow); and a novel system for examining the tumour cells' interactions with normal human tissues in vitro. Misexpression of glypican-4, -5 and -6 on diverse constitutive PG backgrounds modulates the in vitro and in vivo behaviour of the cells. To further define how given PG surface profiles may modify the behavioural traits of the cells, we combined the ectopic/overexpression of the above glypicans with RNAi knockdown of endogenous glypican-1, syndecan-1 and syndecan-4. The outcome of these investigations provide the first evidence that defined surface PGs pattern differentially control tumorigenesis.
doi:10.1016/j.matbio.2008.09.319
107 A central role for decorin during vertebrate convergent extension
105 The bone marrow microenvironment in neuroblastoma progression Yves A. De Clerck The Saban Research Institute of Childrens Hospital Los Angeles, USC Keck School of Medicine, United States It has become increasingly evident that the microenvironment contributes to tumor progression. In this aspect, the bone marrow and the bone are particularly fertile environments. In our laboratory, we have studied the contribution of the bone marrow to the progression of human neuroblastoma, the second most common solid tumor in children and a cancer that frequently metastasizes to the bone and the bone marrow. In the presence of bone marrow stromal cells (BMSC), human neuroblastoma cells stimulate the production of interleukin-6 (IL-6) by these cells. This stimulatory effect does not require cell–cell contact but is mediated by soluble factors like prostaglandin-E2 and galectin-3 binding protein, a glycosylated protein that binds to galectin-3, present at the surface of BMSC. The production of IL-6 in the bone marrow microenvironment has multiple effects that promote tumor progression. It stimulates osteoclast maturation and activity and the formation of osteolytic lesions. It also has a paracrine effect on neuroblastoma cells that express a functional IL-receptor (IL-6R). As a result, neuroblastoma cells proliferate more rapidly and become resistant to druginduced apoptosis. Consistent with IL-6 being a positive contributor to neuroblastoma progression, we found elevated levels of IL-6 in the serum and the bone marrow of patients with neuroblastoma bone metastasis and an increased expression of IL-6 in BMSC obtained from these patients. Thus IL-6 is a critical cytokine produced by BMSC in the bone marrow microenvironment that promotes the proliferation and survival of neuroblastoma cells. doi:10.1016/j.matbio.2008.09.320
106 The proteoglycan metastatic signature of a cancer cell Silvia Rossia, Katia Lacrimaa, Pier Andrea Nicolosib, Sabrina Cattaruzzab, Elena Garusia, Jorge Filmusc, Guido Davidd, Roberto Perrisa,b a Department of Genetics, University of Parma, Italy b National Cancer Institute Aviano, Italy c Sunnybrook and Women's College of Health Sciences, Toronto, Canada d Deparment of Mol. and Devel. Genetics, University of Leuven, Belgium Cell surface proteoglycans PGs are known key regulators of tumor progression and several of them are now emerging as clinically relevant targets. The tumorigenic function of the 11 primary PGs, including syndecans-1-4, glypicans-1-6 and NG2, is only in part resolved for some of these PGs individually, but not cumulatively. We have approached this problem by creating sarcoma cell lines with diverse PGs surface profiles and have comparatively assayed these engineered cells in vitro and for their migratory
doi:10.1016/j.matbio.2008.09.321
Jason J. Zoellera, Wittaya Pimtonga, Helen Corbya, Angela McQuillana, Silvia Goldonia, Alex E. Iozzoa, Rick T. Owensb, David J. McQuillanb, Shiu-Ying Hoa, Renato V. Iozzoa a Thomas Jefferson University, Philadelphia, PA 19107, United States b LifeCell Corporation, Branchburg, NJ 08876, United States Decorin, an archetypal member of the small leucine-rich proteoglycan gene family, regulates collagen fibrillogenesis and cell growth. To further explore its biological function, we examined the role of decorin during zebrafish development. Zebrafish decorin is a chondroitin sulfate proteoglycan which exhibits a high degree of conservation with its mammalian counterparts and displays a unique spatiotemporal expression pattern. Morpholino-mediated knockdown of zebrafish decorin identifies a developmental role for decorin during medial-lateral convergence and anteriorposterior extension of the body plan, as well as in craniofacial cartilage formation. Decorin morphants display a pronounced shortening of the head to tail axis as well as compression, flattening and extension of the jaw cartilages. The morphant phenotype could be rescued by zebrafish decorin mRNA. Unexpectedly, microinjection of excess zebrafish decorin mRNA or proteoglycan/protein core into one-cell stage embryos caused a defect in convergent extension associated with cyclopia. Overall, the results of our investigation indicate a central function for decorin during key early events of body plan development and cartilage formation, and further suggest that decorin may play a role in the Wnt and/or Hedgehog signaling pathways. doi:10.1016/j.matbio.2008.09.322
108 Prolyl 3-hydroxylase 1 null mice have abnormal bones and tendons Janice A. Vrankaa, Elena N. Pokidyshevaa, Kerry Maddoxa, Lauren K. Hayashia, Sara F. Tufaa, Doug R. Keenea, Emily Larsonc, Robert Kleinc, Hans Peter Bächingera,b a Research Department, Shriners Hospitals for Children, Portland, OR 97239, United States b Department of Biochem and Molec Biology, Oregon Health and Sci Uni., Portland, OR 97239, United States c Bone and Mineral Unit, Oregon Health and Science Uni., Portland, OR 97239, United States Osteogenesis imperfecta (OI) is a skeletal disorder primarily caused by mutations in collagen I genes. However, recent investigations have revealed that mutations in cartilage associated protein (CRTAP) or prolyl 3-hydroxylase 1 (P3H1) can cause a severe, recessive form of OI. These reports show minimal 3-hydroxylation of key residues in types I and II collagens as a result of CRTAP or P3H1 deficiency and demonstrate the importance of P3H1 and CRTAP to bone structure and development. P3H1 and CRTAP have previously been shown to form a stable complex with cyclophilin B (CYPB), and P3H1 was shown to catalyze the 3-hydroxylation of specific proline residues in