Limits...
Acquired genetic alterations in tumor cells dictate the development of high-risk neuroblastoma and clinical outcomes.

Khan FH, Pandian V, Ramraj S, Natarajan M, Aravindan S, Herman TS, Aravindan N - BMC Cancer (2015)

Bottom Line: Under serum-free conditions, MSDACs demonstrated profound tumorosphere formation ex vivo.Tissue microarray analysis coupled with automated IHC revealed significant association of RALYL to the tumor grade in a cohort of 25 neuroblastoma patients.Clinical outcome association analysis showed a strong correlation between the expression of CFHR3, CSMD3, MDFIC, FOXP2, RALYL, POLDIP3, SLC25A17, SERHL, MGAT3, TTLL1, or LOC400927 and overall and relapse-free survival in patients with neuroblastoma.

View Article: PubMed Central - PubMed

Affiliation: Department of Radiation Oncology, University of Oklahoma Health Sciences Science Center, 940 Stanton L. Young Blvd., BMSB 737, Oklahoma City, OK, 73104, USA. Faizan-Khan@ouhsc.edu.

ABSTRACT

Background: Determining the driving factors and molecular flow-through that define the switch from favorable to aggressive high-risk disease is critical to the betterment of neuroblastoma cure.

Methods: In this study, we examined the cytogenetic and tumorigenic physiognomies of distinct population of metastatic site- derived aggressive cells (MSDACs) from high-risk tumors, and showed the influence of acquired genetic rearrangements on poor patient outcomes.

Results: Karyotyping in SH-SY5Y and MSDACs revealed trisomy of 1q, with additional non-random chromosomal rearrangements on 1q32, 8p23, 9q34, 15q24, 22q13 (additions), and 7q32 (deletion). Array CGH analysis of individual clones of MSDACs revealed genetic alterations in chromosomes 1, 7, 8, and 22, corresponding to a gain in the copy numbers of LOC100288142, CD1C, CFHR3, FOXP2, MDFIC, RALYL, CSMD3, SAMD12-AS1, and MAL2, and a loss in ADAM5, LOC400927, APOBEC3B, RPL3, MGAT3, SLC25A17, EP300, L3MBTL2, SERHL, POLDIP3, A4GALT, and TTLL1. QPCR analysis and immunoblotting showed a definite association between DNA-copy number changes and matching transcriptional/translational expression in clones of MSDACs. Further, MSDACs exert a stem-like phenotype. Under serum-free conditions, MSDACs demonstrated profound tumorosphere formation ex vivo. Moreover, MSDACs exhibited high tumorigenic capacity in vivo and prompted aggressive metastatic disease. Tissue microarray analysis coupled with automated IHC revealed significant association of RALYL to the tumor grade in a cohort of 25 neuroblastoma patients. Clinical outcome association analysis showed a strong correlation between the expression of CFHR3, CSMD3, MDFIC, FOXP2, RALYL, POLDIP3, SLC25A17, SERHL, MGAT3, TTLL1, or LOC400927 and overall and relapse-free survival in patients with neuroblastoma.

Conclusion: Together, these data highlight the ongoing acquired genetic rearrangements in undifferentiated tumor-forming neural crest cells, and suggest that these alterations could switch favorable neuroblastoma to high-risk aggressive disease, promoting poor clinical outcomes.

No MeSH data available.


Related in: MedlinePlus

Genome wide copy number variations in parental SH-SY5Y and MSDACs. a Array CGH analysis showing digitized copy number variations (CNVs) across the genome plotted for SH-SY5Y cells and MSDACs. b Table showing common copy number gain and/or loss across the clones of MSDACs. Chromosome numbers, regions, and magnitude of CNV variation and corresponding genes are shown
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4496850&req=5

Fig3: Genome wide copy number variations in parental SH-SY5Y and MSDACs. a Array CGH analysis showing digitized copy number variations (CNVs) across the genome plotted for SH-SY5Y cells and MSDACs. b Table showing common copy number gain and/or loss across the clones of MSDACs. Chromosome numbers, regions, and magnitude of CNV variation and corresponding genes are shown

Mentions: Cancer cells are typically characterized by intricate karyotypes, including both structural and numerical changes. To determine and illustrate that the aggressive tumors developing in multiple metastatic sites were derived from the parental human SH-SY5Y cells, we karyotyped MSDACs, with and without characterized CD133+, and compared these with the parental cells. All karyotyping was performed in double blinded fashion. We investigated at least 20 cells per clone. SH-SY5Y cells exhibited the 47,XX, add(1)(q32), +del(7)?(q32), add(8)(p23), add(9)(q34), add(15)(q24), add(22)(q13) [20] karyotype, and served as the positive controls (Fig. 2ai). All investigated clones of MSDACs exhibited an exact match of the parental SH-SY5Y cells. We observed a unique marker composed of a chromosome 1 with a complex insertion of an additional copy of a 1q segment into the long arm, resulting in trisomy of 1q. Karyotyping also revealed six novel non-random chromosomal rearrangements on 1q32, 8p23, 9q34, 15q24, 22q13 (additions), and 7q32 (deletion; Fig. 2aii). Consistently, array CGH analysis corroborated the karyotyping in the clones of parental cells and MSDACs (Fig. 2b) and demonstrated that the developed aggressive metastatic tumors in mice are indeed derived and disseminated from the parental SH-SY5Y cells.Fig. 3


Acquired genetic alterations in tumor cells dictate the development of high-risk neuroblastoma and clinical outcomes.

Khan FH, Pandian V, Ramraj S, Natarajan M, Aravindan S, Herman TS, Aravindan N - BMC Cancer (2015)

Genome wide copy number variations in parental SH-SY5Y and MSDACs. a Array CGH analysis showing digitized copy number variations (CNVs) across the genome plotted for SH-SY5Y cells and MSDACs. b Table showing common copy number gain and/or loss across the clones of MSDACs. Chromosome numbers, regions, and magnitude of CNV variation and corresponding genes are shown
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4496850&req=5

Fig3: Genome wide copy number variations in parental SH-SY5Y and MSDACs. a Array CGH analysis showing digitized copy number variations (CNVs) across the genome plotted for SH-SY5Y cells and MSDACs. b Table showing common copy number gain and/or loss across the clones of MSDACs. Chromosome numbers, regions, and magnitude of CNV variation and corresponding genes are shown
Mentions: Cancer cells are typically characterized by intricate karyotypes, including both structural and numerical changes. To determine and illustrate that the aggressive tumors developing in multiple metastatic sites were derived from the parental human SH-SY5Y cells, we karyotyped MSDACs, with and without characterized CD133+, and compared these with the parental cells. All karyotyping was performed in double blinded fashion. We investigated at least 20 cells per clone. SH-SY5Y cells exhibited the 47,XX, add(1)(q32), +del(7)?(q32), add(8)(p23), add(9)(q34), add(15)(q24), add(22)(q13) [20] karyotype, and served as the positive controls (Fig. 2ai). All investigated clones of MSDACs exhibited an exact match of the parental SH-SY5Y cells. We observed a unique marker composed of a chromosome 1 with a complex insertion of an additional copy of a 1q segment into the long arm, resulting in trisomy of 1q. Karyotyping also revealed six novel non-random chromosomal rearrangements on 1q32, 8p23, 9q34, 15q24, 22q13 (additions), and 7q32 (deletion; Fig. 2aii). Consistently, array CGH analysis corroborated the karyotyping in the clones of parental cells and MSDACs (Fig. 2b) and demonstrated that the developed aggressive metastatic tumors in mice are indeed derived and disseminated from the parental SH-SY5Y cells.Fig. 3

Bottom Line: Under serum-free conditions, MSDACs demonstrated profound tumorosphere formation ex vivo.Tissue microarray analysis coupled with automated IHC revealed significant association of RALYL to the tumor grade in a cohort of 25 neuroblastoma patients.Clinical outcome association analysis showed a strong correlation between the expression of CFHR3, CSMD3, MDFIC, FOXP2, RALYL, POLDIP3, SLC25A17, SERHL, MGAT3, TTLL1, or LOC400927 and overall and relapse-free survival in patients with neuroblastoma.

View Article: PubMed Central - PubMed

Affiliation: Department of Radiation Oncology, University of Oklahoma Health Sciences Science Center, 940 Stanton L. Young Blvd., BMSB 737, Oklahoma City, OK, 73104, USA. Faizan-Khan@ouhsc.edu.

ABSTRACT

Background: Determining the driving factors and molecular flow-through that define the switch from favorable to aggressive high-risk disease is critical to the betterment of neuroblastoma cure.

Methods: In this study, we examined the cytogenetic and tumorigenic physiognomies of distinct population of metastatic site- derived aggressive cells (MSDACs) from high-risk tumors, and showed the influence of acquired genetic rearrangements on poor patient outcomes.

Results: Karyotyping in SH-SY5Y and MSDACs revealed trisomy of 1q, with additional non-random chromosomal rearrangements on 1q32, 8p23, 9q34, 15q24, 22q13 (additions), and 7q32 (deletion). Array CGH analysis of individual clones of MSDACs revealed genetic alterations in chromosomes 1, 7, 8, and 22, corresponding to a gain in the copy numbers of LOC100288142, CD1C, CFHR3, FOXP2, MDFIC, RALYL, CSMD3, SAMD12-AS1, and MAL2, and a loss in ADAM5, LOC400927, APOBEC3B, RPL3, MGAT3, SLC25A17, EP300, L3MBTL2, SERHL, POLDIP3, A4GALT, and TTLL1. QPCR analysis and immunoblotting showed a definite association between DNA-copy number changes and matching transcriptional/translational expression in clones of MSDACs. Further, MSDACs exert a stem-like phenotype. Under serum-free conditions, MSDACs demonstrated profound tumorosphere formation ex vivo. Moreover, MSDACs exhibited high tumorigenic capacity in vivo and prompted aggressive metastatic disease. Tissue microarray analysis coupled with automated IHC revealed significant association of RALYL to the tumor grade in a cohort of 25 neuroblastoma patients. Clinical outcome association analysis showed a strong correlation between the expression of CFHR3, CSMD3, MDFIC, FOXP2, RALYL, POLDIP3, SLC25A17, SERHL, MGAT3, TTLL1, or LOC400927 and overall and relapse-free survival in patients with neuroblastoma.

Conclusion: Together, these data highlight the ongoing acquired genetic rearrangements in undifferentiated tumor-forming neural crest cells, and suggest that these alterations could switch favorable neuroblastoma to high-risk aggressive disease, promoting poor clinical outcomes.

No MeSH data available.


Related in: MedlinePlus