title: Stress-induced isoforms of MDM2 and MDM4 correlate with high-grade disease and an altered splicing network in pediatric rhabdomyosarcoma.
abstract: Pediatric rhabdomyosarcoma (RMS) is a morphologically and genetically heterogeneous malignancy commonly classified into three histologic subtypes, namely, alveolar, embryonal, and anaplastic. An issue that continues to challenge effective RMS patient prognosis is the dearth of molecular markers predictive of disease stage irrespective of tumor subtype. Our study involving a panel of 70 RMS tumors has identified specific alternative splice variants of the oncogenes Murine Double Minute 2 (MDM2) and MDM4 as potential biomarkers for RMS. Our results have demonstrated the strong association of genotoxic-stress inducible splice forms MDM2-ALT1 (91.6% Intergroup Rhabdomyosarcoma Study Group stage 4 tumors) and MDM4-ALT2 (90.9% MDM4-ALT2-positive T2 stage tumors) with high-risk metastatic RMS. Moreover, MDM2-ALT1-positive metastatic tumors belonged to both the alveolar (50%) and embryonal (41.6%) subtypes, making this the first known molecular marker for high-grade metastatic disease across the most common RMS subtypes. Furthermore, our results show that MDM2-ALT1 expression can function by directly contribute to metastatic behavior and promote the invasion of RMS cells through a matrigel-coated membrane. Additionally, expression of both MDM2-ALT1 and MDM4-ALT2 increased anchorage-independent cell-growth in soft agar assays. Intriguingly, we observed a unique coordination in the splicing of MDM2-ALT1 and MDM4-ALT2 in approximately 24% of tumor samples in a manner similar to genotoxic stress response in cell lines. To further explore splicing network alterations with possible relevance to RMS disease, we used an exon microarray approach to examine stress-inducible splicing in an RMS cell line (Rh30) and observed striking parallels between stress-responsive alternative splicing and constitutive splicing in RMS tumors.
authors.affiliation: Center for Childhood Cancer, Research Institute at Nationwide Children's Hospital, Columbus, OH.
authors.forename: Aishwarya G
authors.initials: AG
authors.lastname: Jacob
authors.forename: Dennis
authors.initials: D
authors.lastname: O'Brien
authors.forename: Ravi K
authors.initials: RK
authors.lastname: Singh
authors.forename: Daniel F
authors.initials: DF
authors.lastname: Comiskey
authors.forename: Robert M
authors.initials: RM
authors.lastname: Littleton
authors.forename: Fuad
authors.initials: F
authors.lastname: Mohammad
authors.forename: Jordan T
authors.initials: JT
authors.lastname: Gladman
authors.forename: Maria C
authors.initials: MC
authors.lastname: Widmann
authors.forename: Selvi C
authors.initials: SC
authors.lastname: Jeyaraj
authors.forename: Cheryl
authors.initials: C
authors.lastname: Bolinger
authors.forename: James R
authors.initials: JR
authors.lastname: Anderson
authors.forename: Donald A
authors.initials: DA
authors.lastname: Barkauskas
authors.forename: Kathleen
authors.initials: K
authors.lastname: Boris-Lawrie
authors.forename: Dawn S
authors.initials: DS
authors.lastname: Chandler
mesh_terms: D017398:Alternative Splicing; D014408:Biomarkers, Tumor / Q000235:genetics; D002448:Cell Adhesion / Q000235:genetics; D018797:Cell Cycle Proteins; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D004249:DNA Damage / Q000235:genetics; D006801:Humans; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism; D020033:Protein Isoforms / Q000235:genetics* / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000235:genetics* / Q000378:metabolism; D051736:Proto-Oncogene Proteins c-mdm2 / Q000235:genetics* / Q000378:metabolism; D012208:Rhabdomyosarcoma / Q000235:genetics* / Q000378:metabolism*; D013312:Stress, Physiological / Q000235:genetics
chemical_list: D014408:Biomarkers, Tumor; D018797:Cell Cycle Proteins; C482775:MDM4 protein, human; D009687:Nuclear Proteins; D020033:Protein Isoforms; D011518:Proto-Oncogene Proteins; C497206:MDM2 protein, human; D051736:Proto-Oncogene Proteins c-mdm2
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