PMID- 30575147 OWN - NLM STAT- MEDLINE DCOM- 20190705 LR - 20200415 IS - 1098-2744 (Electronic) IS - 0899-1987 (Print) IS - 0899-1987 (Linking) VI - 58 IP - 5 DP - 2019 May TI - Inter- and intra-tumor heterogeneity of SMAD4 loss in head and neck squamous cell carcinomas. PG - 666-673 LID - 10.1002/mc.22958 [doi] AB - Reports regarding the frequency of SMAD4 loss in human head and neck squamous cell carcinoma (HNSCC) vary significantly. We have shown that SMAD4 deletion contributes to HNSCC initiation and progression. Therefore, accurately detecting genetic SMAD4 loss is critical to determine prognosis and therapeutic interventions in personalized medicine. We developed a SMAD4 fluorescence in situ hybridization (FISH) assay to identify chromosomal SMAD4 loss at the single cell level of primary HNSCC specimens and patient derived xenograft (PDX) tumors derived from HNSCCs. SMAD4 heterozygous loss was detected in 35% of primary HNSCCs and 41.3% of PDX tumors. Additionally, 4.3% of PDX tumors had SMAD4 homozygous loss. These frequencies of SMAD4 loss were similar to those in The Cancer Genome Atlas (TCGA). However, we identified significant heterogeneities of SMAD4 loss (partial or complete) among cells within each tumor. We also found that aneuploidy (monosomy and polysomy) contributed greatly to how to define chromosomal SMAD4 deletion. Furthermore, in cultured PDX tumors, SMAD4 mutant cells outcompeted SMAD4 wildtype cells, resulting in establishing homogenous SMAD4 mutant HNSCC cell lines with partial or complete genomic SMAD4 loss, suggesting a survival advantage of SMAD4 mutant cells. Taken together, our study reveals inter- and intra-tumor heterogeneities of SMAD4 chromosomal loss in HNSCCs. Further, SMAD4 FISH assay provides a platform for future clinical diagnosis of SMAD4 chromosomal loss that potentially serves as a molecular marker for prognosis and therapeutic intervention in cancer patients. CI - (c) 2018 Wiley Periodicals, Inc. FAU - Hernandez, Ariel L AU - Hernandez AL AD - Department of Pathology, School of Medicine, Molecular Biology Program, Medical Scientist Training Program, University of Colorado, Aurora, Colorado. FAU - Wang, Ying AU - Wang Y AD - University of Colorado Cancer Center, Molecular Pathology/Cytogenetics Shared Resource, University of Colorado, Aurora, Colorado. FAU - Somerset, Hilary L AU - Somerset HL AD - Department of Pathology, School of Medicine, Molecular Biology Program, Medical Scientist Training Program, University of Colorado, Aurora, Colorado. FAU - Keysar, Stephen B AU - Keysar SB AD - Department of Medical Oncology, University of Colorado, Aurora, Colorado. FAU - Aisner, Dara L AU - Aisner DL AD - Department of Pathology, School of Medicine, Molecular Biology Program, Medical Scientist Training Program, University of Colorado, Aurora, Colorado. FAU - Marshall, Carrie AU - Marshall C AD - Department of Pathology, School of Medicine, Molecular Biology Program, Medical Scientist Training Program, University of Colorado, Aurora, Colorado. FAU - Bowles, Daniel W AU - Bowles DW AD - Veterans Affairs Medical Center, VA Eastern Colorado Health Care System, Aurora, Colorado. FAU - Karam, Sana D AU - Karam SD AD - Department of Radiation Oncology, University of Colorado, Aurora, Colorado. FAU - Raben, David AU - Raben D AD - Department of Radiation Oncology, University of Colorado, Aurora, Colorado. FAU - Jimeno, Antonio AU - Jimeno A AD - Department of Medical Oncology, University of Colorado, Aurora, Colorado. FAU - Varella-Garcia, Marileila AU - Varella-Garcia M AD - University of Colorado Cancer Center, Molecular Pathology/Cytogenetics Shared Resource, University of Colorado, Aurora, Colorado. AD - Department of Medical Oncology, University of Colorado, Aurora, Colorado. FAU - Wang, Xiao-Jing AU - Wang XJ AUID- ORCID: 0000-0001-8695-7361 AD - Department of Pathology, School of Medicine, Molecular Biology Program, Medical Scientist Training Program, University of Colorado, Aurora, Colorado. AD - Veterans Affairs Medical Center, VA Eastern Colorado Health Care System, Aurora, Colorado. LA - eng GR - (K12, CA086913)/Paul Calabresi Career Development Award/International GR - R01 DE024371/DE/NIDCR NIH HHS/United States GR - I01 BX003232/BX/BLRD VA/United States GR - P30 CA046934/CA/NCI NIH HHS/United States GR - T32 CA174648/CA/NCI NIH HHS/United States GR - R01 DE024371/GF/NIH HHS/United States GR - K12CA086913/GF/NIH HHS/United States GR - P30 CA046934/GF/NIH HHS/United States GR - I01 BX003232/VA/VA/United States GR - R01 CA149456/CA/NCI NIH HHS/United States GR - (T32CA174648)/National Research Service Award Institutional Training/International GR - T32CA174648/GF/NIH HHS/United States GR - K12 CA086913/CA/NCI NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, U.S. Gov't, Non-P.H.S. DEP - 20190116 PL - United States TA - Mol Carcinog JT - Molecular carcinogenesis JID - 8811105 RN - 0 (SMAD4 protein, human) RN - 0 (Smad4 Protein) SB - IM MH - Animals MH - Carcinoma, Squamous Cell/*genetics/pathology MH - *Chromosome Aberrations MH - *Gene Deletion MH - *Gene Expression Regulation, Neoplastic MH - *Genetic Heterogeneity MH - Head and Neck Neoplasms/*genetics/pathology MH - Humans MH - In Situ Hybridization, Fluorescence MH - Mice MH - Smad4 Protein/*genetics MH - Tumor Cells, Cultured MH - Xenograft Model Antitumor Assays PMC - PMC6637962 MID - NIHMS1036595 OTO - NOTNLM OT - Aneuploidy OT - Chromosomal SMAD4 deletion OT - genomic instability EDAT- 2018/12/24 06:00 MHDA- 2019/07/06 06:00 PMCR- 2019/07/18 CRDT- 2018/12/22 06:00 PHST- 2018/09/29 00:00 [received] PHST- 2018/12/04 00:00 [revised] PHST- 2018/12/15 00:00 [accepted] PHST- 2018/12/24 06:00 [pubmed] PHST- 2019/07/06 06:00 [medline] PHST- 2018/12/22 06:00 [entrez] PHST- 2019/07/18 00:00 [pmc-release] AID - 10.1002/mc.22958 [doi] PST - ppublish SO - Mol Carcinog. 2019 May;58(5):666-673. doi: 10.1002/mc.22958. Epub 2019 Jan 16.