Ataxia-Telangiectasia Mutated Loss of Heterozygosity in Melanoma
ATM germline pathogenic variants were recently found enriched in high-risk melanoma patients. However, ATM loss of heterozygosity (LOH) has never been investigated in melanoma and, therefore, a causal association with melanoma development has not been established yet. The purpose of this study was t...
| Main Authors: | , , , , , , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-12-01
|
| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/23/24/16027 |
| _version_ | 1797457210821836800 |
|---|---|
| author | Lorenza Pastorino Bruna Dalmasso Eleonora Allavena Irene Vanni Filippo Ugolini Gianna Baroni Michela Croce Antonio Guadagno Francesco Cabiddu Virginia Andreotti William Bruno Gabriele Zoppoli Lorenzo Ferrando Enrica Teresa Tanda Francesco Spagnolo Chiara Menin Rosaria Gangemi Daniela Massi Paola Ghiorzo |
| author_facet | Lorenza Pastorino Bruna Dalmasso Eleonora Allavena Irene Vanni Filippo Ugolini Gianna Baroni Michela Croce Antonio Guadagno Francesco Cabiddu Virginia Andreotti William Bruno Gabriele Zoppoli Lorenzo Ferrando Enrica Teresa Tanda Francesco Spagnolo Chiara Menin Rosaria Gangemi Daniela Massi Paola Ghiorzo |
| author_sort | Lorenza Pastorino |
| collection | DOAJ |
| description | ATM germline pathogenic variants were recently found enriched in high-risk melanoma patients. However, ATM loss of heterozygosity (LOH) has never been investigated in melanoma and, therefore, a causal association with melanoma development has not been established yet. The purpose of this study was to functionally characterize 13 germline ATM variants found in high-risk melanoma patients—and classified by in silico tools as pathogenic, uncertain significance, or benign—using multiple assays evaluating ATM/pATM expression and/or LOH in melanoma tissues and cell lines. We assessed ATM status by Immunohistochemistry (IHC), Western Blot, Whole-Exome Sequencing/Copy Number Variation analysis, and RNA sequencing, supported by Sanger sequencing and microsatellite analyses. For most variants, IHC results matched those obtained with in silico classification and LOH analysis. Two pathogenic variants (p.Ser1135_Lys1192del and p.Ser1993ArgfsTer23) showed LOH and complete loss of ATM activation in melanoma. Two variants of unknown significance (p.Asn358Ile and p.Asn796His) showed reduced expression and LOH, suggestive of a deleterious effect. This study, showing a classic two-hit scenario in a well-known tumor suppressor gene, supports the inclusion of melanoma in the ATM-related cancer spectrum. |
| first_indexed | 2024-03-09T16:18:55Z |
| format | Article |
| id | doaj.art-8d7b0a16a56d420ca1a13af939ceabd0 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T16:18:55Z |
| publishDate | 2022-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-8d7b0a16a56d420ca1a13af939ceabd02023-11-24T15:31:39ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-12-0123241602710.3390/ijms232416027Ataxia-Telangiectasia Mutated Loss of Heterozygosity in MelanomaLorenza Pastorino0Bruna Dalmasso1Eleonora Allavena2Irene Vanni3Filippo Ugolini4Gianna Baroni5Michela Croce6Antonio Guadagno7Francesco Cabiddu8Virginia Andreotti9William Bruno10Gabriele Zoppoli11Lorenzo Ferrando12Enrica Teresa Tanda13Francesco Spagnolo14Chiara Menin15Rosaria Gangemi16Daniela Massi17Paola Ghiorzo18Department of Internal Medicine and Medical Specialties, University of Genoa, 16132 Genoa, ItalyGenetica dei Tumori Rari, IRCCS Ospedale Policlinico San Martino, 16132 Genoa, ItalyDepartment of Internal Medicine and Medical Specialties, University of Genoa, 16132 Genoa, ItalyGenetica dei Tumori Rari, IRCCS Ospedale Policlinico San Martino, 16132 Genoa, ItalySection of Anatomic Pathology, Department of Health Sciences, University of Florence, 50134 Florence, ItalySection of Anatomic Pathology, Department of Health Sciences, University of Florence, 50134 Florence, ItalyBioterapie, IRCCS Ospedale Policlinico San Martino, 16132 Genoa, ItalyAnatomia Patologica, IRCCS Ospedale Policlinico San Martino, 16132 Genoa, ItalyAnatomia Patologica, IRCCS Ospedale Policlinico San Martino, 16132 Genoa, ItalyGenetica dei Tumori Rari, IRCCS Ospedale Policlinico San Martino, 16132 Genoa, ItalyDepartment of Internal Medicine and Medical Specialties, University of Genoa, 16132 Genoa, ItalyDepartment of Internal Medicine and Medical Specialties, University of Genoa, 16132 Genoa, ItalyDepartment of Internal Medicine and Medical Specialties, University of Genoa, 16132 Genoa, ItalyDepartment of Internal Medicine and Medical Specialties, University of Genoa, 16132 Genoa, ItalyOncologia Medica 2, IRCCS Ospedale Policlinico San Martino, 16132 Genoa, ItalyImmunology and Diagnostic Molecular Oncology Unit, Veneto Institute of Oncology, IOV-IRCCS, 35128 Padua, ItalyBioterapie, IRCCS Ospedale Policlinico San Martino, 16132 Genoa, ItalySection of Anatomic Pathology, Department of Health Sciences, University of Florence, 50134 Florence, ItalyDepartment of Internal Medicine and Medical Specialties, University of Genoa, 16132 Genoa, ItalyATM germline pathogenic variants were recently found enriched in high-risk melanoma patients. However, ATM loss of heterozygosity (LOH) has never been investigated in melanoma and, therefore, a causal association with melanoma development has not been established yet. The purpose of this study was to functionally characterize 13 germline ATM variants found in high-risk melanoma patients—and classified by in silico tools as pathogenic, uncertain significance, or benign—using multiple assays evaluating ATM/pATM expression and/or LOH in melanoma tissues and cell lines. We assessed ATM status by Immunohistochemistry (IHC), Western Blot, Whole-Exome Sequencing/Copy Number Variation analysis, and RNA sequencing, supported by Sanger sequencing and microsatellite analyses. For most variants, IHC results matched those obtained with in silico classification and LOH analysis. Two pathogenic variants (p.Ser1135_Lys1192del and p.Ser1993ArgfsTer23) showed LOH and complete loss of ATM activation in melanoma. Two variants of unknown significance (p.Asn358Ile and p.Asn796His) showed reduced expression and LOH, suggestive of a deleterious effect. This study, showing a classic two-hit scenario in a well-known tumor suppressor gene, supports the inclusion of melanoma in the ATM-related cancer spectrum.https://www.mdpi.com/1422-0067/23/24/16027germline variantloss of heterozygositymelanomaATMsusceptibility |
| spellingShingle | Lorenza Pastorino Bruna Dalmasso Eleonora Allavena Irene Vanni Filippo Ugolini Gianna Baroni Michela Croce Antonio Guadagno Francesco Cabiddu Virginia Andreotti William Bruno Gabriele Zoppoli Lorenzo Ferrando Enrica Teresa Tanda Francesco Spagnolo Chiara Menin Rosaria Gangemi Daniela Massi Paola Ghiorzo Ataxia-Telangiectasia Mutated Loss of Heterozygosity in Melanoma International Journal of Molecular Sciences germline variant loss of heterozygosity melanoma ATM susceptibility |
| title | Ataxia-Telangiectasia Mutated Loss of Heterozygosity in Melanoma |
| title_full | Ataxia-Telangiectasia Mutated Loss of Heterozygosity in Melanoma |
| title_fullStr | Ataxia-Telangiectasia Mutated Loss of Heterozygosity in Melanoma |
| title_full_unstemmed | Ataxia-Telangiectasia Mutated Loss of Heterozygosity in Melanoma |
| title_short | Ataxia-Telangiectasia Mutated Loss of Heterozygosity in Melanoma |
| title_sort | ataxia telangiectasia mutated loss of heterozygosity in melanoma |
| topic | germline variant loss of heterozygosity melanoma ATM susceptibility |
| url | https://www.mdpi.com/1422-0067/23/24/16027 |
| work_keys_str_mv | AT lorenzapastorino ataxiatelangiectasiamutatedlossofheterozygosityinmelanoma AT brunadalmasso ataxiatelangiectasiamutatedlossofheterozygosityinmelanoma AT eleonoraallavena ataxiatelangiectasiamutatedlossofheterozygosityinmelanoma AT irenevanni ataxiatelangiectasiamutatedlossofheterozygosityinmelanoma AT filippougolini ataxiatelangiectasiamutatedlossofheterozygosityinmelanoma AT giannabaroni ataxiatelangiectasiamutatedlossofheterozygosityinmelanoma AT michelacroce ataxiatelangiectasiamutatedlossofheterozygosityinmelanoma AT antonioguadagno ataxiatelangiectasiamutatedlossofheterozygosityinmelanoma AT francescocabiddu ataxiatelangiectasiamutatedlossofheterozygosityinmelanoma AT virginiaandreotti ataxiatelangiectasiamutatedlossofheterozygosityinmelanoma AT williambruno ataxiatelangiectasiamutatedlossofheterozygosityinmelanoma AT gabrielezoppoli ataxiatelangiectasiamutatedlossofheterozygosityinmelanoma AT lorenzoferrando ataxiatelangiectasiamutatedlossofheterozygosityinmelanoma AT enricateresatanda ataxiatelangiectasiamutatedlossofheterozygosityinmelanoma AT francescospagnolo ataxiatelangiectasiamutatedlossofheterozygosityinmelanoma AT chiaramenin ataxiatelangiectasiamutatedlossofheterozygosityinmelanoma AT rosariagangemi ataxiatelangiectasiamutatedlossofheterozygosityinmelanoma AT danielamassi ataxiatelangiectasiamutatedlossofheterozygosityinmelanoma AT paolaghiorzo ataxiatelangiectasiamutatedlossofheterozygosityinmelanoma |