Pyrosequencing-Based Assays for Rapid Detection of HER2 and HER3 Mutations in Clinical Samples Uncover an E332E Mutation Affecting HER3 in Retroperitoneal Leiomyosarcoma
Mutations in Human Epidermal Growth Factor Receptors (HER) are associated with poor prognosis of several types of solid tumors. Although HER-mutation detection methods are currently available, such as Next-Generation Sequencing (NGS), alternative pyrosequencing allow the rapid characterization of sp...
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MDPI AG
2015-08-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | http://www.mdpi.com/1422-0067/16/8/19447 |
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| author | Paula González-Alonso Cristina Chamizo Víctor Moreno Juan Madoz-Gúrpide Nerea Carvajal Lina Daoud Sandra Zazo Ester Martín-Aparicio Ion Cristóbal Raúl Rincón Jesús García-Foncillas Federico Rojo |
| author_facet | Paula González-Alonso Cristina Chamizo Víctor Moreno Juan Madoz-Gúrpide Nerea Carvajal Lina Daoud Sandra Zazo Ester Martín-Aparicio Ion Cristóbal Raúl Rincón Jesús García-Foncillas Federico Rojo |
| author_sort | Paula González-Alonso |
| collection | DOAJ |
| description | Mutations in Human Epidermal Growth Factor Receptors (HER) are associated with poor prognosis of several types of solid tumors. Although HER-mutation detection methods are currently available, such as Next-Generation Sequencing (NGS), alternative pyrosequencing allow the rapid characterization of specific mutations. We developed specific PCR-based pyrosequencing assays for identification of most prevalent HER2 and HER3 mutations, including S310F/Y, R678Q, L755M/P/S/W, V777A/L/M, 774-776 insertion, and V842I mutations in HER2, as well as M91I, V104M/L, D297N/V/Y, and E332E/K mutations in HER3. We tested 85 Formalin Fixed and Paraffin Embbeded (FFPE) samples and we detected three HER2-V842I mutations in colorectal carcinoma (CRC), ovarian carcinoma, and pancreatic carcinoma patients, respectively, and a HER2-L755M mutation in a CRC specimen. We also determined the presence of a HER3-E332K mutation in an urothelial carcinoma sample, and two HER3-D297Y mutations, in both gastric adenocarcinoma and CRC specimens. The D297Y mutation was previously detected in breast and gastric tumors, but not in CRC. Moreover, we found a not-previously-described HER3-E332E synonymous mutation in a retroperitoneal leiomyosarcoma patient. The pyrosequencing assays presented here allow the detection and characterization of specific HER2 and HER3 mutations. These pyrosequencing assays might be implemented in routine diagnosis for molecular characterization of HER2/HER3 receptors as an alternative to complex NGS approaches. |
| first_indexed | 2024-04-11T22:50:22Z |
| format | Article |
| id | doaj.art-f763525e464e42e3a6ae653c146b99ab |
| institution | Directory Open Access Journal |
| issn | 1422-0067 |
| language | English |
| last_indexed | 2024-04-11T22:50:22Z |
| publishDate | 2015-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-f763525e464e42e3a6ae653c146b99ab2022-12-22T03:58:36ZengMDPI AGInternational Journal of Molecular Sciences1422-00672015-08-01168194471945710.3390/ijms160819447ijms160819447Pyrosequencing-Based Assays for Rapid Detection of HER2 and HER3 Mutations in Clinical Samples Uncover an E332E Mutation Affecting HER3 in Retroperitoneal LeiomyosarcomaPaula González-Alonso0Cristina Chamizo1Víctor Moreno2Juan Madoz-Gúrpide3Nerea Carvajal4Lina Daoud5Sandra Zazo6Ester Martín-Aparicio7Ion Cristóbal8Raúl Rincón9Jesús García-Foncillas10Federico Rojo11Group of Cancer Biomarkers, Pathology Department, Health Research Institute Fundación Jiménez Díaz (IIS-FJD, UAM), Avda. Reyes Católicos 2, 28040 Madrid, SpainGroup of Cancer Biomarkers, Pathology Department, Health Research Institute Fundación Jiménez Díaz (IIS-FJD, UAM), Avda. Reyes Católicos 2, 28040 Madrid, SpainTranslational Oncology Division, Oncohealth Institute, Fundación Jiménez Díaz Hospital, Avda. Reyes Católicos 2, 28040 Madrid, SpainGroup of Cancer Biomarkers, Pathology Department, Health Research Institute Fundación Jiménez Díaz (IIS-FJD, UAM), Avda. Reyes Católicos 2, 28040 Madrid, SpainGroup of Cancer Biomarkers, Pathology Department, Health Research Institute Fundación Jiménez Díaz (IIS-FJD, UAM), Avda. Reyes Católicos 2, 28040 Madrid, SpainGroup of Cancer Biomarkers, Pathology Department, Health Research Institute Fundación Jiménez Díaz (IIS-FJD, UAM), Avda. Reyes Católicos 2, 28040 Madrid, SpainGroup of Cancer Biomarkers, Pathology Department, Health Research Institute Fundación Jiménez Díaz (IIS-FJD, UAM), Avda. Reyes Católicos 2, 28040 Madrid, SpainGroup of Cancer Biomarkers, Pathology Department, Health Research Institute Fundación Jiménez Díaz (IIS-FJD, UAM), Avda. Reyes Católicos 2, 28040 Madrid, SpainTranslational Oncology Division, Oncohealth Institute, Fundación Jiménez Díaz Hospital, Avda. Reyes Católicos 2, 28040 Madrid, SpainGroup of Cancer Biomarkers, Pathology Department, Health Research Institute Fundación Jiménez Díaz (IIS-FJD, UAM), Avda. Reyes Católicos 2, 28040 Madrid, SpainTranslational Oncology Division, Oncohealth Institute, Fundación Jiménez Díaz Hospital, Avda. Reyes Católicos 2, 28040 Madrid, SpainGroup of Cancer Biomarkers, Pathology Department, Health Research Institute Fundación Jiménez Díaz (IIS-FJD, UAM), Avda. Reyes Católicos 2, 28040 Madrid, SpainMutations in Human Epidermal Growth Factor Receptors (HER) are associated with poor prognosis of several types of solid tumors. Although HER-mutation detection methods are currently available, such as Next-Generation Sequencing (NGS), alternative pyrosequencing allow the rapid characterization of specific mutations. We developed specific PCR-based pyrosequencing assays for identification of most prevalent HER2 and HER3 mutations, including S310F/Y, R678Q, L755M/P/S/W, V777A/L/M, 774-776 insertion, and V842I mutations in HER2, as well as M91I, V104M/L, D297N/V/Y, and E332E/K mutations in HER3. We tested 85 Formalin Fixed and Paraffin Embbeded (FFPE) samples and we detected three HER2-V842I mutations in colorectal carcinoma (CRC), ovarian carcinoma, and pancreatic carcinoma patients, respectively, and a HER2-L755M mutation in a CRC specimen. We also determined the presence of a HER3-E332K mutation in an urothelial carcinoma sample, and two HER3-D297Y mutations, in both gastric adenocarcinoma and CRC specimens. The D297Y mutation was previously detected in breast and gastric tumors, but not in CRC. Moreover, we found a not-previously-described HER3-E332E synonymous mutation in a retroperitoneal leiomyosarcoma patient. The pyrosequencing assays presented here allow the detection and characterization of specific HER2 and HER3 mutations. These pyrosequencing assays might be implemented in routine diagnosis for molecular characterization of HER2/HER3 receptors as an alternative to complex NGS approaches.http://www.mdpi.com/1422-0067/16/8/19447HER2HER3cancerpyrosequencing |
| spellingShingle | Paula González-Alonso Cristina Chamizo Víctor Moreno Juan Madoz-Gúrpide Nerea Carvajal Lina Daoud Sandra Zazo Ester Martín-Aparicio Ion Cristóbal Raúl Rincón Jesús García-Foncillas Federico Rojo Pyrosequencing-Based Assays for Rapid Detection of HER2 and HER3 Mutations in Clinical Samples Uncover an E332E Mutation Affecting HER3 in Retroperitoneal Leiomyosarcoma International Journal of Molecular Sciences HER2 HER3 cancer pyrosequencing |
| title | Pyrosequencing-Based Assays for Rapid Detection of HER2 and HER3 Mutations in Clinical Samples Uncover an E332E Mutation Affecting HER3 in Retroperitoneal Leiomyosarcoma |
| title_full | Pyrosequencing-Based Assays for Rapid Detection of HER2 and HER3 Mutations in Clinical Samples Uncover an E332E Mutation Affecting HER3 in Retroperitoneal Leiomyosarcoma |
| title_fullStr | Pyrosequencing-Based Assays for Rapid Detection of HER2 and HER3 Mutations in Clinical Samples Uncover an E332E Mutation Affecting HER3 in Retroperitoneal Leiomyosarcoma |
| title_full_unstemmed | Pyrosequencing-Based Assays for Rapid Detection of HER2 and HER3 Mutations in Clinical Samples Uncover an E332E Mutation Affecting HER3 in Retroperitoneal Leiomyosarcoma |
| title_short | Pyrosequencing-Based Assays for Rapid Detection of HER2 and HER3 Mutations in Clinical Samples Uncover an E332E Mutation Affecting HER3 in Retroperitoneal Leiomyosarcoma |
| title_sort | pyrosequencing based assays for rapid detection of her2 and her3 mutations in clinical samples uncover an e332e mutation affecting her3 in retroperitoneal leiomyosarcoma |
| topic | HER2 HER3 cancer pyrosequencing |
| url | http://www.mdpi.com/1422-0067/16/8/19447 |
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