Cell-free DNA next-generation sequencing liquid biopsy as a new revolutionary approach for arteriovenous malformation
Objective: Somatic mosaicism of KRAS gene is currently recognized as the only established molecular basis of arteriovenous malformations (AVM). However, given the limitations of the current technologies, KRAS somatic mutations are detected only in a limited proportion of AVMs and tissue biopsy remai...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2020-01-01
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| Series: | JVS - Vascular Science |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666350320300195 |
| _version_ | 1818835944496693248 |
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| author | Maria Palmieri, MS Aurora Currò, MD Andrea Tommasi, MD Laura Di Sarno, MS Gabriella Doddato, MS Margheria Baldassarri, MD Elisa Frullanti, MS Ann Rita Giliberti, MS Chiara Fallerini, MS Angelo Spinazzola, MD Anna Maria Pinto, MD, PhD Alessandra Renieri, MD, PhD Massimo Vaghi, MD |
| author_facet | Maria Palmieri, MS Aurora Currò, MD Andrea Tommasi, MD Laura Di Sarno, MS Gabriella Doddato, MS Margheria Baldassarri, MD Elisa Frullanti, MS Ann Rita Giliberti, MS Chiara Fallerini, MS Angelo Spinazzola, MD Anna Maria Pinto, MD, PhD Alessandra Renieri, MD, PhD Massimo Vaghi, MD |
| author_sort | Maria Palmieri, MS |
| collection | DOAJ |
| description | Objective: Somatic mosaicism of KRAS gene is currently recognized as the only established molecular basis of arteriovenous malformations (AVM). However, given the limitations of the current technologies, KRAS somatic mutations are detected only in a limited proportion of AVMs and tissue biopsy remains an invasive high risky, sometimes life-threatening, diagnostic procedure. Next-generation sequencing liquid biopsy using cell-free DNA (cfDNA) has emerged as an innovative noninvasive approach for early detection and monitoring of cancer. This approach overcomes the space-time profile constraint of tissue biopsies opens a new scenario for vascular malformations owing to somatic mosaicism. Here, we propose a new approach as a fast noninvasive reliable tool in order to investigate the cfDNA coming from the AVMs. Methods: A group of five patients suffering from AVM were selected. Blood samples from peripheral vein and efferent vein from vascular malformation were collected and cfDNA was extracted. The cfDNA libraries were performed using Oncomine Pan-Cancer Cell-Free Assay. We used Ion Proton for sequencing and Ion Reporter Software for analysis (Life Technologies, Carlsbad, Calif). Results: In all cases, either G12D or G12V mutations in KRAS were identified. The mutational load was higher in the efferent vein than in peripheral blood, confirming the causative role of the identified mutation at a somatic level. Conclusions: We demonstrate that cfDNA next-generation sequencing liquid biopsy is able to identify the KRAS mutation detected in affected tissues. Moreover, we have shown that blood sample withdrawal at the lesion site increases variant allele frequency with an order of magnitude above the limit of detection (usually 0.05%), decreasing the risk of a false negative. Finally, the noninvasiveness of the method avoids any risk of bleeding, being easily performed also in children. We propose this technique as the method of choice to better investigate AVMs and consequently to identify the therapy tailored to the genetic defect. Clinical Relevance: This article highlights the importance of using liquid biopsy as a new method to investigate the molecular profile of AVMs. In view of the frequent inaccessibility of vascular tissues owing to the invasiveness of solid biopsy and the relative high incidence of biopsies with low diagnostic power, here we evaluated the efficacy of detecting cfDNA fragments released into the bloodstream from the affected tissue cells. Through a simple blood draw from the efferent vein at the vascular malformation site, the liquid biopsy allowed us to identify KRAS pathogenic mutations piloting a personalized therapeutic approach and opening a new scenario for new therapeutic strategies. |
| first_indexed | 2024-12-19T02:58:46Z |
| format | Article |
| id | doaj.art-3230d76daea4439ab77cce96d8e31d67 |
| institution | Directory Open Access Journal |
| issn | 2666-3503 |
| language | English |
| last_indexed | 2024-12-19T02:58:46Z |
| publishDate | 2020-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | JVS - Vascular Science |
| spelling | doaj.art-3230d76daea4439ab77cce96d8e31d672022-12-21T20:38:15ZengElsevierJVS - Vascular Science2666-35032020-01-011176180Cell-free DNA next-generation sequencing liquid biopsy as a new revolutionary approach for arteriovenous malformationMaria Palmieri, MS0Aurora Currò, MD1Andrea Tommasi, MD2Laura Di Sarno, MS3Gabriella Doddato, MS4Margheria Baldassarri, MD5Elisa Frullanti, MS6Ann Rita Giliberti, MS7Chiara Fallerini, MS8Angelo Spinazzola, MD9Anna Maria Pinto, MD, PhD10Alessandra Renieri, MD, PhD11Massimo Vaghi, MD12Medical Genetics, University of Siena, Siena, ItalyMedical Genetics, University of Siena, Siena, Italy; Genetica Medica, Azienda Ospedaliera Universitaria Senese, Siena, ItalyMedical Genetics, University of Siena, Siena, Italy; Genetica Medica, Azienda Ospedaliera Universitaria Senese, Siena, ItalyGenetica Medica, Azienda Ospedaliera Universitaria Senese, Siena, ItalyMedical Genetics, University of Siena, Siena, ItalyGenetica Medica, Azienda Ospedaliera Universitaria Senese, Siena, ItalyMedical Genetics, University of Siena, Siena, ItalyMedical Genetics, University of Siena, Siena, ItalyMedical Genetics, University of Siena, Siena, ItalyRadiologia interventistica, Ospedale Maggiore di Crema, Crema, ItalyGenetica Medica, Azienda Ospedaliera Universitaria Senese, Siena, ItalyMedical Genetics, University of Siena, Siena, Italy; Genetica Medica, Azienda Ospedaliera Universitaria Senese, Siena, Italy; Alessandra Renieri, MD, PhD, Medical Genetics Unit, University of Siena, Policlinico “Santa Maria alle Scotte”, Viale Bracci, 2, 53100 Siena, ItalyRadiologia interventistica, Ospedale Maggiore di Crema, Crema, Italy; Chirurgia Vascolare, Ospedale Maggiore di Crema, Largo Ugo Dossena, Crema, Italy; Correspondence: Massimo Vaghi, MD, Chirurgia Vascolare, Ospedale Maggiore di Crema, Largo Ugo Dossena, Cremaa, ItalyObjective: Somatic mosaicism of KRAS gene is currently recognized as the only established molecular basis of arteriovenous malformations (AVM). However, given the limitations of the current technologies, KRAS somatic mutations are detected only in a limited proportion of AVMs and tissue biopsy remains an invasive high risky, sometimes life-threatening, diagnostic procedure. Next-generation sequencing liquid biopsy using cell-free DNA (cfDNA) has emerged as an innovative noninvasive approach for early detection and monitoring of cancer. This approach overcomes the space-time profile constraint of tissue biopsies opens a new scenario for vascular malformations owing to somatic mosaicism. Here, we propose a new approach as a fast noninvasive reliable tool in order to investigate the cfDNA coming from the AVMs. Methods: A group of five patients suffering from AVM were selected. Blood samples from peripheral vein and efferent vein from vascular malformation were collected and cfDNA was extracted. The cfDNA libraries were performed using Oncomine Pan-Cancer Cell-Free Assay. We used Ion Proton for sequencing and Ion Reporter Software for analysis (Life Technologies, Carlsbad, Calif). Results: In all cases, either G12D or G12V mutations in KRAS were identified. The mutational load was higher in the efferent vein than in peripheral blood, confirming the causative role of the identified mutation at a somatic level. Conclusions: We demonstrate that cfDNA next-generation sequencing liquid biopsy is able to identify the KRAS mutation detected in affected tissues. Moreover, we have shown that blood sample withdrawal at the lesion site increases variant allele frequency with an order of magnitude above the limit of detection (usually 0.05%), decreasing the risk of a false negative. Finally, the noninvasiveness of the method avoids any risk of bleeding, being easily performed also in children. We propose this technique as the method of choice to better investigate AVMs and consequently to identify the therapy tailored to the genetic defect. Clinical Relevance: This article highlights the importance of using liquid biopsy as a new method to investigate the molecular profile of AVMs. In view of the frequent inaccessibility of vascular tissues owing to the invasiveness of solid biopsy and the relative high incidence of biopsies with low diagnostic power, here we evaluated the efficacy of detecting cfDNA fragments released into the bloodstream from the affected tissue cells. Through a simple blood draw from the efferent vein at the vascular malformation site, the liquid biopsy allowed us to identify KRAS pathogenic mutations piloting a personalized therapeutic approach and opening a new scenario for new therapeutic strategies.http://www.sciencedirect.com/science/article/pii/S2666350320300195Arteriovenous malformationLiquid biopsycf-DNAKRAS mutationNoninvasive technique |
| spellingShingle | Maria Palmieri, MS Aurora Currò, MD Andrea Tommasi, MD Laura Di Sarno, MS Gabriella Doddato, MS Margheria Baldassarri, MD Elisa Frullanti, MS Ann Rita Giliberti, MS Chiara Fallerini, MS Angelo Spinazzola, MD Anna Maria Pinto, MD, PhD Alessandra Renieri, MD, PhD Massimo Vaghi, MD Cell-free DNA next-generation sequencing liquid biopsy as a new revolutionary approach for arteriovenous malformation JVS - Vascular Science Arteriovenous malformation Liquid biopsy cf-DNA KRAS mutation Noninvasive technique |
| title | Cell-free DNA next-generation sequencing liquid biopsy as a new revolutionary approach for arteriovenous malformation |
| title_full | Cell-free DNA next-generation sequencing liquid biopsy as a new revolutionary approach for arteriovenous malformation |
| title_fullStr | Cell-free DNA next-generation sequencing liquid biopsy as a new revolutionary approach for arteriovenous malformation |
| title_full_unstemmed | Cell-free DNA next-generation sequencing liquid biopsy as a new revolutionary approach for arteriovenous malformation |
| title_short | Cell-free DNA next-generation sequencing liquid biopsy as a new revolutionary approach for arteriovenous malformation |
| title_sort | cell free dna next generation sequencing liquid biopsy as a new revolutionary approach for arteriovenous malformation |
| topic | Arteriovenous malformation Liquid biopsy cf-DNA KRAS mutation Noninvasive technique |
| url | http://www.sciencedirect.com/science/article/pii/S2666350320300195 |
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