Targeted Next-Generation Sequencing of Cell-Free DNA to Detect MYC-Immunoglobulin Translocation and Epstein-Barr Virus DNA in Plasma of Burkitt Lymphoma Patients in East Africa
PURPOSEEpstein-Barr virus (EBV)–positive Burkitt lymphoma (BL) affects children in sub-Saharan Africa, but diagnosis via tissue biopsy is challenging. We explored a liquid biopsy approach using targeted next-generation sequencing to detect the MYC-immunoglobulin (MYC-Ig) translocation and EBV DNA, a...
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| Language: | English |
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American Society of Clinical Oncology
2025-04-01
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| Series: | JCO Global Oncology |
| Online Access: | https://ascopubs.org/doi/10.1200/GO.24.00210 |
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| author | Clara Chamba Daisy Jennings Rehema Shungu Heavenlight Christopher Emmanuel Josephat Kieran Howard Helene Dreau Adam Burns William Mawalla Priscus Mapendo Leah Mnango Ismail Legason Edrick Elias Caroline Achola Anthony Cutts Emmanuel Balandya Anna Schuh |
| author_facet | Clara Chamba Daisy Jennings Rehema Shungu Heavenlight Christopher Emmanuel Josephat Kieran Howard Helene Dreau Adam Burns William Mawalla Priscus Mapendo Leah Mnango Ismail Legason Edrick Elias Caroline Achola Anthony Cutts Emmanuel Balandya Anna Schuh |
| author_sort | Clara Chamba |
| collection | DOAJ |
| description | PURPOSEEpstein-Barr virus (EBV)–positive Burkitt lymphoma (BL) affects children in sub-Saharan Africa, but diagnosis via tissue biopsy is challenging. We explored a liquid biopsy approach using targeted next-generation sequencing to detect the MYC-immunoglobulin (MYC-Ig) translocation and EBV DNA, assessing its potential for minimally invasive BL diagnosis.MATERIALS AND METHODSThe panel included targets for the characteristic MYC-Ig translocation, mutations in intron 1 of MYC, mutations in exon 2 of MYC, and three EBV genes: EBV-encoded RNA (EBER)1, EBER2, and EBV nuclear antigen 2. It was first tested in a small derivation cohort of four precharacterized BL-derived cell lines with known translocation status and eight precharacterized plasma samples with known EBV DNA status by quantitative polymerase chain reaction (qPCR). These different data modalities were combined to assess the accuracy of this approach in the diagnosis of BL in 20 patient plasma samples in Tanzania and Uganda.RESULTSThe next-generation sequencing panel detected three of four MYC-Ig translocations in the BL-derived cell lines. EBV viral load by targeted sequencing correlated strongly with qPCR results (Spearman's rho = 0.94) in precharacterized plasma samples. Using the patient plasma samples, mutations in MYC intron 1 were associated with the presence of a MYC translocation with 25 or more mutations being predictive of a translocation with AUC, sensitivity, and specificity of 1. Overall, liquid biopsy parameters associated with a diagnosis of BL (P < .05) included cell-free DNA concentration, circulating tumor DNA concentration, MYC intron 1 mutations, MYC-Ig translocation, and autosome entropy. Integrating these parameters into a diagnostic model demonstrated excellent performance with an AUC of 0.95, sensitivity of 0.9, and specificity of 1.CONCLUSIONThis analysis demonstrates the potential of liquid biopsy to improve BL diagnosis in settings with limited pathology resources. Validation of our approach in a larger data set is needed. |
| format | Article |
| id | doaj-art-43f2cb883e714f29a991504900c6d016 |
| institution | DOAJ |
| issn | 2687-8941 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | American Society of Clinical Oncology |
| record_format | Article |
| series | JCO Global Oncology |
| spelling | doaj-art-43f2cb883e714f29a991504900c6d0162025-08-20T03:17:58ZengAmerican Society of Clinical OncologyJCO Global Oncology2687-89412025-04-011110.1200/GO.24.00210Targeted Next-Generation Sequencing of Cell-Free DNA to Detect MYC-Immunoglobulin Translocation and Epstein-Barr Virus DNA in Plasma of Burkitt Lymphoma Patients in East AfricaClara Chamba0Daisy Jennings1Rehema Shungu2Heavenlight Christopher3Emmanuel Josephat4Kieran Howard5Helene Dreau6Adam Burns7William Mawalla8Priscus Mapendo9Leah Mnango10Ismail Legason11Edrick Elias12Caroline Achola13Anthony Cutts14Emmanuel Balandya15Anna Schuh16Muhimbili University of Health and Allied Sciences, Dar es Salaam, TanzaniaUniversity of Oxford, Oxford, United KingdomMuhimbili University of Health and Allied Sciences, Dar es Salaam, TanzaniaMuhimbili University of Health and Allied Sciences, Dar es Salaam, TanzaniaMuhimbili University of Health and Allied Sciences, Dar es Salaam, TanzaniaUniversity of Oxford, Oxford, United KingdomUniversity of Oxford, Oxford, United KingdomUniversity of Oxford, Oxford, United KingdomMuhimbili University of Health and Allied Sciences, Dar es Salaam, TanzaniaKilimanjaro Christian Medical Centre, Moshi, TanzaniaMuhimbili National Hospital, Dar es Salaam, TanzaniaSt Mary's Lacor Hospital, Lacor, UgandaBugando Medical Centre, Mwanza, TanzaniaCentral Public Health Laboratory, Kampala, UgandaUniversity of Oxford, Oxford, United KingdomMuhimbili University of Health and Allied Sciences, Dar es Salaam, TanzaniaUniversity of Oxford, Oxford, United KingdomPURPOSEEpstein-Barr virus (EBV)–positive Burkitt lymphoma (BL) affects children in sub-Saharan Africa, but diagnosis via tissue biopsy is challenging. We explored a liquid biopsy approach using targeted next-generation sequencing to detect the MYC-immunoglobulin (MYC-Ig) translocation and EBV DNA, assessing its potential for minimally invasive BL diagnosis.MATERIALS AND METHODSThe panel included targets for the characteristic MYC-Ig translocation, mutations in intron 1 of MYC, mutations in exon 2 of MYC, and three EBV genes: EBV-encoded RNA (EBER)1, EBER2, and EBV nuclear antigen 2. It was first tested in a small derivation cohort of four precharacterized BL-derived cell lines with known translocation status and eight precharacterized plasma samples with known EBV DNA status by quantitative polymerase chain reaction (qPCR). These different data modalities were combined to assess the accuracy of this approach in the diagnosis of BL in 20 patient plasma samples in Tanzania and Uganda.RESULTSThe next-generation sequencing panel detected three of four MYC-Ig translocations in the BL-derived cell lines. EBV viral load by targeted sequencing correlated strongly with qPCR results (Spearman's rho = 0.94) in precharacterized plasma samples. Using the patient plasma samples, mutations in MYC intron 1 were associated with the presence of a MYC translocation with 25 or more mutations being predictive of a translocation with AUC, sensitivity, and specificity of 1. Overall, liquid biopsy parameters associated with a diagnosis of BL (P < .05) included cell-free DNA concentration, circulating tumor DNA concentration, MYC intron 1 mutations, MYC-Ig translocation, and autosome entropy. Integrating these parameters into a diagnostic model demonstrated excellent performance with an AUC of 0.95, sensitivity of 0.9, and specificity of 1.CONCLUSIONThis analysis demonstrates the potential of liquid biopsy to improve BL diagnosis in settings with limited pathology resources. Validation of our approach in a larger data set is needed.https://ascopubs.org/doi/10.1200/GO.24.00210 |
| spellingShingle | Clara Chamba Daisy Jennings Rehema Shungu Heavenlight Christopher Emmanuel Josephat Kieran Howard Helene Dreau Adam Burns William Mawalla Priscus Mapendo Leah Mnango Ismail Legason Edrick Elias Caroline Achola Anthony Cutts Emmanuel Balandya Anna Schuh Targeted Next-Generation Sequencing of Cell-Free DNA to Detect MYC-Immunoglobulin Translocation and Epstein-Barr Virus DNA in Plasma of Burkitt Lymphoma Patients in East Africa JCO Global Oncology |
| title | Targeted Next-Generation Sequencing of Cell-Free DNA to Detect MYC-Immunoglobulin Translocation and Epstein-Barr Virus DNA in Plasma of Burkitt Lymphoma Patients in East Africa |
| title_full | Targeted Next-Generation Sequencing of Cell-Free DNA to Detect MYC-Immunoglobulin Translocation and Epstein-Barr Virus DNA in Plasma of Burkitt Lymphoma Patients in East Africa |
| title_fullStr | Targeted Next-Generation Sequencing of Cell-Free DNA to Detect MYC-Immunoglobulin Translocation and Epstein-Barr Virus DNA in Plasma of Burkitt Lymphoma Patients in East Africa |
| title_full_unstemmed | Targeted Next-Generation Sequencing of Cell-Free DNA to Detect MYC-Immunoglobulin Translocation and Epstein-Barr Virus DNA in Plasma of Burkitt Lymphoma Patients in East Africa |
| title_short | Targeted Next-Generation Sequencing of Cell-Free DNA to Detect MYC-Immunoglobulin Translocation and Epstein-Barr Virus DNA in Plasma of Burkitt Lymphoma Patients in East Africa |
| title_sort | targeted next generation sequencing of cell free dna to detect myc immunoglobulin translocation and epstein barr virus dna in plasma of burkitt lymphoma patients in east africa |
| url | https://ascopubs.org/doi/10.1200/GO.24.00210 |
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