EBV-Associated Lymphoma Consortium Annual Meeting (Abstract): Submission #11
Submission information
Submission Number: 11
Submission ID: 150967
Submission UUID: 0b3b3924-8791-41ae-b8e5-bdab6009af25
Submission URI: /nci/ealv/venue/abstract
Submission Update: /nci/ealv/venue/abstract?token=s9VHZ_l2q0qifF5EYZJ7U_7lg8VqelukZYgI2aSd6Sw
Created: Thu, 09/04/2025 - 18:11
Completed: Thu, 09/04/2025 - 18:11
Changed: Thu, 09/04/2025 - 18:11
Remote IP address: 10.208.24.100
Submitted by: Anonymous
Language: English
Is draft: No
Presenter Information
Rena
R.
Xian
MD
Associate Professor
Johns Hopkins University School of Medicine
Baltimore
Abstract Information
EBV methylation, bisulfite sequencing, plasma, and saliva cfDNA, Wp
Investigating the EBV methylome in PLWH: Discovery and Development of Novel EBV Diagnostics in Plasma and Saliva
Introduction: EBV DNA derived from tumors is consistently methylated with unique methylation profiles. In contrast, virion DNA is never methylated. This project seeks to study the relationship between the EBV epigenome and EBV transcriptome in EBV(+) lymphoma in people with HIV (PWH). We will characterize EBV methylation in cell-free DNA (cfDNA) from plasma and saliva to define methylation patterns in different disease states in order to develop novel diagnostics for HIV-associated lymphoma.
Methods: EBV methylation was evaluated by a bead-based methyl-DNA binding protein capture of the W promoter and bisulfite sequencing of the viral genome (bsEBV-seq). Plasma samples from PWH with Hodgkin lymphoma (HL, n=12) or Plasmablastic lymphoma (PBL, n=21) were analyzed by the bead-based method. Thirty-two samples (tissue biopsy, plasma, and saliva) underwent bsEBV-seq. Lastly, cfDNA from 71 plasma samples and 44 matched saliva samples from PWH without EBV(+) lymphoma were evaluated for the presence of EBV.
Results: Bead-based methylation of Wp shows hypermethylation in HL (median 97%, 83-98%) and variable methylation in PBL (median 47%, 3-96%). bsEBV-seq shows diagnosis-specific and lymphoma subtype-specific patterns of methylation. HL shows consistent methylation across the EBV genome, whereas EBV methylation is more variable in PBL. Regarding OriP, there is either no methylation of the entire region, or there is recurrent site-specific hypermethylation. When evaluating EBV in PWH without EBV(+) lymphoma, the frequency of detecting EBV is higher in saliva (94%, 44/47) than in plasma (67%, 48/71). The level of EBV is also higher in saliva (median 3.8 LogEBVcopies/mL) than in plasma (0.8 LogEBVcopies/mL). bsEBV-seq of select saliva samples show no methylation across the viral genome consistent with presence of virion DNA.
Conclusions: Different methods of analyzing EBV methylation were applied. Findings from the EBV methylome studies will enable the development of novel plasma and saliva diagnostics for lymphoma in PWH.
Methods: EBV methylation was evaluated by a bead-based methyl-DNA binding protein capture of the W promoter and bisulfite sequencing of the viral genome (bsEBV-seq). Plasma samples from PWH with Hodgkin lymphoma (HL, n=12) or Plasmablastic lymphoma (PBL, n=21) were analyzed by the bead-based method. Thirty-two samples (tissue biopsy, plasma, and saliva) underwent bsEBV-seq. Lastly, cfDNA from 71 plasma samples and 44 matched saliva samples from PWH without EBV(+) lymphoma were evaluated for the presence of EBV.
Results: Bead-based methylation of Wp shows hypermethylation in HL (median 97%, 83-98%) and variable methylation in PBL (median 47%, 3-96%). bsEBV-seq shows diagnosis-specific and lymphoma subtype-specific patterns of methylation. HL shows consistent methylation across the EBV genome, whereas EBV methylation is more variable in PBL. Regarding OriP, there is either no methylation of the entire region, or there is recurrent site-specific hypermethylation. When evaluating EBV in PWH without EBV(+) lymphoma, the frequency of detecting EBV is higher in saliva (94%, 44/47) than in plasma (67%, 48/71). The level of EBV is also higher in saliva (median 3.8 LogEBVcopies/mL) than in plasma (0.8 LogEBVcopies/mL). bsEBV-seq of select saliva samples show no methylation across the viral genome consistent with presence of virion DNA.
Conclusions: Different methods of analyzing EBV methylation were applied. Findings from the EBV methylome studies will enable the development of novel plasma and saliva diagnostics for lymphoma in PWH.
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