For the VUMC EBV website
Diagnostic and pathogenic markers
Epstein-Barr virus (EBV) is a ubiquitous human herpesvirus etiologically linked to multiple acute, chronic and malignant disease syndromes. In past years we have identified and validated the use of various (anti-) viral markers in patients with different EBV-linked diseases and developed new methods and guidelines for diagnosis. We have developed a unique series of pathogenic and diagnostic markers. These markers and methods are now being implemented in a worldwide network of collaborations for monitoring transplant recipients, patients with different haematological disease, HIV carriers with lymphadenopathy, patients with chronic fatigue syndrome and autoimmune diseases like SLE and MS and patients with various lymphomas and carcinomas.
Oncogenesis and treatment
EBV is associated with an increasing number of diseases. EBV is classified as a class-I carcinogen causally implicated in multiple lymphomas and carcinomas. During lifelong persistence EBV hides latently in memory B-cells and preferentially replicates in epithelial cells. EBV-driven malignant disease is closely linked to the viral latent state, although serological evidence suggests a role for virus replication in early stages, which can be exploited for diagnosis. Viral DNA load proved useful in diagnosis and monitoring of both carcinomas and lymphomas. Profiling of EBV latent (LMP-1,-2 and EBNA-1,-2, BARF1) and lytic genes (ZEBRA, TK, VCA p18) reveals viral activity in the different malignancies.
Our aims are
1. to further improve the (early) diagnosis by viral load in combination with serology and implementing new methylation markers. The viral load will be used for monitoring therapy in blood and tumour samples
2. to develop diagnostic assays for monitoring the CLVA therapy based upon lytic components (RNA and proteins) and cell free virion quantification
3. characterization of a carcinoma specific oncogene BARF1, function and presences of the secreted protein in body fluids
Exosomes in pathogenesis of cancer and immune disorders
Cancer profiling through accessible and reliable prognostic biomarkers is paramount for early detection and effective therapy. The presence of microRNAs (miRNA) containing exosomes in biofluids opens unique possibilities in using circulating tumor-derived RNAs as 'easily accessible surrogate biomarkers'. However a single RNA biomarker is only in rare cases informative on the disease state and clinically relevant information maybe 'lost' in complex clinical specimens.
Our Previous studies using a viral (EBV) infection as a model, provided the first proof that miRNA-exchange via exosomes is functional and occurs between cells in humans, raising the possibility that this method of cell-cell communication may contribute to and participate in tumorigenesis. To extract clinically relevant information from tumor exosomes we performed deep sequencing analysis and determined which cellular miRNAs are most abundant and which maybe precluded from secretion by cancer cells. The exosomal miRNAome is highly complex and to a large degree similar, but certainly not identical to the smallRNAome from the tumor cells producing them. We also identified several candidate RNA-binding proteins that may drive the incorporation of tumor exosome associated small RNAs "TEASRs" into exosomes that may promote the oncogenic process. Finally using computational analysis it seems possible to construct 'minimal' tumor specific miRNA profiles in tumor tissues and tumor cell-lines and we wish to investigate whether these tumor associated profiles are present in tumor exosomes circulating in patient bodily fluids.
Since the presence of microRNAs (miRNA) containing exosomes in biofluids opens unique possibilities in using circulating tumor-derived RNAs as 'minimally invasive surrogate biomarkers' the aims of our research are
- To characterize the exosomal miRNA communication pathway at the molecular level using RNAomic and proteomic driven approaches to identify novel early-stage cancer biomarkers.
- to combine experimental and computational analysis in constructing tumor specific miRNAs profiles in tumor cell-lines and exosomes and to validate these minimal profiles in tumor tissues and circulating exosomes isolated from patient sera.
- to study the functional significance of exosome-driven secretion of material (i.e. proteins and RNAs) during the various steps of the tumorigenic process and to learn how this process influences post-transcriptional gene expression, oncogenic signalling and immune-escape to support tumor growth.
Chronic inflammatory diseases such as systemic lupus erythematosus (SLE), represent a major challenge for both clinical research and patient care, and evidence indicates that these disorders develop as a result of complex gene-environment interactions. Despite the identification of many potential susceptibility genes by genome-wide association studies they seem to explain only a very small proportion of disease heritability. A better understanding of their cause-and-effect relationship may provide a basis for new diagnostic tools, therapy and prevention. We found a molecular link between Epstein Barr virus infection (EBV) and SLE that may explain the chronic activation of type I IFN responses via innate receptors. Systemic lupus erythematosus (SLE), represent a major challenge for both clinical research and patient care, and evidence indicates that these disorders develop as a result of complex gene-environment interactions. Therefore we aim to exploit circulating RNAs of viral origin as clinical SLE biomarkers and to decipher the molecular pathways by which EBV contributes to the pro-inflammatory release of cytokines in SLE patients as a driver of autoimmunity.
Jaap Middeldorp PhD - program leader
Astrid Greijer PhD - postdoc
Michiel Pegtel PhD - postdoc
Evelien Hoebe MSc - PhD student
Frederik Verweij MSc - PhD student
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