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Experimental Hematology & Oncology
Published in: Experimental Hematology & Oncology 1/2015

Open Access 01-12-2015 | Research

New CD20 alternative splice variants: molecular identification and differential expression within hematological B cell malignancies

Authors: Clémentine Gamonet, Elodie Bole-Richard, Aurélia Delherme, François Aubin, Eric Toussirot, Francine Garnache-Ottou, Yann Godet, Loïc Ysebaert, Olivier Tournilhac, Caroline Dartigeas, Fabrice Larosa, Eric Deconinck, Philippe Saas, Christophe Borg, Marina Deschamps, Christophe Ferrand

Published in: Experimental Hematology & Oncology | Issue 1/2015

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Abstract

Background

CD20 is a B cell lineage–specific marker expressed by normal and leukemic B cells and targeted by several antibody immunotherapies. We have previously shown that the protein from a CD20 mRNA splice variant (D393-CD20) is expressed at various levels in leukemic B cells or lymphoma B cells but not in resting, sorted B cells from the peripheral blood of healthy donors.

Results

Western blot (WB) analysis of B malignancy primary samples showed additional CD20 signals. Deep molecular PCR analysis revealed four new sequences corresponding to in-frame CD20 splice variants (D657-CD20, D618-CD20, D480-CD20, and D177-CD20) matching the length of WB signals. We demonstrated that the cell spliceosome machinery can process ex vivo D480-, D657-, and D618-CD20 transcript variants by involving canonical sites associated with cryptic splice sites. Results of specific and quantitative RT-PCR assays showed that these CD20 splice variants are differentially expressed in B malignancies. Moreover, Epstein–Barr virus (EBV) transformation modified the CD20 splicing profile and mainly increased the D393-CD20 variant transcripts. Finally, investigation of three cohorts of chronic lymphocytic leukemia (CLL) patients showed that the total CD20 splice variant expression was higher in a stage B and C sample collection compared to routinely collected CLL samples or relapsed refractory stage A, B, or C CLL.

Conclusion

The involvement of these newly discovered alternative CD20 transcript variants in EBV transformation makes them interesting molecular indicators, as does their association with oncogenesis rather than non-oncogenic B cell diseases, differential expression in B cell malignancies, and correlation with CLL stage and some predictive CLL markers. This potential should be investigated in further studies.
Appendix
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Metadata
Title
New CD20 alternative splice variants: molecular identification and differential expression within hematological B cell malignancies
Authors
Clémentine Gamonet
Elodie Bole-Richard
Aurélia Delherme
François Aubin
Eric Toussirot
Francine Garnache-Ottou
Yann Godet
Loïc Ysebaert
Olivier Tournilhac
Caroline Dartigeas
Fabrice Larosa
Eric Deconinck
Philippe Saas
Christophe Borg
Marina Deschamps
Christophe Ferrand
Publication date
01-12-2015
Publisher
BioMed Central
Published in
Experimental Hematology & Oncology / Issue 1/2015
Electronic ISSN: 2162-3619
DOI
https://doi.org/10.1186/s40164-016-0036-3

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