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Summary  Cereblon, a member of the cullin 4 ring ligase complex ( CRL 4), is the molecular target of the immunomodulatory drugs ( IM i D s) lenalidomide and pomalidomide and is required for the antiproliferative activity of these agents in multiple myeloma ( MM ) and immunomodulatory activity in  T  cells. Cereblon's central role as a target of lenalidomide and pomalidomide suggests potential utility as a predictive biomarker of response or resistance to  IM i D  therapy. Our studies characterized a cereblon monoclonal antibody  CRBN 65, with high sensitivity and specificity in  W estern analysis and immunohistochemistry that is superior to commercially available antibodies. We identified multiple cereblon splice variants in both  MM  cell lines and primary cells, highlighting challenges with conventional gene expression assays given this gene complexity. Using  CRBN 65 antibody and  T aq M an quantitative reverse transcription polymerase chain reaction assays, we showed lack of correlation between cereblon protein and  mRNA  levels. Furthermore, lack of correlation between cereblon expression in  MM  cell lines and sensitivity to lenalidomide was shown. In cell lines made resistant to lenalidomide and pomalidomide, cereblon protein is greatly reduced. These studies show limitations to the current approaches of cereblon measurement that rely on commercial reagents and assays. Standardized reagents and validated assays are needed to accurately assess the role of cereblon as a predictive biomarker.

british journal of haematology

Measuring cereblon as a biomarker of response or resistance to lenalidomide and pomalidomide requires use of standardized reagents and understanding of gene complexity