RPPA-based protein profiling reveals eIF4G overexpression and 4E-BP1 serine 65 phosphorylation as molecular events that correspond with a pro-survival phenotype in chronic lymphocytic leukemia

// Austin Y. Shull 1,2 , Satish K. Noonepalle 1,2 , Farrukh T. Awan 3 , Jimei Liu 2 , Lirong Pei 2 , Roni J. Bollag 2,4 , Huda Salman 2,5 , Zhiyong Ding 6 and Huidong Shi 1,2 1 Department of Biochemistry & Molecular Biology, Georgia Regents University, Augusta, Georgia, USA 2 GRU Cancer Center, Georgia Regents University, Augusta, Georgia, USA 3 The Ohio State Comprehensive Cancer Center, The Ohio State University, Columbus, Ohio, USA 4 Department of Pathology, Georgia Regents University, Augusta, Georgia, USA 5 Deparment of Medicine, Georgia Regents University, Augusta, Georgia, USA 6 Department of Systems Biology, University of Texas MD Anderson Cancer Center, Houston, Texas, USA Correspondence to: Huidong Shi, email: // Zhiyong Ding, email: // Keywords : CLL, RPPA, EIF4G, 4E-BP1, NVP-BEZ235 Received : January 16, 2015 Accepted : April 08, 2015 Published : May 12, 2015 Abstract Chronic lymphocytic leukemia (CLL), the most common adult leukemia, remains incurable despite advancements in treatment regimens over the past decade. Several expression profile studies have been pursued to better understand CLL pathogenesis. However, these large-scale studies only provide information at the transcriptional level. To better comprehend the differential protein changes that take place in CLL, we performed a reverse-phase protein array (RPPA) analysis using 167 different antibodies on B-cell lysates from 18 CLL patients and 6 normal donors. From our analysis, we discovered an enrichment of protein alterations involved with mRNA translation, specifically upregulation of the translation initiator eIF4G and phosphorylation of the cap-dependent translation inhibitor 4E-BP1 at serine 65. Interestingly, 4E-BP1 phosphorylation occurred independently of AKT phosphorylation, suggesting a disconnect between PI3K/AKT pathway activation and 4E-BP1 phosphorylation. Based on these results, we treated primary CLL samples with NVP-BEZ235, a PI3K/mTOR dual inhibitor, and compared its apoptotic-inducing potential against the BTK inhibitor Ibrutinib and the PI3Kδ inhibitor Idelalisib. We demonstrated that treatment with NVP-BEZ235 caused greater apoptosis, greater apoptotic cleavage of eIF4G, and greater dephosphorylation of 4E-BP1 in primary CLL cells. Taken together, these results highlight the potential dependence of eIF4G overexpression and 4E-BP1 phosphorylation in CLL survival.