TY - JOUR
T1 - Cell lines generated from a chronic lymphocytic leukemia mouse model exhibit constitutive Btk and Akt signaling
AU - Singh, Simar Pal
AU - Pillai, Saravanan Y.
AU - de Bruijn, Marjolein J. W.
AU - Stadhouders, Ralph
AU - Corneth, Odilia B. J.
AU - van den Ham, Henk Jan
AU - Muggen, Alice
AU - van Ijcken, Wilfred
AU - Slinger, Erik
AU - Kuil, Annemieke
AU - Spaargaren, Marcel
AU - Kater, Arnon P.
AU - Langerak, Anton W.
AU - Hendriks, Rudi W.
PY - 2017
Y1 - 2017
N2 - Chronic lymphocytic leukemia (CLL) is characterized by the accumulation of mature CD5(+) B cells in blood. Spontaneous apoptosis of CLL cells in vitro has hampered in-depth investigation of CLL pathogenesis. Here we describe the generation of three monoclonal mouse cell lines, EMC2, EMC4 and EMC6, from the IgH. TE mu CLL mouse model based on sporadic expression of SV40 large T antigen. The cell lines exhibit a stable CD5(+) CD43(+) IgM(+) CD19(+) CLL phenotype in culture and can be adoptively transferred into Rag1(-/-) mice. RNA-seq analysis revealed only minor differences between the cell lines and their primary tumors and suggested that NF-kappa B and mTOR signaling pathways were involved in cell line outgrowth. In vitro survival and proliferation was dependent on constitutive phosphorylation of Bruton's tyrosine kinase (Btk) at Y551/Y223, and Akt(S473). Treatment of the cell lines with small molecule inhibitors specific for Btk (ibrutinib) or PI3K (idelalisib), which is upstream of Akt, resulted in reduced viability, proliferation and fibronectin-dependent cell adhesion. Treatment of cell line-engrafted Rag1(-/-) mice with ibrutinib was associated with transient lymphocytosis, reduced splenomegaly and increased overall survival. Thus, by generating stable cell lines we established a novel platform for in vitro and in vivo investigation of CLL signal transduction and treatment modalities
AB - Chronic lymphocytic leukemia (CLL) is characterized by the accumulation of mature CD5(+) B cells in blood. Spontaneous apoptosis of CLL cells in vitro has hampered in-depth investigation of CLL pathogenesis. Here we describe the generation of three monoclonal mouse cell lines, EMC2, EMC4 and EMC6, from the IgH. TE mu CLL mouse model based on sporadic expression of SV40 large T antigen. The cell lines exhibit a stable CD5(+) CD43(+) IgM(+) CD19(+) CLL phenotype in culture and can be adoptively transferred into Rag1(-/-) mice. RNA-seq analysis revealed only minor differences between the cell lines and their primary tumors and suggested that NF-kappa B and mTOR signaling pathways were involved in cell line outgrowth. In vitro survival and proliferation was dependent on constitutive phosphorylation of Bruton's tyrosine kinase (Btk) at Y551/Y223, and Akt(S473). Treatment of the cell lines with small molecule inhibitors specific for Btk (ibrutinib) or PI3K (idelalisib), which is upstream of Akt, resulted in reduced viability, proliferation and fibronectin-dependent cell adhesion. Treatment of cell line-engrafted Rag1(-/-) mice with ibrutinib was associated with transient lymphocytosis, reduced splenomegaly and increased overall survival. Thus, by generating stable cell lines we established a novel platform for in vitro and in vivo investigation of CLL signal transduction and treatment modalities
KW - B-cell receptor (BCR)
KW - bruton’s tyrosine kinase (Btk)
KW - chronic lymphocytic leukemia (CLL)
KW - ibrutinib
KW - idelalisib
UR - http://www.scopus.com/inward/record.url?scp=85042148875&partnerID=8YFLogxK
U2 - https://doi.org/10.18632/oncotarget.18234
DO - https://doi.org/10.18632/oncotarget.18234
M3 - Article
C2 - 28614781
SN - 1949-2553
VL - 8
SP - 71981
EP - 71995
JO - Oncotarget
JF - Oncotarget
IS - 42
ER -