TY - JOUR
T1 - Development and characterization of prototypes for in vitro and in vivo mouse models of ibrutinib-resistant CLL
AU - Aslan, Burcu
AU - Kismali, Gorkem
AU - Chen, Lisa S.
AU - Iles, Lakesla R.
AU - Mahendra, Mikhila
AU - Peoples, Michael
AU - Gagea, Mihai
AU - Fowlkes, Natalie W.
AU - Zheng, Xiaofeng
AU - Wang, Jing
AU - Vellano, Christopher P.
AU - Marszalek, Joseph R.
AU - Sabrina Bertilaccio, Maria Teresa
AU - Gandhi, Varsha
N1 - Funding Information:
This work was supported in part by The University of Texas MD Anderson Cancer Center Moon Shot Program; by National Institutes of Health, National Cancer Institute grant P30CA016672 (the Flow Cytometry and Cellular Imaging Core Facility, Functional Proteomics RPPA Core Facility, The Sequencing and Non-Coding Program Laboratory, Advanced Technology Genomics Core Cytogenetics, Cell Authentication Core Facility Research Animal Support Facility-Houston, Research Histology Core Laboratory, and Department of Veterinary Medicine and Surgery); and by the Scientific and Technological Research Council of Turkey (TUB€İTAK) Science Fellowships and Grant Programs (BIDEB) International Postdoctoral Research Fellowship Program 2219 (G.K.).
Publisher Copyright:
© 2021 American Society of Hematology. All rights reserved.
PY - 2021/8/24
Y1 - 2021/8/24
N2 - Although ibrutinib improves the overall survival of patients with chronic lymphocytic leukemia (CLL), some patients still develop resistance, most commonly through point mutations affecting cysteine residue 481 (C481) in Bruton s tyrosine kinase (BTKC481S and BTKC481R). To enhance our understanding of the biological impact of these mutations, we established cell lines that overexpress wild-Type or mutant BTK in in vitro and in vivo models that mimic ibrutinib-sensitive and-resistant CLL. MEC-1 cell lines stably overexpressing wild-Type or mutant BTK were generated. All cell lines coexpressed GFP, were CD191 and CD231, and overexpressed BTK. Overexpression of wild-Type or mutant BTK resulted in increased signaling, as evidenced by the induction of p-BTK, p-PLCg2, and p-extracellular signal related kinase (ERK) levels, the latter further augmented upon IgM stimulation. In all cell lines, cell cycle profiles and levels of BTK expression were similar, but the RNA sequencing and reverse-phase protein array results revealed that the molecular transcript and protein profiles were distinct. To mimic aggressive CLL, we created xenograft mouse models by transplanting the generated cell lines into Rag22/2gc 2/2 mice. Spleens, livers, bone marrow, and peripheral blood were collected. All mice developed CLL-like disease with systemic involvement (engraftment efficiency, 100%). We observed splenomegaly, accumulation of leukemic cells in the spleen and liver, and macroscopically evident necrosis. CD191 cells accumulated in the spleen, bone marrow, and peripheral blood. The overall survival duration was slightly lower in mice expressing mutant BTK. Our cell lines and murine models mimicking ibrutinib-resistant CLL will serve as powerful tools to test reversible BTK inhibitors and novel, non BTK-Targeted therapeutics.
AB - Although ibrutinib improves the overall survival of patients with chronic lymphocytic leukemia (CLL), some patients still develop resistance, most commonly through point mutations affecting cysteine residue 481 (C481) in Bruton s tyrosine kinase (BTKC481S and BTKC481R). To enhance our understanding of the biological impact of these mutations, we established cell lines that overexpress wild-Type or mutant BTK in in vitro and in vivo models that mimic ibrutinib-sensitive and-resistant CLL. MEC-1 cell lines stably overexpressing wild-Type or mutant BTK were generated. All cell lines coexpressed GFP, were CD191 and CD231, and overexpressed BTK. Overexpression of wild-Type or mutant BTK resulted in increased signaling, as evidenced by the induction of p-BTK, p-PLCg2, and p-extracellular signal related kinase (ERK) levels, the latter further augmented upon IgM stimulation. In all cell lines, cell cycle profiles and levels of BTK expression were similar, but the RNA sequencing and reverse-phase protein array results revealed that the molecular transcript and protein profiles were distinct. To mimic aggressive CLL, we created xenograft mouse models by transplanting the generated cell lines into Rag22/2gc 2/2 mice. Spleens, livers, bone marrow, and peripheral blood were collected. All mice developed CLL-like disease with systemic involvement (engraftment efficiency, 100%). We observed splenomegaly, accumulation of leukemic cells in the spleen and liver, and macroscopically evident necrosis. CD191 cells accumulated in the spleen, bone marrow, and peripheral blood. The overall survival duration was slightly lower in mice expressing mutant BTK. Our cell lines and murine models mimicking ibrutinib-resistant CLL will serve as powerful tools to test reversible BTK inhibitors and novel, non BTK-Targeted therapeutics.
UR - http://www.scopus.com/inward/record.url?scp=85113879698&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85113879698&partnerID=8YFLogxK
U2 - 10.1182/bloodadvances.2020003821
DO - 10.1182/bloodadvances.2020003821
M3 - Article
C2 - 34424317
AN - SCOPUS:85113879698
SN - 2473-9529
VL - 5
SP - 3134
EP - 3146
JO - Blood Advances
JF - Blood Advances
IS - 16
ER -