Mouse models of mantle cell lymphoma, complex changes in gene expression and phenotype of engrafted MCL cells: implications for preclinical research

Abstract

Mantle cell lymphoma (MCL) is an aggressive type of B-cell non-Hodgkin lymphoma (NHL) associated with poor prognosis. Animal models of MCL are scarce. We established and characterized various in vivo models of metastatic human MCL by tail vein injection of either primary cells isolated from patients with MCL or established MCL cell lines (Jeko-1, Mino, Rec-1, Hbl-2, and Granta-519) into immunodeficient NOD.Cg-Prkdc scid Il2rg tm1Wjl /SzJ mice. MCL infiltration was assessed with immunohistochemistry (tissues) and flow cytometry (peripheral blood). Engraftment of primary MCL cells was observed in 7 out of 12 patient samples. The pattern of engraftment of primary MCL cells varied from isolated involvement of the spleen to multiorgan infiltration. On the other hand, tumor engraftment was achieved in all five MCL cell lines used and lymphoma involvement of murine bone marrow, spleen, liver, and brain was observed. Overall survival of xenografted mice ranged from 22±1 to 54±3 days depending on the cell line used. Subsequently, we compared the gene expression profile (GEP) and phenotype of the engrafted MCL cells compared with the original in vitro growing cell lines (controls). We demonstrated that engrafted MCL cells displayed complex changes of GEP, protein expression, and sensitivity to cytotoxic agents when compared with controls. We further demonstrated that our MCL mouse models could be used to test the therapeutic activity of systemic chemotherapy, monoclonal antibodies, or angiogenesis inhibitors. The characterization of MCL murine models is likely to aid in improving our knowledge in the disease biology and to assist scientists in the preclinical and clinical development of novel agents in relapsed/refractory MCL patients. Mantle cell lymphoma (MCL) is an aggressive type of B-cell non-Hodgkin lymphoma (NHL) characterized by the chromosomal translocation t(11;14)(q13;q32) leading to overexpression of cyclin D1. 1 Apart from this canonical aberration, MCL may harbor a large number of recurrent cytogenetic changes that further deregulate cell cycle machinery (for example, deletion of 9p21 (CDKN2A) and amplification of 12q13 (CDK4)) or interfere with cellular response to DNA damage (for example, alterations of 11q22-q23 (ATM), 17p13 (TP53) and overexpression of MDM2). 2-5 MCL is associated with poor prognosis. 6,7 In order to improve the outcome of MCL patients, the rational development and preclinical/clinical testing of new agents are necessary. Reliable and reproducible in vivo models of MCL are thus urgently needed. In vivo models have several advantages over in vitro approaches. For example, they enable the study of MCL biology in its microenvironment, including engraftment, growth rate, spread patterns, or tumor associated neovascularization.

Targeting of BCL2 Family Proteins with ABT-199 and Homoharringtonine Reveals BCL2- and MCL1-Dependent Subgroups of Diffuse Large B-Cell Lymphoma

“…Primary DLBCL-based mouse models designated KTC (treatment refractory DLBCL) and NOVA-313 (DLBCL transformed from CLL) were established in the same way as previously described (18). KTC or NOVA-313 cells were chosen for in vivo experiments, because they coexpressed BCL2 and MCL1 (Fig.…”

Pevonedistat, a NEDD8-activating enzyme inhibitor, is active in mantle cell lymphoma and enhances rituximab activity in vivo

“…Cytarabine-resistant cell lines (Mino-AraCR, HBL-2-AraCR, Jeko-1-AraCR, and Rec-1-AraCR) were generated and characterized as previously described. 21,22 Neoplastic B cells were isolated from pretreatment biopsy tissue obtained from patients with B-cell non-Hodgkin lymphoma or lymphocyte-predominant Hodgkin lymphoma receiving therapy at Roswell Park Cancer Institute (RPCI) as previously described. 23 …”

Single-agent cytarabine is insufficient for the treatment of human mantle cell lymphoma in mouse xenograft model

“…Mouse xenograft models of human aggressive MCLs were used to compare head-to-head anti-lymphoma efficacies of equally toxic doses of single-agent araC compared to singleagent Pt, and three different combinations of araC and Pt (C1-C3) [9]. Lymphoma-specific survival (defined as the time from lymphoma inoculation to lymphoma-caused demise of experimental animals) was significantly (p<0.05) prolonged in cohorts treated with single-agent Pt, and C1 combination compared to the other cohorts ( Figure 1A, B).…”

“…Jeko-1 and Hbl-2 cell line-based mouse models of MCL were established in the same way as previously described [9]. Briefly, 8 -16 week old female NSG mice were inoculated with 1 x 10 6 Jeko-1 or Hbl-2 cells, and divided into 6 treatment cohorts (8-10 animals/cohort).…”