Notch1 signaling in NOTCH1-mutated mantle cell lymphoma depends on Delta-Like ligand 4 and is a potential target for specific antibody therapy

Abstract

BackgroundNOTCH1 gene mutations in mantle cell lymphoma (MCL) have been described in about 5–10% of cases and are associated with significantly shorter survival rates. The present study aimed to investigate the biological impact of this mutation in MCL and its potential as a therapeutic target.MethodsActivation of Notch1 signaling upon ligand-stimulation and inhibitory effects of the monoclonal anti-Notch1 antibody OMP-52M51 in NOTCH1-mutated and -unmutated MCL cells were assessed by Western Blot and gene expression profiling. Effects of OMP-52M51 treatment on tumor cell migration and tumor angiogenesis were evaluated with chemotaxis and HUVEC tube formation assays. The expression of Delta-like ligand 4 (DLL4) in MCL lymph nodes was analyzed by immunofluorescence staining and confocal microscopy. A MCL mouse model was used to assess the activity of OMP-52M51 in vivo.ResultsNotch1 expression can be effectively stimulated in NOTCH1-mutated Mino cells by DLL4, whereas in the NOTCH1-unmutated cell line JeKo-1, less effect was observed upon any ligand-stimulation. DLL4 was expressed by histiocytes in both, NOTCH1-mutated and –unmutated MCL lymph nodes. Treatment of NOTCH1-mutated MCL cells with the monoclonal anti-Notch1 antibody OMP-52M51 effectively prevented DLL4-dependent activation of Notch1 and suppressed the induction of numerous direct Notch target genes involved in lymphoid biology, lymphomagenesis and disease progression. Importantly, in lymph nodes from primary MCL cases with NOTCH1/2 mutations, we detected an upregulation of the same gene sets as observed in DLL4-stimulated Mino cells. Furthermore, DLL4 stimulation of NOTCH1-mutated Mino cells enhanced tumor cell migration and angiogenesis, which could be abolished by treatment with OMP-52M51. Importantly, the effects observed were specific for NOTCH1-mutated cells as they did not occur in the NOTCH1-wt cell line JeKo-1. Finally, we confirmed the potential activity of OMP-52M51 to inhibit DLL4-induced Notch1-Signaling in vivo in a xenograft mouse model of MCL.ConclusionDLL4 effectively stimulates Notch1 signaling in NOTCH1-mutated MCL and is expressed by the microenvironment in MCL lymph nodes. Our results indicate that specific inhibition of the Notch1-ligand-receptor interaction might provide a therapeutic alternative for a subset of MCL patients.

Time-Resolved scRNA-Seq Tracks the Adaptation of a Sensitive MCL Cell Line to Ibrutinib Treatment

“…Here, a subpopulation of cells showed activity of a HES4 regulated gene network indicating active NOTCH1 signaling [ 34 ]. Unique features such as CXCL10 , SPP1 , and LGALS1 suggested a subtype with aggressive features, higher potential of metastasis, promotion of angiogenesis, and immune escape [ 35 , 36 , 37 , 38 , 39 , 40 ]. These characteristics might determine a specialized MCL population, conserved in the REC-1 cell line, interacting with the microenvironment to build a protective niche.…”

Top Notch Targeting Strategies in Cancer: A Detailed Overview of Recent Insights and Current Perspectives

“…Brontictuzumab (OMP-52M51) is a humanised IgG2 antibody generated by immunising mice with a fragment of the LNR and NRR domains of the Notch 1 receptor. It has shown efficacy in T-ALL and MCL in vitro and in vivo models, and in leukaemia PDX models harbouring two of the common Notch 1 activating mutations, i.e., mutations in the HD and PEST domains [ 304 , 305 ]. A Phase I dose-escalation clinical trial by OncoMed of brontictuzumab in patients with haematological malignancies and known Notch 1 mutation status was completed in 2016, reporting that the therapy was generally well-tolerated but had limited antitumour activity [ 192 ].…”

Targeting Notch to Maximize Chemotherapeutic Benefits: Rationale, Advanced Strategies, and Future Perspectives

“…Consequently, this mAb was evaluated in several CTs in combination with chemotherapeutic drugs, which did not bring any relevant benefit in patients with advanced SCLC or metastatic pancreatic cancer (NCT01859741) [208]. Brontictuzumab (OMP-52M51), a mAb directed against NRR of Notch, was effective in Notch1-mutated T-ALL, CLL, mantle cell lymphoma, and ACC cell and murine models [209][210][211][212]. This agent has been studied in several phase I CTs of relapsed or refractory lymphoid malignancies, solid tumors, and previously treated metastatic colon cancers (NCT01778439, NCT01703572, NCT03031691), with some efficacy signals in patients with ACC associated with Notch1-activating mutations [212,213].…”