Abstract
The first therapeutic proteasome inhibitor bortezomib has clinical efficacy in mantle cell lymphoma (MCL) which resulted in its incorporation in treatment algorithms for this disease. Impairment of proteasomal function by bortezomib is mediated via inhibition of the 20S core particle. However, proteasome function can also be modified by targeting upstream components of the ubiquitin–proteasome system. Recently, b-AP15 has been identified as a small molecule achieving proteasome inhibition by targeting the deubiquitinase (DUB) activity of the 19S regulatory subunit and was found to inhibit cancer cell growth in preclinical analyses. In the present study, both direct antitumor effects and the possibility to induce natural killer group 2 member D ligands (NKG2DL) to reinforce NK cell immunity with b-AP15 were investigated to provide a rational basis for clinical evaluation of this novel DUB inhibitor in MCL. Treatment with b-AP15 resulted in reduced viability as well as induction of apoptosis in a time- and dose-dependent manner, which could be attributed to caspase activation in MCL cells. In addition, treatment with b-AP15 differentially induced NKG2DL expression and subsequent NK cell lysis of MCL cells. These results indicate that the DUB inhibitor b-AP15 displays substantial antitumor activity in human MCL and suggest that b-AP15 might be a novel therapeutic option in the treatment of MCL that warrants clinical investigation.
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Abbreviations
- ATCC:
-
American-Type Culture Collection
- DSMZ:
-
German Collection of Microorganisms and Cell Cultures
- DUB:
-
Deubiquitinase
- FasL:
-
Fas ligand
- FDA:
-
Food and Drug Administration
- MCL:
-
Mantle cell lymphoma
- MIC:
-
Major histocompatibility complex class I chain-related protein
- NKG2D:
-
Natural killer group 2 member D
- NKG2DL:
-
Natural killer group 2 member D ligands
- PARP:
-
Poly-ADP ribose polymerase
- PI:
-
Propidium iodide
- RPL13:
-
Ribosomal protein L13
- UCHL5:
-
Ubiquitin C-terminal hydrolase 5
- ULBP:
-
UL16-binding protein
- USP14:
-
Ubiquitin-specific peptidase 14
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Acknowledgements
We thank Ilona Hagelstein, Sylvia Klein, Melanie Kraft, and Elke Malenke for excellent technical assistance. Flow cytometry sample acquisition was done on shared instruments of the Flow Cytometry Core Facility Tübingen.
Funding
Susanne M. Rittig was supported by the European Social Fund in Baden-Württemberg.
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KNK and SM designed and performed the experiments and analyzed data. KR, BJS, KLC, MS, and BS performed experiments and analyzed data. KNK and DD wrote the initial draft of the manuscript. HGK, AS, and FG analyzed data. SMR and HRS revised the manuscript. DD designed and supervised the study.
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The authors declare that they have no conflict of interest.
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This study was approved by our institutional review board, the ethics committee of the Faculty of Medicine of the Eberhard Karls Universität Tübingen and of the University Hospital Tübingen (reference number 13/2007V) to be in accordance with the Declaration of Helsinki.
Informed consent
Buffy coat preparations were produced by the local blood bank after obtaining informed consent, in accordance with the principles of the Declaration of Helsinki and its later amendments. Peripheral blood samples from MCL patients were obtained after written informed consent.
Cell line authentication
MCL cells were purchased from DSMZ and ATCC for this project.
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Kropp, K.N., Maurer, S., Rothfelder, K. et al. The novel deubiquitinase inhibitor b-AP15 induces direct and NK cell-mediated antitumor effects in human mantle cell lymphoma. Cancer Immunol Immunother 67, 935–947 (2018). https://doi.org/10.1007/s00262-018-2151-y
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DOI: https://doi.org/10.1007/s00262-018-2151-y