Highly specific Immunoproteasome inhibitor M3258 induces proteotoxic stress and apoptosis in KMT2A::AFF1 driven acute lymphoblastic leukemia

Abstract Proteasome inhibitors (PIs) bortezomib, carfilzomib and ixazomib are approved for the treatment of multiple myeloma and mantle cell lymphoma and have clinical activity in acute lymphoblastic leukemia (ALL). The predominant form of proteasome in these hematologic malignancies is the lymphoid...

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Main Authors: Tyler W. Jenkins, Jacquelyn Elise Fitzgerald, Jieun Park, Addison M. Wilson, Kristy L. Berry, Keith S. Wong, Walid A. Houry, Irene Lee, Andrey V. Maksimenko, Peter R. Panizzi, Yulia Y. Maxuitenko, Matthew Shane Loop, Amit K. Mitra, Alexei F. Kisselev
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Language:English
Published: Nature Portfolio 2025-05-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-01657-0
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author Tyler W. Jenkins
Jacquelyn Elise Fitzgerald
Jieun Park
Addison M. Wilson
Kristy L. Berry
Keith S. Wong
Walid A. Houry
Irene Lee
Andrey V. Maksimenko
Peter R. Panizzi
Yulia Y. Maxuitenko
Matthew Shane Loop
Amit K. Mitra
Alexei F. Kisselev
author_facet Tyler W. Jenkins
Jacquelyn Elise Fitzgerald
Jieun Park
Addison M. Wilson
Kristy L. Berry
Keith S. Wong
Walid A. Houry
Irene Lee
Andrey V. Maksimenko
Peter R. Panizzi
Yulia Y. Maxuitenko
Matthew Shane Loop
Amit K. Mitra
Alexei F. Kisselev
author_sort Tyler W. Jenkins
collection DOAJ
description Abstract Proteasome inhibitors (PIs) bortezomib, carfilzomib and ixazomib are approved for the treatment of multiple myeloma and mantle cell lymphoma and have clinical activity in acute lymphoblastic leukemia (ALL). The predominant form of proteasome in these hematologic malignancies is the lymphoid tissue-specific immunoproteasome. FDA-approved PIs inhibit immunoproteasomes and ubiquitously expressed constitutive proteasomes causing on-target toxicities in non-hematological tissues. Replacing PIs with selective immunoproteasome inhibitors (IPIs) should reduce these toxicities. We have previously shown that IPI ONX-0914 causes apoptosis of ALL cells expressing the KMT2A::AFF1 (MLL-AF4) fusion protein but did not elucidate the mechanism. Here we show that a novel, highly specific IPI M3258 induces rapid apoptosis in ALL cells in vitro and is comparable to bortezomib in its ability to reduce tumor growth and to cause tumor regression when combined with chemotherapy in vivo. Treatment of KMT2A::AFF1 ALL cells with M3258, ONX-0914, and bortezomib induced proteotoxic stress that was prevented by the protein synthesis inhibitor cycloheximide, which dramatically desensitized cells to PI-induced apoptosis. Thus, similar to multiple myeloma, ALL cells are sensitive to PIs and IPIs due to increased proteotoxic stress caused by elevated rates of protein synthesis.
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spelling doaj-art-434d3dcaf22442c09686a9738fa7b98f2025-08-20T03:08:25ZengNature PortfolioScientific Reports2045-23222025-05-0115111410.1038/s41598-025-01657-0Highly specific Immunoproteasome inhibitor M3258 induces proteotoxic stress and apoptosis in KMT2A::AFF1 driven acute lymphoblastic leukemiaTyler W. Jenkins0Jacquelyn Elise Fitzgerald1Jieun Park2Addison M. Wilson3Kristy L. Berry4Keith S. Wong5Walid A. Houry6Irene Lee7Andrey V. Maksimenko8Peter R. Panizzi9Yulia Y. Maxuitenko10Matthew Shane Loop11Amit K. Mitra12Alexei F. Kisselev13Department of Drug Discovery and Development, Harrison College of Pharmacy, Auburn UniversityDepartment of Drug Discovery and Development, Harrison College of Pharmacy, Auburn UniversityDivision of Research, Harrison College of Pharmacy, Auburn UniversityDepartment of Drug Discovery and Development, Harrison College of Pharmacy, Auburn UniversityDepartment of Drug Discovery and Development, Harrison College of Pharmacy, Auburn UniversityDepartment of Biochemistry, University of TorontoDepartment of Biochemistry, University of TorontoDepartment of Chemistry, Case Western Reserve UniversityDepartment of Drug Discovery and Development, Harrison College of Pharmacy, Auburn UniversityDepartment of Drug Discovery and Development, Harrison College of Pharmacy, Auburn UniversityDepartment of Drug Discovery and Development, Harrison College of Pharmacy, Auburn UniversityDepartment of Health Outcomes and Research Policy, Harrison College of Pharmacy, Auburn UniversityDepartment of Drug Discovery and Development, Harrison College of Pharmacy, Auburn UniversityDepartment of Drug Discovery and Development, Harrison College of Pharmacy, Auburn UniversityAbstract Proteasome inhibitors (PIs) bortezomib, carfilzomib and ixazomib are approved for the treatment of multiple myeloma and mantle cell lymphoma and have clinical activity in acute lymphoblastic leukemia (ALL). The predominant form of proteasome in these hematologic malignancies is the lymphoid tissue-specific immunoproteasome. FDA-approved PIs inhibit immunoproteasomes and ubiquitously expressed constitutive proteasomes causing on-target toxicities in non-hematological tissues. Replacing PIs with selective immunoproteasome inhibitors (IPIs) should reduce these toxicities. We have previously shown that IPI ONX-0914 causes apoptosis of ALL cells expressing the KMT2A::AFF1 (MLL-AF4) fusion protein but did not elucidate the mechanism. Here we show that a novel, highly specific IPI M3258 induces rapid apoptosis in ALL cells in vitro and is comparable to bortezomib in its ability to reduce tumor growth and to cause tumor regression when combined with chemotherapy in vivo. Treatment of KMT2A::AFF1 ALL cells with M3258, ONX-0914, and bortezomib induced proteotoxic stress that was prevented by the protein synthesis inhibitor cycloheximide, which dramatically desensitized cells to PI-induced apoptosis. Thus, similar to multiple myeloma, ALL cells are sensitive to PIs and IPIs due to increased proteotoxic stress caused by elevated rates of protein synthesis.https://doi.org/10.1038/s41598-025-01657-0ProteasomeProteasome inhibitorUbiquitinProteostasisHeat shock responseUnfolded protein response
spellingShingle Tyler W. Jenkins
Jacquelyn Elise Fitzgerald
Jieun Park
Addison M. Wilson
Kristy L. Berry
Keith S. Wong
Walid A. Houry
Irene Lee
Andrey V. Maksimenko
Peter R. Panizzi
Yulia Y. Maxuitenko
Matthew Shane Loop
Amit K. Mitra
Alexei F. Kisselev
Highly specific Immunoproteasome inhibitor M3258 induces proteotoxic stress and apoptosis in KMT2A::AFF1 driven acute lymphoblastic leukemia
Scientific Reports
Proteasome
Proteasome inhibitor
Ubiquitin
Proteostasis
Heat shock response
Unfolded protein response
title Highly specific Immunoproteasome inhibitor M3258 induces proteotoxic stress and apoptosis in KMT2A::AFF1 driven acute lymphoblastic leukemia
title_full Highly specific Immunoproteasome inhibitor M3258 induces proteotoxic stress and apoptosis in KMT2A::AFF1 driven acute lymphoblastic leukemia
title_fullStr Highly specific Immunoproteasome inhibitor M3258 induces proteotoxic stress and apoptosis in KMT2A::AFF1 driven acute lymphoblastic leukemia
title_full_unstemmed Highly specific Immunoproteasome inhibitor M3258 induces proteotoxic stress and apoptosis in KMT2A::AFF1 driven acute lymphoblastic leukemia
title_short Highly specific Immunoproteasome inhibitor M3258 induces proteotoxic stress and apoptosis in KMT2A::AFF1 driven acute lymphoblastic leukemia
title_sort highly specific immunoproteasome inhibitor m3258 induces proteotoxic stress and apoptosis in kmt2a aff1 driven acute lymphoblastic leukemia
topic Proteasome
Proteasome inhibitor
Ubiquitin
Proteostasis
Heat shock response
Unfolded protein response
url https://doi.org/10.1038/s41598-025-01657-0
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