Systems biology-enabled targeting of NF-κΒ and BCL2 overcomes microenvironment-mediated BH3-mimetic resistance in DLBCL

Abstract In Diffuse Large B-cell Lymphoma (DLBCL), elevated anti-apoptotic BCL2-family proteins (e.g., MCL1, BCL2, BCLXL) and NF-κB subunits (RelA, RelB, cRel) confer poor prognosis. Heterogeneous expression, regulatory complexity, and redundancy offsetting the inhibition of individual proteins, com...

Full description

Saved in:
Bibliographic Details
Main Authors: Aimilia Vareli, Haripriya Vaidehi Narayanan, Heather Clark, Eleanor Jayawant, Hui Zhou, Yi Liu, Emma Kennedy, Lauren Stott, Fabio Simoes, Alexander Hoffmann, Andrea Pepper, Chris Pepper, Simon Mitchell
Format: Article
Language:English
Published: Nature Publishing Group 2025-08-01
Series:Cell Death and Disease
Online Access:https://doi.org/10.1038/s41419-025-07942-0
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849340957136257024
author Aimilia Vareli
Haripriya Vaidehi Narayanan
Heather Clark
Eleanor Jayawant
Hui Zhou
Yi Liu
Emma Kennedy
Lauren Stott
Fabio Simoes
Alexander Hoffmann
Andrea Pepper
Chris Pepper
Simon Mitchell
author_facet Aimilia Vareli
Haripriya Vaidehi Narayanan
Heather Clark
Eleanor Jayawant
Hui Zhou
Yi Liu
Emma Kennedy
Lauren Stott
Fabio Simoes
Alexander Hoffmann
Andrea Pepper
Chris Pepper
Simon Mitchell
author_sort Aimilia Vareli
collection DOAJ
description Abstract In Diffuse Large B-cell Lymphoma (DLBCL), elevated anti-apoptotic BCL2-family proteins (e.g., MCL1, BCL2, BCLXL) and NF-κB subunits (RelA, RelB, cRel) confer poor prognosis. Heterogeneous expression, regulatory complexity, and redundancy offsetting the inhibition of individual proteins, complicate the assignment of targeted therapy. We combined flow cytometry ‘fingerprinting’, immunofluorescence imaging, and computational modeling to identify therapeutic vulnerabilities in DLBCL. The combined workflow predicted selective responses to BCL2 inhibition (venetoclax) and non-canonical NF-κB inhibition (Amgen16). Within the U2932 cell line we identified distinct resistance mechanisms to BCL2 inhibition in cellular sub-populations recapitulating intratumoral heterogeneity. Co-cultures with CD40L-expressing stromal cells, mimicking the tumor microenvironment (TME), induced resistance to BCL2 and BCLXL targeting BH3-mimetics via cell-type specific upregulation of BCLXL or MCL1. Computational models, validated experimentally, showed that basal NF-κB activation determined whether CD40 activation drove BH3-mimetic resistance through upregulation of RelB and BCLXL, or cRel and MCL1. High basal NF-κB activity could be overcome by inhibiting BTK to resensitize cells to BH3-mimetics in CD40L co-culture. Importantly, non-canonical NF-κB inhibition overcame heterogeneous compensatory BCL2 upregulation, restoring sensitivity to both BCL2- and BCLXL-targeting BH3-mimetics. Combined molecular fingerprinting and computational modelling provides a strategy for the precision use of BH3-mimetics and NF-κB inhibitors in DLBCL.
format Article
id doaj-art-60eb283a08ab4d8dba2fe6da69f2a5c5
institution Kabale University
issn 2041-4889
language English
publishDate 2025-08-01
publisher Nature Publishing Group
record_format Article
series Cell Death and Disease
spelling doaj-art-60eb283a08ab4d8dba2fe6da69f2a5c52025-08-20T03:43:45ZengNature Publishing GroupCell Death and Disease2041-48892025-08-0116111210.1038/s41419-025-07942-0Systems biology-enabled targeting of NF-κΒ and BCL2 overcomes microenvironment-mediated BH3-mimetic resistance in DLBCLAimilia Vareli0Haripriya Vaidehi Narayanan1Heather Clark2Eleanor Jayawant3Hui Zhou4Yi Liu5Emma Kennedy6Lauren Stott7Fabio Simoes8Alexander Hoffmann9Andrea Pepper10Chris Pepper11Simon Mitchell12Department of Clinical and Experimental Medicine, Brighton and Sussex Medical School, University of Brighton and University of SussexSignaling Systems Laboratory, Department of Microbiology, Immunology, and Molecular Genetics, and Institute for Quantitative and Computational Biosciences, University of California, Los AngelesDepartment of Clinical and Experimental Medicine, Brighton and Sussex Medical School, University of Brighton and University of SussexDepartment of Clinical and Experimental Medicine, Brighton and Sussex Medical School, University of Brighton and University of SussexDeepKinase BiotechnologiesDeepKinase BiotechnologiesDepartment of Clinical and Experimental Medicine, Brighton and Sussex Medical School, University of Brighton and University of SussexDepartment of Clinical and Experimental Medicine, Brighton and Sussex Medical School, University of Brighton and University of SussexDepartment of Clinical and Experimental Medicine, Brighton and Sussex Medical School, University of Brighton and University of SussexSignaling Systems Laboratory, Department of Microbiology, Immunology, and Molecular Genetics, and Institute for Quantitative and Computational Biosciences, University of California, Los AngelesDepartment of Clinical and Experimental Medicine, Brighton and Sussex Medical School, University of Brighton and University of SussexDepartment of Clinical and Experimental Medicine, Brighton and Sussex Medical School, University of Brighton and University of SussexDepartment of Clinical and Experimental Medicine, Brighton and Sussex Medical School, University of Brighton and University of SussexAbstract In Diffuse Large B-cell Lymphoma (DLBCL), elevated anti-apoptotic BCL2-family proteins (e.g., MCL1, BCL2, BCLXL) and NF-κB subunits (RelA, RelB, cRel) confer poor prognosis. Heterogeneous expression, regulatory complexity, and redundancy offsetting the inhibition of individual proteins, complicate the assignment of targeted therapy. We combined flow cytometry ‘fingerprinting’, immunofluorescence imaging, and computational modeling to identify therapeutic vulnerabilities in DLBCL. The combined workflow predicted selective responses to BCL2 inhibition (venetoclax) and non-canonical NF-κB inhibition (Amgen16). Within the U2932 cell line we identified distinct resistance mechanisms to BCL2 inhibition in cellular sub-populations recapitulating intratumoral heterogeneity. Co-cultures with CD40L-expressing stromal cells, mimicking the tumor microenvironment (TME), induced resistance to BCL2 and BCLXL targeting BH3-mimetics via cell-type specific upregulation of BCLXL or MCL1. Computational models, validated experimentally, showed that basal NF-κB activation determined whether CD40 activation drove BH3-mimetic resistance through upregulation of RelB and BCLXL, or cRel and MCL1. High basal NF-κB activity could be overcome by inhibiting BTK to resensitize cells to BH3-mimetics in CD40L co-culture. Importantly, non-canonical NF-κB inhibition overcame heterogeneous compensatory BCL2 upregulation, restoring sensitivity to both BCL2- and BCLXL-targeting BH3-mimetics. Combined molecular fingerprinting and computational modelling provides a strategy for the precision use of BH3-mimetics and NF-κB inhibitors in DLBCL.https://doi.org/10.1038/s41419-025-07942-0
spellingShingle Aimilia Vareli
Haripriya Vaidehi Narayanan
Heather Clark
Eleanor Jayawant
Hui Zhou
Yi Liu
Emma Kennedy
Lauren Stott
Fabio Simoes
Alexander Hoffmann
Andrea Pepper
Chris Pepper
Simon Mitchell
Systems biology-enabled targeting of NF-κΒ and BCL2 overcomes microenvironment-mediated BH3-mimetic resistance in DLBCL
Cell Death and Disease
title Systems biology-enabled targeting of NF-κΒ and BCL2 overcomes microenvironment-mediated BH3-mimetic resistance in DLBCL
title_full Systems biology-enabled targeting of NF-κΒ and BCL2 overcomes microenvironment-mediated BH3-mimetic resistance in DLBCL
title_fullStr Systems biology-enabled targeting of NF-κΒ and BCL2 overcomes microenvironment-mediated BH3-mimetic resistance in DLBCL
title_full_unstemmed Systems biology-enabled targeting of NF-κΒ and BCL2 overcomes microenvironment-mediated BH3-mimetic resistance in DLBCL
title_short Systems biology-enabled targeting of NF-κΒ and BCL2 overcomes microenvironment-mediated BH3-mimetic resistance in DLBCL
title_sort systems biology enabled targeting of nf κβ and bcl2 overcomes microenvironment mediated bh3 mimetic resistance in dlbcl
url https://doi.org/10.1038/s41419-025-07942-0
work_keys_str_mv AT aimiliavareli systemsbiologyenabledtargetingofnfkbandbcl2overcomesmicroenvironmentmediatedbh3mimeticresistanceindlbcl
AT haripriyavaidehinarayanan systemsbiologyenabledtargetingofnfkbandbcl2overcomesmicroenvironmentmediatedbh3mimeticresistanceindlbcl
AT heatherclark systemsbiologyenabledtargetingofnfkbandbcl2overcomesmicroenvironmentmediatedbh3mimeticresistanceindlbcl
AT eleanorjayawant systemsbiologyenabledtargetingofnfkbandbcl2overcomesmicroenvironmentmediatedbh3mimeticresistanceindlbcl
AT huizhou systemsbiologyenabledtargetingofnfkbandbcl2overcomesmicroenvironmentmediatedbh3mimeticresistanceindlbcl
AT yiliu systemsbiologyenabledtargetingofnfkbandbcl2overcomesmicroenvironmentmediatedbh3mimeticresistanceindlbcl
AT emmakennedy systemsbiologyenabledtargetingofnfkbandbcl2overcomesmicroenvironmentmediatedbh3mimeticresistanceindlbcl
AT laurenstott systemsbiologyenabledtargetingofnfkbandbcl2overcomesmicroenvironmentmediatedbh3mimeticresistanceindlbcl
AT fabiosimoes systemsbiologyenabledtargetingofnfkbandbcl2overcomesmicroenvironmentmediatedbh3mimeticresistanceindlbcl
AT alexanderhoffmann systemsbiologyenabledtargetingofnfkbandbcl2overcomesmicroenvironmentmediatedbh3mimeticresistanceindlbcl
AT andreapepper systemsbiologyenabledtargetingofnfkbandbcl2overcomesmicroenvironmentmediatedbh3mimeticresistanceindlbcl
AT chrispepper systemsbiologyenabledtargetingofnfkbandbcl2overcomesmicroenvironmentmediatedbh3mimeticresistanceindlbcl
AT simonmitchell systemsbiologyenabledtargetingofnfkbandbcl2overcomesmicroenvironmentmediatedbh3mimeticresistanceindlbcl