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Updated: Dec 23, 2025
Modeling Chemotherapy Resistant Leukemia In Vitro
Published on: February 9, 2016
A20 and RBX1 Regulate Brentuximab Vedotin Sensitivity in Hodgkin Lymphoma Models
Wei Wei1, Yuquan Lin2, Zhihui Song1
1Blood Cell Development and Function Program, Fox Chase Cancer Center, Philadelphia, Pennsylvania.
Purpose:
For patients with refractory/relapsed Hodgkin lymphoma (roughly 20% of total cases), few effective therapeutic options exist. Currently, brentuximab vedotin (BV), a drug-conjugated anti-CD30 antibody, is one of the most effective approved therapy agents for these patients. However, many patients do not achieve complete remission and ultimately develop BV-resistant disease, necessitating a more detailed understanding of the molecular circuitry that drives BV sensitivity and the mechanism of BV resistance.
Experimental Design:
Here, we established a ubiquitin regulator-focused CRISPR library screening platform in Hodgkin lymphoma and carried out a drug sensitization screen against BV to identify genes regulating BV treatment sensitivity.
Results:
Our CRISPR library screens revealed the ubiquitin-editing enzymes A20 and RBX1 as key molecule effectors that regulate BV sensitivity in Hodgkin lymphoma line L428. A20 negatively regulates NF-κB activity which is required to prevent BV cytotoxicity. In line with these results, the RNA-seq analysis of the BV-resistant single-cell clones demonstrated a consistent upregulation of NF-κB signature genes, as well as the ABC transporter gene ABCB1. Mechanically, NF-κB regulates BV treatment sensitivity through mediating ABCB1 expression. Targeting NF-κB activity synergized well with BV in killing Hodgkin lymphoma cell lines, augmented BV sensitivity, and overcame BV resistance in vitro and in Hodgkin lymphoma xenograft mouse models.
Conclusions:
Our identification of this previously unrecognized mechanism provides novel knowledge of possible BV responsiveness and resistance mechanisms in Hodgkin lymphoma, as well as leads to promising hypotheses for the development of therapeutic strategies to overcome BV resistance in this disease.
Insights
Brentuximab vedotin (BV) resistance in Hodgkin lymphoma involves the ubiquitin system and NF-κB signaling. Targeting NF-κB enhances BV efficacy and overcomes resistance, offering new therapeutic strategies.
Area of Science:
- Oncology
- Molecular Biology
- Immunology
Background:
- Refractory/relapsed Hodgkin lymphoma has limited treatment options.
- Brentuximab vedotin (BV) is an effective therapy but resistance develops.
- Understanding BV resistance mechanisms is crucial for improving patient outcomes.
Purpose of the Study:
- To identify genes regulating BV sensitivity using a CRISPR library screen.
- To elucidate the molecular mechanisms underlying BV resistance in Hodgkin lymphoma.
Main Methods:
- Established a ubiquitin regulator-focused CRISPR library screening platform.
- Performed drug sensitization screens against BV in Hodgkin lymphoma cell lines.
- Utilized RNA-sequencing to analyze BV-resistant clones.
- Validated findings in vitro and in vivo using xenograft mouse models.
Main Results:
- CRISPR screens identified A20 and RBX1 as key regulators of BV sensitivity.
- A20 negatively regulates NF-κB activity, which is essential for BV cytotoxicity.
- BV-resistant clones showed upregulation of NF-κB and ABCB1.
- NF-κB mediates BV sensitivity via ABCB1 expression.
- Targeting NF-κB synergized with BV, enhancing sensitivity and overcoming resistance.
Conclusions:
- Identified a novel mechanism of BV responsiveness and resistance in Hodgkin lymphoma.
- Revealed the role of the ubiquitin system and NF-κB signaling in BV resistance.
- Provides a basis for developing new therapeutic strategies to overcome BV resistance.
