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Plasma Membrane Channel TRPM4 Mediates Immunogenic Therapy-Induced Necrosis
Santanu Ghosh1, Rachel Yang1, Darjan Duraki1
1Departments of Biochemistry, Molecular and Integrative Physiology and Chemistry, University of Illinois at Urbana-Champaign, Urbana, Illinois.
Abstract:
Several emerging therapies kill cancer cells primarily by inducing necrosis. As necrosis activates immune cells, potentially, uncovering the molecular drivers of anticancer therapy-induced necrosis could reveal approaches for enhancing immunotherapy efficacy. To identify necrosis-associated genes, we performed a genome-wide CRISPR-Cas9 screen with negative selection against necrosis-inducing preclinical agents BHPI and conducted follow-on experiments with ErSO. The screen identified transient receptor potential melastatin member 4 (TRPM4), a calcium-activated, ATP-inhibited, sodium-selective plasma membrane channel. Cancer cells selected for resistance to BHPI and ErSO exhibited robust TRPM4 downregulation, and TRPM4 reexpression restored sensitivity to ErSO. Notably, TRPM4 knockout (TKO) abolished ErSO-induced regression of breast tumors in mice. Supporting a broad role for TRPM4 in necrosis, knockout of TRPM4 reversed cell death induced by four additional diverse necrosis-inducing cancer therapies. ErSO induced anticipatory unfolded protein response (a-UPR) hyperactivation, long-term necrotic cell death, and release of damage-associated molecular patterns that activated macrophages and increased monocyte migration, all of which was abolished by TKO. Furthermore, loss of TRPM4 suppressed the ErSO-induced increase in cell volume and depletion of ATP. These data suggest that ErSO triggers initial activation of the a-UPR but that it is TRPM4-mediated sodium influx and cell swelling, resulting in osmotic stress, which sustains and propagates lethal a-UPR hyperactivation. Thus, TRPM4 plays a pivotal role in sustaining lethal a-UPR hyperactivation that mediates the anticancer activity of diverse necrosis-inducing therapies.
Significance:
A genome-wide CRISPR screen reveals a pivotal role for TRPM4 in cell death and immune activation following treatment with diverse necrosis-inducing anticancer therapies, which could facilitate development of necrosis-based cancer immunotherapies.
Insights
Researchers identified Transient Receptor Potential Melastatin 4 (TRPM4) as crucial for necrosis-induced cancer cell death. Targeting TRPM4 may enhance cancer immunotherapies by promoting immune cell activation against tumors.
Area of Science:
- Oncology
- Molecular Biology
- Immunology
Background:
- Emerging cancer therapies induce cell death via necrosis, activating immune cells.
- Understanding necrosis drivers could enhance immunotherapy effectiveness.
Purpose of the Study:
- Identify genes associated with anticancer therapy-induced necrosis.
- Investigate the role of identified genes in cancer cell death and immune response.
Main Methods:
- Genome-wide CRISPR-Cas9 screen using necrosis-inducing agents (BHPI, ErSO).
- Functional validation of identified genes (TRPM4) through knockout and reexpression studies.
- In vivo studies using mouse tumor models.
Main Results:
- CRISPR screen identified Transient Receptor Potential Melastatin 4 (TRPM4) as critical for necrosis.
- TRPM4 knockout abolished cancer cell death and tumor regression induced by multiple necrosis-inducing agents.
- TRPM4 mediates sodium influx, cell swelling, and sustained unfolded protein response (a-UPR) hyperactivation, crucial for therapy efficacy.
- TRPM4 loss suppressed release of damage-associated molecular patterns, impairing immune cell activation.
Conclusions:
- TRPM4 is a pivotal mediator of necrosis-inducing anticancer therapy efficacy.
- TRPM4 plays a key role in sustaining lethal a-UPR hyperactivation and immune cell activation.
- Targeting TRPM4 presents a potential strategy for enhancing necrosis-based cancer immunotherapies.
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