GO:0160020 positive regulation of ferroptosis: Signaling Pathways, Genes, Functions and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0160020 (positive regulation of ferroptosis) describes any process that activates or increases the frequency, rate or extent of ferroptosis, an iron-dependent form of regulated cell death.
• Positive regulation of ferroptosis is driven by excessive lipid peroxidation, often through ACSL4-mediated incorporation of polyunsaturated fatty acids into membrane phospholipids.
• GPX4 is the central negative regulator of ferroptosis; its inactivation or degradation removes the brake on ferroptosis and constitutes positive regulation.
• Multiple signaling pathways, including CD36-mediated fatty acid uptake, PD-1 signaling, and redox-sensitive SUMOylation, can positively regulate ferroptosis in immune and neuronal contexts.
• Dysregulated positive regulation of ferroptosis contributes to cancer, sepsis-associated lung injury, and neurodegeneration, making it a therapeutic target.
• CRISPR knockout, point-mutation, knock-in, and overexpression models are essential to dissect causal roles of specific genes in positive regulation of ferroptosis.
Description
Ferroptosis is an iron-dependent form of regulated cell death characterized by the accumulation of lipid peroxides to lethal levels. The Gene Ontology term GO:0160020, positive regulation of ferroptosis, captures any process that activates or increases the frequency, rate or extent of this cell death modality. This term is critical for researchers because ferroptosis is implicated in a wide range of physiological and pathological contexts, from tumor immunity to neurodegeneration. Understanding the positive regulators of ferroptosis provides mechanistic insight into how cells commit to this death pathway and offers potential targets for therapeutic intervention. The positive regulation of ferroptosis is not a single linear pathway but a network of metabolic, redox, and signaling events. Key nodes include the availability of polyunsaturated fatty acids (PUFAs), the activity of the lipid repair enzyme GPX4, and the abundance of labile iron. For example, CD36-mediated fatty acid uptake can sensitize cells to ferroptosis, and this process dampens intratumoral CD8+ T cell effector function. Conversely, GPX4 palmitoylation by ZDHHC8 regulates its stability and thus controls ferroptosis sensitivity. These findings highlight the importance of precise regulation. Given the growing interest in ferroptosis as a therapeutic strategy, especially in cancer and inflammatory diseases, a clear understanding of GO:0160020 is essential. This article synthesizes current knowledge on the mechanisms, key genes, disease relevance, and research methods used to study positive regulation of ferroptosis, with a focus on CRISPR-based models that enable causal interrogation of candidate regulators.
positive regulation of ferroptosis At A Glance
| GO ID | GO:0160020 |
|---|---|
| GO term | positive regulation of ferroptosis |
| Ontology | biological_process |
| Synonym | None |
| Major function | Activates or increases the frequency, rate or extent of ferroptosis, an iron-dependent form of regulated cell death driven by lipid peroxidation. |
| Related processes | Lipid peroxidation, iron homeostasis, glutathione metabolism, oxidative stress response. |
| Key positive regulators | ACSL4, CD36, METTL3, TRIM28, PD-1 signaling, ZDHHC8 (via GPX4 destabilization). |
| Key negative regulators | GPX4, FASN, USP5, OPTN. |
| Disease relevance | Cancer, sepsis-associated lung injury, neurodegeneration, immune regulation. |
What Is GO:0160020?
GO:0160020, positive regulation of ferroptosis, is defined by the Gene Ontology as any process that activates or increases the frequency, rate or extent of ferroptosis. In other words, it encompasses all molecular events and pathways that promote the execution of iron-dependent lipid peroxidation-driven cell death. This term is a biological process and does not have synonyms in the current ontology. It is distinct from negative regulation of ferroptosis, which includes mechanisms that suppress ferroptosis, such as GPX4-mediated lipid peroxide reduction.
Why Is positive regulation of ferroptosis Important in Cell Biology?
Positive regulation of ferroptosis is critically important because it determines whether cells undergo this form of cell death, which has profound implications for human health and disease. In cancer, inducing ferroptosis can overcome therapy resistance and enhance antitumor immunity, as shown by CD36-mediated ferroptosis impairing CD8+ T cell function and GPX4 palmitoylation influencing antitumor immunity. In sepsis-associated lung injury, histone lactylation-regulated METTL3 promotes ferroptosis via m6A modification of ACSL4, highlighting a role in inflammatory tissue damage. In neurodegeneration, redox regulation of TRIM28 facilitates neuronal ferroptosis by promoting SUMOylation and inhibiting OPTN-selective autophagic degradation of ACSL4. Thus, understanding positive regulation of ferroptosis offers opportunities for therapeutic intervention across diverse pathologies.
• Cancer therapy: Inducing ferroptosis in tumor cells can bypass apoptosis resistance and enhance immunotherapy efficacy.
• Immune regulation: Ferroptosis in CD8+ T cells dampens antitumor immunity, and PD-1 signaling promotes T cell ferroptosis.
• Inflammatory diseases: Sepsis-associated lung injury involves METTL3-mediated ferroptosis via ACSL4 m6A modification.
• Neurodegeneration: TRIM28-driven neuronal ferroptosis contributes to neuronal loss in degenerative conditions.
• Metabolic homeostasis: Ferroptosis is intertwined with lipid and redox metabolism, influencing cellular stress responses.
• Therapeutic targeting: Key regulators such as GPX4, ACSL4, and CD36 are potential drug targets.
• CRISPR screening: Genome-wide screens can identify novel positive regulators of ferroptosis, accelerating target discovery.
• Biomarker development: Expression levels of ferroptosis regulators may predict disease outcomes or treatment responses.
• Fundamental cell biology: Understanding how cells commit to ferroptosis sheds light on regulated cell death networks.
• Translational research: Modulating ferroptosis sensitivity could improve outcomes in cancer, sepsis, and neurodegeneration.
What Happens During positive regulation of ferroptosis?
Initiation by lipid peroxidation
In simple terms: The process starts when fats in cell membranes are attacked by reactive oxygen species, creating lipid peroxides.
Positive regulation of ferroptosis is initiated by the accumulation of lipid peroxides in cellular membranes. This requires the availability of polyunsaturated fatty acids (PUFAs), which are incorporated into phospholipids by enzymes such as ACSL4. ACSL4-mediated remodeling of membrane lipids sensitizes cells to ferroptosis, and its expression or activity is often increased in contexts of positive regulation. For instance, in sepsis-associated lung injury, METTL3 promotes ferroptosis via m6A modification on ACSL4, increasing its stability and lipid peroxidation. Similarly, redox regulation of TRIM28 facilitates neuronal ferroptosis by promoting SUMOylation and inhibiting OPTN-selective autophagic degradation of ACSL4, leading to ACSL4 accumulation.
Loss of GPX4-mediated defense
In simple terms: The cell's main defense against lipid peroxides, an enzyme called GPX4, is weakened or removed, allowing peroxides to build up.
GPX4 is a glutathione peroxidase that reduces lipid peroxides to alcohols, thereby preventing ferroptosis. Positive regulation of ferroptosis often involves inhibition or degradation of GPX4. For example, palmitoylation of GPX4 via ZDHHC8 determines ferroptosis sensitivity; targeting ZDHHC8 can destabilize GPX4 and promote ferroptosis. In breast cancer, FASN inhibits ferroptosis via USP5 palmitoylation-dependent regulation of GPX4 deubiquitination, meaning that when FASN is low, GPX4 is degraded and ferroptosis is promoted. Thus, any process that reduces GPX4 activity or levels positively regulates ferroptosis.
Iron-dependent oxidative stress
In simple terms: Iron helps create highly reactive molecules that damage lipids, and more iron means more damage.
Iron is essential for ferroptosis because it catalyzes the Fenton reaction, generating hydroxyl radicals that attack membrane lipids. Positive regulation of ferroptosis can occur through increased labile iron pools, either by enhanced iron uptake or reduced iron storage. Signaling pathways that increase iron availability, such as those involving CD36-mediated fatty acid uptake, can indirectly promote ferroptosis by supplying both iron and PUFAs. The redox regulation of TRIM28 also ties into iron-dependent oxidative stress, as TRIM28 promotes SUMOylation and inhibits OPTN-selective autophagic degradation of ACSL4, enhancing lipid peroxidation in neurons.
Signaling pathways that amplify ferroptosis
In simple terms: Various cellular signals act like accelerators, pushing the cell further toward ferroptosis.
Multiple signaling pathways positively regulate ferroptosis. CD36-mediated fatty acid uptake promotes ferroptosis in intratumoral CD8+ T cells, dampening their effector function and antitumor ability. PD-1 signaling limits expression of phospholipid phosphatase 1 (PLPP1) and promotes intratumoral CD8+ T cell ferroptosis, linking immune checkpoint pathways to ferroptosis induction. Additionally, histone lactylation-regulated METTL3 promotes ferroptosis via m6A modification on ACSL4 in sepsis-associated lung injury, demonstrating epigenetic control. These pathways converge on lipid peroxidation and GPX4 inactivation to execute ferroptosis.
Execution of ferroptotic cell death
In simple terms: Once lipid peroxides reach a critical level, the cell membrane breaks down and the cell dies.
The final step of positive regulation of ferroptosis is the execution phase, where overwhelming lipid peroxidation leads to membrane rupture and cell death. This phase is characterized by loss of plasma membrane integrity, release of damage-associated molecular patterns, and cellular collapse. The process is regulated by the balance between pro-ferroptotic factors like ACSL4 and anti-ferroptotic factors like GPX4. Positive regulation tips this balance toward ferroptosis, often through post-translational modifications such as palmitoylation, ubiquitination, and SUMOylation that alter the stability or activity of key regulators.
Key Genes Involved in GO:0160020 positive regulation of ferroptosis
The following genes and proteins are central to the positive regulation of ferroptosis, as supported by recent literature.
| Gene | Major Role | Research Relevance |
|---|---|---|
| GPX4 | Negative regulator; reduces lipid peroxides | Target for inducing ferroptosis; palmitoylation by ZDHHC8 regulates stability |
| ACSL4 | Promotes lipid peroxidation by incorporating PUFAs into phospholipids | Key positive regulator; regulated by METTL3 and TRIM28 |
| CD36 | Mediates fatty acid uptake, promoting ferroptosis | Dampens CD8+ T cell antitumor function |
| ZDHHC8 | Palmitoyltransferase that modifies GPX4 | Targetable regulator of ferroptosis sensitivity |
| FASN | Inhibits ferroptosis via USP5-dependent GPX4 deubiquitination | Breast cancer context; links lipid synthesis to ferroptosis |
| USP5 | Deubiquitinase stabilizing GPX4 | Modulated by FASN palmitoylation |
| METTL3 | m6A methyltransferase promoting ACSL4 expression | Histone lactylation-regulated in sepsis lung injury |
| TRIM28 | Promotes SUMOylation and inhibits OPTN-mediated ACSL4 degradation | Neuronal ferroptosis regulator |
| OPTN | Selective autophagy receptor for ACSL4 degradation | Inhibited by TRIM28, leading to ACSL4 accumulation |
| PD-1 | Immune checkpoint receptor that limits PLPP1 and promotes T cell ferroptosis | Links immune signaling to ferroptosis |
| PLPP1 | Phospholipid phosphatase 1, negatively regulates ferroptosis | Downregulated by PD-1 signaling |
| SLC7A11 | Cystine/glutamate antiporter, supports glutathione synthesis | Indirect negative regulator; not directly cited in provided list but commonly studied |
| NFS1 | Iron-sulfur cluster biosynthesis, affects iron homeostasis | Potential regulator; not directly cited in provided list |
| FSP1 | Ferroptosis suppressor protein 1, reduces CoQ10 | Negative regulator; not directly cited in provided list |
| GCH1 | GTP cyclohydrolase 1, synthesizes BH4 | Negative regulator; not directly cited in provided list |
| HMOX1 | Heme oxygenase 1, releases labile iron | Can promote ferroptosis; not directly cited in provided list |
| TFRC | Transferrin receptor, mediates iron uptake | Positive regulator via iron loading; not directly cited in provided list |
How Is positive regulation of ferroptosis Regulated?
Positive regulation of ferroptosis is controlled at multiple levels, including transcriptional, post-transcriptional, translational, and post-translational mechanisms. Epigenetic regulation via histone lactylation can drive METTL3 expression, which in turn m6A-modifies ACSL4 mRNA to promote ferroptosis. Post-translational modifications such as palmitoylation, ubiquitination, and SUMOylation directly affect the stability and activity of key regulators like GPX4 and ACSL4. Signaling pathways, including CD36-mediated fatty acid uptake and PD-1 signaling, can amplify ferroptosis in specific cell types. Additionally, redox-sensitive proteins such as TRIM28 integrate oxidative stress signals to promote neuronal ferroptosis. These layers of regulation ensure that ferroptosis is tightly controlled and can be rapidly induced in response to cellular stress.
positive regulation of ferroptosis and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| GPX4 | Cancer, ferroptosis sensitivity | GPX4 knockout or point mutant cell lines; xenograft models |
| ACSL4 | Sepsis-associated lung injury, neurodegeneration | ACSL4 knockout mice or cells; METTL3/TRIM28 manipulation |
| CD36 | Cancer immunotherapy, T cell exhaustion | CD36 knockout T cells; adoptive transfer models |
| PD-1 | Cancer immunotherapy, T cell ferroptosis | PD-1 knockout mice; anti-PD-1 treatment models |
| TRIM28 | Neurodegeneration | TRIM28 knockout or SUMOylation-deficient mutants in neurons |
Cancer and antitumor immunity
Positive regulation of ferroptosis plays a dual role in cancer. On one hand, inducing ferroptosis in tumor cells can suppress tumor growth and overcome drug resistance. On the other hand, ferroptosis in immune cells can impair antitumor immunity. CD36-mediated ferroptosis dampens intratumoral CD8+ T cell effector function and impairs their antitumor ability. PD-1 signaling limits expression of phospholipid phosphatase 1 and promotes intratumoral CD8+ T cell ferroptosis, suggesting that checkpoint blockade may inadvertently induce T cell ferroptosis. GPX4 palmitoylation via ZDHHC8 determines ferroptosis sensitivity and antitumor immunity, highlighting a targetable axis. In breast cancer, FASN inhibits ferroptosis via USP5 palmitoylation-dependent regulation of GPX4 deubiquitination, linking lipid metabolism to ferroptosis evasion.
Sepsis-associated lung injury
In sepsis-associated lung injury, histone lactylation-regulated METTL3 promotes ferroptosis via m6A modification on ACSL4. This pathway exemplifies how metabolic and epigenetic changes during inflammation can positively regulate ferroptosis, leading to tissue damage. Targeting this axis may provide therapeutic benefit in sepsis-induced lung injury.
Neurodegeneration
Redox regulation of TRIM28 facilitates neuronal ferroptosis by promoting SUMOylation and inhibiting OPTN-selective autophagic degradation of ACSL4. This mechanism links oxidative stress to neuronal loss, suggesting that positive regulation of ferroptosis contributes to neurodegenerative diseases. Inhibiting this pathway could protect neurons.
From positive regulation of ferroptosis-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does loss of GPX4 increase ferroptosis? | GPX4 knockout cell lines (CRISPR) |
| Does ACSL4 m6A modification promote ferroptosis? | ACSL4 point mutant (m6A site) knock-in cells |
| Does CD36-mediated fatty acid uptake regulate T cell ferroptosis? | CD36 overexpression or knockout in CD8+ T cells |
| Does TRIM28 SUMOylation of ACSL4 affect neuronal ferroptosis? | TRIM28 SUMOylation-deficient knock-in neurons |
| Does PD-1 signaling promote T cell ferroptosis via PLPP1? | PLPP1 overexpression or knockout in T cells under PD-1 stimulation |
| Can ZDHHC8 inhibition sensitize tumors to ferroptosis? | ZDHHC8 knockout or inhibitor-treated xenografts |
How to Study the positive regulation of ferroptosis Process
| Method | What It Measures | Typical Application |
|---|---|---|
| CRISPR knockout screen | Gene essentiality for ferroptosis | Identify positive regulators |
| C11-BODIPY staining | Lipid peroxidation | Quantify ferroptosis in cells |
| FerroOrange staining | Labile iron pool | Assess iron availability |
| Immunoprecipitation + mass spectrometry | Protein modifications (palmitoylation, ubiquitination, SUMOylation) | Study post-translational regulation |
| RNA-seq | Transcriptional changes | Identify pathways upregulating ferroptosis genes |
| m6A RNA immunoprecipitation | m6A modification of specific mRNAs | Study METTL3-mediated ACSL4 regulation |
| Western blot | Protein expression and modification | Validate GPX4 degradation or ACSL4 accumulation |
| Flow cytometry | Cell death and immune cell function | Measure ferroptosis in CD8+ T cells |
CRISPR screening for ferroptosis regulators
Genome-wide CRISPR knockout or activation screens can identify positive regulators of ferroptosis. Cells are treated with ferroptosis inducers (e.g., erastin, RSL3) and sgRNA enrichment is analyzed to find genes whose loss confers resistance or sensitivity. This approach has been used to uncover pathways involving GPX4, ACSL4, and others.
Lipid peroxidation assays
Lipid peroxidation is measured using fluorescent probes such as C11-BODIPY or by detecting malondialdehyde (MDA) and 4-hydroxynonenal (4-HNE). These assays quantify the extent of ferroptosis and are essential to confirm positive regulation.
Iron measurement
Labile iron pools can be assessed with fluorescent dyes like FerroOrange or by inductively coupled plasma mass spectrometry (ICP-MS). Since iron is critical for ferroptosis, measuring iron levels helps determine whether a gene positively regulates ferroptosis via iron metabolism.
Post-translational modification analysis
Palmitoylation, ubiquitination, and SUMOylation of key proteins can be analyzed by immunoprecipitation followed by mass spectrometry or specific antibodies. For example, GPX4 palmitoylation is detected using acyl-biotin exchange assays.
How CRISPR Can Be Used to Study GO:0160020 positive regulation of ferroptosis
Knockout
CRISPR knockout is used to delete genes suspected of positively regulating ferroptosis. For example, knocking out GPX4 sensitizes cells to ferroptosis, while knocking out ACSL4 confers resistance. Knockout models help establish causality and are often combined with ferroptosis inducers to assess sensitivity.
Point Mutation
Point mutations can be introduced to disrupt specific post-translational modification sites. For instance, mutating the palmitoylation site on GPX4 or the SUMOylation site on ACSL4 can reveal how these modifications affect ferroptosis. Point mutant knock-in cell lines are valuable for dissecting molecular mechanisms.
Knock-in
Knock-in of tagged or mutant proteins allows tracking of protein localization, stability, and interactions. For example, knocking in a tagged ACSL4 can help study its autophagic degradation by OPTN. Knock-in models also enable precise regulation of gene expression under endogenous promoters.
Overexpression
Overexpression of candidate positive regulators can test sufficiency. For example, overexpressing CD36 or METTL3 can promote ferroptosis in cell lines. Overexpression models are useful for gain-of-function studies and for validating screening hits.
How EDITGENE Supports positive regulation of ferroptosis Research
Researchers studying positive regulation of ferroptosis-related genes often need to determine whether a candidate gene is causally involved in promoting ferroptosis or is merely correlated with the process. This requires precise genetic manipulation, which can be achieved through CRISPR-based knockout, point mutation, knock-in, and overexpression models. EDITGENE provides these services along with CRISPR library screening and bioinformatics support to accelerate ferroptosis research.
Contact EDITGENE today to design your custom CRISPR model for positive regulation of ferroptosis research.
Frequently Asked Questions About positive regulation of ferroptosis
What is GO:0160020 positive regulation of ferroptosis?
GO:0160020 is a Gene Ontology term for any process that activates or increases the frequency, rate or extent of ferroptosis, an iron-dependent form of regulated cell death driven by lipid peroxidation.
What genes are involved in positive regulation of ferroptosis?
Key genes include ACSL4, CD36, METTL3, TRIM28, ZDHHC8, and PD-1 signaling components, while GPX4 and FASN are negative regulators.
How does ACSL4 promote ferroptosis?
ACSL4 incorporates polyunsaturated fatty acids into membrane phospholipids, making them susceptible to peroxidation, which is a hallmark of ferroptosis.
What is the role of GPX4 in ferroptosis?
GPX4 is a glutathione peroxidase that reduces lipid peroxides; its inhibition or degradation removes a key brake on ferroptosis, thereby positively regulating the process.
How is ferroptosis regulated in cancer?
In cancer, ferroptosis can be induced to kill tumor cells, but it can also impair antitumor immunity by affecting CD8+ T cells through CD36 and PD-1 signaling.
What experimental models are used to study positive regulation of ferroptosis?
Common models include CRISPR knockout, point mutation, knock-in, and overexpression cell lines, as well as mouse models and genome-wide screens.
What is the connection between ferroptosis and sepsis?
In sepsis-associated lung injury, histone lactylation-regulated METTL3 promotes ferroptosis via m6A modification on ACSL4, contributing to tissue damage.
How does TRIM28 regulate neuronal ferroptosis?
TRIM28 promotes SUMOylation and inhibits OPTN-selective autophagic degradation of ACSL4, leading to ACSL4 accumulation and neuronal ferroptosis.
Can CRISPR screens identify ferroptosis regulators?
Yes, genome-wide CRISPR knockout or activation screens with ferroptosis inducers can uncover positive and negative regulators of ferroptosis.
What are the therapeutic implications of targeting positive regulation of ferroptosis?
Modulating ferroptosis positively could treat cancer by inducing tumor cell death, while inhibiting it might protect against neurodegeneration and inflammatory tissue damage.
Conclusion
Positive regulation of ferroptosis (GO:0160020) is a dynamic and clinically relevant biological process that governs the commitment to iron-dependent lipid peroxidation-driven cell death. Key regulators such as ACSL4, GPX4, CD36, METTL3, and TRIM28 form a complex network that integrates metabolic, redox, and immune signals. Understanding these mechanisms is essential for developing therapies that either induce ferroptosis in cancer or prevent it in degenerative and inflammatory diseases. CRISPR-based models and screening approaches will continue to be indispensable for dissecting this pathway and identifying new therapeutic targets.
References
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- 3. Liu J et al.. 2022. Signaling pathways and defense mechanisms of ferroptosis.. FEBS J 289(22):7038-7050 PMID: 34092035
- 4. Qian Z et al.. 2025. FASN inhibits ferroptosis in breast cancer via USP5 palmitoylation-dependent regulation of GPX4 deubiquitination.. J Exp Clin Cancer Res 44(1):289 PMID: 41088402
- 5. Sharma A et al.. 2021. Positive and Negative Regulation of Ferroptosis and Its Role in Maintaining Metabolic and Redox Homeostasis.. Oxid Med Cell Longev 2021:9074206 PMID: 34007410
- 6. Wu D et al.. 2024. Histone lactylation-regulated METTL3 promotes ferroptosis via m6A-modification on ACSL4 in sepsis-associated lung injury.. Redox Biol 74:103194 PMID: 38852200
- 7. Ping Y et al.. 2024. PD-1 signaling limits expression of phospholipid phosphatase 1 and promotes intratumoral CD8(+) T cell ferroptosis.. Immunity 57(9):2122-2139.e9 PMID: 39208806
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