GO:0045821 positive regulation of glycolysis: Metabolic Reprogramming Pathway, Genes, Functions and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0045821 (positive regulation of glycolysis) describes any process that activates or increases the frequency, rate, or extent of glycolysis, the cytosolic conversion of glucose to pyruvate with net ATP production.
• This GO term is central to cancer metabolic reprogramming, immune cell activation, and fibrotic disease, where increased glycolytic flux supports biosynthesis, redox balance, and signaling [1, 4, 8].
• Multiple positive feedback loops amplify glycolysis: histone lactylation, O-GlcNAcylation, and HIF-1α-driven transcription create self-reinforcing circuits in pancreatic, gastric, and hepatocellular cancers [1, 3, 5, 6].
• Key regulatory nodes include PFKFB3, PIM2, Zeb1, NAT10, SEPT9, HIF-1α, SIRT1, and YBX1, which modulate glycolytic enzyme expression or activity [1, 3, 4, 5, 6, 8].
• Dysregulated positive regulation of glycolysis contributes to paclitaxel resistance, oncogenesis, fibroblast activation, and altered macrophage polarization [1, 2, 4, 8].
• CRISPR-based knockout, point mutation, knock-in, and overexpression models enable causal dissection of glycolytic regulatory networks in disease-relevant cell types.
Description
Glycolysis is the ancient cytosolic pathway that converts glucose to pyruvate, generating ATP and NADH while feeding biosynthetic intermediates into anabolic routes. The Gene Ontology term GO:0045821, positive regulation of glycolysis, captures any process that activates or increases the frequency, rate, or extent of this pathway. In biomedical research, this term is indispensable because enhanced glycolytic flux is a hallmark of activated immune cells, cancer cells, and fibrotic fibroblasts, and it is frequently driven by oncogenic signaling or metabolic stress [1, 4, 8]. Understanding how glycolysis is positively regulated at the molecular level has direct implications for identifying therapeutic targets in oncology, immunology, and fibrosis [2, 6, 8]. Mechanistically, positive regulation of glycolysis often involves transcriptional induction of glycolytic enzymes, allosteric activation of rate-limiting steps, and post-translational modifications that stabilize glycolytic regulators. For example, the PIM2 kinase phosphorylates and stabilizes PFKFB3, increasing glycolytic flux and contributing to paclitaxel resistance in breast cancer. In pancreatic ductal adenocarcinoma, glycolysis promotes histone lactylation, which in turn drives transcription of glycolytic genes, forming a positive feedback loop that sustains oncogenesis. Similar feedback circuits involving H19, SIRT1, and histone lactylation have been described in gastric cancer. These examples illustrate that positive regulation of glycolysis is not a simple linear cascade but a network of interconnected loops that can be targeted at multiple nodes. For researchers, GO:0045821 provides a structured framework to annotate and interrogate genes, pathways, and experimental perturbations that enhance glycolytic flux. Whether studying macrophage polarization in breast cancer, colitis-associated immune regulation, or TGF-β1-mediated fibroblast activation, the term helps unify diverse observations under a common functional umbrella. This article reviews the definition, mechanisms, key genes, disease relevance, and research methods for positive regulation of glycolysis, with a focus on publication-ready evidence from real PubMed literature.
positive regulation of glycolysis At A Glance
| GO ID | GO:0045821 |
|---|---|
| GO term | positive regulation of glycolysis |
| Ontology | biological_process |
| Synonym | activation of glycolysis; positive regulation of glycolytic process; stimulation of glycolysis; up regulation of glycolysis; up-regulation of glycolysis; upregulation of glycolysis |
| Major function | Increases the rate, frequency, or extent of glycolysis, often in response to metabolic, oncogenic, or immune signals |
| Definition | Any process that activates or increases the frequency, rate or extent of glycolysis. |
| Related terms | glycolysis (GO:0006096); negative regulation of glycolysis (GO:0045820); regulation of glycolysis (GO:0045820) |
| Aspect | biological_process |
What Is GO:0045821?
GO:0045821, positive regulation of glycolysis, is defined by QuickGO as any process that activates or increases the frequency, rate, or extent of glycolysis. Glycolysis itself is the breakdown of glucose into pyruvate, producing ATP and NADH. Positive regulation therefore encompasses molecular events such as transcriptional activation of glycolytic genes, allosteric activation of glycolytic enzymes, stabilization of glycolytic regulators, and signaling cascades that elevate glycolytic flux. This term is a biological process and is distinct from negative regulation of glycolysis (GO:0045820) and from glycolysis itself (GO:0006096).
Why Is positive regulation of glycolysis Important in Cell Biology?
Positive regulation of glycolysis is a central node in metabolic reprogramming, linking oncogenic signaling, immune activation, and fibrotic responses to altered glucose metabolism. Its dysregulation is implicated in cancer progression, therapy resistance, and inflammatory diseases, making it a high-value target for mechanistic studies and therapeutic intervention [1, 2, 4, 6, 8].
• Drives oncogenic metabolic reprogramming in pancreatic, gastric, breast, and hepatocellular cancers [1, 3, 4, 5, 6, 8].
• Supports immune cell activation and macrophage polarization, influencing tumor microenvironment and inflammation [4, 7].
• Contributes to paclitaxel resistance in breast cancer through PFKFB3 stabilization.
• Mediates TGF-β1-induced fibroblast activation and fibrosis, with alamandine/MrgD axis counteracting this process.
• Creates positive feedback loops via histone lactylation and O-GlcNAcylation that sustain glycolytic gene expression [1, 3, 5].
• Provides mechanistic links between metabolism and gene regulation through epigenetic modifications [1, 3, 5].
• Offers multiple druggable nodes, including PFKFB3, PIM2, HIF-1α, and NAT10 [6, 8].
• Serves as a functional annotation for CRISPR screens and transcriptomic studies of metabolic pathways.
• Enables cross-species and cross-cell-type comparisons of glycolytic regulation in health and disease.
• Underpins the development of metabolic biomarkers and combination therapies targeting glycolysis.
What Happens During positive regulation of glycolysis?
Transcriptional activation of glycolytic genes
In simple terms: Cells increase the production of glycolytic enzymes by turning on their genes.
Positive regulation of glycolysis frequently begins with transcriptional induction of glycolytic enzymes such as hexokinase 2, PFKFB3, and lactate dehydrogenase A. In pancreatic ductal adenocarcinoma, glycolysis promotes histone lactylation, which enhances transcription of glycolytic genes, forming a positive feedback loop that drives oncogenesis. Similarly, in gastric cancer, the NAT10/SEPT9/HIF-1α axis induces glycolysis addiction through a positive feedback loop involving N4-acetylcytidine modification. These examples demonstrate that transcriptional activation is a core mechanism for increasing glycolytic capacity.
Post-translational stabilization of glycolytic regulators
In simple terms: Chemical modifications can make glycolytic enzymes more stable or active.
Beyond transcription, post-translational modifications stabilize key glycolytic regulators. PIM2 kinase positively regulates PFKFB3 by phosphorylation, promoting glycolysis and paclitaxel resistance in breast cancer. In hepatocellular carcinoma, O-GlcNAcylation of YBX1 drives a glycolysis-histone lactylation feedback loop, linking nutrient sensing to epigenetic regulation. These modifications allow rapid, reversible increases in glycolytic flux without new gene expression.
Epigenetic feedback via histone lactylation
In simple terms: Lactate produced by glycolysis modifies histones, which then turn on more glycolytic genes.
Histone lactylation is a recently described epigenetic mark that connects glycolytic flux to gene expression. In pancreatic ductal adenocarcinoma, a positive feedback loop between glycolysis and histone lactylation drives oncogenesis. In gastric cancer, SIRT1 acts as a delactylase to modulate the H19-glycolysis-histone lactylation loop, fine-tuning glycolytic output. This feedback mechanism exemplifies how positive regulation of glycolysis can be self-reinforcing.
Signaling pathways that amplify glycolysis
In simple terms: External signals can switch on glycolysis through specific signaling cascades.
Several signaling pathways positively regulate glycolysis. In breast cancer, Zeb1-induced metabolic reprogramming of glycolysis is essential for macrophage polarization, linking epithelial-mesenchymal transition programs to immune cell metabolism. In colitis, GPR120 signaling regulates CD4+ T cell interleukin 10 production, indirectly influencing glycolytic activity in immune cells. The alamandine/MrgD axis prevents TGF-β1-mediated fibroblast activation by regulating aerobic glycolysis and mitophagy, showing that counter-regulatory signals can suppress glycolysis.
Key Genes Involved in GO:0045821 positive regulation of glycolysis
The following genes and proteins are experimentally validated participants in positive regulation of glycolysis, as supported by the cited literature.
| Gene | Major Role | Research Relevance |
|---|---|---|
| PFKFB3 | Fructose-2,6-bisphosphatase 3; allosteric activator of PFK-1 | Phosphorylated by PIM2; promotes glycolysis and paclitaxel resistance in breast cancer |
| PIM2 | Serine/threonine kinase | Positively regulates PFKFB3, enhancing glycolysis |
| Zeb1 | Transcription factor | Induces metabolic reprogramming of glycolysis in breast cancer macrophages |
| NAT10 | N-acetyltransferase | Drives N4-acetylcytidine modification in NAT10/SEPT9/HIF-1α loop in gastric cancer |
| SEPT9 | Septin family GTPase | Part of NAT10/SEPT9/HIF-1α positive feedback loop |
| HIF-1α | Hypoxia-inducible factor 1 alpha | Transcriptionally activates glycolytic genes; central to feedback loops |
| SIRT1 | NAD-dependent deacetylase and delactylase | Modulates H19-glycolysis-histone lactylation loop in gastric cancer |
| H19 | Long non-coding RNA | Involved in glycolysis-histone lactylation feedback in gastric cancer |
| YBX1 | Y-box binding protein 1 | O-GlcNAcylation drives glycolysis-histone lactylation feedback in HCC |
| GPR120 | Free fatty acid receptor 4 | Regulates CD4+ T cell IL-10 production and colitis |
| MrgD | Mas-related G protein-coupled receptor D | Alamandine/MrgD axis regulates aerobic glycolysis in fibroblasts |
| Alamandine | Bioactive peptide | Counteracts TGF-β1-mediated fibroblast activation via glycolysis regulation |
| TGF-β1 | Transforming growth factor beta 1 | Induces fibroblast activation and aerobic glycolysis |
| IL-10 | Interleukin 10 | Anti-inflammatory cytokine regulated by GPR120 in CD4+ T cells |
| LDHA | Lactate dehydrogenase A | Converts pyruvate to lactate; supports glycolytic flux and lactylation [1, 3, 5] |
| HK2 | Hexokinase 2 | Rate-limiting glycolytic enzyme; often upregulated in cancer [1, 6] |
| PKM2 | Pyruvate kinase M2 | Glycolytic enzyme with regulatory roles in cancer metabolism [1, 4] |
| GLUT1 | Glucose transporter 1 | Facilitates glucose uptake, supporting increased glycolysis [4, 6] |
How Is positive regulation of glycolysis Regulated?
Positive regulation of glycolysis is controlled by a multilayered regulatory network. Transcriptional regulators such as HIF-1α and Zeb1 induce glycolytic gene expression in response to hypoxia or oncogenic signals [4, 6]. Post-translational modifications, including phosphorylation by PIM2 and O-GlcNAcylation of YBX1, stabilize or activate glycolytic enzymes and regulators [5, 8]. Epigenetic feedback via histone lactylation creates self-reinforcing loops that sustain glycolytic flux [1, 3]. Counter-regulatory pathways, such as the alamandine/MrgD axis, can suppress glycolysis in fibroblasts, highlighting the balance between positive and negative regulation. Additionally, immune signals like GPR120 activation modulate glycolysis indirectly through cytokine production.
positive regulation of glycolysis and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| PFKFB3 | Breast cancer paclitaxel resistance | Knockout or point mutation in breast cancer cell lines; overexpression for resistance studies |
| NAT10 | Gastric cancer glycolysis addiction | Knockout in gastric cancer organoids; knock-in of acetylation-deficient mutants |
| SIRT1 | Gastric cancer histone lactylation | Delactylase-dead point mutant knock-in; knockout for feedback loop analysis |
| YBX1 | Hepatocellular carcinoma | O-GlcNAcylation site point mutants; knockout for glycolysis-lactylation loop |
| Zeb1 | Breast cancer macrophage polarization | Conditional knockout in macrophage co-culture models; overexpression in epithelial cells |
Cancer metabolic reprogramming
Positive regulation of glycolysis is a hallmark of many cancers. In pancreatic ductal adenocarcinoma, a positive feedback loop between glycolysis and histone lactylation drives oncogenesis. Gastric cancer exhibits glycolysis addiction via the NAT10/SEPT9/HIF-1α loop, and SIRT1 modulates a H19-glycolysis-histone lactylation feedback loop. Hepatocellular carcinoma features O-GlcNAcylation of YBX1 that sustains a glycolysis-histone lactylation feedback loop. Breast cancer cells utilize Zeb1-induced glycolytic reprogramming for macrophage polarization and PIM2-mediated PFKFB3 stabilization for paclitaxel resistance. These findings underscore the therapeutic potential of targeting glycolytic regulatory nodes.
Fibrosis and fibroblast activation
In fibrotic diseases, TGF-β1 promotes fibroblast activation and aerobic glycolysis. The alamandine/MrgD axis counteracts this process by regulating glycolysis and mitophagy, suggesting a protective mechanism that could be harnessed therapeutically. This highlights the importance of positive regulation of glycolysis in non-malignant proliferative disorders.
Immune regulation and colitis
GPR120 signaling in CD4+ T cells regulates interleukin 10 production and protects against colitis, indirectly influencing glycolytic activity in immune cells. Macrophage polarization in breast cancer is dependent on Zeb1-induced glycolytic reprogramming, linking metabolism to immune cell function in the tumor microenvironment. These examples illustrate the role of glycolysis regulation in inflammatory and immune-mediated diseases.
From positive regulation of glycolysis-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Is PFKFB3 phosphorylation by PIM2 required for glycolysis and paclitaxel resistance? | Point mutation of PFKFB3 phosphorylation sites via CRISPR knock-in; PIM2 knockout |
| Does histone lactylation feedback drive pancreatic cancer oncogenesis? | Knockout of lactyltransferases or delactylases; histone point mutants; overexpression of glycolytic enzymes |
| What is the role of NAT10/SEPT9/HIF-1α loop in gastric cancer? | Knockout of NAT10 or SEPT9; knock-in of acetylation-deficient HIF-1α; overexpression of loop components |
| How does SIRT1 delactylase activity modulate H19-glycolysis feedback? | SIRT1 catalytic-dead knock-in; H19 knockout; delactylase activity assays |
| Does O-GlcNAcylation of YBX1 regulate glycolysis in HCC? | YBX1 O-GlcNAc site point mutants; OGT knockout; overexpression of wild-type vs. mutant YBX1 |
| Can alamandine/MrgD axis suppress fibroblast glycolysis? | MrgD knockout fibroblasts; alamandine treatment; TGF-β1 stimulation; glycolysis flux assays |
How to Study the positive regulation of glycolysis Process
| Method | What It Measures | Typical Application |
|---|---|---|
| ECAR (Seahorse) | Extracellular acidification rate as proxy for glycolysis | Real-time glycolytic flux in live cells [1, 8] |
| Lactate assay | Lactate production | Quantifying glycolytic output [1, 8] |
| RNA-seq | Transcriptional changes | Identifying glycolytic gene expression changes |
| ChIP-seq | Histone modifications and transcription factor binding | Mapping histone lactylation and HIF-1α binding [1, 3, 6] |
| Phosphoproteomics | Phosphorylation sites on proteins | Detecting PIM2-mediated PFKFB3 phosphorylation |
| O-GlcNAc proteomics | O-GlcNAcylation sites | Mapping YBX1 modification in HCC |
| CRISPR knockout screen | Gene essentiality or fitness | Discovering positive regulators of glycolysis [1, 4, 6] |
| CRISPR activation screen | Gene overexpression effects | Identifying genes that increase glycolytic flux [1, 4, 6] |
Metabolic flux analysis
Measuring glycolytic flux is essential to confirm positive regulation. Extracellular acidification rate (ECAR) and lactate production assays quantify glycolytic activity. In pancreatic cancer, glycolysis-driven histone lactylation was linked to oncogenesis using such methods. Similarly, PFKFB3-mediated glycolysis in breast cancer was assessed by lactate production and glucose consumption.
Transcriptomics and epigenomics
RNA-seq and ChIP-seq can identify transcriptional changes and histone modifications associated with increased glycolysis. In gastric cancer, the NAT10/SEPT9/HIF-1α loop was dissected using RNA-seq and ac4C modification mapping. Histone lactylation was profiled by ChIP-seq in pancreatic cancer and gastric cancer.
Proteomics and post-translational modification analysis
Mass spectrometry-based proteomics detects phosphorylation, O-GlcNAcylation, and lactylation of glycolytic regulators. PIM2-mediated PFKFB3 phosphorylation was identified by phosphoproteomics, and YBX1 O-GlcNAcylation was mapped in HCC.
CRISPR screens and functional genomics
Genome-wide CRISPR knockout or activation screens can identify positive regulators of glycolysis. Such screens are powerful for discovering novel genes that increase glycolytic flux under specific conditions, complementing candidate-based studies [1, 4, 6].
How CRISPR Can Be Used to Study GO:0045821 positive regulation of glycolysis
Knockout
CRISPR knockout of candidate positive regulators (e.g., PFKFB3, PIM2, NAT10, SEPT9, YBX1) can abolish increased glycolysis and reverse disease phenotypes. For example, PIM2 knockout reduces PFKFB3 phosphorylation and glycolysis, sensitizing breast cancer cells to paclitaxel. NAT10 knockout disrupts the NAT10/SEPT9/HIF-1α loop and reduces gastric cancer glycolysis.
Point Mutation
Point mutations can dissect specific post-translational modification sites. Knocking in phosphorylation-deficient PFKFB3 mutants can test whether PIM2-mediated phosphorylation is required for glycolysis and drug resistance. Similarly, O-GlcNAcylation site mutants of YBX1 can clarify its role in the glycolysis-lactylation feedback loop.
Knock-in
Knock-in of tagged or mutant alleles enables precise tracking and functional analysis. For instance, knock-in of a delactylase-dead SIRT1 mutant can test its role in modulating the H19-glycolysis-histone lactylation loop. Knock-in of acetylation-deficient HIF-1α can disrupt the NAT10/SEPT9/HIF-1α feedback loop.
Overexpression
CRISPR activation or cDNA overexpression can drive positive regulation of glycolysis. Overexpressing Zeb1 in breast cancer cells induces glycolytic reprogramming and macrophage polarization. Overexpression of wild-type versus mutant YBX1 can reveal the impact of O-GlcNAcylation on glycolysis.
How EDITGENE Supports positive regulation of glycolysis Research
Researchers studying positive regulation of glycolysis-related genes often need to determine whether a candidate gene is causally involved in increasing glycolytic flux, and whether specific domains or post-translational modification sites are required. EDITGENE provides end-to-end CRISPR services to generate precisely engineered cell models that answer these questions.
Contact EDITGENE today to design your custom CRISPR model for positive regulation of glycolysis research.
Frequently Asked Questions About positive regulation of glycolysis
What is GO:0045821?
GO:0045821 is the Gene Ontology term for positive regulation of glycolysis, defined as any process that activates or increases the frequency, rate, or extent of glycolysis.
What genes are involved in positive regulation of glycolysis?
Key genes include PFKFB3, PIM2, Zeb1, NAT10, SEPT9, HIF-1α, SIRT1, H19, YBX1, GPR120, MrgD, and LDHA, among others [1, 2, 3, 4, 5, 6, 7, 8].
How is glycolysis positively regulated in cancer?
Cancer cells often upregulate glycolysis through transcriptional activation (e.g., HIF-1α), post-translational stabilization (e.g., PIM2-PFKFB3), and epigenetic feedback loops (e.g., histone lactylation) [1, 3, 5, 6, 8].
What is the role of histone lactylation in glycolysis regulation?
Histone lactylation is a lactate-derived epigenetic mark that can enhance transcription of glycolytic genes, creating a positive feedback loop that sustains glycolysis in cancers [1, 3, 5].
Which diseases are associated with positive regulation of glycolysis?
Cancer (pancreatic, gastric, breast, hepatocellular), fibrosis, and inflammatory conditions such as colitis are associated with dysregulated positive regulation of glycolysis [1, 2, 3, 4, 5, 6, 7, 8].
How can CRISPR be used to study positive regulation of glycolysis?
CRISPR knockout, point mutation, knock-in, and overexpression can test the causal role of specific genes and modification sites in increasing glycolytic flux [1, 3, 5, 6, 8].
What methods measure glycolytic flux?
ECAR, lactate production assays, glucose consumption, and stable isotope tracing are commonly used to quantify glycolytic flux [1, 8].
What is the PIM2-PFKFB3 axis?
PIM2 phosphorylates PFKFB3, stabilizing it and promoting glycolysis, which contributes to paclitaxel resistance in breast cancer.
How does the NAT10/SEPT9/HIF-1α loop work?
NAT10-mediated ac4C modification of SEPT9 enhances HIF-1α signaling, which in turn drives glycolysis, forming a positive feedback loop in gastric cancer.
What is the H19-glycolysis-histone lactylation loop?
In gastric cancer, the long non-coding RNA H19 promotes glycolysis, leading to lactate production and histone lactylation, which further activates glycolytic genes; SIRT1 modulates this loop.
Conclusion
Positive regulation of glycolysis (GO:0045821) is a fundamental biological process that integrates transcriptional, post-translational, and epigenetic mechanisms to increase glycolytic flux. Its dysregulation is central to cancer, fibrosis, and immune disorders, making it a rich area for mechanistic and therapeutic research. The genes and pathways highlighted here, supported by real PubMed literature, provide a foundation for designing CRISPR-based experiments to dissect causal relationships and identify new targets.
References
- 1. Li F et al.. 2024. Positive feedback regulation between glycolysis and histone lactylation drives oncogenesis in pancreatic ductal adenocarcinoma.. Mol Cancer 23(1):90 PMID: 38711083
- 2. Wang W et al.. 2023. Alamandine/MrgD axis prevents TGF-β1-mediated fibroblast activation via regulation of aerobic glycolysis and mitophagy.. J Transl Med 21(1):24 PMID: 36635651
- 3. Tsukihara S et al.. 2025. Delactylase effects of SIRT1 on a positive feedback loop involving the H19-glycolysis-histone lactylation in gastric cancer.. Oncogene 44(11):724-738 PMID: 39658647
- 4. Jiang H et al.. 2022. Zeb1-induced metabolic reprogramming of glycolysis is essential for macrophage polarization in breast cancer.. Cell Death Dis 13(3):206 PMID: 35246504
- 5. Ji Y et al.. 2025. O-GlcNAcylation of YBX1 drives a glycolysis-histone lactylation feedback loop in hepatocellular carcinoma.. Cancer Lett 631:217957 PMID: 40721081
- 6. Yang Q et al.. 2023. N4-Acetylcytidine Drives Glycolysis Addiction in Gastric Cancer via NAT10/SEPT9/HIF-1α Positive Feedback Loop.. Adv Sci (Weinh) 10(23):e2300898 PMID: 37328448
- 7. Yang W et al.. 2022. GPR120 Inhibits Colitis Through Regulation of CD4(+) T Cell Interleukin 10 Production.. Gastroenterology 162(1):150-165 PMID: 34536451
- 8. Lu C et al.. 2021. Positive regulation of PFKFB3 by PIM2 promotes glycolysis and paclitaxel resistance in breast cancer.. Clin Transl Med 11(4):e400 PMID: 33931981
