GO:0035689 chemokine (C-C motif) ligand 5 signaling pathway: Immune Recruitment Pathway, Genes, Functions and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0035689 describes the molecular signaling cascade triggered when CCL5 (RANTES) binds its receptor on a target cell, culminating in regulation of downstream cellular processes such as transcription.
• CCL5 signals primarily through the G-protein-coupled receptor CCR5, and this axis is a central mediator of immune cell recruitment, tumor microenvironment remodeling, and inflammatory disease.
• The CCL5/CCR5 axis drives cancer progression by recruiting immunosuppressive macrophages and maintaining glioma stem cells in glioblastoma.
• CCL5 signaling is implicated in non-cancer pathologies including vascular aging, atherosclerosis, and osteoarthritis, though its role in osteoarthritis is context-dependent.
• CCL5 expression is regulated at multiple levels, including m6A RNA modification by ALKBH5, which affects macrophage senescence and atherosclerosis.
• CRISPR-based knockout, knock-in, and overexpression models are essential tools for dissecting causal roles of CCL5 signaling components in disease.
Description
The chemokine (C-C motif) ligand 5 signaling pathway (GO:0035689) is a biological process defined as the series of molecular signals initiated by CCL5 binding to its receptor on the surface of a target cell, ending with regulation of a downstream cellular process such as transcription. CCL5, also known as RANTES, is a secreted chemokine that plays a pivotal role in immune cell trafficking and activation. This pathway is of intense research interest because it bridges innate and adaptive immunity and is frequently dysregulated in cancer, chronic inflammation, and cardiovascular disease. Understanding GO:0035689 is therefore critical for researchers investigating tumor immunology, inflammatory disorders, and tissue remodeling.
chemokine (C-C motif) ligand 5 signaling pathway At A Glance
| GO ID | GO:0035689 |
|---|---|
| GO term | chemokine (C-C motif) ligand 5 signaling pathway |
| Ontology | biological_process |
| Synonym | CCL5-mediated signaling pathway; RANTES-mediated signaling pathway |
| Major function | Transduces extracellular CCL5 signals to regulate immune cell recruitment, activation, and gene transcription |
| Key receptor | CCR5 (primary), with possible crosstalk with other chemokine receptors |
| Key ligand | CCL5 (RANTES) |
| Downstream effects | Chemotaxis, integrin activation, cytokine production, transcriptional reprogramming |
| Disease relevance | Cancer, atherosclerosis, vascular aging, inflammatory diseases |
What Is GO:0035689?
GO:0035689 is the biological process comprising the molecular signals initiated when the chemokine CCL5 binds to its receptor on the surface of a target cell, leading to regulation of downstream cellular processes such as transcription. It is synonymous with CCL5-mediated signaling pathway and RANTES-mediated signaling pathway.
Why Is chemokine (C-C motif) ligand 5 signaling pathway Important in Cell Biology?
GO:0035689 is important because CCL5 signaling is a master regulator of immune cell dynamics in health and disease. It controls the recruitment of T cells, macrophages, and other leukocytes to sites of inflammation and tumors. In cancer, the CCL5/CCR5 axis promotes an immunosuppressive tumor microenvironment and supports stem cell maintenance in glioblastoma. In cardiovascular disease, CCL5 from adipose tissue contributes to vascular aging, and its m6A modification drives macrophage senescence in atherosclerosis. Thus, understanding this pathway offers therapeutic opportunities across oncology and inflammatory disorders.
• Mediates T cell engraftment and immune attack in solid tumors.
• Drives macrophage-induced immunosuppression and angiogenesis in cancer.
• Supports glioma stem cell maintenance in glioblastoma.
• Links subcutaneous adipose tissue to vascular aging.
• Regulated by m6A modification via ALKBH5 in atherosclerosis.
• Context-dependent role in osteoarthritis: CCL2/CCR2 but not CCL5/CCR5 mediates monocyte recruitment.
• Serves as a target for therapeutic blockade in cancer and inflammatory diseases.
• Provides biomarkers for immune checkpoint blockade sensitivity.
What Happens During chemokine (C-C motif) ligand 5 signaling pathway?
CCL5 Secretion and Receptor Binding
In simple terms: CCL5 is released by cells and docks onto a receptor on target cells.
CCL5 is secreted by various cell types, including T cells, macrophages, and tumor cells. It binds primarily to the G-protein-coupled receptor CCR5 on the surface of target cells, initiating the signaling cascade. This binding is a prerequisite for all downstream events in GO:0035689.
G-Protein Activation and Intracellular Signaling
In simple terms: The receptor activates G-proteins inside the cell, triggering a chain of molecular signals.
Upon CCL5 binding, CCR5 undergoes conformational changes that activate heterotrimeric G-proteins. This leads to dissociation of Gα and Gβγ subunits, which modulate effectors such as adenylyl cyclase and phospholipase C, resulting in calcium flux and activation of kinases including PI3K/AKT and MAPK pathways. These events propagate the signal toward transcriptional regulation.
Cytoskeletal Rearrangement and Chemotaxis
In simple terms: The cell changes shape and moves toward the CCL5 signal.
CCL5 signaling induces actin polymerization and integrin activation, enabling directed cell migration (chemotaxis). This is critical for recruiting immune cells to tumor sites and inflammatory lesions. In glioblastoma, CCL5/CCR5 signaling promotes glioma stem cell maintenance through similar cytoskeletal and survival cues.
Transcriptional Reprogramming and Immune Modulation
In simple terms: The signal reaches the nucleus and changes which genes are turned on or off.
Downstream of kinase cascades, transcription factors such as NF-κB and STATs are activated, leading to expression of pro-inflammatory cytokines, chemokines, and immune checkpoint molecules. This transcriptional reprogramming can foster an immunosuppressive microenvironment, as seen in NSCLC where CCL5/CCR5 axis provokes macrophage-mediated immunosuppression. In breast cancer, cooperation between constitutive and inducible chemokines including CCL5 enables T cell engraftment.
Key Genes Involved in GO:0035689 chemokine (C-C motif) ligand 5 signaling pathway
The following genes and proteins are central to the chemokine (C-C motif) ligand 5 signaling pathway (GO:0035689).
| Gene | Major Role | Research Relevance |
|---|---|---|
| CCL5 | Ligand that initiates signaling | Knockout reduces immune cell recruitment; overexpression models tumor microenvironment |
| CCR5 | Primary receptor for CCL5 | Knockout blocks signaling; point mutations affect ligand binding |
| CCR2 | Receptor for CCL2, not CCL5 | Context-dependent; CCL2/CCR2 mediates monocyte recruitment in osteoarthritis |
| ALKBH5 | m6A demethylase regulating CCL5 mRNA | Knockout increases CCL5 m6A and macrophage senescence |
| CD147 | Tumor cell surface protein crosstalk with macrophages | CD147-K148me2 drives CCL5/CCR5 axis in NSCLC |
| ACE2 | Modulates macrophage immunosuppression and angiogenesis | ACE2 enhances PD-L1 blockade sensitivity via CCL5-related pathways |
| NF-κB | Transcription factor downstream of CCL5 | Knockout or knockdown blocks inflammatory gene expression |
| STAT3 | Transcription factor activated by CCL5 | Involved in immunosuppressive macrophage polarization |
| PI3K | Kinase in CCL5 signaling | Inhibitors reduce chemotaxis and survival |
| AKT | Downstream kinase | Phosphorylation status reflects pathway activation |
| MAPK | Kinase cascade | Regulates transcriptional responses |
| RANTES | Alternative name for CCL5 | Same as CCL5 |
| GNAI | G-protein subunit | Mediates receptor signaling |
| ARRB1 | Beta-arrestin | Regulates receptor desensitization |
| GRK | G-protein-coupled receptor kinase | Phosphorylates activated CCR5 |
| CXCL9 | Inducible chemokine cooperating with CCL5 | Cooperation enables T cell engraftment |
| CXCL10 | Inducible chemokine cooperating with CCL5 | Cooperation enables T cell engraftment |
| PD-L1 | Immune checkpoint | CCL5 signaling influences PD-L1 blockade response |
How Is chemokine (C-C motif) ligand 5 signaling pathway Regulated?
CCL5 signaling is regulated at multiple levels. Epigenetic regulation via m6A modification of CCL5 mRNA by ALKBH5 affects macrophage senescence and atherosclerosis. Receptor desensitization by GRK-mediated phosphorylation and beta-arrestin recruitment modulates signal duration. In the tumor microenvironment, cooperation between constitutive and inducible chemokines such as CXCL9 and CXCL10 enhances CCL5-mediated T cell engraftment. Additionally, CD147-K148me2-driven crosstalk between tumor cells and macrophages amplifies CCL5/CCR5 signaling in NSCLC.
chemokine (C-C motif) ligand 5 signaling pathway and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| CCL5 | Breast cancer, tumor immunology | Knockout mouse models, syngeneic tumor transplantation |
| CCR5 | NSCLC, glioblastoma | CRISPR knockout in cancer cell lines, xenograft models |
| ALKBH5 | Atherosclerosis | Macrophage-specific knockout, ApoE-/- background |
| CD147 | NSCLC immunosuppression | Point mutation at K148, macrophage co-culture |
| ACE2 | Cancer immunotherapy | Overexpression in tumor cells, PD-L1 blockade combination |
Cancer and Tumor Immunology
The CCL5/CCR5 axis is a key driver of tumor progression. In solid tumors, CCL5 cooperates with inducible chemokines to enable T cell engraftment and immune attack. In NSCLC, CD147-K148me2-driven tumor cell-macrophage crosstalk provokes immunosuppression via the CCL5/CCR5 axis. In glioblastoma, reactive oligodendrocytes promote tumor progression through CCL5/CCR5-mediated glioma stem cell maintenance. ACE2 enhances sensitivity to PD-L1 blockade by inhibiting macrophage-induced immunosuppression and angiogenesis, partly through CCL5-related mechanisms.
Cardiovascular and Metabolic Diseases
CCL5 from subcutaneous adipose tissue plays a central role in vascular aging. In atherosclerosis, ALKBH5 regulates macrophage senescence by promoting CCL5 m6A modification, accelerating disease progression. These findings highlight CCL5 signaling as a therapeutic target in cardiovascular aging and metabolic disorders.
Inflammatory and Musculoskeletal Diseases
In osteoarthritis, CCL2/CCR2 but not CCL5/CCR5 mediates monocyte recruitment, inflammation, and cartilage destruction, indicating a context-dependent role for CCL5 signaling. This underscores the need for careful model selection when studying CCL5 in inflammatory joint diseases.
From chemokine (C-C motif) ligand 5 signaling pathway-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does CCL5 loss affect tumor immune infiltration? | CCL5 knockout mouse or CRISPR KO in tumor cells |
| Does CCR5 point mutation alter ligand binding? | Knock-in of CCR5 variants in cell lines |
| Does ALKBH5 regulate CCL5 m6A modification? | ALKBH5 knockout macrophages, m6A IP |
| Does CD147-K148me2 drive CCL5/CCR5 crosstalk? | Point mutation at K148 in NSCLC cells, co-culture with macrophages |
| Does CCL5 overexpression promote glioma stem cell maintenance? | Overexpression in glioblastoma cells, stem cell assays |
| Does ACE2 modulate PD-L1 blockade sensitivity? | ACE2 overexpression in tumor models, anti-PD-L1 treatment |
How to Study the chemokine (C-C motif) ligand 5 signaling pathway Process
| Method | What It Measures | Typical Application |
|---|---|---|
| RNA-seq | Global transcriptional changes | Identify CCL5-induced gene expression programs |
| Proteomics | Protein abundance and modifications | Detect m6A-modified CCL5 or signaling proteins |
| Transwell migration | Chemotaxis of immune cells | Assess CCL5-mediated recruitment |
| Phospho-kinase array | Activation of signaling kinases | Map downstream pathways |
| CRISPR knockout screen | Gene essentiality and modifiers | Discover regulators of CCL5 signaling |
| Flow cytometry | Immune cell populations and activation | Evaluate tumor microenvironment changes |
| Immunohistochemistry | Tissue localization of CCL5/CCR5 | Correlate with disease progression |
Transcriptomic and Proteomic Profiling
RNA-seq and proteomics can identify global changes in gene expression and protein abundance following CCL5 stimulation or CCR5 knockout. These methods reveal downstream transcriptional programs and signaling networks.
Chemotaxis and Immune Cell Migration Assays
Transwell migration and live imaging assays measure the functional impact of CCL5 signaling on immune cell recruitment. These are critical for validating GO:0035689 in tumor and inflammation models.
Phospho-Signaling Arrays and Western Blot
Phospho-kinase arrays and immunoblotting detect activation of PI3K/AKT, MAPK, and STAT pathways downstream of CCL5/CCR5, providing mechanistic insights.
CRISPR Screening and Functional Genomics
Genome-wide CRISPR knockout screens can identify modifiers of CCL5 signaling and resistance mechanisms. These screens are powerful for discovering novel pathway components.
How CRISPR Can Be Used to Study GO:0035689 chemokine (C-C motif) ligand 5 signaling pathway
Knockout
CRISPR knockout of CCL5 or CCR5 is used to abolish signaling and assess loss-of-function phenotypes in cancer, inflammation, and cardiovascular models. For example, CCR5 knockout in glioblastoma cells reduces stem cell maintenance.
Point Mutation
Point mutations can be introduced to dissect specific residues critical for ligand binding or post-translational modifications. For instance, mutation of CD147 at K148 prevents methylation and blocks CCL5/CCR5-driven immunosuppression.
Knock-in
Knock-in of tagged or reporter versions of CCL5 or CCR5 allows real-time tracking of signaling dynamics and protein localization. This is useful for imaging and biochemical studies.
Overexpression
Overexpression of CCL5 or CCR5 in cell lines or mouse models can mimic pathological activation and test therapeutic interventions. For example, ACE2 overexpression enhances PD-L1 blockade sensitivity.
How EDITGENE Supports chemokine (C-C motif) ligand 5 signaling pathway Research
Researchers studying chemokine (C-C motif) ligand 5 signaling pathway-related genes often need to determine whether a candidate gene is causally involved in immune recruitment, tumor progression, or inflammatory disease. EDITGENE provides comprehensive CRISPR-based services to accelerate this discovery.
Contact EDITGENE today to design your custom CRISPR model for chemokine (C-C motif) ligand 5 signaling pathway research.
Frequently Asked Questions About chemokine (C-C motif) ligand 5 signaling pathway
What is GO:0035689?
GO:0035689 is the Gene Ontology term for the chemokine (C-C motif) ligand 5 signaling pathway, describing the molecular signals initiated by CCL5 binding to its receptor on a target cell, leading to regulation of downstream cellular processes such as transcription.
What genes are involved in chemokine (C-C motif) ligand 5 signaling pathway?
Key genes include CCL5 (ligand), CCR5 (receptor), and downstream effectors such as NF-κB, STAT3, PI3K, and AKT. Other modulators include ALKBH5, CD147, and ACE2.
What is CCL5 and what does it do?
CCL5, also known as RANTES, is a chemokine that binds to CCR5 and other receptors to recruit immune cells to sites of inflammation and tumors, and to modulate immune responses.
How does CCL5 signaling contribute to cancer?
CCL5 signaling recruits immunosuppressive macrophages and T cells, promotes angiogenesis, and supports cancer stem cell maintenance, contributing to tumor progression and resistance to immunotherapy.
What diseases are associated with CCL5 signaling?
CCL5 signaling is implicated in breast cancer, NSCLC, glioblastoma, atherosclerosis, vascular aging, and osteoarthritis, though its role in osteoarthritis is context-dependent.
How is CCL5 signaling regulated?
CCL5 signaling is regulated by m6A RNA modification via ALKBH5, receptor desensitization by GRK and beta-arrestin, and cooperation with other chemokines such as CXCL9 and CXCL10.
What research methods are used to study CCL5 signaling?
Common methods include RNA-seq, proteomics, chemotaxis assays, phospho-kinase arrays, CRISPR screens, flow cytometry, and immunohistochemistry.
Can CRISPR be used to study CCL5 signaling?
Yes, CRISPR knockout, point mutation, knock-in, and overexpression models are widely used to dissect the causal roles of CCL5, CCR5, and related genes in disease.
What is the role of CCR5 in CCL5 signaling?
CCR5 is the primary receptor for CCL5. Upon binding, it activates G-proteins and downstream signaling cascades that regulate chemotaxis, transcription, and immune cell function.
How does ALKBH5 affect CCL5 signaling?
ALKBH5 is an m6A demethylase that regulates CCL5 mRNA modification. Its loss increases CCL5 m6A levels, promoting macrophage senescence and accelerating atherosclerosis.
Conclusion
The chemokine (C-C motif) ligand 5 signaling pathway (GO:0035689) is a central mediator of immune cell recruitment and activation, with profound implications for cancer, cardiovascular disease, and inflammation. Understanding its molecular mechanisms and regulation offers opportunities for therapeutic intervention. CRISPR-based models are indispensable for dissecting the causal roles of CCL5, CCR5, and their regulators in human disease.
References
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- 2. Xie P et al.. 2025. ACE2 Enhances Sensitivity to PD-L1 Blockade by Inhibiting Macrophage-Induced Immunosuppression and Angiogenesis.. Cancer Res 85(2):299-313 PMID: 39495239
- 3. Wang K et al.. 2024. CD147-K148me2-Driven Tumor Cell-Macrophage Crosstalk Provokes NSCLC Immunosuppression via the CCL5/CCR5 Axis.. Adv Sci (Weinh) 11(29):e2400611 PMID: 38873823
- 4. Le Pelletier L et al.. 2025. C-C chemokine ligand 5 from women subcutaneous adipose tissue has a central role in vascular aging.. Cardiovasc Diabetol 24(1):295 PMID: 40682056
- 5. Ghasemi K. 2025. C-C motif glycoprotein ligand 5 (CCL5) and its GPCR CCR5: Macromolecular game-changers in cancer biology.. Int J Biol Macromol 329(Pt 1):147737 PMID: 40976299
- 6. Gao R et al.. 2025. ALKBH5 Regulates Macrophage Senescence and Accelerates Atherosclerosis by Promoting CCL5 m(6)A Modification.. Arterioscler Thromb Vasc Biol 45(6):928-944 PMID: 40177773
- 7. Raghu H et al.. 2017. CCL2/CCR2, but not CCL5/CCR5, mediates monocyte recruitment, inflammation and cartilage destruction in osteoarthritis.. Ann Rheum Dis 76(5):914-922 PMID: 27965260
- 8. Mikolajewicz N et al.. 2026. Reactive oligodendrocytes promote glioblastoma progression through CCL5/CCR5-mediated glioma stem cell maintenance.. Neuron 114(2):237-249.e10 PMID: 41570802
