| 2011 |
CCL18 released by breast tumor-associated macrophages promotes cancer cell invasiveness by triggering integrin clustering and enhancing adherence to extracellular matrix. PITPNM3 was identified as a functional receptor for CCL18 that mediates these effects and activates intracellular calcium signaling. Suppressing PITPNM3 abrogated CCL18-induced invasion and metastasis of breast cancer xenografts. |
Co-immunoprecipitation, calcium signaling assays, siRNA knockdown of PITPNM3, in vitro invasion assays, xenograft mouse models |
Cancer cell |
High |
21481794
|
| 2013 |
Human CCR8 is a functional receptor for CCL18. CCL18 induced chemotaxis and calcium flux in CCR8-transfected cells, bound CCR8 with high affinity, and induced CCR8 internalization. CCL18 and CCL1 (the known CCR8 ligand) competed for CCR8 binding and induced heterologous cross-desensitization. CCL18 induced chemotaxis and calcium flux of human Th2 cells through CCR8, and wild-type but not Ccr8-deficient mouse Th2 cells migrated in response to CCL18. |
Receptor-transfected cell chemotaxis and calcium flux assays, radioligand competition binding, receptor internalization assay, cross-desensitization assay, Ccr8-knockout mouse cells |
The Journal of experimental medicine |
High |
23999500
|
| 2015 |
Crystal structure of human CCL18 was determined. CCL18 adopts an α-helical conformation at its N-terminus that weakens dimerization, explaining its preference for the monomeric state. CCL18 can reversibly form a unique open-ended oligomer distinct from CCL3, CCL4, and CCL5 oligomers. Lysine at position 8 (replacing the conserved proline 8 of CCL3/CCL4) is the molecular determinant for these differences. Insulin-degrading enzyme degrades CCL3 and CCL4 but not CCL18, explained by structural differences. |
X-ray crystallography, small-angle X-ray scattering (SAXS), mutational analysis, biochemical degradation assays |
Journal of molecular biology |
High |
25636406
|
| 2006 |
CCL18 stimulates collagen production in primary human pulmonary fibroblasts through a PKCα-dependent pathway. CCL18 stimulation causes PKCα (but not PKCδ or PKCε) to undergo rapid phosphorylation and nuclear translocation. PKCα activates ERK2 phosphorylation, which drives collagen production. CCL18 also increases intracellular calcium, which is required for PKCα nuclear translocation and downstream collagen production. This signaling is independent of autocrine TGF-β. |
Dominant-negative PKC mutant transfection, pharmacologic PKC inhibitor (Gö6976), intracellular calcium chelation (BAPTA), collagen promoter reporter assay, ERK2 phosphorylation Western blot, TGF-β neutralizing antibodies |
American journal of respiratory cell and molecular biology |
High |
16601239
|
| 2006 |
CCL18-stimulated upregulation of collagen production in lung fibroblasts requires Sp1 signaling and basal Smad3 activity. CCL18 increased Sp1 phosphorylation, Sp1 DNA-binding, and Sp1-dependent reporter activity. Dominant-negative mutants of Sp1 and Smad3 abrogated CCL18-dependent collagen production. CCL18 did not increase TGF-β levels, and TGF-β neutralization did not block CCL18-induced collagen production, indicating TGF-β-independent signaling. |
Dominant-negative Sp1 and Smad3 transfection, Sp1 DNA-binding assay, reporter gene assay, TGF-β neutralizing antibodies, collagen mRNA and protein measurement |
Journal of cellular physiology |
High |
16021625
|
| 2006 |
Overexpression of CCL18 in mouse lungs via adenoviral gene transfer triggered selective, prolonged perivascular and peribronchial T-lymphocyte infiltration and collagen accumulation. The collagen deposition was T-lymphocyte-dependent (not directly CCL18-dependent), as depletion with antilymphocyte serum abrogated it. CCL18 prestimulation of T lymphocytes in vitro upregulated TGF-β1 and enhanced collagen production in T lymphocyte/fibroblast co-cultures through a TGF-β1-dependent mechanism. |
Adenoviral gene transfer in vivo, antilymphocyte serum depletion, flow cytometry, immunohistochemistry, T lymphocyte/fibroblast co-culture |
Arthritis and rheumatism |
Medium |
16868995
|
| 2007 |
In a bleomycin injury model, CCL18 overexpression had additive effects on T-cell infiltration but unexpectedly attenuated bleomycin-induced collagen accumulation. CCL18-stimulated T-lymphocytic infiltration drove MMP-2 and MMP-9 elevation. Pharmacological inhibition of MMPs (GM6001) or T-cell depletion abrogated collagen accumulation and changes in TNF-α, IFN-γ, and active TGF-β1, demonstrating that in a pro-fibrotic milieu, CCL18-driven T-cell infiltration and MMP induction can paradoxically reduce fibrosis. |
Adenoviral CCL18 overexpression combined with bleomycin instillation, antilymphocyte serum depletion, MMP inhibitor GM6001, collagen and cytokine measurement |
The American journal of pathology |
Medium |
17569779
|
| 2006 |
Alveolar macrophages from IPF patients produce CCL18 that stimulates collagen production by normal lung fibroblasts, partly mediated via CCL18. Native collagen significantly up-regulated CCL18 expression in normal AMs activated with Th2 cytokines via a mechanism mediated by β2-integrin/scavenger receptor(s), creating a positive feedback loop. |
Bronchoalveolar lavage cell culture, co-culture experiments, cytokine stimulation, flow cytometry, in situ hybridization, ELISA |
American journal of respiratory and critical care medicine |
Medium |
16415274
|
| 2003 |
CCL18 is constitutively produced by immature dendritic cells and is selectively down-regulated during DC maturation induced by LPS, TNF, CD40L, and microbial stimuli — opposite to all other chemokines tested. IL-10 and vitamin D3 strongly promoted CCL18 secretion, while IFN-γ inhibited it. CCL18 was found to be a chemotactic factor for immature DCs. |
DC maturation assays with various stimuli, ELISA, chemotaxis assays with immature DCs |
Journal of immunology |
Medium |
12646652
|
| 2001 |
CCL18/DC-CK1 produced by germinal center dendritic cells is a pertussis toxin-dependent chemoattractant for B lymphocytes with a preference for attracting CD38-negative mantle zone B cells. |
Chemotaxis assay with B lymphocyte subsets, pertussis toxin inhibition, immunohistochemistry, RNA analysis |
Journal of immunology |
Medium |
11207283
|
| 1999 |
Recombinant or synthetic CCL18 (MIP-4) induced calcium mobilization in naive and activated T lymphocyte subpopulations in vitro and, upon intraperitoneal injection in mice, led to the selective accumulation of CD4+ and CD8+ T lymphocytes but not monocytes or granulocytes, demonstrating T-cell-selective chemotactic activity in vivo. |
Calcium mobilization assay, in vivo peritoneal injection chemotaxis model, flow cytometry |
Genomics |
Medium |
10087196
|
| 2014 |
Upon CCL18 binding to PITPNM3 in breast cancer cells, Pyk2 translocates from the cytoplasm to the plasma membrane to form a stable complex with PITPNM3, subsequently activating Src kinase. Pyk2 and Src are essential for integrin α5/β1 clustering-dependent adherence, migration, and invasion induced by CCL18. |
Co-immunoprecipitation, immunofluorescence localization, Boyden chamber migration/invasion assay, Western blot, siRNA knockdown |
Journal of cellular biochemistry |
Medium |
24142406
|
| 2015 |
CCL18 promotes angiogenesis in breast cancer through PITPNM3 on endothelial cells. Silencing PITPNM3 on HUVECs abrogated CCL18-mediated pro-migration and tube formation independently of VEGFR signaling. CCL18 induced endothelial-mesenchymal transformation and activated ERK and Akt/GSK-3β/Snail signaling in HUVECs. |
PITPNM3 siRNA knockdown in HUVECs, tube formation assay, migration assay, Western blot for ERK/Akt/GSK-3β/Snail, in vivo angiogenesis model, double immunohistochemistry |
Oncotarget |
Medium |
26416449
|
| 2015 |
CCL18 enhances HCC cell migration, invasion, and epithelial-mesenchymal transition through PITPNM3 and activation of NF-κB signaling (phosphorylation of IKK and IκBα, p65 nuclear translocation). Silencing PITPNM3 by siRNA inhibited all these CCL18-induced effects. |
siRNA knockdown of PITPNM3, migration/invasion assays, Western blot for IKK/IκBα phosphorylation and p65 nuclear translocation, EMT marker analysis |
Tumour biology |
Medium |
26449829
|
| 2022 |
CCL18 secreted by TAMs activates NF-κB signaling via PITPNM3 in normal breast fibroblasts, driving IL-6 and IL-8 production and conversion to CD10+GPR77+ cancer-associated fibroblasts. These CAFs enrich cancer stem cells and induce chemoresistance. Anti-CCL18 antibody blocked CAF formation and restored chemosensitivity in vivo. |
Co-culture assays, PITPNM3 siRNA knockdown, NF-κB signaling Western blot, flow cytometry for CAF markers, in vivo xenograft with intratumoral CCL18 injection, anti-CCL18 antibody treatment |
Oncogene |
Medium |
36418470
|
| 2016 |
CCL18 in ovarian cancer ascites promotes cancer cell migration through activation of proline-rich tyrosine kinase 2 (Pyk2). CCL18 stimulated phosphorylation of Pyk2 in OC cell lines; siRNA-mediated downregulation of Pyk2 markedly inhibited CCL18-induced cell migration. CCL18 blocking antibodies attenuated ascites-induced cell migration. |
Boyden chamber migration assay, siRNA knockdown of Pyk2, Western blot for Pyk2 phosphorylation, CCL18 neutralizing antibody, exogenous gene expression of Pyk2 |
Molecular cancer |
Medium |
27613122
|
| 2016 |
CCL18 promotes migration and invasion of non-small cell lung cancer cells by binding to PITPNM3/Nir1 receptor. CCL18 modulates cell migration and invasion by activating RAC1 in an ELMO1-dependent manner, causing cytoskeleton reorganization, and enhances adhesion via ELMO1-integrin β1 signaling. |
Receptor binding assay (Nir1/PITPNM3), siRNA knockdown, RAC1 activation assay, cytoskeleton imaging, integrin β1 functional assay |
Molecular carcinogenesis |
Medium |
26756176
|
| 2010 |
CCL18 attenuates CXCR4-mediated responses (calcium mobilization, chemotaxis, pseudo-emperipolesis, proliferation) in pre-B ALL cells via interactions with GPR30. CCL18 bound to GPR30-expressing cells, and anti-GPR30 antibodies abolished this binding and CCL18-mediated functional effects. Direct effects on CXCR4 were excluded. |
Calcium mobilization assay, chemotaxis assay, anti-GPR30 antibody blocking, binding assay with GPR30-expressing cells, CXCR4 binding exclusion |
Journal of cellular physiology |
Medium |
20568229
|
| 2013 |
CCL18 has regulatory activity inhibiting CCR1, CCR2, CCR3, CCR4, and CCR5-mediated chemotaxis. CCL18 is a competitive functional antagonist at CCR3 but only binds a subset of CCR3 receptor populations and does not bind the other CCR receptors it inhibits. Instead, CCL18 displaces glycosaminoglycan (GAG)-bound chemokines from heparin, preventing their presentation to receptors. Chemokines from all four chemokine sub-classes displace cell-bound CCL18, indicating a GAG displacement mechanism. |
Schild plot analysis for receptor antagonism, chemotaxis assays with multiple CCR receptors, heparin competition binding assays, cell-bound CCL18 displacement assays |
PloS one |
Medium |
23951310
|
| 2012 |
CCL18 promotes chemotaxis of CD4+CD25+CD127low regulatory T cells in a dose-dependent manner. CCL18-recruited memory T cells produce IL-10 and TGF-β1, and inhibit effector T-cell proliferation through an IL-10-dependent mechanism. In vivo intradermal injection of CCL18 in SCID mice grafted with human skin recruited CD4+, CD25+, and IL-10+ cells. |
Chemotaxis assays with different Treg subsets, cytokine production analysis, T-cell proliferation suppression assay, in vivo SCID mouse model with human skin graft |
Journal of immunology |
Medium |
22649201
|
| 2011 |
CCL18 differentiates monocytes into tolerogenic dendritic cells (semimature phenotype with increased CCR7 expression) that induce IL-10 production in allogenic naive T cells and generate type 1 T regulatory cells. This tolerogenic DC differentiation by CCL18 is dependent on induction of indoleamine 2,3-dioxygenase through an IL-10-mediated mechanism. The tolerogenic effect was absent when DCs originated from allergic patients. |
Monocyte DC differentiation assay with CCL18, flow cytometry for DC phenotype, co-culture with naive T cells, T-regulatory function assay, IDO inhibition, IL-10 blocking antibody |
Blood |
Medium |
21803856
|
| 2010 |
CCL18 converts memory CD4+CD25- T cells from healthy subjects into CD4+CD25+Foxp3+ regulatory T cells capable of suppressing effector T-cell proliferation by both cytokine and cell contact-dependent mechanisms. This regulatory effect was lost with T cells from allergic patients, correlated with decreased CCL18 binding to these cells. |
T-cell polarization assay, Foxp3 and cytokine expression analysis, T-cell suppression assay, CCL18 binding assay (MFI), healthy versus allergic donor comparison |
FASEB journal |
Medium |
20702776
|
| 2012 |
CCL18 causes monocytes to mature into M2-spectrum macrophages expressing CD14, CD206, and 15-lipoxygenase but not mature DC markers. CCL18-stimulated macrophages showed high capacity for unspecific phagocytosis and pinocytosis without oxidative burst, and produced CXCL8, CCL2, CCL3, CCL22, IL-10, and PDGF but not M1 cytokines IL-1β or IL-12. |
Monocyte culture with CCL18, morphological analysis, flow cytometry for macrophage markers, cytokine ELISA, phagocytosis and pinocytosis functional assays |
Immunology |
Medium |
22117697
|
| 2008 |
Thrombin stimulates the Rho-dependent release of CCL18 from mature LPS-matured dendritic cells via PAR1 and PAR3 receptors. CCL18 release was dependent on ERK1/2 and Rho kinase activation, and was not triggered by TNF-α-matured DCs. CCL18 was colocalized with CD83+ DCs in human atherosclerotic plaques. |
DC maturation assays, thrombin and PAR-activating peptide stimulation, ERK and Rho kinase pharmacological inhibitors, ELISA for CCL18, immunofluorescence in atherosclerotic plaques |
Journal of immunology |
Medium |
18606675
|
| 2018 |
CCL18 promotes migration, invasion, and EMT in bladder cancer cells by binding CCR8. Blocking CCR8 via small molecule inhibitor or shRNA reversed rCCL18-induced decreases in E-cadherin and increases in MMP-2 and VEGF-C, and reversed CCL18-induced invasion, migration, and EMT. |
CCR8 shRNA knockdown, CCR8 small molecule inhibitor, invasion/migration assays, Western blot for EMT markers (E-cadherin, MMP-2, VEGF-C) |
Molecular medicine reports |
Medium |
30592282
|
| 2019 |
CCL18 promotes breast cancer invasion and metastasis through Annexin A2 (AnxA2), which acts as a downstream molecule of Nir1 (PITPNM3). AnxA2 promotes CCL18-induced F-actin polymerization through phosphorylation of integrin β1 and facilitates EMT via the PI3K/Akt/GSK3β/Snail signaling pathway. LY294002 inhibited phosphorylation of AnxA2, linking PI3K to AnxA2 function. |
Western blot for AnxA2 expression, invasion/migration assays, F-actin measurement, xenograft lung metastasis in SCID mice, immunofluorescence, PI3K inhibitor LY294002 |
Oncology reports |
Medium |
31894281
|
| 2018 |
ACAP4 (ARF6 GTPase-activating protein) regulates CCL18-elicited breast cancer cell migration via PCAF acetyltransferase-mediated acetylation at Lys311. Dynamic ACAP4 acetylation is essential for CCL18-induced migration; persistent acetylation-mimicking or non-acetylatable ACAP4 mutants blocked CCL18-elicited migration. Mechanistically, acetylation of ACAP4 at Lys311 reduced lipid-binding activity, enabling dynamic ARF6-ACAP4 cycling with the plasma membrane. |
Mass spectrometry acetylation site mapping, PCAF co-immunoprecipitation, acetylation-mimicking/non-acetylatable ACAP4 mutants, lipid-binding assay, migration/invasion assays |
Journal of molecular cell biology |
Medium |
30395269
|
| 2020 |
Ezrin is acetylated by PCAF acetyltransferase in breast cancer cells in response to CCL18 stimulation. The acetylation site was mapped by mass spectrometry. Dynamic ezrin acetylation is essential for CCL18-induced cell migration; both persistent acetylation-mimicking and non-acetylatable mutants blocked migration. Mechanistically, ezrin acetylation reduces lipid-binding activity, prevents phosphorylation at Thr567, and induces an unfolded dominant structure as shown by atomic force microscopy. |
Mass spectrometry acetylation site mapping, PCAF co-immunoprecipitation, acetylation mutants (gain/loss of function), lipid-binding assay, atomic force microscopy, migration/invasion assays |
Journal of molecular cell biology |
Medium |
31638145
|
| 2015 |
CCL18 promotes metastasis of epithelial ovarian cancer cells through the mTORC2 pathway. Overexpression of CCL18 enhanced migration and invasion; proteomics analysis identified mTORC2 pathway correlation with CCL18-mediated invasiveness. |
CCL18 overexpression, in vitro migration/invasion assays, in vivo xenograft, proteomics analysis |
Molecular carcinogenesis |
Low |
26457987
|
| 2021 |
M2-TAMs induce EMT of ovarian cancer cells by releasing CCL18. CCL18 induces macrophage colony-stimulating factor (M-CSF) transcription in OvCa cells through zinc finger E-box-binding homeobox 1 (ZEB1), forming a CCL18-ZEB1-M-CSF feedback loop. M-CSF secreted by OvCa cells drives M2-TAM polarization. Blocking ZEB1 impeded spheroid formation and transcoelomic metastasis in mouse models. |
3D co-culture system, transwell co-culture, migration/invasion assays, Western blot for EMT markers, flow cytometry for TAM polarization, RNA-seq, in vivo intraperitoneal OvCa mouse model, ZEB1 knockdown/overexpression |
Journal for immunotherapy of cancer |
Medium |
34969774
|
| 2022 |
CCL18 promotes glioma cell growth and invasion via CCR8 as its functional receptor on glioma cells. ACP5 (acid phosphatase 5) is identified as an important downstream signaling component mediating glioma growth in response to CCL18-CCR8 signaling. Validated in a humanized ex vivo brain slice model with iPSC-derived human microglia and in vivo GBM model. |
iPSC-derived human microglia transplantation into mouse brain slices (humanized model), CCR8 receptor identification, in vitro and ex vivo invasion/growth assays, in vivo GBM model |
Cell reports |
Medium |
35417708
|
| 2018 |
Tumor-associated macrophages promote progression and the Warburg effect in PDAC via the CCL18/PITPNM3/NF-κB/VCAM-1 regulatory pathway. CCL18 from TAMs induces VCAM-1 expression in pancreatic cancer cells via NF-κB; VCAM-1-driven lactate production from cancer cells with enhanced aerobic glycolysis reciprocally activates macrophages to a TAM-like phenotype. |
VCAM-1 siRNA knockdown, flow cytometry for cell cycle analysis, migration/invasion/colony formation assays, mouse xenograft model, ELISA for CCL18 |
Cell death & disease |
Medium |
29670110
|
| 2024 |
Lactate regulates CCL18 expression in macrophages via H3K18 lactylation. Lactate-treated macrophages undergo M2 polarization and secrete CCL18; silencing of Gpr132 in macrophages or treatment with anti-CCL18 antibody reversed lactate-driven promotion of ovarian cancer cell proliferation and migration. H3K18 lactylation at the CCL18 locus was confirmed by ChIP-qPCR and luciferase reporter assays. |
ChIP-qPCR for H3K18 lactylation, luciferase reporter assay, Gpr132 siRNA knockdown, anti-CCL18 antibody treatment, co-culture assay, xenograft model |
Acta biochimica et biophysica Sinica |
Medium |
39010846
|
| 2022 |
CCL18 promotes breast cancer metastasis through exosomal miR-760 activating the ARF6/AMAP1/Src/PI3K/Akt signaling pathway. CCL18 increased ARF6 expression and p-AMAP1, activating downstream Src/PI3K/Akt. ARF6 knockdown impaired CCL18-induced malignant behaviors. CCL18-stimulated high-metastatic cells released miR-760-rich exosomes targeting ARF6 in recipient cells, promoting EMT and chemoresistance. |
ARF6 siRNA knockdown, Western blot for ARF6/p-AMAP1/Src/PI3K/Akt, exosome isolation and uptake assay, miR-760 target validation, in vivo tumor growth and metastasis |
Molecular therapy oncolytics |
Medium |
35399607
|
| 2000 |
The CCL18 (AMAC-1) gene consists of three exons. The promoter contains putative STAT6 and STAT1 binding sites, as well as AP-1 and C/EBP elements. A combined STAT6/STAT1 binding element near the first transcription start point suggests competitive binding of IL-4-induced STAT6 versus IFN-γ-induced STAT1 may explain antagonistic cytokine regulation of CCL18 expression. |
Genomic library screening, PCR amplification, sequencing, reporter gene construct with 2.7-kb promoter fragment |
Cytokine |
Low |
10671296
|
| 2022 |
VHL-deficient kidney epithelial cells secrete IL-6 that induces macrophage infiltration and M2 polarization; these macrophages secrete CCL18 (and TGF-β1) to stimulate EMT of kidney tubule cells. In a human ccRCC xenograft model, exogenous macrophages promoted tumor growth and metastasis in a CCL18-dependent manner. |
Proteomic and genomic analyses of tumor microenvironment, Vhlh conditional knockout mouse model, IL-6-neutralizing antibody treatment, human ccRCC xenograft with human macrophages, CCL18 functional dependency assays |
Cancer research |
Medium |
35666812
|