Gastroenterology

Gastroenterology

Volume 154, Issue 5, April 2018, Pages 1524-1537.e6
Gastroenterology

Original Research
Full Report: Basic and Translational—Pancreas
Galectin-3 Mediates Tumor Cell–Stroma Interactions by Activating Pancreatic Stellate Cells to Produce Cytokines via Integrin Signaling

https://doi.org/10.1053/j.gastro.2017.12.014Get rights and content

Background & Aims

Pancreatic ductal adenocarcinoma (PDAC) is characterized by activated pancreatic stellate cells (PSCs), abundance of extracellular matrix (ECM), and production of cytokines and chemokines. Galectin 3 (GAL3), a β-galactoside–specific lectin, contributes to PDAC development but its effects on the stroma and cytokine production are unclear.

Methods

The effect of recombinant human GAL3 (rGAL3) on activation of PSCs, production of cytokines, and ECM proteins was determined by proliferation, invasion, cytokine array, and quantitative polymerase chain reaction. We assessed co-cultures of PDAC cells with GAL3 genetic alterations with PSCs. Production of interleukin 8 (IL8) and activities of nuclear factor (NF)-κB were determined by enzyme-linked immunosorbent assay and luciferase reporter analyses. We studied the effects of inhibitors of NF-κB and integrin-linked kinase (ILK) on pathways activated by rGAL3.

Results

In analyses of the Gene Expression Omnibus database and our dataset, we observed higher levels of GAL3, IL8, and other cytokines in PDAC than in nontumor tissues. Production of IL8, granulocyte-macrophage colony-stimulating factor, chemokine ligand 1, and C-C motif chemokine ligand 2 increased in PSCs exposed to rGAL3 compared with controls. Culture of PSCs with PDAC cells that express different levels of GAL3 resulted in proliferation and invasion of PSCs that increased with level of GAL3. GAL3 stimulated transcription of IL8 through integrin subunit beta 1 (ITGB1) on PSCs, which activates NF-κB through ILK. Inhibitors of ILK or NF-κB or a neutralizing antibody against ITGB1 blocked transcription and production of IL8 from PSCs induced by rGAL3. The GAL3 inhibitor significantly reduced growth and metastases of orthotopic tumors that formed from PDAC and PSC cells co-implanted in mice.

Conclusion

GAL3 activates PSC cells to produce inflammatory cytokines via ITGB1signaling to ILK and activation of NF-κB. Inhibition of this pathway reduced growth and metastases of pancreatic orthotopic tumors in mice.

Section snippets

Materials and Methods

Please refer to the online Supplementary Materials for detailed additional Methods.

Activation of Human PSCs by GAL3

To assess the expression of GAL3 in PDAC tissues, we observed that GAL3 was expressed in both PDAC cells and tumor-associated stromal cells (Supplementary Figure 1A and C) of human PDAC tissues. There was weak expression of GAL3 in normal tissues and human pancreatic ductal epithelial cells (Supplementary Figure 1A and D). In addition, GAL3 was up-regulated in mouse PDAC tissues isolated from several genetic mouse models (LSL-K-RasG12D and other genetic mouse models LSL-p53 and cyclo-oxygenase

Discussion

PDAC is characterized by extreme desmoplastic infiltration that may impede effectiveness of therapy leading to poor patient survival. PSCs are thought to manipulate the TME, producing dense stroma and an abundance of cytokine/chemokines that are increasingly recognized to impart an aggressive tumor phenotype by promoting proliferation, invasion, and metastases. In this study, using cytokine array and co-culture of PSCs and PDAC cells with genetically altered GAL3 levels, we identified that GAL3

References (49)

  • S. Acharyya et al.

    A CXCL1 paracrine network links cancer chemoresistance and metastasis

    Cell

    (2012)
  • S. Wan et al.

    Tumor-associated macrophages produce interleukin 6 and signal via STAT3 to promote expansion of human hepatocellular carcinoma stem cells

    Gastroenterology

    (2014)
  • R.T. Abraham

    Chemokine to the rescue: interleukin-8 mediates resistance to PI3K-pathway-targeted therapy in breast cancer

    Cancer Cell

    (2012)
  • J. Zhao et al.

    Down-regulation of osteopontin suppresses growth and metastasis of hepatocellular carcinoma via induction of apoptosis

    Gastroenterology

    (2008)
  • Y. Nishi et al.

    Role of galectin-3 in human pulmonary fibrosis

    Allergol Int

    (2007)
  • A. Andoh et al.

    Cytokine regulation of chemokine (IL-8, MCP-1, and RANTES) gene expression in human pancreatic periacinar myofibroblasts

    Gastroenterology

    (2000)
  • J. Friedrichs et al.

    Galectin-3 regulates integrin alpha2beta1-mediated adhesion to collagen-I and -IV

    J Biol Chem

    (2008)
  • J.M. Herman et al.

    Analysis of fluorouracil-based adjuvant chemotherapy and radiation after pancreaticoduodenectomy for ductal adenocarcinoma of the pancreas: results of a large, prospectively collected database at the Johns Hopkins Hospital

    J Clin Oncol

    (2008)
  • K. Nakajima et al.

    Galectin-3 cleavage alters bone remodeling: different outcomes in breast and prostate cancer skeletal metastasis

    Cancer Res

    (2016)
  • V.G. Prieto et al.

    Galectin-3 expression is associated with tumor progression and pattern of sun exposure in melanoma

    Clin Cancer Res

    (2006)
  • J.W. Valle et al.

    Optimal duration and timing of adjuvant chemotherapy after definitive surgery for ductal adenocarcinoma of the pancreas: ongoing lessons from the ESPAC-3 study

    J Clin Oncol

    (2014)
  • S. Holter et al.

    Germline BRCA mutations in a large clinic-based cohort of patients with pancreatic adenocarcinoma

    J Clin Oncol

    (2015)
  • F.T. Liu

    Regulatory roles of galectins in the immune response

    Int Arch Allergy Immunol

    (2005)
  • S. Song et al.

    Galectin-3 mediates nuclear beta-catenin accumulation and Wnt signaling in human colon cancer cells by regulation of glycogen synthase kinase-3beta activity

    Cancer Res

    (2009)
  • Cited by (81)

    • The microbiota and aging microenvironment in pancreatic cancer: Cell origin and fate

      2022, Biochimica et Biophysica Acta - Reviews on Cancer
      Citation Excerpt :

      M2 macrophages and Th2 cells are recruited to the TME by microbiota, secrete TGF-β [131,133,145], effectively activate PSCs, mediate the pro-fibroblast proliferation response, secrete a large amount of fibronectin and type I and III collagens, and promote pancreatic fibrosis and the metastasis of cancer cells in the stromal region [77,146]. Microbe-mediated ROS production by immune cells induces senescence in PSCs and promotes SASP secretion [147], including IL8, granulocyte-macrophage colony-stimulating factor, chemokine ligand 1, C-C motif chemokine ligand 2, and dickkopf-3 (DKK3), promoting cancer cell proliferation, invasion and migration, enhancing PDAC resistance to gemcitabine and inhibiting CD8+ T-cell activity [92,148,149]. Furthermore, activated PSCs can be significantly stimulated by factors such as PDGF, VEGF, and TGF-β1 secreted by tumor cells and can proliferate and differentiate into cancer-associated fibroblasts (CAFs) under hypoxic conditions.

    • The interactive role of inflammatory mediators and metabolic reprogramming in pancreatic cancer

      2022, Trends in Cancer
      Citation Excerpt :

      Inflammatory mediators play a central role in PDAC progression by modulating the TME and contributing to PDAC desmoplastic stroma [3]. Additionally, through secretion of inflammatory mediators, growth factors, and enzymes, CAFs contribute to desmoplastic stroma, recruitment of inflammatory and immune cells into TME, and metabolic alterations [3,15–19]. Furthermore, adipocytes, muscle cells, and hepatocytes interact with PDAC cells through IL-6 signaling [20,21], and neurons support PDAC under serine-deprived conditions [22].

    • Strategies targeting tumor immune and stromal microenvironment and their clinical relevance

      2022, Advanced Drug Delivery Reviews
      Citation Excerpt :

      A phase I trial of pamrevlumab (anti-CTGF mAb, FG-3019) combined with chemotherapy in patients with advanced PDAC has demonstrated improved OS [228]. Gal3 is a β-galactoside–binding protein often over-expressed in tumor cells and TME that has been implicated in TAF activation and immune regulation [229]. The combination of belapectin (small molecule Gal3 inhibitor) and anti-PD-1 mAb showed ORR of 50% and 33% in patients with melanoma and SCCHN, associated with increased effector memory T-cell activation and reduced M−MDSCs in responders in a phase I trial [230].

    View all citing articles on Scopus

    Conflicts of interest The authors declare no competing interests in the present study.

    Funding Grant support was provided by the following: American Gastroenterological Association Research Scholar Award (Shumei Song) and Public Health Service Grant DF56338, which supports the Texas Medical Center Digestive Diseases Center (Shumei Song). University of Texas M.D. Anderson Cancer Center IRG (3-0026317; Shumei Song); CA160433 from DOD (Shumei Song). National Key Research & Development Program of China (2016 YFA 0500303), National Natural Science Foundation of China (grant numbers 81330051, 81372594), the ‘‘863’’ Project (2014AA021606 and 2015AA020403), Beijing Natural Science Foundation (7182030) and CA138671, CA129906, CA127672, CA172741, CA150334, and CA160445 from the National Cancer Institute and DOD (Jaffer A. Ajani).

    Author names in bold designate shared co-first authorship.

    View full text