Fgfr 2 in gastrointestinal cancer
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Fibroblast Growth Factor Receptor 2 (FGFR-2) in gastrointestinal cancer Departments of Pathology and Integrative Oncological Pathology Nippon Medical School
Transcript of Fgfr 2 in gastrointestinal cancer
Fibroblast Growth Factor Receptor 2 (FGFR-2) in
gastrointestinal cancer
Departments of Pathology and Integrative Oncological PathologyNippon Medical School
Characteristics of FGFR-2
• Located in chromosome 10q26
• Single transmembrane receptor
• Alternative splicing isoforms
* IIIb and IIIc isoforms (extracellular domain)
* C1, C2 and C3 variants (intracellular domain)
FGFR-2 has more than 20 alternative splicing isoforms. Expressions and roles of these two types of splicing in extracellular and intracellular domains are closely examined in cancers.
FGFR-2
Gene amplification
Overexpression of C3
Missense mutation
Increase of IIIb or IIIc
FGFR-2 in Various CancersGastric Ca, Breast Ca Lung Ca, Endometrial Ca, Melanoma
Risk of Breast and Endometrial Ca
Gastric Ca
Progression of Prostate Ca.EMT of Bladder Ca.
Cervical Ca, Colorectal Ca, Esophageal Ca, Gastric CaPancreatic Ca
SNPs in intron 2
Splicing switch from IIIb to IIIc
Endometrial Ca
Activating mutation
Genetic alterations and overexpression of FGFR-2 have been reported in various. Abnormalities of FGFR-2 are often reported in gastrointestinal cancers.
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FGFR-2 Expression in Colorectal Cancer Tissues
Matsuda et al., Cancer Lett. 309: 209-219, 2011
In colorectal cancer, FGFR2 immunoreactivity was localized at the peripheral lesion of the cancer cell nests, as shown by the arrows.
Inhibitory effects of FGFR-2 in Colorectal Cancer
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Sh-6 and Sh-12: FGFR-2 shRNA stably transfected LoVo cells
Cell proliferation assay in vitro Subcutaneous tumor in nude mice
Matsuda et al., Cancer Lett. 309: 209-219, 2011
Mice injected with FGFR2 shRNA-transected LoVo cells had smaller subcutaneous tumors than mice injected with control cells.These findings suggest that FGFR2 is a therapeutic target for colorectal cancer.
FGFR-2 IIIc isoform in pancreatic cancer
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FGFR-2 IIIc in Human Pancreatic Cancer Cases
anti-sense sense
Ca: pancreatic cancerArrows: fibroblasts
Immuohistochemistry In situ hybridization
In human pancreatic cancer tissues, FGFR2 IIIc and its mRNA were expressed in pancreatic cancer cells and in some fibroblasts close to the cancer cells, as shown by the arrows.
Correlation of Clinicopathological Features and FGFR-2 IIIc Expression in Pancreatic Cancers
FGFR-2 IIIc (-)
FGFR-2 IIIc (+)
FGFR-2 IIIc (-)
FGFR-2 IIIc (+)
Overall survival curves Duration to development of liver metastasis after surgery
Ishiwata et al., Am J Pathol 180: 1928-1941, 2012
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Overall survival curves showed a tendency for poorer prognoses in the FGFR2 IIIc-positive groups. The duration of the development of liver metastasis after surgery was significantly shorter in IIIc-positive groups than in those of the negative groups.
Preparation of FGFR-2 IIIc Stably Transfected KLM-1 Cells
Ishiwata et al., Am J Pathol 2012
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FGFR-2 IIIc mRNA Flow cytometry
As shown here, IIIc mRNA levels were markedly higher in two IIIc-transfected clones, but not in mock cells. Flow cytometry revealed that higher IIIc-expressing cells on the cell surface were more common in IIIc-transfected cells than in mock cells.
Orthotopic Implantation Model of FGFR-2 IIIc-transfected cells
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In the orthotopic implantation model, IIIc-transfected cells also formed larger primary tumors compared to the mock cells. Moreover, pancreatic tumor weights were significantly elevated in IIIc-transfected cells compared to those with mock cells.
Liver Metastasis in Orthotopic Implantation Model
Ishiwata et al., Am J Pathol 2012
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Both the amount and weight of liver metastases were increased in the IIIc-transfected cells.
Effects of siRNA targeting FGFR-2 IIIc on PANC-1 cell growth
qRT-PCR
Flow cytometry
Cell proliferation rates
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The siRNA decreased IIIc mRNA and IIIc protein expression, and the proliferation rates of these cells were lower than those in control siRNA transfected cells.
Effects of Polyclonal anti-FGFR-2 IIIc on Pancreatic Cancer Cell Growth and Migration
PANC-1: High FGFR-2 IIIc expression cellKLM-1: Low FGFR-2 IIIc expression cell
Inhibition of cell proliferation Inhibition of migration
Ishiwata et al., Am J Pathol 2012
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A significant inhibition of proliferation was observed after the addition of the anti-FGFR2 IIIcantibody in PANC-1 cells. The anti-FGFR2 IIIc antibody also inhibited the migration of PANC-1 cells.
FGFR-2 IIIc isoform in colorectal cancer
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Immunoreactivity of FGFR-2 IIIc Survival Curves of FGFR-2IIIchigh and low colorectal cancer
FGFR-2 IIIc (-)
FGFR-2 IIIc (+)
The percentages of FGFR2 IIIc-positive cells in these lesions increased in the order shown here. The group in which FGFR2 IIIc was highly expressed in colorectal cancer cells, showed shorter survival rates.
Matsuda et al., Mol Cancer Ther 11: 2010-2020, 2012
FGFR-2 IIIc-transfected DLD-1 cells in vivo
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Fgfr-9
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In subcutaneous transplantation studies, FGFR2 IIIc transfected colorectal cancer cells formed larger tumors than mock cells. Furthermore, FGFR2 IIIc transfected cells formed larger tumors in the orthotopic implantation model, as shown by the arrows. One out of three animals with IIIc transfected cells showed metastasis in their small intestines, as shown by the white arrow.
Matsuda et al., Mol Cancer Ther, 2012
Inhibition of CRC cell growth using human monoclonal anti-FGFR-2 IIIc antibody
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Human monoclonal anti-FGFR2 IIIc antibody was prepared by phage display method.
Cell proliferation
rhIIIb rhIIIc
FGFR-2 IIIc
human IgG
Western blot
For possible clinical applications in the future, we prepared a human monoclonal anti-FGFR2 IIIcantibody using the phage-display method. The antibody reacted with recombinant human FGFR2 IIIc, but it did not react with recombinant FGFR2 IIIb. Cell proliferation was significantly inhibited by the addition of the human monoclonal anti-FGFR2 IIIc antibodies.
Matsuda et al., Mol Cancer Ther, 2012
Inhibition of CRC cell migration using human monoclonal anti-FGFR-2 IIIc antibody
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In both Boyden chamber and time lapse analyses, cell migration was also significantly inhibited by the addition of the human monoclonal anti-FGFR2 IIIc antibodies.These findings suggest that FGFR2 IIIc plays important roles in the carcinogenesis of colorectal cancer and tumor progression. Matsuda et al., Mol Cancer Ther, 2012
Summary of FGFR-2 IIIc
IHC analysis of FGFR-2 IIIc
Pancreatic cancer Liver metastasis
Colorectal cancer Overall survival↓
FGFR-2 IIIc-transfected cell Growth Migration Invasion Metastasis Attachment
PancreaticCancer ↑ ↑ ↑ ↑ →
ColorectalCancer ↑ ↑ ↑ ↑ ↓
Clinicopathological studies
Experimental studies
Clinicopathologically, FGFR2-IIIc correlated with liver metastasis in pancreatic cancer and overall survival in colorectal cancers. Experimental studies using transplantation models showed that FGFR-2 IIIc contributes to the aggressiveness of these cancers.
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