BCL2 Regulates Differentiation of Intestinal Fibroblasts.
- Weder B Department of Gastroenterology and Hepatology, University Hospital Zurich, Zurich, Switzerland.
- Mamie C Department of Gastroenterology and Hepatology, University Hospital Zurich, Zurich, Switzerland.
- Rogler G Department of Gastroenterology and Hepatology, University Hospital Zurich, Zurich, Switzerland.
- Clarke S AbbVie Bioresearch Center, AbbVie, Worcester, Massachusetts.
- McRae B AbbVie Bioresearch Center, AbbVie, Worcester, Massachusetts.
- Ruiz PA Department of Gastroenterology and Hepatology, University Hospital Zurich, Zurich, Switzerland.
- Hausmann M Department of Gastroenterology and Hepatology, University Hospital Zurich, Zurich, Switzerland.
- 2018-05-26
Published in:
- Inflammatory bowel diseases. - 2018
BCL2 antagonist ABT-737
apoptosis
intestinal fibrosis
myofibroblast differentiation
transplantation
Aged
Animals
Biphenyl Compounds
Cell Culture Techniques
Cell Differentiation
Dextran Sulfate
Female
Fibroblasts
Fibrosis
Humans
Intestinal Mucosa
Intestines
Mice
Mice, Inbred C57BL
Middle Aged
Nitrophenols
Piperazines
Proto-Oncogene Proteins c-bcl-2
Sulfonamides
English
BACKGROUND
Fibrosis in patients with Crohn's disease (CD) results from an imbalance toward excessive fibrous tissue formation driven by fibroblasts. Activation of fibroblasts is linked to the B-cell lymphoma 2 (BCL2) family, which is involved in the induction of apoptosis. We investigated the impact of BCL2 repression on fibrogenesis.
METHODS
The model of dextran sodium sulfate (DSS)-induced chronic colitis and the heterotopic transplantation model of fibrosis were used. Following the administration of the BCL2 antagonist (ABT-737, 50 mg/kg/d), collagen layer thickness and hydroxyproline (HYP) content were determined. Fibroblasts were stimulated with the BCL2 antagonist (0.01-100 µM). BCL2, alpha smooth muscle actin (αSMA), and collagen I (COL1A1) were determined by quantitative polymerase chain reaction (qPCR), immunofluorescence microscopy (IF), and western blot (WB). mRNA expression pattern was determined by next-generation sequencing (NGS).
RESULTS
Collagen layer thickness was significantly decreased in both DSS-induced chronic colitis and the transplantation model of fibrosis upon BCL2 antagonist administration compared with vehicle. Decreased HYP content confirmed the preventive effects of the BCL2 antagonist on fibrosis. In vitro, a significant increase in PI+/annexin V+ human colonic fibroblasts was determined by fluorescence-activated cell sorting upon treatment with high-dose BCL2 antagonist; at a lower dose, αSMA, COL1A1, and TGF were decreased. NGS, IF, and qPCR revealed decreased expression and nuclear translocation of GATA6 and SOX9, known for reprogramming fibroblasts.
CONCLUSION
BCL2 antagonist administration partially prevented fibrogenesis in both fibrosis models. The BCL2 antagonist reduced the expression of TGFβ-induced factors involved in differentiation of myofibroblasts, and therefore might represent a potential treatment option against CD-associated fibrosis.
Fibrosis in patients with Crohn's disease (CD) results from an imbalance toward excessive fibrous tissue formation driven by fibroblasts. Activation of fibroblasts is linked to the B-cell lymphoma 2 (BCL2) family, which is involved in the induction of apoptosis. We investigated the impact of BCL2 repression on fibrogenesis.
METHODS
The model of dextran sodium sulfate (DSS)-induced chronic colitis and the heterotopic transplantation model of fibrosis were used. Following the administration of the BCL2 antagonist (ABT-737, 50 mg/kg/d), collagen layer thickness and hydroxyproline (HYP) content were determined. Fibroblasts were stimulated with the BCL2 antagonist (0.01-100 µM). BCL2, alpha smooth muscle actin (αSMA), and collagen I (COL1A1) were determined by quantitative polymerase chain reaction (qPCR), immunofluorescence microscopy (IF), and western blot (WB). mRNA expression pattern was determined by next-generation sequencing (NGS).
RESULTS
Collagen layer thickness was significantly decreased in both DSS-induced chronic colitis and the transplantation model of fibrosis upon BCL2 antagonist administration compared with vehicle. Decreased HYP content confirmed the preventive effects of the BCL2 antagonist on fibrosis. In vitro, a significant increase in PI+/annexin V+ human colonic fibroblasts was determined by fluorescence-activated cell sorting upon treatment with high-dose BCL2 antagonist; at a lower dose, αSMA, COL1A1, and TGF were decreased. NGS, IF, and qPCR revealed decreased expression and nuclear translocation of GATA6 and SOX9, known for reprogramming fibroblasts.
CONCLUSION
BCL2 antagonist administration partially prevented fibrogenesis in both fibrosis models. The BCL2 antagonist reduced the expression of TGFβ-induced factors involved in differentiation of myofibroblasts, and therefore might represent a potential treatment option against CD-associated fibrosis.
- Language
-
- English
- Open access status
- green
- Identifiers
-
- DOI 10.1093/ibd/izy147
- PMID 29796658
- Persistent URL
- https://sonar.ch/global/documents/38259
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