Epithelial-Myeloid cell crosstalk regulates acinar cell plasticity and pancreatic remodeling in mice
- University of Michigan, United States
- Fourth Military Medical University, China
Figures
Figure 1 with 1 supplement
Myeloid cells are required for PanIN maintenance.
(A) Genetic makeup of the iKras*;CD11b-DTR mouse model. (B) Experimental design, n = 7 mice/cohort. (C) H&E staining of iKras* and iKras*;CD11b-DTR pancreata 3 weeks post pancreatitis induction and …
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Figure 1—source data 1
Histopathological analysis (related to Figure 1D).
- https://doi.org/10.7554/eLife.27388.005
Figure 1—figure supplement 1
Depletion of multiple myeloid subsets results in redifferentiation of epithelial cells and remodeling of the stroma.
(A) Experimental design. (B) Flow Cytometry analysis of myeloid cells in wild type (WT) and CD11b-DTR pancreata one day following acute pancreatitis induction. (C) Immunohistochemistry for EGFP of …
Figure 2 with 1 supplement
Myeloid cell depletion induces PanIN regression and stroma inactivation.
(A) Periodic acid–Schiff (PAS) staining; (B) immunohistochemistry staining for Cleaved caspase three and (C) F4/80 in iKras* and iKras*;CD11b-DTR pancreata 3 week post pancreatitis induction and in …
Figure 2—figure supplement 1
CD8 T cell depletion does not affect progression of low-grade PanIN lesions, nor PanIN regression upon myeloid cell depletion.
(A) Experimental design, n = 3 ~ 4 mice/cohort. Oncogenic Kras was activated via Dox adminitration in iKras* and iKras*-CD11b-DTR mice; acute pancreatitis was then induced. Three weeks following the …
Figure 3 with 1 supplement
Oncogenic Kras in epithelial cells regulates macrophages polarization and function.
(A) Experimental design, n = 4 ~ 8 mice/cohort. (B) Immunohistochemistry staining for F4/80 in iKras* pancreata 3 weeks post pancreatitis and 3 days, 1 week upon Kras* inactivation. Scale bar 50 µm. …
Figure 3—figure supplement 1
Changes in expression of EGFR family receptors in epithelial cells following Kras inactivation.
qRT-PCR for Egfr and Erbb4 expression in epithelial cells (EGFP+CD45-) freshly sorted from control normal pancreata, iKras* pancreata 3 weeks post pancreatitis, iKras* and iKras*;CD11b-DTR pancreata …
Figure 4 with 1 supplement
Myeloid cells are required for tissue remodeling upon Kras* inactivation.
(A) Experimental design (n = 4 ~ 8 mice/cohort). (B) H&E staining; (C) Gomori Trichrome staining in iKras* and iKras*;CD11b-DTR pancreata 3 weeks post pancreatitis induction and 3 days, 1 week …
Figure 4—figure supplement 1
Myeloid cells are required for pancreatic acinar cell re-differentiation upon Kras* inactivation.
(A) Periodic acid–Schiff (PAS) staining; (B) EGFP immunohistochemistry staining in iKras* and iKras*;CD11b-DTR pancreata 3 weeks post pancreatitis induction and 3 days, 1 week upon Kras* …
Figure 5 with 2 supplements
Myeloid cells regulate acinar re-differentiation, cell survival and stromal activation of EGFR/MAPK signaling.
(A) Co-immunofluorescent staining for CK19 (green), Amylase (red), SMA (magenta) and DAPI (blue); (B) co-immunofluorescent staining for Cleaved caspase 3 (green), E-cadherin (magenta) and DAPI …
Figure 5—figure supplement 1
Acinar cell regeneration upon Kras* inactivation requires stromal activation of MAPK signaling.
(A) Immunohistochemistry for Ki67 in iKras* and iKras*;CD11b-DTR pancreata 3 weeks post pancreatitis induction and 3 days, 1 week upon Kras* inactivation and DT treatment. Scale bar 25 µm. (B) …
Figure 5—figure supplement 2
Cell death induced by myeloid cell depletion upon Kras* inactivation is not mediated by CD8+T cells.
(A) Experimental design, n = 2 ~ 7 mice/cohort. (B) H&E staining and immunohistochemistry staining for Cleaved caspase three in iKras* or iKras*;CD11b-DTR pancreata 3 weeks post pancreatitis …
Figure 6 with 1 supplement
Oncogenic Kras regulates EGFR ligands and MMPs production.
(A) qRT-PCR for Egf, Tgfα, Hbegf, Areg, Ereg, Mmp1, Mmp2, Mmp9, Mmp12 and Mmp14 expression in littermate control, iKras* pancreata 3 weeks post pancreatitis, iKras* and iKras*;CD11b-DTR pancreata 3 …
Figure 6—figure supplement 1
Changes in expression of EGFR ligands in epithelial cells following Kras inactivation.
qRT-PCR for Egf, Tgfα and Hbegf expression in epithelial cells (EGFP+CD45-) freshly sorted from control normal pancreata, iKras* pancreata 3 weeks post pancreatitis, iKras* and iKras*;CD11b-DTR …
Figure 7
EGFR inhibition impairs ECM remodeling and tissue repair upon Kras* inactivation.
(A) Experimental design, n = 4 ~ 6 mice/cohort. (B) H&E staining, Gomori Trichrome staining (inset), co-immunofluorescent staining for p-ERK1/2 (green), CK19 (red), SMA (magenta) and DAPI (blue), …
Figure 8
Stroma remodeling upon Kras* inactivation depends on MEK/ERK activity in stroma.
(A) Experimental design, n = 4 ~ 5 mice/cohort. (B) H&E staining, Gomori Trichrome staining (inset), co-immunofluorescent staining for p-ERK1/2 (green), CK19 (red), SMA (magenta) and DAPI (blue) and …
Figure 9
Diagram depicting our working model.
(A) In the presence of epithelial oncogenic Kras expression, myeloid cells are required to maintain high MAPK activity in the epithelial cells themselves, and thus maintain de-differentiation and …
Additional files
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Supplementary file 1
Primary antibodies used in this study
- https://doi.org/10.7554/eLife.27388.020
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Supplementary file 2
Primer sequences for quantitative RT-PCR
- https://doi.org/10.7554/eLife.27388.021
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Transparent reporting form
- https://doi.org/10.7554/eLife.27388.022
