Type 2 diabetes alters quiescent pancreatic stellate cells to tumor-prone state
Yutaro Hara, Hiroki Mizukami, Takahiro Yamada, Shuji Shimoyama, Keisuke Yamazaki, Takanori Sasaki, Zhenchao Wang, Hanae Kushibiki, Masaki Ryuzaki, Saori Ogasawara, Hiroaki Tamba, Akiko Itaya, Norihisa Kimura, Keinosuke Ishido, Shinya Ueno, Kenichi Hakamada
Yutaro Hara, Hiroki Mizukami, Takahiro Yamada, Shuji Shimoyama, Keisuke Yamazaki, Takanori Sasaki, Zhenchao Wang, Hanae Kushibiki, Masaki Ryuzaki, Saori Ogasawara, Hiroaki Tamba, Akiko Itaya, Norihisa Kimura, Keinosuke Ishido, Shinya Ueno, Kenichi Hakamada
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Research Article Endocrinology Gastroenterology

Type 2 diabetes alters quiescent pancreatic stellate cells to tumor-prone state

Abstract

Pancreatic stellate cells (PSCs) are the origin of cancer-associated fibroblasts. Type 2 diabetes mellitus (T2D) may promote pancreatic ductal adenocarcinoma (PDAC), eliciting changes in the quiescent PSC (qPSC) population from the precancerous stage. However, the details are unknown. We evaluated the subpopulations of qPSCs and the impact of T2D. PSCs isolated from 8-week-old C57BL/6J mice and diabetic db/db mice were analyzed by single-cell RNA-seq. Sorted qPSCs and PDAC cells were transplanted into allogenic mice. The isolated qPSCs were broadly classified into mesothelial cell and pancreatic fibroblast (Paf) populations by single-cell RNA-seq. Pafs were subclassified into inflammatory Pafs, myofibroblastic Pafs (myPafs) and a small population named tumor immunity- and angiogenesis-promoting Pafs (tapPafs), expressing Cxcl13. In the subcutaneous transplantation model, the tumors transplanted with myPafs were significantly larger than the tumors transplanted with tapPafs. An increase in myPafs and a decrease in tapPafs were observed from the precancerous stage in human T2D, indicating the effects of tumor progression. This study revealed the subpopulation changes in qPSCs in T2D. A therapy that increases the number of tapPafs could be a therapeutic option for patients with PDAC and T2D and even those in a precancerous stage of T2D.

Authors

Yutaro Hara, Hiroki Mizukami, Takahiro Yamada, Shuji Shimoyama, Keisuke Yamazaki, Takanori Sasaki, Zhenchao Wang, Hanae Kushibiki, Masaki Ryuzaki, Saori Ogasawara, Hiroaki Tamba, Akiko Itaya, Norihisa Kimura, Keinosuke Ishido, Shinya Ueno, Kenichi Hakamada

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Figure 1

PSCs are quiescent in the pancreas of nondiabetic and diabetic mice.

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PSCs are quiescent in the pancreas of nondiabetic and diabetic mice. (A)...
(A) Schematic overview depicting the approach used for the experiments. (B) H&E and silver staining showing pathological findings in the pancreas of WT and db/db mice (original magnification, ×10 and ×20). (C) Immunofluorescence image showing FABP4-positive and vWF-negative PSCs in the stroma of the pancreas from WT and db/db mice (original magnification, ×20). (D) Semiquantification of fibrosis in WT (n = 5) and db/db (n = 5) mice. (E) The density of FABP4-positive and vWF-negative cells in WT (n = 5) and db/db mice (n = 5). (F) Morphology of PSCs isolated from the pancreas. Lipid droplets detected by oil red O staining (arrows) in WT mice (n = 8). Immunofluorescence analysis of FABP4 expression in isolated PSCs (original magnification, ×20). (G) Time course of Acta2 mRNA expression in PSCs after isolation evaluated by qPCR (n = 4 per each time point). (H) Representative flow cytometry plots showing retinol-positive cell clusters evaluated with indo-1 and EPCAM antibody, or indo-1 and CD45 antibody. The data are presented as the means ± SDs. Statistical analysis was performed by 2-way ANOVA with post hoc multiple-comparison tests. PSCs, pancreatic stellate cells; vWF, von Willebrand factor. *P < 0.01 and †P < 0.05. Scale bars: 100 μm (B, H&E staining) or 50 μm (others).