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 6

Effects of DIO on tumor immunity in allogeneic transplantation–model mice with normal immunity.

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Effects of DIO on tumor immunity in allogeneic transplantation–model mic...
(A) Schematic representation showing the protocol for the subcutaneous injection of 1.0 × 105 KPCY tumor cells cotransformed with 1.0 × 105 myPafs, tapPafs, or total PSCs in syngeneic B6 mice fed a high-fat diet to generate DIO. (B) The tumor growth rate was evaluated in DIO B6 mice after subcutaneous tumor transplantation, with n = 4 mice per condition. (C) Acta2 and (D) Cxcl13 mRNA expression was quantitatively evaluated in the tumors (n = 4 per each group). (E) Representative images of pathological H&E staining and IHC analysis of immune cell infiltration of CD3, CD8, and B220 in tumors transplanted with myPafs and tapPafs (original magnification, ×20). (F) The densities of total cells and CD3-, CD8- and B220-positive cells quantitatively evaluated in IHC sections of the transplanted tumors (n = 4 per each group). The data are presented as the mean ± SD. Statistical analysis was performed by 2-way ANOVA with post hoc multiple-comparison tests. TV, tumor volume; HFD, high-fat diet; OGTT, oral glucose tolerance test; ITT, insulin tolerance test; DIO, diet-induced obesity; CD, control diet; Pafs, pancreatic fibroblasts; myPafs, myofibroblastic Pafs; tapPafs, tumor immunity- and angiogenesis-promoting Pafs; PSCs, pancreatic stellate cells: MK, mPafs+KPCY; DMK, DIO/mPafs+KPCY; TK, tapPafs+KPCY; DTK, DIO/tapPafs+KPCY; Fd, field. *P < 0.05, **P < 0.01, ***P < 0.001. Scale bar: 100 μm.