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Combined blockade of TGf-β1 and GM-CSF improves chemotherapeutic effects for pancreatic cancer by modulating tumor microenvironment

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Abstract

The interactions between tumor immune microenvironment (TIME) and pancreatic cancer cells can affect chemotherapeutic efficacy; however, the mechanisms still remain largely unknown. Thirty items in TIME were comprehensively screened by using tissue microarray from pancreatic cancer patients. Their expressions, interconnections and predictive roles for survival were analyzed. Twenty-one of 30 items could stratify the survival of the patients; however, multivariate analysis found that only 5 independent risk factors could predict worse survival (M2-polarized tumor-associated macrophages (TAMs), IgG4 positive cells, TGF-β1, GM-CSF and lymphangiogenesis). They had a much higher expression levels in tumoral tissue, compared to peritumoral tissue. The Spearman analysis showed that M2-polarized TAM, TGF-β1 and GM-CSF were positively correlated with pancreatic cancer stem cells (PCSC), angiogenesis and lymphangiogenesis. Both human and murine pancreatic cancer cells could induce M2-polarized TAM, which showed substantial roles to decease chemotherapeutic effects. After treated by gemcitabine, both human and murine pancreatic cancer cell lines expressed higher level of immune check points, PCSC markers and varieties of immunosuppressive factors; however, TGF-β1 and GM-CSF had the highest increase. Based on the above results, TGF-β1 and GM-CSF were proposed to be the optimal potential targets to improve chemotherapeutic effects. In immunocompetent murine models, we demonstrated that combined blockade of TGF-β1 and GM-CSF improved the chemotherapeutic effects by inhibition of M2-polarized TAM and induction of CD8 positive T cells. This study presents a novel promising combined strategy to improve the chemotherapeutic effects for pancreatic cancer.

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Abbreviations

ACT:

Adjuvant chemotherapy

ARG1:

Arginase1

CSF:

Colony stimulating factor

CTL:

Cytotoxic T lymphocytes

Gem:

Gemcitabine

GM/M-CSF:

Granulocyte macrophage/macrophage colony stimulating factor

IFN-γ:

Interferon-γ

IL:

Interleukin

iNOS:

Inducible nitric oxide synthase

LPS:

Lipopolysaccharides

MDSC:

Myeloid-derived suppressor cells

MLVD:

Microlymphatic vascular density

MVD:

Microvascular density

PCSC:

Pancreatic cancer stem cells

PDAC:

Pancreatic ductal adenocarcinoma

PD-1:

Programmed death 1

PDL-1:

Programmed death ligand-1

STAT:

Signal transducer and activator of transcription

TAM:

Tumor-associated macrophages

TGF-β1:

Transforming growth factor beta-1

Th1/2:

T helper 1/2

TIME:

Tumor immune microenvironment

TNF-α:

Tumor necrosis factor-α

Treg:

Regulatory T cells

VEGF:

Vascular endothelial growth factor

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Acknowledgements

We sincerely expressed our thanks to Junyi Pang, Liangrui Zhou, Xiaolong Liang and Xuguang Liu for their excellent pathological work.

Funding

This work was supported by National Natural Science Foundation of China (81673023, 81872501, 81502068, 81272573 and 81560387) and Beijing Natural Science Foundation of China (7172177) and CAMS innovation Fun for Medical Sciences (CIFMS), NWA0564-3.

Author information

Author notes

  1. Qiaofei Liu, Huanwen Wu, Yuan Li and Ronghua Zhang have contributed equally to this work.

Authors and Affiliations

  1. Department of General Surgery, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, 100730, China

    Qiaofei Liu, Ronghua Zhang, Xiang Zhang, Ming Cui, Jingkai Liu, Tong Li, Junyi Gao, Wenming Wu, Weibin Wang, Li Zhou, Junchao Guo, Menghua Dai, Taiping Zhang, Quan Liao & Yupei Zhao

  2. Department of Pathology, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, 100730, China

    Huanwen Wu, Yuan Li, Boju Pan & Zhaohui Lu

  3. Department of Medicine, Tsinghua University, No. 1, Shuai Fu Yuan, Dong Chen District, Beijing, 100730, China

    Qiaofei Liu, Huanwen Wu, Yuan Li, Ronghua Zhang, Xiang Zhang, Ming Cui, Jingkai Liu, Tong Li, Junyi Gao, Boju Pan, Wenming Wu, Weibin Wang, Li Zhou, Junchao Guo, Menghua Dai, Taiping Zhang, Quan Liao, Zhaohui Lu & Yupei Zhao

  4. National Translational Medicine Center, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, 100730, China

    Qiaofei Liu, Huanwen Wu, Yuan Li, Ronghua Zhang, Xiang Zhang, Ming Cui, Jingkai Liu, Tong Li, Junyi Gao, Boju Pan, Wenming Wu, Weibin Wang, Li Zhou, Junchao Guo, Menghua Dai, Taiping Zhang, Quan Liao, Zhaohui Lu & Yupei Zhao

  5. Department of Visceral, Vascular and Endocrine Surgery, Martin Luther University Halle-Wittenberg, 06108, Halle (Saale), Saxony-Anhalt, Germany

    Jorg Kleeff

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  1. Qiaofei Liu
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Contributions

QL, YZ, QL and ZL designed the study. QL, WW, YL, RZ, XG, MC, XZ, JL, TL and BP collected and analyzed the data. RZ, JL and TL performed the in vitro experiments. QL, QL, YZ and JK discussed and wrote the manuscript. WW, WW, JG, MD, TZ, QL and YZ performed the operations and analyzed the data. ZL, HW and YL performed the pathological work. JK, QL and YZ revised the manuscript.

Corresponding authors

Correspondence to Quan Liao, Zhaohui Lu or Yupei Zhao.

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Conflict of interest

All of the authors declared there was no conflict of interest.

Ethical approval and ethical standards

This study was a retrospective study and was approved by the Ethics Committee of Peking Union Medical College Hospital and was registered at ClinicalTrials.org (NCT02654288).

Informed consent

All of the patients signed the informed consent to donate samples for scientific research. In this study, only resected tumor and peritumor sample were used; therefore, there were nearly no potential harms to the patients.

Human and animal rights

Male 4-week-old or 8-week-old C57BL/6 J mice were purchased from VITAL RIVER Animal Center (Beijing, China). Animal studies were performed in accordance with the institutional guidelines of Peking Union Medical College Hospital and was approved by animal study committee of Peking Union Medical College Hospital on December 1, 2014.

Cell line authentication

All cell lines were purchased from Cell Line Bank of Chinese Academy of Medicine Sciences.

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Liu, Q., Wu, H., Li, Y. et al. Combined blockade of TGf-β1 and GM-CSF improves chemotherapeutic effects for pancreatic cancer by modulating tumor microenvironment. Cancer Immunol Immunother 69, 1477–1492 (2020). https://doi.org/10.1007/s00262-020-02542-7

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