Characterizing tumor nicroenvironment features and identifying potential therapeutic targets in patients with diabetes mellitus and pancreatic cancer based on single-cell transcriptome sequencing

PENG Han; PAN Yuwei; WANG Xuesong; SUN Yixin; HUANG Shuo; LIANG Houjie

doi:10.16016/j.2097-0927.202501022

您当前的位置：

首页 >

文章列表页 >

Characterizing tumor nicroenvironment features and identifying potential therapeutic targets in patients with diabetes mellitus and pancreatic cancer based on single-cell transcriptome sequencing

Basic Medicine | 更新时间：2025-05-26

- Characterizing tumor nicroenvironment features and identifying potential therapeutic targets in patients with diabetes mellitus and pancreatic cancer based on single-cell transcriptome sequencing
- Journal of Army Medical University Vol. 47, Issue 10, Pages: 1069-1080(2025)
- 作者机构：
  
  1.陆军军医大学（第三军医大学）第一附属医院
  2.肿瘤科，西南癌症中心
  3.放射治疗中心，重庆
  3.重庆大学医学院，重庆
- 作者简介：
  
  LIANG Houjie， E-mail： lianghoujie@sina.com；
- 基金信息：
  
  the Young Scientist Fund of National Nature Science Foundation of China(81902927)
- DOI：10.16016/j.2097-0927.202501022
  CLC： R735.9;R730.23;R730.51
- Received：08 January 2025，
  
  Revised：30 March 2025，
  
  Published：30 May 2025
- 稿件说明：
移动端阅览
彭　涵，潘雨薇，王雪松，等.基于单细胞转录组测序解析胰腺癌合并糖尿病患者肿瘤微环境特征及潜在的治疗靶点［J］.陆军军医大学学报，2025，47（10）：1069-1080.DOI：10.16016/j.2097-0927.202501022.

‍：‍PENG H，PAN Y W，WANG X S，et al.Characterizing tumor nicroenvironment features and identifying potential therapeutic targets in patients with diabetes mellitus and pancreatic cancer based on single-cell transcriptome sequencing［J］.J Army Med Univ，2025，47（10）：1069-1080.DOI：10.16016/j.2097-0927.202501022.
彭　涵，潘雨薇，王雪松，等.基于单细胞转录组测序解析胰腺癌合并糖尿病患者肿瘤微环境特征及潜在的治疗靶点［J］.陆军军医大学学报，2025，47（10）：1069-1080.DOI：10.16016/j.2097-0927.202501022. DOI：

‍：‍PENG H，PAN Y W，WANG X S，et al.Characterizing tumor nicroenvironment features and identifying potential therapeutic targets in patients with diabetes mellitus and pancreatic cancer based on single-cell transcriptome sequencing［J］.J Army Med Univ，2025，47（10）：1069-1080.DOI：10.16016/j.2097-0927.202501022. DOI：

摘要

目的

通过分析转录组和单细胞转录组数据，建立高糖及胰腺癌原位小鼠模型探究胰腺癌（pancreatic cancer，PDAC）合并糖尿病（diabetes mellitus，DM）肿瘤微环境变化并寻找潜在治疗靶点。

方法

整合单细胞转录组和普通转录组数据，采用生物信息学分析比较PDAC合并DM（DM组）以及PDAC不合并DM组（non-DM组）的肿瘤细胞和肿瘤免疫微环境特征。将20只6周龄雄性C57BL/6小鼠（体质量18~20 g），用随机数字表法分为2组（

=10）：高糖组（STZ组），连续腹腔注射溶于柠檬酸盐缓冲液的链脲佐菌素（streptozocin，STZ）（50 mg/kg，终浓度1%）；对照组（Control组），同步注射等量不含STZ的柠檬酸盐缓冲液。采用尾尖血糖检测小鼠血糖水平。将胰腺肿瘤细胞原位接种于对照组和STZ组小鼠后，收集肿瘤浸润免疫细胞和肿瘤细胞。采用流式细胞术评估高糖环境对以下指标的影响：总CD8

T细胞和调节性T细胞（regulatory T cells，Treg）数量；表达Ki67、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、颗粒酶B（granzyme B，GZMB）和干扰素γ（interferon-γ，IFN-γ）的CD8

T细胞亚群数量；CD8

T细胞表面程序性死亡受体1（Programmed Death-1，PD-1）、淋巴细胞活化基因3（lymphocyte activation gene-3，LAG-3）和T细胞免疫球蛋白黏蛋白-3（T cell immunoglobulin and mucin domain-3，Tim-3）的表达水平；肿瘤细胞表面程序性死亡配体1（programmed death-ligand 1，PD-L1）的表达情况；肿瘤相关巨噬细胞表面主要组织相容性复合体Ⅰ类分子（major histocompatibility complex class I，MHC-Ⅰ）和甘露糖受体（cluster of differentiation 206，CD206）的表达情况。

结果

生物信息学分析揭示了，与non-DM组相比，DM组中显著上调基因与预后不良相关（

0.001），2型导管细胞比例增加，具有更高的拷贝数变异水平（

0.001）；在肿瘤免疫微环境中，DM组Treg细胞的比例增加（

0.05），CD8

T细胞的耗竭得分升高（

0.001），并伴随效应分子的表达下调，抑制性检查点的表达上升，M2样巨噬细胞的M2得分显著增高（

0.001）。动物实验及流式细胞术实验发现，与Control组相比，STZ组小鼠生存时间更短（

0.001），总体CD8

T细胞比例减少（

0.01），Treg细胞比例增加（

0.001），表达Ki67、TNF-α、GZMB、IFN-γ的CD8

T细胞比例降低（

0.001），CD8

T细胞表面PD-1（

0.001）、LAG-3（

0.001）和Tim-3（

0.01）表达上升，肿瘤细胞表面PD-L1表达上调（

0.001），肿瘤相关巨噬细胞表面CD206分子表达上升（

0.001），而MHC-Ⅰ分子表达下调（

0.001）。

结论

高糖促进PDAC免疫抑制性微环境形成；针对2型导管细胞及肿瘤微环境中的免疫抑制细胞，并结合免疫检查点双重抗体治疗，可能改善患者的预后情况。

Abstract

‍Objective‍　‍To investigate the changes in the tumor microenvironment of pancreatic cancer (PDAC) complicated with diabetes mellitus (DM) in a mouse model of hyperglycemia and orthotopic pancreatic cancer by analyzing transcriptome and single-cell transcriptome data in order to identify potential therapeutic targets. Method‍　‍By integrating single-cell transcriptome and bulk transcriptome data

bioinformatics analysis was conducted to compare the characteristics of tumor cells and tumor immune microenvironment between PDAC patients with DM (DM group) and those without DM (non-DM group). Twenty male C57BL/6 mice (6 weeks old

weighing 18~20 g) were randomly divided into a hyperglycemic group [STZ group

continuous intraperitoneal injection of 50 mg/kg streptozocin (STZ) (final concentration of 1%) dissolved in citrate buffer

]

and a control group (Control group

an equivalent volume of citrate buffer without STZ at the same time points)

with 10 mice in each group. Tail-tip blood glucose level was measured to monitor glycemic status. After orthotopic inoculation of pancreatic cancer cells in both Control and STZ groups

tumor-infiltrating immune cells were harvested. Flow cytometry was employed to determine the effects of hyperglycemia on: total CD8

T cell and Treg cell populations; CD8

T cell subsets expressing Ki67

TNF-α

granzyme B (GZMB) and IFN-γ; surface expression of PD-1

lymphocyte activation gene-3 (LAG-3) and T cell immunoglobulin and mucin domain-3 (Tim-3) on CD8

T cells; programmed death-ligand 1 (PD-L1) expression on tumor cells; and tumor-associated macrophage surface expression of major histocompatibility complex

class I (MHC-Ⅰ) and cluster of differentiation 206 (CD206). Results‍　‍Bioinformatics analysis revealed that

compared to the non-DM group

the genes significantly up-regulated in the DM group were associated with poor prognosis (

0.001). The proportion of type 2 ductal cells was increased in the DM group

exhibiting higher levels of copy number variation (

0.001). In the tumor immune microenvironment of the DM group

there was an increase in the proportion of Treg cells (

0.05) and an elevated exhaustion score for CD8

T cells (

0.001)

accompanied by down-regulated expression of effector molecules

up-regulated expression of inhibitory checkpoints

and a significant increase in the M2 score of M2-like macrophages (

0.001). Animal experiments and flow cytometry found that

compared to the Control group

the STZ group had a shorter survival time (P

0.001)

with decreased proportions of total CD8

T cells (

0.01) and CD8

T cells expressing Ki67

TNF-α

GZMB and IFN-γ (

0.01)

increased proportion of Treg cells (

0.001)

up-regulated expression of PD-1

LAG-3 and Tim-3 on the surface of CD8

T cells (

0.001)

and up-regulation of PD-L1 on tumor cell surface (

0.001) and enhanced expression of CD206 on the surface of tumor-associated macrophages

while down-regulated expression of MHC-I (

0.001). Conclusion‍　‍High glucose promotes the formation of an immunosuppressive microenvironment in PDAC

and targeting type 2 ductal cells and immunosuppressive cells in the tumor microenvironment

combined with dual immune checkpoint antibody therapy

may improve patient prognosis.

关键词

Keywords

references

Risk Factoir Collaboration （NCD-RISC） N C D . Worldwide trends in diabetes prevalence and treatment from 1990 to 2022： a pooled analysis of 1108 population-representative studies with 141 million participants ［J］. Lancet ， 2024 ， 404 （ 10467 ）： 2077 - 2093 . DOI： 10.1016/S0140-6736（24）02317-1 http://dx.doi.org/10.1016/S0140-6736（24）02317-1 .

LAWLER T ， WALTS Z L ， STEINWANDEL M ， et al . Type 2 diabetes and colorectal cancer risk ［J］. JAMA Netw Open ， 2023 ， 6 （ 11 ）： e2343333 . DOI： 10.1001/jamanetworkopen. 2023.43333 http://dx.doi.org/10.1001/jamanetworkopen.2023.43333 .

SAMUEL S M ， VARGHESE E ， VARGHESE S ， et al . Challenges and perspectives in the treatment of diabetes associated breast cancer ［J］. Cancer Treat Rev ， 2018 ， 70 ： 98 - 111 . DOI： 10.1016/j.ctrv.2018.08.004 http://dx.doi.org/10.1016/j.ctrv.2018.08.004 .

HATLEN P ， GRØNBERG B H ， LANGHAMMER A ， et al . Prolonged survival in patients with lung cancer with diabetes mellitus ［J］. J Thorac Oncol ， 2011 ， 6 （ 11 ）： 1810 - 1817 . DOI： 10.1097/jto.0b013e31822a75be http://dx.doi.org/10.1097/jto.0b013e31822a75be .

PANG Y J ， KARTSONAKI C ， GUO Y ， et al . Diabetes， plasma glucose and incidence of pancreatic cancer： a prospective study of 0.5 million Chinese adults and a meta-analysis of 22 cohort studies ［J］. Int J Cancer ， 2017 ， 140 （ 8 ）： 1781 - 1788 . DOI： 10.1002/ijc.30599 http://dx.doi.org/10.1002/ijc.30599 .

SHARMA A ， SMYRK T C ， LEVY M J ， et al . Fasting blood glucose levels provide estimate of duration and progression of pancreatic cancer before diagnosis ［J］. Gastroenterology ， 2018 ， 155 （ 2 ）： 490 - 500.e2 . DOI： 10.1053/j.gastro. 2018.04.025 http://dx.doi.org/10.1053/j.gastro.2018.04.025 .

KPOSSOU A R ， GBESSI D G ， GNANGNON F R ， et al . Epidemiology of primitive digestive cancers in adult in three specialized health centers in Cotonou （Benin Republic）［J］. Bull Soc Pathol Exot ， 2020 ， 113 （ 5 ）： 254 - 257 . DOI： 10.3166/bspe-2020-0152 http://dx.doi.org/10.3166/bspe-2020-0152 .

HAN B ， ZHENG R ， ZENG H ， et al . Cancer incidence and mortality in China， 2022 ［J］. J Natl Cancer Cent ， 2024 ， 4 （ 1 ）： 47 - 53 . DOI： 10.1016/j.jncc.2024.01.006 http://dx.doi.org/10.1016/j.jncc.2024.01.006 .

SIEGEL R L ， GIAQUINTO A N ， JEMAL A . Cancer statistics， 2024 ［J］. CA A Cancer J Clinicians ， 2024 ， 74 （ 1 ）： 12 - 49 . DOI： 10.3322/caac.21820 http://dx.doi.org/10.3322/caac.21820 .

MU X F ， XIANG Z ， XU Y ， et al . Glucose metabolism controls human γδ T-cell-mediated tumor immunosurveillance in diabetes ［J］. Cell Mol Immunol ， 2022 ， 19 （ 8 ）： 944 - 956 . DOI： 10.1038/s41423-022-00894-x http://dx.doi.org/10.1038/s41423-022-00894-x .

GUO D ， TONG Y Y ， JIANG X M ， et al . Aerobic glycolysis promotes tumor immune evasion by hexokinase2-mediated phosphorylation of IκBα ［J］. Cell Metab ， 2022 ， 34 （ 9 ）： 1312 - 1324.e6 . DOI： 10.1016/j.cmet.2022.08.002 http://dx.doi.org/10.1016/j.cmet.2022.08.002 .

CORTELLINI A ， D’ALESSIO A ， CLEARY S ， et al . Type 2 diabetes mellitus and efficacy outcomes from immune checkpoint blockade in patients with cancer ［J］. Clin Cancer Res ， 2023 ， 29 （ 14 ）： 2714 - 2724 . DOI： 10.1158/1078-0432.ccr-22-3116 http://dx.doi.org/10.1158/1078-0432.ccr-22-3116 .

YOUSUF S ， QIU M J ， VOITH VON VOITHENBERG L ， et al . Spatially resolved multi-omics single-cell analyses inform mechanisms of immune dysfunction in pancreatic cancer ［J］. Gastroenterology ， 2023 ， 165 （ 4 ）： 891 - 908.e14 . DOI： 10.1053/j.gastro.2023.05.036 http://dx.doi.org/10.1053/j.gastro.2023.05.036 .

STEELE N G ， CARPENTER E S ， KEMP S B ， et al . Multimodal mapping of the tumor and peripheral blood immune landscape in human pancreatic cancer ［J］. Nat Cancer ， 2020 ， 1 （ 11 ）： 1097 - 1112 . DOI： 10.1038/s43018-020-00121-4 http://dx.doi.org/10.1038/s43018-020-00121-4 .

潘雨薇，谭玉婷，薛锐，等 . 基于普通转录组和单细胞转录组测序解析间质型胃癌复发风险相关的肿瘤细胞及微环境特征［J］. 陆军军医大学学报， 2025 ， 47 （ 05 ）： 443 - 452 . DOI： 10.16016/j.2097-0927.202411050 http://dx.doi.org/10.16016/j.2097-0927.202411050 .

‍ PAN Y W ， TAN Y T ， XUE R ， et al . Features of tumor cells and microenvironment associated with recurrence risk of mesenchymal-subtype gastric cancer based on bulk RNA-seq and scRNA-seq ［J］. J Army Med Univ ， 2025 ， 47 （ 05 ）： 443 - 452 . DOI： 10.16016/j.2097-0927.202411050 http://dx.doi.org/10.16016/j.2097-0927.202411050 .

赵思洁，王猛，高源，等 . 高原低氧改善肿瘤免疫微环境并抑制结直肠癌皮下肿瘤生长［J］. 陆军军医大学学报， 2025 ， 47 （ 1 ）： 38 - 50 . DOI： 10.16016/j.2097-0927.202410011 http://dx.doi.org/10.16016/j.2097-0927.202410011 .

ZHAO S J ， WANG M ， GAO Y ， et al . Plateau hypoxia improves tumor immune microenvironment and inhibits subcutaneous tumor growth of colorectal cancer ［J］. J Army Med Univ ， 2025 ， 47 （ 1 ）： 38 - 50 . DOI： 10.16016/j.2097-0927.202410011 http://dx.doi.org/10.16016/j.2097-0927.202410011 .

SUN H ， SAEEDI P ， KARURANGA S ， et al . IDF Diabetes Atlas： Global， regional and country-level diabetes prevalence estimates for 2021 and projections for 2045 ［J］. Diabetes Res Clin Pract ， 2022 ， 183 ： 109119 . DOI： 10.1016/j.diabres.2021.109119 http://dx.doi.org/10.1016/j.diabres.2021.109119 .

YUAN C ， BABIC A ， KHALAF N ， et al . Diabetes， weight change， and pancreatic cancer risk ［J］. JAMA Oncol ， 2020 ， 6 （ 10 ）： e202948 . DOI： 10.1001/jamaoncol. 2020. 2948 http://dx.doi.org/10.1001/jamaoncol.2020.2948 .

RUZE R ， SONG J ， YIN X ， et al . Mechanisms of obesity- and diabetes mellitus-related pancreatic carcinogenesis： a comprehensive and systematic review ［J］. Signal Transduct Target Ther ， 2023 ， 8 （ 1 ）： 139 . DOI： 10.1038/s41392-023-01376-w http://dx.doi.org/10.1038/s41392-023-01376-w .

ROSS J S ， RUSSO S B ， CHAVIS G C ， et al . Sphingolipid regulators of cellular dysfunction in Type 2 diabetes mellitus： a systems overview ［J］. Clin Lipidol ， 2014 ， 9 （ 5 ）： 553 - 569 .

HUA H ， KONG Q B ， YIN J ， et al . Insulin-like growth factor receptor signaling in tumorigenesis and drug resistance： a challenge for cancer therapy ［J］. J Hematol Oncol ， 2020 ， 13 （ 1 ）： 64 . DOI： 10.1186/s13045-020-00904-3 http://dx.doi.org/10.1186/s13045-020-00904-3 .

PENG J Y ， SUN B F ， CHEN C Y ， et al . Author Correction： Single-cell RNA-seq highlights intra-tumoral heterogeneity and malignant progression in pancreatic ductal adenocarcinoma ［J］. Cell Res ， 2019 ， 29 （ 9 ）： 777 . DOI： 10.1038/s41422-019-0212-1 http://dx.doi.org/10.1038/s41422-019-0212-1 .

MILLER B C ， SEN D R ， SEN D R ， et al . Subsets of exhausted CD8 + T cells differentially mediate tumor control and respond to checkpoint blockade ［J］. Nat Immunol ， 2024 ， 20 ： 326 - 336 . DOI： 10.1038/s41590-019-0312-6 http://dx.doi.org/10.1038/s41590-019-0312-6 .

MCLANE L M ， ABDEL-HAKEEM M S ， JOHN WHERRY E . CD8 T cell exhaustion during chronic viral infection and cancer ［J］. Annu Rev Immunol ， 2019 ， 37 ： 457 - 495 . DOI： 10.1146/annurev-immunol-041015-055318 http://dx.doi.org/10.1146/annurev-immunol-041015-055318 .

POLANIA J W ， HOYT-MIGGELBRINK A ， TOMASZEWSKI W H ， et al . Antigen presentation by tumor-associated macrophages drives T cells from a progenitor exhaustion state to terminal exhaustion ［J］. Immunity ， 2025 ， 58 （ 1 ）： 232 - 246.e6 . DOI： 10.1016/j.immuni.2024.11.026 http://dx.doi.org/10.1016/j.immuni.2024.11.026 .

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Features of tumor cells and microenvironment associated with recurrence risk of mesenchymal-subtype gastric cancer based on bulk RNA-seq and scRNA-seq

Oncolytic influenza virus enhances killing effect of gemcitabine against pancreatic cancer cells—MENG Zihe^1,2, XU Yongru², DENG Zhaoda², ZHANG Yuxin^1,2, YANG Penghui^1,2, HU Ruiping¹ ¹College of Basic Medical Sciences, Inner Mongolia Medical University, Hohhot, Inner Mongolia;²Institute of Hepato-Pancreato-Biliary Surgery, First Medical Center, Chinese PLA General Hospital, Beijing, China

Plateau hypoxia improves tumor immune microenvironment and inhibits subcutaneous tumor growth of colorectal cancer

Starvation microenvironment activates Yes-associated protein 1 to regulate autophagy and promote proliferation of bladder cancer cells

M2 macrophages promotes bladder cancer EMT through CLDN10 involved signaling pathway in tumor micro-environment

Related Author

PAN Yuwei

TAN Yuting

XUE Rui

CHU Zhaole

LIU Biying

LI Xianfeng

WANG Tao

WANG Bin

Related Institution

Department of Oncology， Chongqing Hospital of Traditional Chinese Medicine

Department of Gastroenterology， Chongqing Key Laboratory of Precise Prevention and Treatment of Digestive Malignancies， Army Medical Center of PLA/Daping Hospital of Third Military Medical University

Department of Gastroenterology， Chongqing University Cancer Hospital

School of Medicine， Chongqing University

中国人民解放军总医院第一医学中心肝胆胰外科医学部研究所

⁰