Effect and mechanism of HER2 antibody-drug conjugate combined with anti-PD-1 antibody in mouse bladder cancer models

HE Shuo; TAO Lu; WU Yue; SHI Mengting; ZHANG Tiantian; GUO Yuanyuan; WANG Rui

doi:10.16016/j.2097-0927.202504097

您当前的位置：

首页 >

文章列表页 >

Effect and mechanism of HER2 antibody-drug conjugate combined with anti-PD-1 antibody in mouse bladder cancer models

Basic Medicine | 更新时间：2025-07-25

- Effect and mechanism of HER2 antibody-drug conjugate combined with anti-PD-1 antibody in mouse bladder cancer models
- Journal of Army Medical University Vol. 47, Issue 14, Pages: 1623-1631(2025)
- 作者机构：
  
  1.蚌埠医科大学第一附属医院：，肿瘤内科，安徽蚌埠
  2.蚌埠医科大学第一附属医院：，泌尿外科，安徽蚌埠
  3.蚌埠医科大学第一附属医院：，安徽省肿瘤演进及智能诊疗重点实验室，安徽蚌埠
- 作者简介：
  
  WANG Rui， E-mail： ruiwang1011@163.com
- 基金信息：
  
  the Key Medical Scientific Research Project of Anhui Provincial Health Commission(AHWJ2023A10096)
- DOI：10.16016/j.2097-0927.202504097
  CLC： R737.14;R969.4;R979.19
- Received：26 April 2025，
  
  Revised：18 June 2025，
  
  Published：30 July 2025
- 稿件说明：
移动端阅览
何硕，陶璐，武越，等.抗HER2抗体偶联药物联合抗PD-1单抗对小鼠膀胱肿瘤的作用及机制研究［J］.陆军军医大学学报，2025，47（14）：1623-1631.DOI：10.16016/j.2097-0927.202504097.

HE S，TAO L，WU Y，et al.Effect and mechanism of HER2 antibody-drug conjugate combined with anti-PD-1 antibody in mouse bladder cancer models［J］.J Army Med Univ，2025，47（14）：1623-1631.DOI：10.16016/j.2097-0927.202504097.
何硕，陶璐，武越，等.抗HER2抗体偶联药物联合抗PD-1单抗对小鼠膀胱肿瘤的作用及机制研究［J］.陆军军医大学学报，2025，47（14）：1623-1631.DOI：10.16016/j.2097-0927.202504097. DOI：

HE S，TAO L，WU Y，et al.Effect and mechanism of HER2 antibody-drug conjugate combined with anti-PD-1 antibody in mouse bladder cancer models［J］.J Army Med Univ，2025，47（14）：1623-1631.DOI：10.16016/j.2097-0927.202504097. DOI：

摘要

目的

探究抗HER2抗体偶联药物（HER2 antibody-drug conjugate，HER2-ADC）联合抗PD-1单抗（anti-PD-1 antibody，anti-PD-1）对HER2表达膀胱癌的协同治疗效果，并阐明其对肿瘤免疫微环境的调控机制。

方法

选取40只6~8周龄（体质量18~22 g）的C57BL/6雌性小鼠构建过表达人HER2的MB49膀胱癌细胞C57BL/6小鼠异位肿瘤模型。当肿瘤体积达100 mm³时，采用随机数字表法分为4组（

=10）：对照组（腹腔注射PBS 1.0 mL），anti-PD-1组（腹腔注射anti-PD-1，每只小鼠200 μg/次，间隔3 d给药），HER2-ADC组（腹腔注射HER2-ADC，2.5 mg/kg，1次/7 d）和联合组（HER2-ADC和anti-PD-1给药方案同单药组）。连续给药28 d，期间每3天测量肿瘤体积与体质量，比较各组异位肿瘤生长速度与荷瘤小鼠生存率。停药后持续观察小鼠生存状态至自然死亡，记录中位生存时间（

=5）。药物干预28 d后留取各组部分荷瘤鼠外周血及肿瘤样本（

=5），流式细胞术检测外周血MDSCs细胞（CD11b

、Gr1

）和Treg细胞（CD4

、CD25

、FOXP3

），免疫组化分析肿瘤内CD3

T、CD8

T、FOXP3

T细胞浸润，全自动生化仪评估肝肾功能：谷丙转氨酶（ALT）、谷草转氨酶（AST）、血尿素氮（BUN）、肌酐（CRE）及组织损伤指标：乳酸脱氢酶（LDH-L）的水平。

结果

治疗28 d时，联合组肿瘤体积显著低于对照组（

0.01）及单药组（

0.01）。单药组（anti-PD-1组中位生存期53 d；HER2-ADC组55 d）及联合组（65 d）均较对照组（41 d）显著延长（

0.01）。药物干预28 d后，单药组和联合组小鼠外周血中MDSCs细胞和Treg细胞比率低于对照组，联合组较单药组降低（

0.05）。肿瘤微环境中，联合组CD3

T和CD8

T细胞浸润水平较对照组和单药组明显增加（

0.05），FOXP3

T细胞浸润水平较对照组和单药组显著减少（

0.05）。各组血清ALT、AST、BUN、CRE、LDH-L水平差异无统计学意义，且均在正常参考范围内。

结论

HER2-ADC联合anti-PD-1可抑制异位膀胱肿瘤生长，可能通过下调MDSCs/Treg并促进CD8

T细胞浸润发挥协同作用。

Abstract

Objective‍　‍To investigate the synergistic therapeutic effects of HER2 antibody-drug conjugate (HER2-ADC) combined with anti-PD-1 antibody (anti-PD-1) on HER2-expressing bladder cancer and elucidate its regulatory mechanisms on the tumor immune microenvironment. Methods‍　‍Orthotopic tumor models were established in 40 female C57BL/6 mice (6~

8 weeks old

body mass 18~22 g) using MB49 bladder cancer cells overexpressing human HER2. When tumors reached 100 mm³

the mice were randomized into (

=10) control (intraperitoneal injection of 1.0 mL PBS)

anti-PD-1 (200 µg per mouse every 3 d)

HER2-ADC (2.5 mg/kg once weekly)

and combination groups (same regimens as above monotherapy). Tumor volume and body mass were measured every 3 d during 28-day treatment. Tumor growth kinetics and survival rates were analyzed. Post-treatment survival was monitored until natural death to determine median survival time (

=5). At day 28

blood and tumor samples (

=5) were collected to detect myeloid-derived suppressor cells (MDSCs; CD11b⁺Gr1⁺) and regulatory T cells (Tregs; CD4⁺CD25⁺FOXP3⁺) with flow cytometry

tumor-infiltrating CD3⁺T

CD8⁺T

and FOXP3⁺T cells with immunohistochemical assasy

and liver/kidney functions [alanine aminotransferase (ALT)

aspartate aminotransferase (AST)

blood urea nitrogen (BUN)

creatinine (CRE)

]

and tissue damage indicators [lactate dehydrogenase isoenzyme (LDH-L)

]

. Results‍　‍In 28 d after treatment

the combination group obtained significantly smallest tumor volume than the control group and the 2 monotherapy groups (all

0.01). The longest median survival was observed in the combination group (65 d

0.01)

followed by the HER2-ADC group (55 d)

anti-PD-1 group (53 d) and control group (41 d). After 28 d of treatment

the combination group exhibited obviously the smallest peripheral proportions of MDSCs/Tregs

most tumor-infiltrating CD3⁺T/CD8⁺T cells

and less FOXP3⁺T cells when compared with the 2 monotherapy groups and control group (all

0.05). While

the 2 monotherapy groups had smaller MDSCs/Tregs proportions than the control group (

0.05). No significant differences were observed among the 4 groups in serum ALT

AST

BUN

CRE

or LDH-L levels

and all of them were within normal ranges. Conclusion‍

　‍HER2-ADC combined with anti-PD-1 suppresses the growth of orthotopic bladder tumor

probably through their synergic effects on down-regulating MDSCs/Treg and enhancing CD8⁺T cell infiltration.

关键词

Keywords

references

LOPEZ-BELTRAN A ， COOKSON M S ， GUERCIO B J ， et al . Advances in diagnosis and treatment of bladder cancer ［J］. BMJ ， 2024 ， 384 ： e076743 . DOI： 10.1136/bmj-2023-076743 http://dx.doi.org/10.1136/bmj-2023-076743 .

ZOUEIN J ， BOUERI M ， KOURIE H R ， et al . Advanced bladder cancer management： history in the making ［J］. Future Oncol ， 2023 ， 19 （ 39 ）： 2565 - 2567 . DOI： 10.2217/fon-2023-0908 http://dx.doi.org/10.2217/fon-2023-0908 .

CERSOSIMO R J . Targeted and immunotherapy for the management of advanced urothelial carcinoma of the bladder ［J］. Am J Health Syst Pharm ， 2024 ， 81 （ 22 ）： 1109 - 1123 . DOI： 10.1093/ajhp/zxae090 http://dx.doi.org/10.1093/ajhp/zxae090 .

LI X H ， LIU J ， LI A A ， et al . Analysis of the relationship between bladder cancer gene mutation and clinical prognosis by high-throughput sequencing ［J］. Lab Med ， 2023 ， 54 （ 2 ）： 142 - 152 . DOI： 10.1093/labmed/lmac083 http://dx.doi.org/10.1093/labmed/lmac083 .

NADAL R ， VALDERRAMA B P ， BELLMUNT J . Progress in systemic therapy for advanced-stage urothelial carcinoma ［J］. Nat Rev Clin Oncol ， 2024 ， 21 （ 1 ）： 8 - 27 . DOI： 10.1038/s41571-023-00826-2 http://dx.doi.org/10.1038/s41571-023-00826-2 .

ALBARRÁN V ， ROSERO D I ， CHAMORRO J ， et al . Her-2 targeted therapy in advanced urothelial cancer： from monoclonal antibodies to antibody-drug conjugates ［J］. Int J Mol Sci ， 2022 ， 23 （ 20 ）： 12659 . DOI： 10.3390/ijms232012659 http://dx.doi.org/10.3390/ijms232012659 .

LEARY J B ， ENRIGHT T ， BAKALOUDI D R ， et al . Frequency and nature of genomic alterations in ERBB2-altered urothelial bladder cancer ［J］. Target Oncol ， 2024 ， 19 （ 3 ）： 447 - 458 . DOI： 10.1007/s11523-024-01056-x http://dx.doi.org/10.1007/s11523-024-01056-x .

MERIC-BERNSTAM F ， MAKKER V ， OAKNIN A ， et al . Efficacy and safety of trastuzumab deruxtecan in patients with HER2-expressing solid tumors： primary results from the DESTINY-PanTumor02 phase Ⅱ trial ［J］. J Clin Oncol ， 2024 ， 42 （ 1 ）： 47 - 58 . DOI： 10.1200/JCO.23.02005 http://dx.doi.org/10.1200/JCO.23.02005 .

SHENG X N ， WANG L ， HE Z S ， et al . Efficacy and safety of disitamab vedotin in patients with human epidermal growth factor receptor 2-positive locally advanced or metastatic urothelial carcinoma： a combined analysis of two phase Ⅱ clinical trials ［J］. J Clin Oncol ， 2024 ， 42 （ 12 ）： 1391 - 1402 . DOI： 10.1200/JCO.22.02912 http://dx.doi.org/10.1200/JCO.22.02912 .

DE VRIES E G E ， RÜSCHOFF J ， LOLKEMA M ， et al . Phase Ⅱ study （KAMELEON） of single-agent T-DM1 in patients with HER2-positive advanced urothelial bladder cancer or pancreatic cancer/cholangiocarcinoma ［J］. Cancer Med ， 2023 ， 12 （ 11 ）： 12071 - 12083 . DOI： 10.1002/cam4.5893 http://dx.doi.org/10.1002/cam4.5893 .

QU W ， FU C ， HAN W Q ， et al . A single-arm， multicenter， phase 2 clinical study of recombinant humanized anti-HER2 monoclonal antibody-MMAE conjugate （MRG002） in HER2-positive unresectable locally advanced or metastatic urothelial carcinoma ［J］. Eur J Cancer ， 2024 ， 205 ： 114096 . DOI： 10.1016/j.ejca.2024.114096 http://dx.doi.org/10.1016/j.ejca.2024.114096 .

WEI Y B ， ZHANG R C ， YU C B ， et al . Disitamab vedotin in combination with immune checkpoint inhibitors for locally and locally advanced bladder urothelial carcinoma： a two-center’s real-world study ［J］. Front Pharmacol ， 2023 ， 14 ： 1230395 . DOI： 10.3389/fphar.2023.1230395 http://dx.doi.org/10.3389/fphar.2023.1230395 .

MANDREKAR S J ， SARGENT D J . Clinical trial designs for predictive biomarker validation： theoretical considerations and practical challenges ［J］. J Clin Oncol ， 2009 ， 27 （ 24 ）： 4027 - 4034 . DOI： 10.1200/JCO.2009. 22.3701 http://dx.doi.org/10.1200/JCO.2009.22.3701 .

FUENTES-ANTRÁS J ， GENTA S ， VIJENTHIRA A ， et al . Antibody-drug conjugates： in search of partners of choice ［J］. Trends Cancer ， 2023 ， 9 （ 4 ）： 339 - 354 . DOI： 10.1016/j.trecan.2023.01.003 http://dx.doi.org/10.1016/j.trecan.2023.01.003 .

ZHOU L ， YANG K W ， ZHANG S ， et al . Disitamab vedotin plus toripalimab in patients with locally advanced or metastatic urothelial carcinoma （RC48-C014）： a phase Ib/Ⅱ dose-escalation and dose-expansion study ［J］. Ann Oncol ， 2025 ， 36 （ 3 ）： 331 - 339 . DOI： 10.1016/j.annonc. 2024.12.002 http://dx.doi.org/10.1016/j.annonc.2024.12.002 .

李森林，李心远，陈勇，等 . 维迪西妥单抗联合替雷利珠单抗在膀胱癌新辅助治疗中的疗效和安全性分析［J］. 中华泌尿外科杂志， 2023 ， 44 （ 11 ）： 812 - 817 .

LI S L ， LI X Y ， CHEN Y ， et al . Efficacy and safety of disitamab vedotin combined with tislelizumab in the neoadjuvant treatment of bladder cancer ［J］. Chin J Urol ， 2023 ， 44 （ 11 ）： 812 - 817 .

邢金春，赵仲杰，刘正升 . ADC药物联合免疫治疗在膀胱癌的应用［J］. 临床泌尿外科杂志， 2024 ， 39 （ 9 ）： 755 - 759 . DOI： 10.13201/j.issn.1001-1420.2024.09.001 http://dx.doi.org/10.13201/j.issn.1001-1420.2024.09.001 .

XING J C ， ZHAO Z J ， LIU Z S . Application of ADC and immunotherapy in bladder cancer ［J］. J Clin Urol ， 2024 ， 39 （ 9 ）： 755 - 759 . DOI： 10.13201/j.issn.1001-1420.2024.09.001 http://dx.doi.org/10.13201/j.issn.1001-1420.2024.09.001 .

NICOLÒ E ， GIUGLIANO F ， ASCIONE L ， et al . Combining antibody-drug conjugates with immunotherapy in solid tumors： current landscape and future perspectives ［J］. Cancer Treat Rev ， 2022 ， 106 ： 102395 . DOI： 10.1016/j.ctrv.2022.102395 http://dx.doi.org/10.1016/j.ctrv.2022.102395 .

FU Z W ， LI S J ， HAN S F ， et al . Antibody drug conjugate： the “biological missile” for targeted cancer therapy ［J］. Signal Transduct Target Ther ， 2022 ， 7 ： 93 . DOI： 10.1038/s41392-022-00947-7 http://dx.doi.org/10.1038/s41392-022-00947-7 .

中国抗癌协会肿瘤药物临床研究专业委员会，国家抗肿瘤药物临床应用监测专家委员会，国家肿瘤质控中心乳腺癌专家委员会，等 . 抗体药物偶联物治疗恶性肿瘤临床应用中国专家共识（2023版）［J］. 中华肿瘤杂志， 2023 ， 45 （ 9 ）： 741 - 762 . DOI： 10.3760/cma.j.cn112152-20220713-00489 http://dx.doi.org/10.3760/cma.j.cn112152-20220713-00489 .

Professional Committee on Clinical Research of Oncology Drugs of Chinese AntiCancer Association ， Expert Committee for Monitoring the Clinical Application of Antitumor Drugs ， Breast Cancer Expert Committee ， et al . Expert consensus on the clinical application of antibody drug conjugates in the treatment of malignant tumors（2023 edition）［J］. Chin J Oncol ， 2023 ， 45 （ 9 ）： 741 - 762 . DOI： 10.3760/cma.j.cn112152-20220713-00489 http://dx.doi.org/10.3760/cma.j.cn112152-20220713-00489 .

NUCERA S ， CONTI C ， MARTORANA F ， et al . Antibody-drug conjugates to promote immune surveillance： lessons learned from breast cancer ［J］. Biomedicines ， 2024 ， 12 （ 7 ）： 1491 . DOI： 10.3390/biomedicines12071491 http://dx.doi.org/10.3390/biomedicines12071491 .

GUO Q S ， GAO B ， SONG R W ， et al . FZ-AD005， a novel DLL3-targeted antibody-drug conjugate with topoisomerase I inhibitor， shows potent antitumor activity in preclinical models ［J］. Mol Cancer Ther ， 2024 ， 23 （ 10 ）： 1367 - 1377 . DOI： 10.1158/1535-7163.MCT-23-0701 http://dx.doi.org/10.1158/1535-7163.MCT-23-0701 .

SARANGI S C ， SOPORY P ， PATTNAIK S S ， et al . Antibody-drug conjugates， cancer immunotherapy， and metronomic chemotherapy as novel approaches in cancer management ［J］. Indian J Pharmacol ， 2020 ， 52 （ 5 ）： 402 - 413 . DOI： 10.4103/ijp.IJP_475_18 http://dx.doi.org/10.4103/ijp.IJP_475_18 .

HINGORANI D V ， ALLEVATO M M ， CAMARGO M F ， et al . Monomethyl auristatin antibody and peptide drug conjugates for trimodal cancer chemo-radio-immunotherapy ［J］. Nat Commun ， 2022 ， 13 （ 1 ）： 3869 . DOI： 10.1038/s41467-022-31601-z http://dx.doi.org/10.1038/s41467-022-31601-z .

WU X H ， XU L Y ， LI X F ， et al . A HER2-targeting antibody-MMAE conjugate RC48 sensitizes immunotherapy in HER2-positive colon cancer by triggering the cGAS-STING pathway ［J］. Cell Death Dis ， 2023 ， 14 （ 8 ）： 550 . DOI： 10.1038/s41419-023-06073-8 http://dx.doi.org/10.1038/s41419-023-06073-8 .

JIANG Z M ， ZHANG Y M ， ZHANG Y ， et al . Cancer derived exosomes induce macrophages immunosuppressive polarization to promote bladder cancer progression ［J］. Cell Commun Signal ， 2021 ， 19 （ 1 ）： 93 . DOI： 10.1186/s12964-021-00768-1 http://dx.doi.org/10.1186/s12964-021-00768-1 .

LI K ， SHI H H ， ZHANG B X ， et al . Myeloid-derived suppressor cells as immunosuppressive regulators and therapeutic targets in cancer ［J］. Signal Transduct Target Ther ， 2021 ， 6 （ 1 ）： 362 . DOI： 10.1038/s41392-021-00670-9 http://dx.doi.org/10.1038/s41392-021-00670-9 .

LI Q ， XIANG M . Metabolic reprograming of MDSCs within tumor microenvironment and targeting for cancer immunotherapy ［J］. Acta Pharmacol Sin ， 2022 ， 43 （ 6 ）： 1337 - 1348 . DOI： 10.1038/s41401-021-00776-4 http://dx.doi.org/10.1038/s41401-021-00776-4 .

LASSER S A ， OZBAY KURT F G ， ARKHYPOV I ， et al . Myeloid-derived suppressor cells in cancer and cancer therapy ［J］. Nat Rev Clin Oncol ， 2024 ， 21 （ 2 ）： 147 - 164 . DOI： 10.1038/s41571-023-00846-y http://dx.doi.org/10.1038/s41571-023-00846-y .

HUANG L ， WANG R Q ， XIE K ， et al . A HER2 target antibody drug conjugate combined with anti-PD-（L） 1 treatment eliminates hHER2+ tumors in hPD-1 transgenic mouse model and contributes immune memory formation ［J］. Breast Cancer Res Treat ， 2022 ， 191 （ 1 ）： 51 - 61 . DOI： 10.1007/s10549-021-06384-4 http://dx.doi.org/10.1007/s10549-021-06384-4 .

IWATA T N ， ISHII C ， ISHIDA S ， et al . A HER2-targeting antibody-drug conjugate， trastuzumab deruxtecan （DS-8201a）， enhances antitumor immunity in a mouse model ［J］. Mol Cancer Ther ， 2018 ， 17 （ 7 ）： 1494 - 1503 . DOI： 10.1158/1535-7163.MCT-17-0749 http://dx.doi.org/10.1158/1535-7163.MCT-17-0749 .

BAUZON M ， DRAKE P M ， BARFIELD R M ， et al . Maytansine-bearing antibody-drug conjugates induce in vitro hallmarks of immunogenic cell death selectively in antigen-positive target cells ［J］. Oncoimmunology ， 2019 ， 8 （ 4 ）： e1565859 . DOI： 10.1080/2162402X.2019.1565859 http://dx.doi.org/10.1080/2162402X.2019.1565859 .

BOSHUIZEN J ， PENCHEVA N ， KRIJGSMAN O ， et al . Cooperative targeting of immunotherapy-resistant melanoma and lung cancer by an AXL-targeting antibody-drug conjugate and immune checkpoint blockade ［J］. Cancer Res ， 2021 ， 81 （ 7 ）： 1775 - 1787 . DOI： 10.1158/0008-5472.CAN-20-0434 http://dx.doi.org/10.1158/0008-5472.CAN-20-0434 .

BARTOK O ， PATASKAR A ， NAGEL R ， et al . Anti-tumour immunity induces aberrant peptide presentation in melanoma ［J］. Nature ， 2021 ， 590 （ 7845 ）： 332 - 337 . DOI： 10.1038/s41586-020-03054-1 http://dx.doi.org/10.1038/s41586-020-03054-1 .

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Transfection of wild type hepaCAM gene inhibits cell growth and VEGF expression in BIU-87 cells

Silencing integrin-linked kinase by small interfering RNA inhibits metastasis and invasion of human EJ bladder cancer cells

Related Author

LI Xuemei

LI Yuwei

LI Zhenyan

CHEN Zhiwen

Yang Shuzhe

Luo Chunli

Wu Xiaohou

Pan Cuicui

Related Institution

Department of Urology, First Affiliated Hospital, Army Medical University (Third Military Medical University)

Key Laboratory of Laboratory Medical Diagnostics of Ministry of Education, Faculty of Laboratory Medicine, Department of Urinary Surgery, First Affiliated Hospital, Chongqing Medical University

Department of Cell Biology and Genetics, Center of Molecular Medicine and Cancer Research, College of Basic Medical Sciences, Chongqing Medical University

⁰