{"title":"CD64+成纤维细胞靶向的维兰特罗和 STING 激动剂通过减少脱落细胞和富集干样 CD8+ T 细胞,增强了 CLDN18.2 BiTEs 对胰腺癌的疗效。","authors":"Tianxing Zhou, Xupeng Hou, Jingrui Yan, Lin Li, Yongjie Xie, Weiwei Bai, Wenna Jiang, Yiping Zou, Xueyang Li, Ziyun Liu, Zhaoyu Zhang, Bohang Xu, Guohua Mao, Yifei Wang, Song Gao, Xiuchao Wang, Tiansuo Zhao, Hongwei Wang, Hongxia Sun, Xiufeng Zhang, Jun Yu, Chongbiao Huang, Jing Liu, Jihui Hao","doi":"10.1136/gutjnl-2024-332371","DOIUrl":null,"url":null,"abstract":"<p><strong>Objective: </strong>The objective of this study is to improve the efficacy of CLDN18.2/CD3 bispecific T-cell engagers (BiTEs) as a promising immunotherapy against pancreatic ductal adenocarcinoma (PDAC).</p><p><strong>Design: </strong>Humanised hCD34<sup>+</sup>/hCD3e<sup>+</sup>, Trp53<sup>R172H</sup>Kras<sup>G12D</sup>Pdx1-Cre (KPC), pancreas-specific Cldn18.2 knockout (KO), fibroblast-specific Fcgr1 KO and patient-derived xenograft/organoid mouse models were constructed. Flow cytometry, Masson staining, Cell Titer Glo assay, virtual drug screening, molecular docking and chromatin immunoprecipitation were conducted.</p><p><strong>Results: </strong>CLDN18.2 BiTEs effectively inhibited early tumour growth, but late-stage efficacy was significantly diminished. Mechanically, the Fc fragment of BiTEs interacted with CD64<sup>+</sup> cancer-associated fibroblasts (CAFs) via activation of the SYK-VAV2-RhoA-ROCK-MLC2-MRTF-A-α-SMA/collagen-I pathway, which enhanced desmoplasia and limited late-stage infiltration of T cells. Molecular docking analysis found that vilanterol suppressed BiTEs-induced phosphorylation of VAV2 (Y172) in CD64<sup>+</sup> CAFs and weakened desmoplasia. Additionally, decreased cyclic guanosine-adenosine monophosphate synthase/stimulator of interferon genes (STING) activity reduced proliferation of TCF-1<sup>+</sup>PD-1<sup>+</sup> stem-like CD8<sup>+</sup> T cells, which limited late-stage effects of BiTEs. Finally, vilanterol and the STING agonist synergistically boosted the efficacy of BiTEs by inhibiting the activation of CD64<sup>+</sup> CAFs and enriching proliferation of stem-like CD8<sup>+</sup> T cells, resulting in sustained anti-tumour activity.</p><p><strong>Conclusion: </strong>Vilanterol plus the STING agonist sensitised PDAC to CLDN18.2 BiTEs and augmented efficacy as a potential novel strategy.</p>","PeriodicalId":12825,"journal":{"name":"Gut","volume":null,"pages":null},"PeriodicalIF":23.0000,"publicationDate":"2024-08-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"CD64<sup>+</sup> fibroblast-targeted vilanterol and a STING agonist augment CLDN18.2 BiTEs efficacy against pancreatic cancer by reducing desmoplasia and enriching stem-like CD8<sup>+</sup> T cells.\",\"authors\":\"Tianxing Zhou, Xupeng Hou, Jingrui Yan, Lin Li, Yongjie Xie, Weiwei Bai, Wenna Jiang, Yiping Zou, Xueyang Li, Ziyun Liu, Zhaoyu Zhang, Bohang Xu, Guohua Mao, Yifei Wang, Song Gao, Xiuchao Wang, Tiansuo Zhao, Hongwei Wang, Hongxia Sun, Xiufeng Zhang, Jun Yu, Chongbiao Huang, Jing Liu, Jihui Hao\",\"doi\":\"10.1136/gutjnl-2024-332371\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Objective: </strong>The objective of this study is to improve the efficacy of CLDN18.2/CD3 bispecific T-cell engagers (BiTEs) as a promising immunotherapy against pancreatic ductal adenocarcinoma (PDAC).</p><p><strong>Design: </strong>Humanised hCD34<sup>+</sup>/hCD3e<sup>+</sup>, Trp53<sup>R172H</sup>Kras<sup>G12D</sup>Pdx1-Cre (KPC), pancreas-specific Cldn18.2 knockout (KO), fibroblast-specific Fcgr1 KO and patient-derived xenograft/organoid mouse models were constructed. Flow cytometry, Masson staining, Cell Titer Glo assay, virtual drug screening, molecular docking and chromatin immunoprecipitation were conducted.</p><p><strong>Results: </strong>CLDN18.2 BiTEs effectively inhibited early tumour growth, but late-stage efficacy was significantly diminished. Mechanically, the Fc fragment of BiTEs interacted with CD64<sup>+</sup> cancer-associated fibroblasts (CAFs) via activation of the SYK-VAV2-RhoA-ROCK-MLC2-MRTF-A-α-SMA/collagen-I pathway, which enhanced desmoplasia and limited late-stage infiltration of T cells. Molecular docking analysis found that vilanterol suppressed BiTEs-induced phosphorylation of VAV2 (Y172) in CD64<sup>+</sup> CAFs and weakened desmoplasia. Additionally, decreased cyclic guanosine-adenosine monophosphate synthase/stimulator of interferon genes (STING) activity reduced proliferation of TCF-1<sup>+</sup>PD-1<sup>+</sup> stem-like CD8<sup>+</sup> T cells, which limited late-stage effects of BiTEs. Finally, vilanterol and the STING agonist synergistically boosted the efficacy of BiTEs by inhibiting the activation of CD64<sup>+</sup> CAFs and enriching proliferation of stem-like CD8<sup>+</sup> T cells, resulting in sustained anti-tumour activity.</p><p><strong>Conclusion: </strong>Vilanterol plus the STING agonist sensitised PDAC to CLDN18.2 BiTEs and augmented efficacy as a potential novel strategy.</p>\",\"PeriodicalId\":12825,\"journal\":{\"name\":\"Gut\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":23.0000,\"publicationDate\":\"2024-08-30\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Gut\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.1136/gutjnl-2024-332371\",\"RegionNum\":1,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"GASTROENTEROLOGY & HEPATOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Gut","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1136/gutjnl-2024-332371","RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"GASTROENTEROLOGY & HEPATOLOGY","Score":null,"Total":0}
引用次数: 0
摘要
研究目的本研究的目的是提高CLDN18.2/CD3双特异性T细胞吞噬体(BiTEs)的疗效,将其作为一种很有前景的针对胰腺导管腺癌(PDAC)的免疫疗法:设计:构建了人源化 hCD34+/hCD3e+、Trp53R172HKrasG12DPdx1-Cre(KPC)、胰腺特异性 Cldn18.2 基因敲除(KO)、成纤维细胞特异性 Fcgr1 KO 和患者来源的异种移植/类器官小鼠模型。进行了流式细胞术、Masson 染色、细胞滴度 Glo 检测、虚拟药物筛选、分子对接和染色质免疫沉淀:结果:CLDN18.2 BiTEs 能有效抑制早期肿瘤生长,但晚期疗效明显降低。从机理上讲,BiTEs 的 Fc 片段通过激活 SYK-VAV2-RhoA-ROCK-MLC2-MRTF-A-α-SMA/collagen-I 通路与 CD64+ 癌相关成纤维细胞(CAFs)相互作用,从而增强了脱钙作用并限制了晚期 T 细胞的浸润。分子对接分析发现,维兰特罗抑制了BiTEs诱导的CD64+CAFs中VAV2(Y172)的磷酸化,并削弱了脱钙作用。此外,环鸟苷腺苷单磷酸合成酶/干扰素基因刺激因子(STING)活性的降低减少了TCF-1+PD-1+干样CD8+T细胞的增殖,从而限制了BiTEs的晚期效应。最后,维兰特罗和STING激动剂通过抑制CD64+ CAFs的活化和丰富干样CD8+ T细胞的增殖,协同提高了BiTEs的疗效,从而产生了持续的抗肿瘤活性:Vilanterol加STING激动剂可使PDAC对CLDN18.2 BiTEs敏感并增强疗效,是一种潜在的新策略。
CD64+ fibroblast-targeted vilanterol and a STING agonist augment CLDN18.2 BiTEs efficacy against pancreatic cancer by reducing desmoplasia and enriching stem-like CD8+ T cells.
Objective: The objective of this study is to improve the efficacy of CLDN18.2/CD3 bispecific T-cell engagers (BiTEs) as a promising immunotherapy against pancreatic ductal adenocarcinoma (PDAC).
Design: Humanised hCD34+/hCD3e+, Trp53R172HKrasG12DPdx1-Cre (KPC), pancreas-specific Cldn18.2 knockout (KO), fibroblast-specific Fcgr1 KO and patient-derived xenograft/organoid mouse models were constructed. Flow cytometry, Masson staining, Cell Titer Glo assay, virtual drug screening, molecular docking and chromatin immunoprecipitation were conducted.
Results: CLDN18.2 BiTEs effectively inhibited early tumour growth, but late-stage efficacy was significantly diminished. Mechanically, the Fc fragment of BiTEs interacted with CD64+ cancer-associated fibroblasts (CAFs) via activation of the SYK-VAV2-RhoA-ROCK-MLC2-MRTF-A-α-SMA/collagen-I pathway, which enhanced desmoplasia and limited late-stage infiltration of T cells. Molecular docking analysis found that vilanterol suppressed BiTEs-induced phosphorylation of VAV2 (Y172) in CD64+ CAFs and weakened desmoplasia. Additionally, decreased cyclic guanosine-adenosine monophosphate synthase/stimulator of interferon genes (STING) activity reduced proliferation of TCF-1+PD-1+ stem-like CD8+ T cells, which limited late-stage effects of BiTEs. Finally, vilanterol and the STING agonist synergistically boosted the efficacy of BiTEs by inhibiting the activation of CD64+ CAFs and enriching proliferation of stem-like CD8+ T cells, resulting in sustained anti-tumour activity.
Conclusion: Vilanterol plus the STING agonist sensitised PDAC to CLDN18.2 BiTEs and augmented efficacy as a potential novel strategy.
期刊介绍:
Gut is a renowned international journal specializing in gastroenterology and hepatology, known for its high-quality clinical research covering the alimentary tract, liver, biliary tree, and pancreas. It offers authoritative and current coverage across all aspects of gastroenterology and hepatology, featuring articles on emerging disease mechanisms and innovative diagnostic and therapeutic approaches authored by leading experts.
As the flagship journal of BMJ's gastroenterology portfolio, Gut is accompanied by two companion journals: Frontline Gastroenterology, focusing on education and practice-oriented papers, and BMJ Open Gastroenterology for open access original research.