Ectopic expression of GDF15 in cancer-associated fibroblasts enhances melanoma immunosuppression via the GFRAL/RET cascade

Background A key aspect of tumor biology is the involvement of cancer-associated fibroblasts (CAFs) in shaping the immunosuppressive microenvironment. However, the dynamic and complex key roles of CAFs in the melanoma immune microenvironment have not been elucidated.Methods The CAFs landscape in mel...

Full description

Saved in:
Bibliographic Details
Main Authors: Zhiheng Lin, Li Lin, Yisheng Chen, Yantao Ding, Xiaojing Wang, Zhiwen Luo, Zhenzhen Zhao, Zhijie Zhao, Huabao Cai, Wenyang Nie, Fu Zhao
Format: Article
Language:English
Published: BMJ Publishing Group 2025-06-01
Series:Journal for ImmunoTherapy of Cancer
Online Access:https://jitc.bmj.com/content/13/6/e011036.full
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1850164192698433536
author Zhiheng Lin
Li Lin
Yisheng Chen
Yantao Ding
Xiaojing Wang
Zhiwen Luo
Zhenzhen Zhao
Zhijie Zhao
Huabao Cai
Wenyang Nie
Fu Zhao
author_facet Zhiheng Lin
Li Lin
Yisheng Chen
Yantao Ding
Xiaojing Wang
Zhiwen Luo
Zhenzhen Zhao
Zhijie Zhao
Huabao Cai
Wenyang Nie
Fu Zhao
author_sort Zhiheng Lin
collection DOAJ
description Background A key aspect of tumor biology is the involvement of cancer-associated fibroblasts (CAFs) in shaping the immunosuppressive microenvironment. However, the dynamic and complex key roles of CAFs in the melanoma immune microenvironment have not been elucidated.Methods The CAFs landscape in melanoma was characterized using single-cell RNA-seq and spatial transcriptomics. Molecular dynamics simulations were employed to validate the interactions between CAFs and melanoma cells. Bulk RNA-seq was used to establish a prognostic model. To validate the expression of key targets, western blotting, quantitative real-time PCR, and ELISA were performed. The molecular interactions were confirmed via co-immunoprecipitation, chromatin immunoprecipitation, and luciferase gene reporter assays. In-depth molecular mechanisms were explored using lentiviral transfection, cell co-culture experiments, recombinant protein rescue experiments, flow cytometry, knockout mice, and Cre-loxP system mice.Results This study identified a unique group of CAFs expressing high levels of growth differentiation factor 15 (GDF15). The paracrine secretion of GDF15 was regulated by the transcription factor FOXP1, which subsequently binds to the TGFBR2 receptor on melanoma cells, driving their proliferation and metastatic capacity. In addition, CAFs-derived GDF15 interacts with the GFRAL receptor on melanoma cells, thereby promoting RET phosphorylation and triggering downstream signaling axes, inducing increased tumor cell stemness and secretion of inflammatory factors CCL18 and TGF-β. This cascade reaction ultimately induces macrophage polarization to the immunosuppressive M2 phenotype, assists in the establishment of an immunosuppressive microenvironment, and leads to accelerated melanoma lung metastasis.Conclusion By integrating single-cell RNA-seq, spatial transcriptomics, bulk RNA-seq, molecular dynamics simulation and complete experimental design, this study comprehensively characterized that ectopic expression of CAFs-derived GDF15 can act as an accomplice in melanoma progression by inducing increased tumor cell stemness and macrophage M2 polarization, reshaping the immune landscape of melanoma, and providing new ideas and new targets for precision immunotherapy of melanoma.
format Article
id doaj-art-d076deaf9fb246788314383efe6cf5e1
institution OA Journals
issn 2051-1426
language English
publishDate 2025-06-01
publisher BMJ Publishing Group
record_format Article
series Journal for ImmunoTherapy of Cancer
spelling doaj-art-d076deaf9fb246788314383efe6cf5e12025-08-20T02:22:02ZengBMJ Publishing GroupJournal for ImmunoTherapy of Cancer2051-14262025-06-0113610.1136/jitc-2024-011036Ectopic expression of GDF15 in cancer-associated fibroblasts enhances melanoma immunosuppression via the GFRAL/RET cascadeZhiheng Lin0Li Lin1Yisheng Chen2Yantao Ding3Xiaojing Wang4Zhiwen Luo5Zhenzhen Zhao6Zhijie Zhao7Huabao Cai8Wenyang Nie9Fu Zhao10Department of Gynecology, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, ChinaDepartment of Plastic and Reconstructive Surgery, Shanghai Ninth People`s Hospital, School of Medicine, Shanghai JiaoTong University, Shanghai, ChinaFujian Key Laboratory of Toxicant and Drug Toxicology, Medical College, Ningde Normal University, Ningde, Fujian, ChinaDepartment of Dermatology, The First Affiliated Hospital, Anhui, Medical University, Hefei, Anhui, ChinaDepartment of Rheumatology and Immunology, Tongren Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, ChinaDepartment of Sport Medicine, Huashan Hospital, Fudan University, Shanghai, ChinaThe First Clinical Medical College of Shandong University of Traditional Chinese Medicine, Jinan, Shandong, ChinaDepartment of Plastic and Reconstructive Surgery, Shanghai Ninth People`s Hospital, School of Medicine, Shanghai JiaoTong University, Shanghai, ChinaDepartment of Neurosurgery, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, ChinaThe First Clinical Medical College of Shandong University of Traditional Chinese Medicine, Jinan, Shandong, ChinaSchool of Traditional Chinese Medicine, Jinan University, Guangzhou, Guangdong, ChinaBackground A key aspect of tumor biology is the involvement of cancer-associated fibroblasts (CAFs) in shaping the immunosuppressive microenvironment. However, the dynamic and complex key roles of CAFs in the melanoma immune microenvironment have not been elucidated.Methods The CAFs landscape in melanoma was characterized using single-cell RNA-seq and spatial transcriptomics. Molecular dynamics simulations were employed to validate the interactions between CAFs and melanoma cells. Bulk RNA-seq was used to establish a prognostic model. To validate the expression of key targets, western blotting, quantitative real-time PCR, and ELISA were performed. The molecular interactions were confirmed via co-immunoprecipitation, chromatin immunoprecipitation, and luciferase gene reporter assays. In-depth molecular mechanisms were explored using lentiviral transfection, cell co-culture experiments, recombinant protein rescue experiments, flow cytometry, knockout mice, and Cre-loxP system mice.Results This study identified a unique group of CAFs expressing high levels of growth differentiation factor 15 (GDF15). The paracrine secretion of GDF15 was regulated by the transcription factor FOXP1, which subsequently binds to the TGFBR2 receptor on melanoma cells, driving their proliferation and metastatic capacity. In addition, CAFs-derived GDF15 interacts with the GFRAL receptor on melanoma cells, thereby promoting RET phosphorylation and triggering downstream signaling axes, inducing increased tumor cell stemness and secretion of inflammatory factors CCL18 and TGF-β. This cascade reaction ultimately induces macrophage polarization to the immunosuppressive M2 phenotype, assists in the establishment of an immunosuppressive microenvironment, and leads to accelerated melanoma lung metastasis.Conclusion By integrating single-cell RNA-seq, spatial transcriptomics, bulk RNA-seq, molecular dynamics simulation and complete experimental design, this study comprehensively characterized that ectopic expression of CAFs-derived GDF15 can act as an accomplice in melanoma progression by inducing increased tumor cell stemness and macrophage M2 polarization, reshaping the immune landscape of melanoma, and providing new ideas and new targets for precision immunotherapy of melanoma.https://jitc.bmj.com/content/13/6/e011036.full
spellingShingle Zhiheng Lin
Li Lin
Yisheng Chen
Yantao Ding
Xiaojing Wang
Zhiwen Luo
Zhenzhen Zhao
Zhijie Zhao
Huabao Cai
Wenyang Nie
Fu Zhao
Ectopic expression of GDF15 in cancer-associated fibroblasts enhances melanoma immunosuppression via the GFRAL/RET cascade
Journal for ImmunoTherapy of Cancer
title Ectopic expression of GDF15 in cancer-associated fibroblasts enhances melanoma immunosuppression via the GFRAL/RET cascade
title_full Ectopic expression of GDF15 in cancer-associated fibroblasts enhances melanoma immunosuppression via the GFRAL/RET cascade
title_fullStr Ectopic expression of GDF15 in cancer-associated fibroblasts enhances melanoma immunosuppression via the GFRAL/RET cascade
title_full_unstemmed Ectopic expression of GDF15 in cancer-associated fibroblasts enhances melanoma immunosuppression via the GFRAL/RET cascade
title_short Ectopic expression of GDF15 in cancer-associated fibroblasts enhances melanoma immunosuppression via the GFRAL/RET cascade
title_sort ectopic expression of gdf15 in cancer associated fibroblasts enhances melanoma immunosuppression via the gfral ret cascade
url https://jitc.bmj.com/content/13/6/e011036.full
work_keys_str_mv AT zhihenglin ectopicexpressionofgdf15incancerassociatedfibroblastsenhancesmelanomaimmunosuppressionviathegfralretcascade
AT lilin ectopicexpressionofgdf15incancerassociatedfibroblastsenhancesmelanomaimmunosuppressionviathegfralretcascade
AT yishengchen ectopicexpressionofgdf15incancerassociatedfibroblastsenhancesmelanomaimmunosuppressionviathegfralretcascade
AT yantaoding ectopicexpressionofgdf15incancerassociatedfibroblastsenhancesmelanomaimmunosuppressionviathegfralretcascade
AT xiaojingwang ectopicexpressionofgdf15incancerassociatedfibroblastsenhancesmelanomaimmunosuppressionviathegfralretcascade
AT zhiwenluo ectopicexpressionofgdf15incancerassociatedfibroblastsenhancesmelanomaimmunosuppressionviathegfralretcascade
AT zhenzhenzhao ectopicexpressionofgdf15incancerassociatedfibroblastsenhancesmelanomaimmunosuppressionviathegfralretcascade
AT zhijiezhao ectopicexpressionofgdf15incancerassociatedfibroblastsenhancesmelanomaimmunosuppressionviathegfralretcascade
AT huabaocai ectopicexpressionofgdf15incancerassociatedfibroblastsenhancesmelanomaimmunosuppressionviathegfralretcascade
AT wenyangnie ectopicexpressionofgdf15incancerassociatedfibroblastsenhancesmelanomaimmunosuppressionviathegfralretcascade
AT fuzhao ectopicexpressionofgdf15incancerassociatedfibroblastsenhancesmelanomaimmunosuppressionviathegfralretcascade