Abstract
Lung cancer, primarily non-small cell lung cancer (NSCLC), causes the highest cancer-related mortality. Although PD-1/PD-L1 inhibitors have improved survival in advanced NSCLC, they can cause immune-related adverse events. Cordyceps sinensis (C. sinensis), a traditional Chinese medicine used for tonifying the lung and kidney and enhancing immune function, has shown therapeutic promise in combination with anti-PD-1 therapy for NSCLC. This study aimed to explore the anti-tumor effect of wild C. sinensis combined with anti-mouse PD-1 in the treatment of Lewis lung adenocarcinoma (LLC) and to elucidate the underlying pharmacodynamic mechanism. LLC mouse model was established via inoculation with LLC cells, followed by treatment with anti-mouse PD-1, C. sinensis, or their combination. The tumor volume, weight, and histological changes of LLC mice were evaluated. The proportions of tumor-infiltrating immune cells in blood and tumors were evaluated by flow cytometry, immunohistochemistry, and immunofluorescence. The underlying mechanisms of the combination of C. sinensis and anti-mouse PD-1 therapy in LLC mice were investigated using an integrated transcriptomics and metabolomics analysis. Treatment with anti-mouse PD-1, C. sinensis, or their combination significantly reduced tumor volume and weight, and attenuated the histopathological changes of LLC mice tumors. Among which, medium-dose C. sinensis combination exhibited significant improvements. Furthermore, the combination of C. sinensis and anti-mouse PD-1 significantly increased the proportion of CD8+ T cells and decreased the abundance of Tregs and PMN-MDSCs. Integrated transcriptomics and metabolomics analysis revealed that the combination of C. sinensis and anti-mouse PD-1 can enhance anti-tumor immunity in LLC mice by acting on key immune-related genes, including DGKA, PLA2G7, AMPD1, ATP8B4, and BST1, thereby modulating glycerophospholipid metabolism, the TCA cycle, purine metabolism, and nicotinate-nicotinamide metabolism. Wild C. sinensis combined with anti-mouse PD-1 therapy exerts therapeutic effects against LLC by targeting immune-related genes, modulating associated pathways, increasing the proportion of CD8+ T cells, and reducing the infiltration of Tregs and PMN-MDSCs, thereby suppressing tumor growth and inhibiting LLC progression. Further research and clinical studies are needed to validate and expand upon these promising findings.
Data availability
The datasets used and/or analyzed during the current study are available from the corresponding author upon reasonable request.
Abbreviations
- NSCLC:
-
Non-small cell lung cancer
- ICIs:
-
Immune checkpoint inhibitors
- PD-1:
-
Programmed cell death-1
- PD-L1:
-
Programmed cell death ligand 1
- C. sinensis :
-
Cordyceps sinensis
- DAB:
-
Diaminobenzidine
- DEGs:
-
Differentially expressed genes
- FC:
-
Flow cytometry
- LLC:
-
Lewis lung adenocarcinoma
- LC:
-
Lung cancer
- TCMs:
-
Traditional Chinese medicines
- TGI:
-
Tumor growth inhibition rate
- H&E:
-
Hematoxylin-eosin
- ES:
-
Enrichment scores
- QC:
-
Quality control
- OPLS-DA:
-
Orthogonal partial least squares discriminant analysis
- PCA:
-
Principal component analysis
- PLS-DA:
-
Partial least squares discriminant analysis
- VIP:
-
Variable importance in projection
- IHC:
-
Immunohistochemistry
- IF:
-
Immunofluorescence
- TME:
-
Tumor microenvironment
- PMN-MDSCs:
-
Polymorphonuclear myeloid-derived suppressor cells
- Tregs:
-
Regulatory T cells
- RNA-Seq:
-
RNA sequencing
- NLRs:
-
NOD-like receptors
- NK:
-
Natural killer
- FMN:
-
Flavin mononucleotide
- FAD:
-
Flavin adenine dinucleotide
- SD:
-
Standard deviation
References
Pan, L., Wang, X., Long, F. & Tang, A. Clinical efficacy and safety evaluation of traditional Chinese medicine for nourishing Yin and replenishing Qi in combination with PD-1/PD-L1 inhibitors in the treatment of NSCLC patients: a meta-analysis. Toxicol. Res. (Camb). 14 (1), tfaf013. https://doi.org/10.1093/toxres/tfaf013 (2025).
Li, Y., Yan, B. & He, S. Advances and challenges in the treatment of lung cancer. Biomed. Pharmacother. 169, 115891. https://doi.org/10.1016/j.biopha.2023.115891 (2023).
Lv, B. et al. Immunotherapy: reshape the tumor immune microenvironment. Front. Immunol. 13, 844142. https://doi.org/10.3389/fimmu.2022.844142 (2022).
Rui, R., Zhou, L. & He, S. Cancer immunotherapies: advances and bottlenecks. Front. Immunol. 14, 1212476. https://doi.org/10.3389/fimmu.2023.1212476 (2023).
Shang, S. et al. Combined treatment of non-small cell lung cancer using radiotherapy and immunotherapy: challenges and updates. Cancer Commun. (Lond). 41 (11), 1086–1099. https://doi.org/10.1002/cac2.12226 (2021).
Huang, M. Y., Jiang, X. M., Wang, B. L., Sun, Y. & Lu, J. J. Combination therapy with PD-1/PD-L1 Blockade in non-small cell lung cancer: strategies and mechanisms. Pharmacol. Ther. 219, 107694. https://doi.org/10.1016/j.pharmthera.2020.107694 (2021).
Lv, J. et al. Gegen Qinlian Decoction enhances the effect of PD-1 Blockade in colorectal cancer with microsatellite stability by remodelling the gut microbiota and the tumour microenvironment. Cell. Death Dis. 10 (6), 415. https://doi.org/10.1038/s41419-019-1638-6 (2019).
Kong, Q. et al. Modified Bushen Yiqi formula enhances antitumor immunity by reducing the chemotactic recruitment of M2-TAMs and PMN-MDSCs in Lewis lung cancer-bearing mice. J. Ethnopharmacol. 319 (Pt 1), 117183. https://doi.org/10.1016/j.jep.2023.117183 (2024).
Li, B. B. et al. Qingrehuoxue formula enhances anti-PD-1 immunotherapy in NSCLC by remodeling the tumor immune microenvironment via TREM2 signaling. BMC Complement. Med. Ther. 25 (1), 270. https://doi.org/10.1186/s12906-025-05020-8 (2025).
Chen, J. et al. Huaier overcomes tumor-induced immunosuppression in colorectal cancer by activating MHC I and CD8 + T cells. Phytomedicine 147, 157157. https://doi.org/10.1016/j.phymed.2025.157157 (2025).
Zhou, W. B. et al. Integrating proteomics, metabolomics, and network Pharmacology to investigate the mechanism of Cordyceps sinensis in the treatment of COPD rats. J. Pharm. Biomed. Anal. 266, 117076. https://doi.org/10.1016/j.jpba.2025.117076 (2025).
Li, C. X., Yang, S., Qiu, S. J., Lian, Y. B. & Qiu, Y. N. Study on the efficacy of bailing capsule on chemotherapy-associated fatigue in lung cancer. Chin. Med. Res. 14 (20), 51–52. https://doi.org/10.14033/j.cnki.cfmr.2016.20.026 (2016).
Guo, J. Observation on the therapeutic effect of Jiawei lifting and dispersing combined with bailing capsule in the treatment of chemotherapy complications of lung cancer. Shaanxi J. Tradit Chin. Med. 38 (6), 699–700. https://doi.org/10.3969/j.issn.10.3969/j.issn.1000-7369.2017.06.008 (2017).
Tang, L. Regulatory effect of Yupingfeng granules combined with bailing capsule on immune function of lung cancer chemotherapy patients. Mod. J. Integr. Tradit Chin. West. Med. 24 (12), 1318–1319. https://doi.org/10.3969/j.issn.1008-8849.2015.12.024 (2015).
Wang, C., Wang, J. & Qi, Y. Adjuvant treatment with Cordyceps sinensis for lung cancer: A systematic review and meta-analysis of randomized controlled trials. J. Ethnopharmacol. 327, 118044. https://doi.org/10.1016/j.jep.2024.118044 (2024).
Lu, T. et al. Cordyceps sinensis relieves non-small cell lung cancer by inhibiting the MAPK pathway. Chin. Med. 19 (1), 54. https://doi.org/10.1186/s13020-024-00895-0 (2024).
Zhou, J. et al. Comparison of widely targeted metabolomics and untargeted metabolomics of wild Ophiocordyceps sinensis. Molecules 27 (11), 3645. https://doi.org/10.3390/molecules27113645 (2022).
Liu, Y. Y. et al. Therapeutic potential of Cordyceps sinensis targeting oxidative stress and inflammatory response in the treatment of COPD rats: insights from metabolomics analysis. J. Asian Nat. Prod. Res. 27 (3), 460–477. https://doi.org/10.1080/10286020.2024.2403611 (2025).
Koh, C. H., Lee, S., Kwak, M., Kim, B. S. & Chung, Y. CD8 T-cell subsets: heterogeneity, functions, and therapeutic potential. Exp. Mol. Med. 55 (11), 2287–2299. https://doi.org/10.1038/s12276-023-01105-x (2023).
Huang, Z. et al. Signatures of multi-omics reveal distinct tumor immune microenvironment contributing to immunotherapy in lung adenocarcinoma. Front. Immunol. 12, 723172. https://doi.org/10.3389/fimmu.2021.723172 (2021).
Fu, L. et al. DGKA interacts with SRC/FAK to promote the metastasis of non-small cell lung cancer. Cancer Lett. 532, 215585. https://doi.org/10.1016/j.canlet.2022.215585 (2022).
Peng, D. et al. PLA2G7 promotes immune evasion of bladder cancer through the JAK-STAT-PDL1 axis. Cell. Death Dis. 16 (1), 234. https://doi.org/10.1038/s41419-025-07593-1 (2025).
Zhang, F. et al. Inhibiting PLA2G7 reverses the immunosuppressive function of intratumoral macrophages and augments immunotherapy response in hepatocellular carcinoma. J. Immunother Cancer. 12 (1), e008094. https://doi.org/10.1136/jitc-2023-008094 (2024).
Wang, G. et al. Lung cancer scRNA-seq and lipidomics reveal aberrant lipid metabolism for early-stage diagnosis. Sci. Transl Med. 14 (630), eabk2756. https://doi.org/10.1126/scitranslmed.abk2756 (2022).
Jiang, X. et al. Tumor metabolome remolded by low dose mitochondrial uncoupler elicites robust CD8+ T cell response. Cell. Death Discov. 11 (1), 291. https://doi.org/10.1038/s41420-025-02584-9 (2025).
Lan, Y., Du, W., Ma, Y. & Cao, J. Prognostic value and immune infiltration of novel markers TNRC6C/AMPD1 in pancreatic cancer microenvironment. Biochem. Biophys. Rep. 43, 102185. https://doi.org/10.1016/j.bbrep.2025.102185 (2025).
Yang, L. et al. Prognostic tumor microenvironment gene and the relationship with immune infiltration characteristics in metastatic breast cancer. BIOCELL 46 (5), 1215–1243. https://doi.org/10.32604/biocell.2022.018221 (2022).
Yang, L. et al. Blockade of purine metabolism reverses macrophage immunosuppression and enhances anti-tumor immunity in non-small cell lung cancer. Drug Resist. Updat. 78, 101175. https://doi.org/10.1016/j.drup.2024.101175 (2025).
Wo, Y. J. et al. The roles of CD38 and CD157 in the solid tumor microenvironment and cancer immunotherapy. Cells 9 (1), 26. https://doi.org/10.3390/cells9010026 (2019).
Ortolan, E., Augeri, S., Fissolo, G., Musso, I. & Funaro, A. CD157: from immunoregulatory protein to potential therapeutic target. Immunol. Lett. 205, 59–64. https://doi.org/10.1016/j.imlet.2018.06.007 (2019).
Agliano, F., Karginov, T. A., Ménoret, A., Provatas, A. & Vella, A. T. Nicotinamide breaks effector CD8 T cell responses by targeting mTOR signaling. iScience 25 (3), 103932. https://doi.org/10.1016/j.isci.2022.103932 (2022).
Acknowledgements
This work has been supported by Major Science and Technology Project of Qinghai Province (NO. 2021-SF-A4), Chinese Academy of Sciences - People’s Government of Qinghai Province on Sanjiangyuan National Park (NO. LHZX-2022-01), and Qinghai Association for Science and Technology Youth and Middle-aged Talent Support Program (NO. 2022QHSKXRCTJ35). We also acknowledge our colleagues for their work on this paper.
Funding
This work has been supported by Major Science and Technology Project of Qinghai Province (NO. 2021-SF-A4), Chinese Academy of Sciences - People’s Government of Qinghai Province on Sanjiangyuan National Park (NO. LHZX-2022-01), and Qinghai Association for Science and Technology Youth and Middle-aged Talent Support Program (NO. 2022QHSKXRCTJ35).
Author information
Authors and Affiliations
Contributions
Conceptualization: W.Z. Data curation: Y.L. Formal analysis: Y.L., Y.G., S.L., Y.M., and W.Z. Funding acquisition: L.W., and W.Z. Investigation: Y.L., Y.G., S.L., Y.M., Y.X., and W.Z. Methodology: Y.L. Resources: L.W. Supervision: L.W. Writing – original draft: Y.L., and W.Z. Writing – review & editing: L.W., and W.Z.
Corresponding authors
Ethics declarations
Competing interests
The authors declare no competing interests.
Ethics approval and consent for publication
This study was approved by the Animal Experimentation Committee of Northwest Plateau Biology Institute, Chinese Academy of Sciences (allowance number NWIPB2023–19).
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.
About this article
Cite this article
Liu, Y., Gao, Y., Suonanlamao et al. Cordyceps sinensis enhances the efficacy of anti-PD-1 immunotherapy in Lewis lung adenocarcinoma. Sci Rep (2026). https://doi.org/10.1038/s41598-026-37878-0
Received:
Accepted:
Published:
DOI: https://doi.org/10.1038/s41598-026-37878-0