Background: Neoadjuvant chemotherapy combined with immunotherapy (NACI) has shown promise in oesophageal squamous cell carcinoma (ESCC). However, a significant proportion of patients exhibit resistance to NACI, and the underlying mechanisms remain unresolved.
Methods: We integrated single-cell RNA sequencing data, including seven patients with ESCC treated with NACI and 69 patients with ESCC treated with surgery alone. Bulk RNA sequencing data were obtained from a public database. Immunohistochemistry and multiplexed immunofluorescence staining were performed to verify the role of important immune cells and molecules in clinical treatment outcomes.
Results: Here, we profiled the transcriptomes of 512 736 cells from 76 patients with ESCC, revealing that the nonresponder baseline tumour microenvironment exhibited a relative absence of major histocompatibility complex II molecules expressed on CD20+B cells and a low expression of CXCL13 on CD4_Tfh and CD8_Tex cells. We also identified CD68+CD163+ macrophages that highly expressed the immunosuppressive LGALS9 gene and preferentially accumulated in the nonresponders after NACI treatment. In addition, nonresponders had a higher baseline fraction of POSTN+fibroblasts, which is associated with higher infiltration of CD68+CD163+ macrophages and lower infiltration of germinal centre B cells. Finally, we described the different characteristics of malignant epithelial cells from different pathological responses to tumours.
Conclusions: This study has unveiled a potential regulatory network among immune cells, stromal cells and malignant epithelial cells under different pathological response conditions and provides a valuable resource for discovering novel targeted therapies for ESCC.
| [1] |
Sung H, Ferlay J, Siegel RL, et al. Global Cancer Statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: a cancer journal for clinicians. 2021; 71: 209-249.
|
| [2] |
Thrift AP. Global burden and epidemiology of Barrett oesophagus and oesophageal cancer. Nat Rev Gastroenterol Hepatol. 2021; 18: 432-443.
|
| [3] |
Yi M, Zheng XL, Niu M, et al. Combination strategies with PD-1/PD-L1 blockade: current advances and future directions. Mol Cancer. 2022; 21: 28.
|
| [4] |
Wu HX, Pan YQ, He Y, et al. Clinical benefit of first-line programmed death-1 antibody plus chemotherapy in low programmed cell death ligand 1-expressing esophageal squamous cell carcinoma: a post hoc analysis of JUPITER-06 and meta-analysis. J Clin Oncol. 2022; 41: 1735-1746.
|
| [5] |
Liu J, Yang Y, Liu ZC, et al. Multicenter, single-arm, phase II trial of camrelizumab and chemotherapy as neoadjuvant treatment for locally advanced esophageal squamous cell carcinoma. J Immunother Cancer. 2022; 10:e004291.
|
| [6] |
Chen XF, Xu X, Wang D, et al. Neoadjuvant sintilimab and chemotherapy in patients with potentially resectable esophageal squamous cell carcinoma (KEEP-G 03): an open-label, single-arm, phase 2 trial. J Immunother Cancer. 2023; 11:e005830.
|
| [7] |
Bejarano L, Jordāo MJC, Joyce JA. Therapeutic targeting of the tumor microenvironment. Cancer Discov. 2021; 11: 933-959.
|
| [8] |
Kim R An M, Lee H, et al. Early tumor-immune microenvironmental remodeling and response to first-line fluoropyrimidine and platinum chemotherapy in advanced gastric cancer. Cancer Discov. 2021; 12: 984-1001.
|
| [9] |
Qin P, Chen H, Wang Y, et al. Cancer-associated fibroblasts undergoing neoadjuvant chemotherapy suppress rectal cancer revealed by single-cell and spatial transcriptomics. Cell Rep Med. 2023; 4:101231.
|
| [10] |
Liu ZC, Zhang Y, Ma N, et al. Progenitor-like exhausted SPRY1(+)CD8(+) T cells potentiate responsiveness to neoadjuvant PD-1 blockade in esophageal squamous cell carcinoma. Cancer Cell. 2023; 41: 1852-1870.e9.
|
| [11] |
Zhang D, Jiang DX, Jiang L, et al. HLA-A(+) tertiary lymphoid structures with reactivated tumor infiltrating lymphocytes are associated with a positive immunotherapy response in esophageal squamous cell carcinoma. Br J Cancer. 2024; 131: 184-195.
|
| [12] |
Mao X, Xu J, Wang W, et al. Crosstalk between cancer-associated fibroblasts and immune cells in the tumor microenvironment: new findings and future perspectives. Mol Cancer. 2021; 20: 131.
|
| [13] |
Niu B, Liao K, Zhou Y, et al. Application of glutathione depletion in cancer therapy: enhanced ROS-based therapy, ferroptosis, and chemotherapy. Biomaterials. 2021; 277:121110.
|
| [14] |
Zhang W, Dai J, Hou G, et al. SMURF2 predisposes cancer cell toward ferroptosis in GPX4-independent manners by promoting GSTP1 degradation. Mol Cell. 2023; 83: 4352-4369.e8.
|
| [15] |
Ji G, Yang Q, Wang S, et al. Single-cell profiling of response to neoadjuvant chemo-immunotherapy in surgically resectable esophageal squamous cell carcinoma. Genome Med. 2024; 16: 49.
|
| [16] |
Wang Z, Zhao Y, Wo Y, et al. The single cell immunogenomic landscape after neoadjuvant immunotherapy combined chemotherapy in esophageal squamous cell carcinoma. Cancer Lett. 2024; 593:216951.
|
| [17] |
Butler A, Hoffman P, Smibert P, et al. Integrating single-cell transcriptomic data across different conditions, technologies, and species. Nat Biotechnol. 2018; 36: 411-420.
|
| [18] |
McGinnis CS, Murrow L, and Gartner Z. DoubletFinder: doublet detection in single-cell RNA sequencing data using artificial nearest neighbors. Cell Syst. 2019; 8: 329-337.e4.
|
| [19] |
Korsunsky I, Millard N, Fan J, et al. Fast, sensitive and accurate integration of single-cell data with Harmony. Nat Methods. 2019; 16: 1289-1296.
|
| [20] |
Yang Y, Li Y, Yu H, et al. Comprehensive landscape of resistance mechanisms for neoadjuvant therapy in esophageal squamous cell carcinoma by single-cell transcriptomics. Signal Transduct Target Ther. 2023; 8: 298.
|
| [21] |
Qiu X, Mao Q, Tang Y, et al. Reversed graph embedding resolves complex single-cell trajectories. Nat Methods. 2017; 14: 979-982.
|
| [22] |
Mandard AM, Dalibard F, Mandard JC, et al. Pathologic assessment of tumor regression after preoperative chemoradiotherapy of esophageal carcinoma. Clinicopathologic correlations. Cancer. 1994; 73: 2680-2686.
|
| [23] |
Wu XX, Zhang HD, Sui ZL, et al. CXCR4 promotes the growth and metastasis of esophageal squamous cell carcinoma as a critical downstream mediator of HIF-1α. Cancer Sci. 2022; 113: 926-939.
|
| [24] |
Long H, Jia Q, Wang L, et al. Tumor-induced erythroid precursor-differentiated myeloid cells mediate immunosuppression and curtail anti-PD-1/PD-L1 treatment efficacy. Cancer Cell. 2022; 40: 674-693.e7.
|
| [25] |
Li Y, Azmi AS, Mohammad RM. Deregulated transcription factors and poor clinical outcomes in cancer patients. Semin Cancer Biol. 2022; 86: 122-134.
|
| [26] |
Yan R, Lin B, Jin W, et al. NRF2, a Superstar of Ferroptosis. Antioxidants (Basel). 2023: 12: 1739.
|
| [27] |
Yang W, Xing X, Yeung SC, et al. Neoadjuvant programmed cell death 1 blockade combined with chemotherapy for resectable esophageal squamous cell carcinoma. J Immunother Cancer. 2022; 10:e003497.
|
| [28] |
Yan X, Duan H, Ni Y, et al. Tislelizumab combined with chemotherapy as neoadjuvant therapy for surgically resectable esophageal cancer: a prospective, single-arm, phase II study (TD-NICE). Int J Surg. 2022; 103:106680.
|
| [29] |
Wu Z, Wu C, Zhao J, et al. Camrelizumab, chemotherapy and apatinib in the neoadjuvant treatment of resectable oesophageal squamous cell carcinoma: a single-arm phase 2 trial. EClinicalMedicine. 2024; 71:102579.
|
| [30] |
Liu B, Zhang Y, Wang D, et al. Single-cell meta-analyses reveal responses of tumor-reactive CXCL13(+) T cells to immune-checkpoint blockade. Nat Cancer. 2022; 3: 1123-1136.
|
| [31] |
Yue P, Bie F, Zhu J, et al. Minimal residual disease profiling predicts pathological complete response in esophageal squamous cell carcinoma. Mol Cancer. 2024; 23: 96.
|
| [32] |
Nakamura S, Ohuchida K, Ohtsubo Y, et al. Single-cell transcriptome analysis reveals functional changes in tumour-infiltrating B lymphocytes after chemotherapy in oesophageal squamous cell carcinoma. Clin Transl Med. 2023; 13:e1181.
|
| [33] |
Zhang H, Wen H, Zhu Q, et al. Genomic profiling and associated B cell lineages delineate the efficacy of neoadjuvant anti-PD-1-based therapy in oesophageal squamous cell carcinoma. EBioMedicine. 2024; 100:104971.
|
| [34] |
Rudy GB, Lew AM. The nonpolymorphic MHC class II isotype, HLA-DQA2, is expressed on the surface of B lymphoblastoid cells. J Immunol. 1997; 158: 2116-2125.
|
| [35] |
Kropshofer H, Hämmerling GJ, Vogt AB. The impact of the non-classical MHC proteins HLA-DM and HLA-DO on loading of MHC class II molecules. Immunol Rev. 2000; 172: 267-278.
|
| [36] |
Coulie PG, Van den Eynde BJ, van der Bruggen P, et al. Tumour antigens recognized by T lymphocytes: at the core of cancer immunotherapy. Nat Rev Cancer. 2014; 14: 135-146.
|
| [37] |
Paijens ST, Vledder A, de Bruyn M, et al. Tumor-infiltrating lymphocytes in the immunotherapy era. Cell Mol Immunol. 2021; 18(4): 842-859.
|
| [38] |
Ghosh D, Jiang W, Mukhopadhyay D, et al. New insights into B cells as antigen presenting cells. Curr Opin Immunol. 2021; 70: 129-137.
|
| [39] |
Silina K. B cell-rich niches support stem-like CD8(+) T cells in cancer microenvironment. Cancer Cell. 2023; 41: 824-825.
|
| [40] |
Wang X, Waschke BC, Woolaver RA, et al. HDAC inhibitors overcome immunotherapy resistance in B-cell lymphoma. Protein Cell. 2020; 11: 472-482.
|
| [41] |
Liu Z, Zhang Y, Ma N, et al. Progenitor-like exhausted SPRY1(+)CD8(+) T cells potentiate responsiveness to neoadjuvant PD-1 blockade in esophageal squamous cell carcinoma. Cancer Cell. 2023; 41: 1852-1870.
|
| [42] |
Li JP, Wu CY, Chen MY, et al. PD-1CXCR5CD4 Th-CXCL13 cell subset drives B cells into tertiary lymphoid structures of nasopharyngeal carcinoma. J Immunother Cancer. 2021; 9:e002101.
|
| [43] |
Yang M, Lu J, Zhang G, et al. CXCL13 shapes immunoactive tumor microenvironment and enhances the efficacy of PD-1 checkpoint blockade in high-grade serous ovarian cancer. J Immunother Cancer. 2021; 9:e001136.
|
| [44] |
Ukita M, Hamanishi J, Yoshitomi H, et al. CXCL13-producing CD4+ T cells accumulate in the early phase of tertiary lymphoid structures in ovarian cancer. JCI Insight. 2022; 7:e157215.
|
| [45] |
Li JP, Wu CY, Chen MY, et al. PD-1(+)CXCR5(-)CD4(+) Th-CXCL13 cell subset drives B cells into tertiary lymphoid structures of nasopharyngeal carcinoma. J Immunother Cancer. 2021; 9:e002101.
|
| [46] |
Magen A, Hamon P, Fiaschi N, et al. Intratumoral dendritic cell-CD4(+) T helper cell niches enable CD8(+) T cell differentiation following PD-1 blockade in hepatocellular carcinoma. Nat Med. 2023; 29: 1389-1399.
|
| [47] |
Yang M, Lu J, Zhang G, et al. CXCL13 shapes immunoactive tumor microenvironment and enhances the efficacy of PD-1 checkpoint blockade in high-grade serous ovarian cancer. J Immunother Cancer. 2021; 9.
|
| [48] |
Rimal R, Desai P, Daware R, et al. Cancer-associated fibroblasts: origin, function, imaging, and therapeutic targeting. Adv Drug Deliv Rev. 2022; 189:114504.
|
| [49] |
You T, Tang H, Wu W, et al. POSTN secretion by extracellular matrix cancer-associated fibroblasts (eCAFs) correlates with poor ICB response via macrophage chemotaxis activation of Akt signaling pathway in gastric cancer. Aging Dis. 2023; 14: 2177-2192.
|
| [50] |
Kundu M, Butti R, Panda VK, et al. Modulation of the tumor microenvironment and mechanism of immunotherapy-based drug resistance in breast cancer. Mol Cancer. 2024; 23: 92.
|
| [51] |
Li J, Yang J, Jiang S, et al. Targeted reprogramming of tumor-associated macrophages for overcoming glioblastoma resistance to chemotherapy and immunotherapy. Biomaterials. 2024; 311:122708.
|
| [52] |
Wang H, Yao L, Chen J, et al. The dual role of POSTN in maintaining glioblastoma stem cells and the immunosuppressive phenotype of microglia in glioblastoma. J Exp Clin Cancer Res. 2024; 43: 252.
|
| [53] |
Lin J, Zhang X, Cheng A, et al. Delivery of peptide-LYTAC via polyporus polysaccharide microneedles for targeted CD47 degradation and enhanced tumor immunotherapy. J Am Chem Soc. 2025; 147: 25004-25016.
|
| [54] |
Vadevoo SMP, Kang Y, Rangaswamy G, et al. IL4 receptor targeting enables nab-paclitaxel to enhance reprogramming of M2-type macrophages into M1-like phenotype via ROS-HMGB1-TLR4 axis and inhibition of tumor growth and metastasis. Theranostics. 2024; 14: 2605-2621.
|
| [55] |
Kang N, Son S, Min S, et al. Stimuli-responsive ferroptosis for cancer therapy. Chem Soc Rev. 2023; 52.
|
RIGHTS & PERMISSIONS
2025 The Author(s). Clinical and Translational Discovery published by John Wiley & Sons Australia, Ltd on behalf of Shanghai Institute of Clinical Bioinformatics.
PDF
