Effect of immune infiltration intensity on the efficacy of neoadjuvant immunotherapy for esophageal cancer

Yong Zhang¹Xinyao Xu^2,3Xiaorong Mu^4,5Juzheng Wang⁶Jipeng Zhang³Guangyu Xiang⁷Jiahe Li⁷Chunlong Zheng³Huaiyu Wang⁸Qiang Lu^3*

• ¹Tangdu Hospital, Air Force Medical University, Xi'an, China
• ²College of Life Sciences, Northwest University, Xi'an, Shaanxi, China
• ³Department of Thoracic Surgery, Tangdu Hospital, Air Force Medical University, Xi’an, China
• ⁴Department of Pharmacy, Tangdu Hospital, Air Force Medical University, Xi’an, China
• ⁵Department of Pathology, Tangdu Hospital, Air Force Medical University, Xi’an, China
• ⁶Department of Thoracic Surgery,The First People’s Hospital of Xianyang, Xianyang, China
• ⁷Basic Medical College, Air Force Medical University, Xi’an, China
• ⁸Department of Thoracic Surgery, Airforce Medical Center, Beijing, China

Background: Esophageal squamous cell carcinoma (ESCC) treatment often involves neoadjuvant therapy combining chemotherapy and immune checkpoint inhibitors.However, the effectiveness of these treatments is limited by immune infiltration in the tumor microenvironment.We analyzed single-cell transcriptomic data from 22 patients with resectable ESCC, collected before and after neoadjuvant therapy. Differences in gene expression between patients achieving a complete pathological response (pCR) and those who did not were assessed. We further validated our findings using RNAseq data from The Cancer Genome Atlas (TCGA), and conducted quantitative qRT-PCR and Western blot analyses on tumor tissues from a clinical cohort.Results: Significant differences in gene expression related to T cell activation, natural killer cell activity, and cytokine signaling were observed between pCR and non-pCR patients. Notable genes included CXCL10, CXCL11, ME1, MT1X, FAT1, OAS2, and MT2A. TCGA data confirmed a correlation between high gene expression and increased tumor mutational burden as well as improved survival rates, particularly for CXCL10. qRT-PCR revealed significant upregulation of CXCL10, CXCL11, ME1, MT1X, FAT1, OAS2, and MT2A in tumor tissues compared to normal tissues.Western blot analysis showed increased protein levels of CXCL10, CXCL11, OAS2, MT1E, and MT1X, while FAT1 was downregulated.Our study highlights the critical role of immune infiltration and associated molecular pathways in the efficacy of neoadjuvant immunotherapy for ESCC. Specific genes, such as CXCL10, are promising as predictive markers for treatment response and survival.