Development of a monoclonal antibody to ITPRIPL1 for immunohistochemical diagnosis of non-small cell lung cancers: accuracy and correlation with CD8+ T cell infiltration

Shouyan Deng; Jiawei Shi; Yufan Sun; Yingfei Quan; Zan Shen; Yonggang Wang; Hai Li; Jie Xu

doi:10.3389/fcell.2023.1297211

Development of a monoclonal antibody to ITPRIPL1 for immunohistochemical diagnosis of non-small cell lung cancers: accuracy and correlation with CD8⁺ T cell infiltration

Front Cell Dev Biol. 2023 Dec 19:11:1297211. doi: 10.3389/fcell.2023.1297211. eCollection 2023.

Authors

Shouyan Deng^{1

2}, Jiawei Shi², Yufan Sun³, Yingfei Quan³, Zan Shen⁴, Yonggang Wang⁴, Hai Li¹, Jie Xu²

Affiliations

¹ Division of Gastroenterology and Hepatology, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
² Shanghai Key Laboratory of Medical Epigenetics, International Co-Laboratory of Medical Epigenetics and Metabolism (Ministry of Science and Technology), Institutes of Biomedical Sciences, Fudan University, Shanghai, China.
³ BioTroy Therapeutics, Shanghai, China.
⁴ Department of Oncology, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China.

Abstract

Introduction: Cancer biomarkers are substances or processes highly associated with the presence and progression of cancer, which are applicable for cancer screening, progression surveillance, and prognosis prediction in clinical practice. In our previous studies, we discovered that cancer cells upregulate inositol 1,4,5-triphosphate receptor-interacting protein-like 1 (ITPRIPL1), a natural CD3 ligand, to evade immune surveillance and promote tumor growth. We also developed a monoclonal ITPRIPL1 antibody with high sensitivity and specificity. Here, we explored the application of anti-ITPRIPL1 antibody for auxiliary diagnosis of non-small cell lung cancer (NSCLC). Methods: NSCLC patient tissue samples (n = 75) were collected and stained by anti-ITPRIPL1 or anti-CD8 antibodies. After excluding the flaked samples (n = 15), we evaluated the expression by intensity (0-3) and extent (0-100%) of staining to generate an h-score for each sample. The expression status was classified into negative (h-score < 20), low-positive (20-99), and high-positive (≥ 100). We compared the h-scores between the solid cancer tissue and stroma and analyzed the correlation between the h-scores of the ITPRIPL1 and CD8 expression in situ in adjacent tissue slices. Results: The data suggested ITPRIPL1 is widely overexpressed in NSCLC and positively correlates with tumor stages. We also found that ITPRIPL1 expression is negatively correlated with CD8 staining, which demonstrates that ITPRIPL1 overexpression is indicative of poorer immune infiltration and clinical prognosis. Therefore, we set 50 as the cutoff point of ITPRIPL1 expression H scores to differentiate normal and lung cancer tissues, which is of an excellent sensitivity and specificity score (100% within our sample collection). Discussion: These results highlight the potential of ITPRIPL1 as a proteomic immunohistochemical NSCLC biomarker with possible advantages over the existing NSCLC biomarkers, and the ITPRIPL1 antibody can be applied for accurate diagnosis and prognosis prediction.

Keywords: CD8; ITPRIPL1; T cell infiltration; cancer biomarker; tumor stage.

Grants and funding

This work was supported by the National Natural Science Foundation of China (Nos. 82030104, 81874050, and 81572326), Leading Talent of “Ten Thousand Plan” of China Basic Research Projects of Shanghai Science and Technology Innovation Action Plan (20JC1410700), National Key R&D Program of China (2016YFC0906002), Tang Scholar (XJ), and Startup Research Funding of Fudan University.