CRABP2 (Cellular Retinoic Acid Binding Protein 2D): A novel biomarker for the diagnosis and prognosis involved in immune infiltration of lung adenocarcinoma.

Objective: Overexpressed CRABP2 (Cellula Retinoi Aci Bindin Protei 2D) can promote progression of various tumors. However, there are few comprehensive analysis studies on CRABP2 in lung adenocarcinoma (LUAD). Methods: Several large public databases and online analysis tools such as TCGA, GEO, GEPIA2, UALCAN, Kaplan Meier plotter, LinkedOmics, TIMER, CCLE and Metascape were used for big data mining analysis. RNA interference technology, CCK8 assay, flow cytometry and apoptosis detection, and western blot were used for in vitro experiments. Results: The study revealed that the expression level of CRABP2 in plasma were higher (mean level 31.6587 ±13.8541 ng/mL vs. 13.9328 ± 5.5805 ng/mL, p<0.0001) in patients with early stage (stage IA) LUAD compared to the control group based on analysis of 640 LUAD patients and 640 matched healthy control plasma samples from Lishui Central Hospital. Receiver Operating Characteristic curve showed that CRABP2 had certain accuracy in predicting early LUAD, with a sensitivity of 70.98%, a specificity of 94.53%, a cut-off value of 0.6551 ng/mL, and an Area Under the Curve of 0.839 (95%CI: 0.817 - 0.859, p<0.0001). Compared with normal lung tissue, CRABP2 was significantly overexpressed in LUAD (p<0.05). High CRABP2 expression in LUAD predicts poor prognosis both in Overall Survival (95%CI: 1.04-1.46, HR:1.23, p=0.018) and FP (First Progression, 95%CI: 1.10-1.65, HR = 1.35, p=0.0032) in LUAD patients. CRABP2 can promote the progression of LUAD by promoting the G2/M phase transition, inhibiting the apoptosis and participating in the regulation of immune microenvironment. The high expression of CRABP2 will inhibit the recruitment of immune effector cells and promote the proportion of immuno-suppressive cells, thus promoting the progression of LUAD. The low expression of CRABP2 may enhance the expression of CD274(PD-L1), HAVCR2 and PDCD1LG2(PD-L2) in LUAD. While, the high expression of CRABP2 may enhance the expression of CTLA4, LAG3, PDCD1(PD-1), TIGIT and IGSF8 in LUAD. Conclusions: CRABP2 may be a valuable biomarker for diagnosis, treatment and prognosis of LUAD. Patients with high expression of CRABP2 in LUAD may have suboptimal efficacy when treated with inhibitors targeting CD274, HAVCR2, and PDCD1LG2, whereas they may experience better efficacy with inhibitors targeting CTLA4, LAG3, PDCD1, TIGIT, and IGSF8. Most of cancer patients with high CRABP2 expression may benefit from immune checkpoint inhibitor therapy. Our study results have laid a positive foundation for LUAD diagnosis and therapy.