Determination of proteomic profiles of mononuclear cells in adult severe asthma patients and their relationship with treatment sensitivity

Biological treatments are becoming increasingly important for adult patients with severe asthma, but these treatments, which are quite expensive today, do not yield the expected response in some patients. A marker that can predict sensitivity or resistance to biological treatments is of great importance for both the patient and the national economy. This research aims to discover proteins that can determine the patients' response to treatment before the treatment starts. Blood samples of 10 cc with EDTA were collected for proteomic analysis before the treatment from patients who applied to the Chest Diseases outpatient clinic of Çukurova University Hospital between November 2022 and November 2023, diagnosed with severe asthma, and agreed to participate in the research for biological agent treatment. Mononuclear cell isolation from blood samples was performed using Ficoll solution. Proteins were isolated from mononuclear cells. The obtained protein samples were divided into their peptides using a commercial kit and analyzed proteomically with LC-MS/MS. Then, the raw proteomic data were analyzed with the MaxQuant program. Proteins obtained as a result of the analysis have been identified with the Perseus program to which biological pathway they belong and their KEGG numbers have been determined. Their interactions were then examined from the STRING database. 20 severe asthma patients aged between 18 and 65 were included in the study, with 14 starting on mepolizumab and 6 on omalizumab. Patients were re-evaluated at the start of treatment and in the 1st year of treatment, and proteomic analyses were conducted on 3 resistant and 5 sensitive patients. In proteomic analyses, a total of 2033 proteins were detected in 8 patients. 11 of the detected proteins showed significant differences. Proteins coded by FHL1, GLUD1, GLUD2, RPL13, IMMT, DAAM1 genes in resistant patients and HLA-A, ACTB, USP14, DAZAP1, MYO1F, TUBB4B, TUBB4A in sensitive patients showed a significant increase. All proteins with changed expression, when evaluated from the STRING and Reactome database, have been seen to have the potential to affect 39 different pathways. Many of the detected proteins play roles in the acquired and hereditary immune system. Among the proteins with changed expression, FHL1, ANXA2, VAMP8, ACTB, MMP9, NME1, NME2 are predicted to be used as biomarkers when examined from the literature. It is thought that FHL1 and ANXA2 can predict smooth muscle hypertrophy and remodeling, and VAMP8 can change the course of the disease by increasing secretion amount through goblet hyperplasia. The proteins discovered in the research have the potential to be used as biomarkers after determining their sensitivity and specificity through comprehensive screening studies. Keywords: Asthma, Severe asthma, Biological treatments, Proteomics, Biomarker discovery