Quantitative proteomic analysis of human plasma using tandem mass tags to identify novel biomarkers for herpes zoster

Herpes zoster (HZ), commonly called shingles, it is a distinctive syndrome caused by reactivation of varicella zoster virus (VZV). A better understanding of the biological characteristics of HZ patients can help develop new targeted therapies to improve the prognosis. High-throughput proteomics technology can deeply study the molecular changes in the development and progression of HZ disease and integrate different levels of information, this is important to help make clinical decisions. Circulating blood contains a lot of biological information, we conducted a proteomics study of patient plasma, hoping to identify key proteins that could indicate the development of HZ. Compared to healthy human plasma, we found 44 differentially expressed proteins in the plasma of HZ patients, the main pathways involved in these molecules are MAPK signaling pathway, Neuroactive ligand-receptor interaction, Acute myeloid leukemia, Transcriptional misregulation in cancer. We found that 27 proteins have direct protein-protein interactions. Based on the comprehensive score, we identified six key molecules as candidate molecules for further study, and then validated another 80 plasma samples (40 HZ patient plasma and 40 healthy human plasma) using enzyme-linked immunosorbent assay (ELISA), immunoblot assay and receiver operating characteristic (ROC) curve analysis. Finally, we found that the expression levels of these three proteins (PLG, F2, VTN) were significantly lower than those of healthy controls (P<.05). To the best of our knowledge, we first used tandem mass tag (TMT) combined with liquid chromatography-mass spectrometry (LC-MS/MS) to screen for differentially expressed proteins in plasma between HZ patients and healthy individuals. It is preliminarily proved that the plasma protein expression profile of HZ patients is different from that of uninfected patients, it has also been found that these three altered key proteins may be used as biomarkers to test early HZ infection. This study reveals new insights into HZ that help to more accurately identify early HZ patients and to find new therapeutic targets. Significance: Varicella-zoster virus (VZV; termed human alphaherpesvirus 3 by the International Committee on Taxonomy of Viruses) is a herpesvirus that is ubiquitous in humans and can cause chickenpox and herpes zoster (HZ). After the initial infection of varicella, the VZV goes into a dormant state in the sensory ganglia and cranial nerves. As age or immunosuppression increases, the cellular immunity to VZV decreases, and the virus reactivates and spreads along the sensory nerves to the skin, causing a unique prodromal pain followed by a rash. About one in five people around the world may be infected with VZV at some point in their lives. According to statistics, about one-third of infected people will develop HZ in their lifetime, and an estimated 1 million cases of herpes zoster occur in the United States each year. Herpes zoster can occur at any age and is usually less severe in children and young adults, but the greatest morbidity and mortality are observed in elderly and immunocompromised patients. 20% of patients with HZ have complications including vasculitis, increased risk of myocardial infarction, or postherpetic neuralgia, the overall mortality rate of patients with HZ in the United States is close to 5%. Considering the wide clinical severity and complications of this disease, there is a great need for biomarkers that contribute to early diagnosis, classification of risks, and prediction of outcomes, which will help elucidate the mechanisms underlying their clinical development. As a useful tool in biology, quantitative proteomics can repeatedly identify and accurately quantify proteins in a variety of biological samples. Proteomic analysis focuses on translational proteins, which play a direct role in most biological processes. Although a small number of proteins can be studied simultaneously with traditional methods, such as ELISA and Western blotting, typical proteomics studies can simultaneously analyze thousands of proteins for a more comprehensive identification. Proteomics has been successfully applied to human-based disease research, Analysis of exposed and unexposed subjects based on mass spectrometry (MS) has been found to reveal altered expression of proteins that can be identified as intermediate biomarkers of early disease effects. Tandem mass tags (TMTs) are chemical labels used for MS-based identification and quantification of biological molecules. TMTs play an important role in proteomic analysis in a variety of samples such as cells, tissues, and body fluids. The body fluids that are often detected clinically are blood, which are easy to obtain and contain abundant biological information related to physiological and pathological processes, we hope to develop protein biomarkers from these blood. Therefore, in order to better characterize the pathological process of HZ patients, we performed proteomic analysis of HZ patients and healthy human plasma using the TMT method. This comparison aims to identify specific processes in the development of HZ disease through protein profiling, which may help to improve our biological understanding of HZ.