Detection of Epstein-Barr Virus Infection in Non-Small Cell Lung Cancer

Abstract

Previous investigations proposed a link between the Epstein-Barr virus (EBV) and lung cancer (LC), but the results are highly controversial largely due to the insufficient sample size and the inherent limitation of the traditional viral screening methods such as PCR. Unlike PCR, current next-generation sequencing (NGS) utilizes an unbiased method for the global assessment of all exogenous agents within a cancer sample with high sensitivity and specificity. In our current study, we aim to resolve this long-standing controversy by utilizing our unbiased NGS-based informatics approaches in conjunction with traditional molecular methods to investigate the role of EBV in a total of 1127 LC. In situ hybridization analysis of 110 LC and 10 normal lung samples detected EBV transcripts in 3 LC samples. Comprehensive virome analyses of RNA sequencing (RNA-seq) data sets from 1017 LC and 110 paired adjacent normal lung specimens revealed EBV transcripts in three lung squamous cell carcinoma and one lung adenocarcinoma samples. In the sample with the highest EBV coverage, transcripts from the BamHI A region accounted for the majority of EBV reads. Expression of EBNA-1, LMP-1 and LMP-2 was observed. A number of viral circular RNA candidates were also detected. Thus, we for the first time revealed a type II latency-like viral transcriptome in the setting of LC in vivo. The high-level expression of viral BamHI A transcripts in LC suggests a functional role of these transcripts, likely as long non-coding RNA. Analyses of cellular gene expression and stained tissue sections indicated an increased immune cell infiltration in the sample expressing high levels of EBV transcripts compared to samples expressing low EBV transcripts. Increased level of immune checkpoint blockade factors was also detected in the sample with higher levels of EBV transcripts, indicating an induced immune tolerance. Lastly, inhibition of immune pathways and activation of oncogenic pathways were detected in the sample with high EBV transcripts compared to the EBV-low LC indicating the direct regulation of cancer pathways by EBV. Taken together, our data support the notion that EBV likely plays a pathological role in a subset of LC.

T-Lymphocyte Activation Is Correlated With the Presence of Anti-EBV in Patients With Laryngeal Squamous Cell Carcinoma

“…It was also shown that EBV infection is associated with a reduction of the number of CD4 + Tlymphocytes in patients with EBV-related cancer (20). Despite the involvement of EBV infection in the development of cancer having been extensively studied (5,8,9,(21)(22)(23)(24)(25)(26)(27), reports analyzing the initial phase of immune response and lymphocyte activation affected by viral infection are rare. During primary EBV infection, antibodies to VCA IgM are generated and persist from weeks to months.…”

<p>Possible Oncogenic Viruses Associated with Lung Cancer</p>

“…158,159 The association of EBV and lung cancer presents significant differences according to tumor histotype and geographical site. 15,160 The EBV is often detected in pulmonary LELC occurring in patients from east and southeast Asia where nasopharyngeal carcinoma is highly prevalent, 13,[161][162][163][164] but rarely detected or even undetected in other types of lung cancer such as adenocarcinoma, squamous cell carcinoma, and SCLC. 42,165,166 Recently, Wang et al explored the EBV genomic variations in lung carcinoma and reported four newly sequenced EBV genomes isolated from primary lung carcinomas with apparent genomic diversity among these EBV genomes.…”

SARS-CoV-2 Infection and Lung Cancer: Potential Therapeutic Modalities

“…It is believed, that both genetic, as in family history and polymorphisms, and environmental risk factors are responsible for lung cancer; smoking being the major risk factor for this disease [3,4]. Several recent studies have highlighted infectious agents such as bacteria [5][6][7] and viruses [8][9][10][11] as high-risk factors of the disease.…”

“…It is projected that at least 20% of all human cancers are attributed to microorganisms' infections including viral [12]; viruses are linked with the onset of both solid and hematological malignancies in humans [13]. Some of the frequently linked oncoviruses with lung cancer include Epstein Barr virus (EBV), hepatitis viruses B and C (HBV and HCV), and human papillomaviruses (HPVs) [8,9,11]. However, their causal relationship to lung cancer has not been established yet.…”