Exosomes derived miR-362 exacerbates pneumonia by increasing Interleukin-6 via targeting VENTX

Abstract

Pneumonia is a condition characterized by lung damage resulting from a robust immune response by the host. While the defense and immunity against bacterial lung infections have been extensively studied, little is known about the specific immune factors involved in the progression of bacterial pneumonia. To address this knowledge gap, our study aimed to compare normal lung tissues with pneumonia tissues using various techniques, including HE staining, RNA sequencing, RT-PCR, and Elisa assay. Our analysis revealed a significant increase in the levels of interleukin-6 (IL-6) in pneumonia tissues compared to normal lung tissues. To further investigate the underlying mechanism, we extracted exosomes from both pneumonia and normal lung tissues using ultracentrifugation. The exosomes were then examined using electron microscopy, diameter analysis, and western blot assay. RNA sequencing of the exosomes revealed an upregulation of several microRNAs (miRNAs), with miR-362 exhibiting the most significant change. This finding was confirmed through RT-PCR analysis conducted on lung tissues and alveolar lavage fluid. To gain insights into the specific target genes of miR-362, we employed bioinformatics analysis, which identified VENTX as a potential target gene. This finding was further validated through RT-PCR, western blot, and luciferase assay. Our experimental evidence demonstrated that miR-362 regulates VENTX expression, as evidenced by the use of miR-362 mimics or inhibitors on lung cells. Furthermore, we discovered that exosomes derived from pneumonia tissues upregulate IL-6 production through the miR-362/VENTX axis. Importantly, the blocking of IL-6 generation, which is facilitated by miR-362 inhibitor and VENTX overexpression lentivirus, can be achieved by treating exosomes. Moreover, we conducted in vivo experiments using pneumonia models. Rats were treated with IL-6, miR-362 mimics, or VENTX knock-down lentivirus. The results demonstrated a worse prognosis for rats treated with these factors, indicating their potential as prognostic markers. Taken together, our study suggests that exosomes facilitate IL-6 generation by transferring miR-362, thereby suppressing VENTX transcription. Consequently, the IL-6/miR-362/VENTX axis emerges as a promising therapeutic target for pneumonia.

Keywords: Interleukin-6; VENTX; exosomes; miR-362; pneumonia.