Genetic Exchange Of Pulmonary Exosomes To The Brain Causes Neuronal Changes in COVID-19 Infection And Cistanche Treatment

Abstract

The novel coronavirus pandemic has brought chaos to normal human life around the world. Although COVID-19 symptoms are common, variability in clinical phenotypes has been reported worldwide. Reports of the novel coronavirus suggest neurological symptoms. In addition, the susceptibility and complexity of SARS-CoV-2 in patients with neurodegenerative diseases are largely unknown. Here, we aim to demonstrate that exosomes may transport from SARS-CoV-2-infected lungs to brain regions associated with neurodegenerative diseases, using multiple transcriptome data sets from SARs-CoV-2-infected lungs, RNA fragments from pulmonary exosomes, and gene expression fragments from human brains. During transport, exosome-localized transcription factors regulate genes in the lateral substantia nigra, medial substantia nigra, and superior frontal gyrus regions in Parkinson's disease (PD) and the frontal cortex, hippocampus, and temporal cortex in Alzheimer's disease (AD). During SARS-CoV-2 infection, the transcription factors BCL3, JUND, MXD1, IRF2, IRF9 and STAT1 in exosomes affect neuronal gene regulatory networks and accelerate neurodegeneration. The STAT1 transcription factor regulates 64 PD genes in the lateral substantia nigra, 65 in the superior frontal gyrus and 19 in the medial substantia nigra. Similarly, in AD, STAT1 regulates 74 AD genes in the temporal cortex, 40 genes in the hippocampus, and 16 genes in the frontal cortex. We further confirmed that the disorder of neuronal genes is involved in immune response, signal transduction, apoptosis and stress response. Meanwhile, the inhibitory effect of cistanche on neuronal apoptosis was tested. In conclusion, SARS-CoV-2 may reduce the severity of neurodegenerative diseases by regulating neuronal gene regulatory networks through exosomes, and cistanche may inhibit neuropathic diseases (PD) by stimulating neuronal reproduction and reducing neuronal apoptosis.

Keywords

Cistanche SARS-CoV-2 ·Covid-19· Exosome · Neurodegeneration · Parkinson's Disease · Alzheimer's disease

Introduction

Exosomes are one of the extracellular vesicles secreted by most multicellular organisms for intercellular communication. Exosomes carry a variety of biomolecules, such as nucleotide molecules, including RNA, MRNA, lncRNA, proteins, and metabolites, from primitive cells to tar cells. Initially, exosomes were thought to be involved in removing waste molecules from cells. Research advances have shown that exosomes transfer micromolecules and macromolecules to communicate and regulate receptor cells during physiological and pathological processes. Studies have shown that exosomes are involved in cancer progression, cardiovascular disease, neurodegenerative diseases, and even microbial and viral infections. Interestingly, exosomes released by virus-infected cells carry viral particles to reprogram target cells and transmit pathogenic mechanisms.

More recently, SARS-CoV-2 (COVID-19) infection has become a pandemic affecting millions of people worldwide. Increased vulnerability has been reported in patients with chronic diseases such as diabetes and hypertension. SARS-CoV-2 infects the lower respiratory tract and progresses to multiple organ failure. Currently, there is no specific treatment for COVID-19. Social distancing and vaccinations help prevent the disease from spreading among people. However, there is an urgent need to develop a drug to treat the disease. Understanding the disease mechanism could help develop a potential drug to treat SARS-CoV-2 infection. Many viruses, including the RNA of SARS-CoV-2, are known to enter exosomes for intra-host transmission. Studvies found the presence of SARS-CoV-2 virus particles in exosomes. In addition, SARS-CoV-2 was found in the vacuoles of host cells by histopathological analysis. These findings suggest that SARS-CoV-2 infection may involve interexosomatic communication from the primary lung site to other communication organs in the human system.

Among the various affected organs, there has been limited research into the pathogenesis of COVID-19 in the brain. In addition to the main symptoms, 36.4% of COVID-19 cases also reported neurological changes. It is worth noting that some patients who have recovered from COVID-19 show significant memory loss and cognitive impairment. More recently, Smith et al. have proposed that COVID-19 may affect the central nervous system. In particular, some studies suggest that COVID-19 may accelerate neurodegeneration in Parkinson's disease (PD) and Alzheimer's disease (AD). A study by John et al. showed that COVID-19 leads to selective vulnerability to Parkinson's disease by activating alpha-synaptic nucleoprotein lesions in the CNS. In addition, investigations conducted by JR-Lin et al. reported worsening of Parkinson's disease symptoms following COVID-19 infection. Similarly, Adam et al. demonstrated a possible role for COVID-19 infection in patients with AD. In addition, Tom et al. suggest that COVID-19 patients are at high risk of developing AD.

In this study, we developed a systems biology framework to demonstrate the role of exosom-carried mrnas that encode transcription factors from infected lungs to the brain. We studied multiple transcriptome data sets in SARS-CoV-2-infected lungs, RNA fragments of pulmonary exosomes, and gene expression fragments in different regions of the human brain. In addition, an exosomatic intercellular network between lung and brain regions in neurodegenerative diseases was constructed based on literature evidence. Our data identified transcription factors (TF) from the lungs that regulate gene expression in brain regions, which accelerate PD and AD in COVID-19 infection.

Pic: Faw Cistanche

Materials and Methods

Data Collection

Using the keyword combination of "SARS-CoV-2" and "human host", the RNA-Seq dataset of pulmonary SARS-CoV-2 infection was retrieved from sequence reading files and NCBI database. This dataset contains RNA-Seq profiles of 110 Sam-Dples under various experimental conditions of SARS-CoV-2 infection. Out of 110 samples, 2 healthy lung biopsies and 2 SARS-CoV-2-infected lung biopsies were selected for our downstream analysis. The selected samples had no prior history of disease, including neuropathology.

RNA-Seq Expression Analysis

FastQC v0.11.7 was used to evaluate data quality. The reads are then trimmed using the Trimmomatic v0.35. The eligible reads were consistent with the HG19 human genome. The Reads count for each transcript was normalized and upregulated genes in SARS-CoV-2 infection compared with healthy lungs were captured using the DESeq method. With false discovery rate, p< 0.05 was the standard identification of significant up-regulated genes.

Lung Exosome Profle

The GSE121307 microarray data set describing respiratory-derived exosomes and exosome shuttle Rnas was derived from NCBI, Gene Expression Omnibus database. The exosome shuttle mRNA mainly expressed in GSE121307 lung data was identified and mapped with the overexpressed gene of SARS-CoV-2 infection. We speculate that during SARS-CoV-2 infection, these mapped overexpressed genes are packaged into exosomes in the form of esRNA. Among them, mRNA coding transcription factors were identified by database.

Results

RNA-Seq Analysis and esRNA

Lung RNA-Seq data of SARS-CoV-2 infection and its control group were analyzed. Among 18956 transcripts, 437 were differentially regulated with FDR on SARS-CoV-2 infection, p< 0.05. We then mapped 431 overexpressed genes using GSE121307 esRNA data to identify their pulmonary exosomes. We suggest that the feeding of pulmonary overexpressed genes in SARS-CoV-2 infection may be packaged into exosomes and transported to target cells. At the same time, total glycosides of Cistanche tubulosa can also protect the hippocampal ultrastructure of D-galactose model mice.

Pic: Effects of Cistanche anti Parkinson’s Disease

It is suggested that total glycosides of cistanche may play a role in delaying senescence and preventing Alzheimer's disease through antioxidant mechanism. Of the 362 overexpressed genes, 217 were classified into exosomes. For example, GSK3B, DPP4, SMAD3, PARP1, and IKBKB are inflammatory genes that are the result of a lung cytokine storm and may be carried to target cells by exosomes as esRnas. Of the 196 overexpressed esRnas, 13 encoded TFS identified using molecular signatures and the TcoF-DB v2 database.

Pic: Cistanches Benefits

Exosomal Communication Network

Next, we constructed an exosome communication network based on the literature evidence. This network demonstrated exosomal connectivity between lung and blood-brain barrier microendothelial cells. Similarly, exosome communication between blood-brain barrier endothelial cells and various nerve regions is based on literature evidence. Overall, the exosome network confirms exosome connectivity between the lung and neuronal regions associated with neurodegenerative diseases through the blood-brain barrier. It was found that cistanche tubulosa can improve the learning and memory level of AD mice induced by quinolinic acid, reduce the malondialdehyde content of brain tissue induced by quinolinic acid and increase glutathione

The activity of peptide peroxidase and superoxide dismutase can reduce the activity of acetylcholinesterase, apoptosis rate of brain cells and calcium accumulation in brain tissue. The mechanism may be related to the enhancement of free radical scavenging enzyme activity, the reduction of lipid peroxidation, the decrease of calcium content in brain tissue and the inhibition of QA induced apoptosis of brain cells. Secondly, the total glycosides of cistanche Improve the learning and memory level of AD mice induced by β-AP and aluminum trichloride, decrease the content of MDA in brain tissue, and increase the level of SOD GSH-Px activity can improve some pathological changes in brain tissue and reduce the apoptosis rate of brain cells.

Conclusions

In summary, our study explored the behavior of exosome TF and Cistanche tubulosa neuronal regions in regulating PD and AD pathogenic genes in SARS-CoV-2 infection. Although using public repository data, we implemented several curatorial levels to implement a pathological TF modulation network related to neurodegenerative diseases. More specifically, the step-by-step procedure implemented in this study has strategic confirmation at each methodological stage of biological and documentary evidence. With the rapid expansion of the COVID-19 pandemic, the research sector and access to laboratory work have been affected like never before. As a result, many studies are under way to use publicly available data on SARS-CoV-2 to expand research efforts in response to the current critical situation. On this basis, this study has enhanced the understanding of the neuronal pathogenesis of COVID-19, described the previously unknown COVID-19 pandemic mechanism, and provided a new channel for the treatment of neuropathic diseases (AD and PD) with cistanche tubulosa.

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