Identification function and prediction of brain traumatic exosomes

Recently, the neurological trauma team led by Prof. Zhang Jianning, the general manager of Tianjin Medical University's General Hospital of Tianjin Medical University, has studied brain trauma. The research group brought together two research hotspots of exosomes and circular RNAs . The whole transcriptome sequencing service provided by Yunzoo Biotechnology was successfully used in the study of circular RNA expression profiles in brain traumatic exosomes. An article in the world's first mouse brain trauma model exosomal circular RNA was published in the Journal of Neurotrauma (influence factor 5.19 ) .

Research Background:

Brain trauma (TBI) has a very high morbidity and mortality, which can harm the health of the body to varying degrees. At present, the molecular mechanism of its pathogenesis, especially damage to the nervous system, is unknown. It is known that exosomes are rich in circular RNA, which is a newly discovered circular non-coding RNA, and most of them use miRNA sponge to regulate the expression of downstream functional genes. The authors of this study are the two research hotspots of exosomes and circular RNA , which use high-throughput sequencing to reveal the changes of circular RNA in brain traumatic exosomes and their potential biological functions.
Research ideas:
The authors first isolated mouse brain tissue, extracted periplasmic exosomes, and detected the expression of circular RNA in exosomes of TBI brain tissue by whole-transcriptome sequencing of exosomes. As a result, 231 differentially expressed circular RNAs were found, of which 155 were up-regulated and 76 were down-regulated. Next, functional analysis of the differentially expressed circular RNA of exosomes was carried out by biosignal analysis such as GO and KEGG. GO analysis indicated that these RNA molecules are mainly involved in nerve damage repair, nervous system development and nerve signaling, and these molecules are mainly enriched in the cGMP-PKG signaling pathway. The CircRNA-miRNA interaction network map reveals the role of circular RNA in targeting miRNAs and possible functions. This paper first studied the changes of circular RNA in brain traumatic exosomes, provided many target molecules for detailed study of subsequent molecular mechanisms, and also provided potential targets for interventional treatment of brain trauma.
Research roadmap:

The results show:
The extracted exosomes were identified by transmission electron microscopy TEM and Western Blot. TEM showed that the extracted exosomes conformed to the irregular spherical shape with a particle size of 30-100 nm. WB results also showed that exogenous marker proteins such as CD63, TSG101 and HSP70 were expressed, and astrocyte marker protein GFAP was also expressed. The above results confirm the successful extraction of exosomes from brain tissue.

Illustration: exosome electron microscopy and WB detection

The expression of circular RNA in TBI vs normal (3v3) exosomes was detected by circular RNA sequencing. It was found that 231 circular RNAs were significantly differentially expressed, of which 155 were up-regulated and 76 were down-regulated.

Schematic: Circular RNA cluster analysis and scatter plots for differential expression of exosomes

Through GO analysis of differentially expressed circular RNA-derived genes, it was found that from the perspective of biological processes, differentially expressed circular RNA was more significantly enriched in biological processes such as sudden triggering and transportation. From the perspective of cell components, differentially expressed circular RNA is more significantly enriched in the cytoplasm, organelles, neurons, synapses and other parts. From a molecular function perspective, differentially expressed circular RNA is more significantly enriched in binding to those specific substrates such as proteins, enzymes, kinases, and the like.
KEGG signaling pathway analysis showed that differentially expressed circular RNA was more significantly enriched in the cGMP-PKG signaling pathway.

Schematic: GO analysis of the differentially expressed cDNA of the exosome

Schematic: KEGG analysis of the differentially expressed cDNA of the exosome

Illustration: Top10 significantly up-and-down CRC-related KEGG pathway network diagram

In 2013, two Nature articles revealed that circular RNA has the function of miRNA sponge, which can adsorb miRNA to regulate the expression of related genes. Since then, the ceRNA functional mechanism of circular RNA is being widely studied. The researchers predicted through bioinformatics that there are multiple miRNA binding sites in the circular RNA, and selected 33 circular RNAs and 35 miRNAs to construct a circRNA-miRNA network map. The results showed that mmu-miR-883a-3p, mmu- miR-3057-5p, mmu-miR-6980-3p, mmu-miR-6968-5p and mum-miR-9768-3p are more regulated by circular RNA than other miRNAs.

Illustration: circRNA-miRNA network map

about the author:

Zhang Jianning, male, doctor, doctoral tutor. He is currently the Dean of the General Hospital of Tianjin Medical University, Director of Neurosurgery, Director of the Tianjin Institute of Neurology, Senior Consultant of the Institute of Neurology, Montreal, Canada, and a leader in neurosurgery. He has published more than 100 academic papers and participated in several monographs. He is the deputy editor of the "Eleventh" Higher Education Department textbook "Surgery". He has been awarded the Outstanding Young Professional and Technical Personnel of the Ministry of Health and the Tianjin Municipal Health Bureau, the Cross-Century Talents of Tianjin and Medical University, the Talents of the New Century, the “Five-Year” Key Construction Subjects of Tianjin, and the “131” Project of Tianjin. Level talent. In recent years, he has undertaken many scientific research projects of the Ministry of Science and Technology (973 Project), the National Natural Science Foundation of China, the Ministry of Health, the Tianjin Science and Technology Commission, and the Education Commission.

References: Circular RNA expression alteration in exosomes from the brain extracellular space after traumatic brain injury in mice

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m6A circular RNA methylation sequencing

M5c circular RNA methylation sequencing

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Lithium CAS No.7439-93-2

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