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Identification of HSPA5 targets involved in non-alcoholic fatty liver disease

ABSTRACT: Background: It has been reported that HSPA5 is an endoplasmic reticulum chaperone, which regulates cell metabolism, especially lipid metabolism. There are many reports about the interaction between HSPA5 and protein regulating cell function, but there is no report about HSPA5 binding RNA and its biological function. However, it may have RNA binding ability by using interaction capture. Method: In the present study, the ability of HSPA5 to modulate the AS of cellular genes was assessed using a high-throughput RT-PCR approach to examine AS in lipid metabolism-associated genes. RNA immunoprecipitation coupled to RNA sequencing (RIP-Seq) assays were also performed to identify cellular mRNA sassessment. Results: Upon HSPA5 expression, we detected modifications to the AS profiles of numerou genes involved in pathology of nonalcoholic fatty liver disease. Moreover, we show that HSPA5 modulates the expression levels of various splicing factors such as NEAT1, LRP1, and EGFR. Finally, RNA immunoprecipitation coupled to RIP-Seq assays demonstrate that HSPA5 immunoprecipitates specific cellular mRNAs. Conclusion: The HSPA5 protein can modulate the AS profiles of numerous cellular genes and it binds lncRNA and mRNA, and the related genes are links to nonalcoholic fatty liver disease, which is also the innovation of this project.

ORGANISM(S): Homo sapiens

PROVIDER: GSE169538 | GEO | 2023/09/08

REPOSITORIES: GEO

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Project description:To explore the underlying mechanism for the regulatory role of branched-chain amino acids (BCAA) catabolism in hypothalamic RIP-Cre neurons to metabolic organs under normal chow diet feeding, we established RIP-Cre neurons selective BCAA catabolic enzyme branched-chain ketoacid dehydrogenase E1 subunit alpha(Bckdha) knockout mice, we conducted RNA sequencing on the inguinal adipose tissue (iWAT), epididymal adipose tissues (eWAT), and liverfrom wild type (WT, Bckdhaf/f) and knockout (KO, Bckdhaf/f;RIP-Cre) mice (two groups in total). Three biological replicates were performed for each group. Methods: RNA sequencing (RNA-Seq) and data analysis were performed by Gene Denovo Biotechnology (Guangzhou, China). Results: In comparison with findings in the Bckdhaf/f group, 393 up-regulated and 214 down-regulated differentially expressed genes (DEGs) were identified in eWAT from Bckdhaf/f;RIP-Cre group . Similarly, 41 up-regulated genes and 152 down-regulated DEGs were identified in iWAT from Bckdhaf/f;RIP-Cre compared to the Bckdhaf/f group. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed that DEGs in eWAT were enriched in the categories including all subclass pathways in carbohydrate metabolism class, insulin resistance, PPAR signaling, oxidative phosphorylation, thermogenesis, carbon metabolism, fatty acid metabolism, biosynthesis of amino acids and degenerative disease pathways. Similarly, DEGs in iWATwere clustered in the pathways, including the TCA cycle, pyruvate metabolism, glucagon signaling, glycolysis and gluconeogenesis, and carbon metabolism. These changes indicated broad disturbance in WAT energy metabolism in Bckdhaf/f;RIP-Cre mice. Liver transcriptomes showed minor perturbances in subclasses in lipid metabolism, advanced glycation end products and their receptor (AGE-RAGE), PPAR, estrogen, prolactin signaling pathways. Notably, all 13 mitochondrial DNA encoded mRNA were all significantly down-regulated in eWAT of Bckdhaf/f;RIP-Cre mice, along with nucleus DNA encoded mitochondrial located mRNA. Conclusions: Our results using RNA-Seq indicated universal defects of mitochondrial gene expression in eWAT and iWAT.

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Identification of HSPA5 targets involved in non-alcoholic fatty liver disease

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