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Autophagic Regulation of Adipogenesis Through TP53INP2: Insights from In Silico and In Vitro Analysis

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Abstract

Obesity is an epidemic disease associated with multimorbidity resulting in higher mortality risk. The imbalance between energy storage and expenditure is the prime factor in the prognosis of the disease. Specifically, excessive lipid storage through adipogenesis leads to obesity. Adipogenesis is the process that converts preadipocytes into mature adipocytes by regulating major transcription factors like PPARγ and C/EBPα, contributes to lipid storage in adipose tissue. On the contrary, autophagy is a self-degradative process that maintains homeostasis in adipose tissue by regulating adipogenesis and lipolysis. TP53INP2 is a key player that regulates the autophagy process, and it negatively regulates adipogenesis and lipid storage. The gene expression profile GSE93637 was retrieved from the GEO database and analyzed using an integrated bioinformatics approach. The differentially expressed genes (DEGs) were analyzed using R-Bioconductor for TP53INP2 knockdown microarray dataset of 3T3L1 cells, and the DEGs were analyzed for the functional enrichment analysis. Further, the genes involved in the potential biological and molecular functions were evaluated for pathway enrichment analysis by KEGG (Kyoto Encyclopedia of Genes and Genomes). A total of 726 DEGs were found including 391 upregulated and 335 downregulated genes. Further, the functional and pathway enrichment analysis was employed to identify the highly interacting genes, and we identified a total of 56 genes that are highly interacting through a protein–protein interaction network. The DEGs mainly regulate the Peroxisome proliferator-activated receptor (PPAR) signaling pathway, lipolysis, and autophagy. Further, we investigated the associated Hub genes for enriched pathway genes and found the involvement of two autophagic genes ATG7 and sequestosome 1 (p62). In addition, in vitro studies of qRT-PCR (Quantitative real-time polymerase chain reaction) and Western blot analysis revealed that increased autophagy resulted in reduced lipid storage through down-regulation of the adipogenic gene. Moreover, increased expression of autophagic gene TP53INP2 and ATG7 facilitates the down-regulation of p62 and PPARγ gene resulting in lipolysis in mature adipocytes through autophagy. There is no specific treatment to reduce obesity other than a caloric diet and exercise. Hence, this study provides sufficient evidence to conclude that TP53INP2 negatively regulates adipogenesis and increases the degradation of lipids in mature adipocytes which is crucial for reducing obesity. Therefore, it is plausible to consider TP53INP2 as a promising therapeutic target for managing adipogenesis and obesity. However, further studies are necessary to validate their functional and molecular pathway analysis in the regulation of adipogenesis and obesity.

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Data Availability

The GEO (Gene Expression Omnibus) database from NCBI was used to access the GSE93637 dataset (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE93637).

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Acknowledgements

We would like to express our acknowledgment to the ICMR and Administration of VIT for their provision of the essential resources and support that enabled the successful execution of this study.

Funding

This work was financially supported by the Indian Council of Medical Research (ICMR) Senior Research fellowship [File. No: No. 3/1/2(20)/OBS/2022-NCD-II] and the Vellore Institute of Technology (VIT).

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  1. Structural Biology Lab, #412, Pearl Research Park, School of Biosciences & Technology, Vellore Institute of Technology, Vellore, Tamil Nadu, 632014, India

    Mouliganesh Sekar & Kavitha Thirumurugan

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MS and KT design the study. MS did the collection, analysis, and interpretation of the data. MS and KT drafted the manuscript. KT supervised the entire study.

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Sekar, M., Thirumurugan, K. Autophagic Regulation of Adipogenesis Through TP53INP2: Insights from In Silico and In Vitro Analysis. Mol Biotechnol 66, 1188–1205 (2024). https://doi.org/10.1007/s12033-023-01020-6

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