Interpretation of Next Generation Sequencing (NGS) Application in Insulin Resistance Zebrafish Larvae Model

Irman Kang, Lisa Amira (2021) Interpretation of Next Generation Sequencing (NGS) Application in Insulin Resistance Zebrafish Larvae Model. [Project Paper] (Submitted)

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Abstract

Type 2 diabetes mellitus (T2DM) is a persistent metabolic disorder that progressively rising around the world. Insulin insensitivity is a symptom of T2DM, which is caused by insulin resistance, decreasing the insulin action towards cells, and eventually leads to pancreatic β-cell being impaired. Over time, the ability of pancreas to release insulin declined, causing cells to stop responding normally to the hormone. Insulin-resistant rodent models have firmly established themselves as a paradigm for insulin resistance and diabetes drug discovery but have some limitations to high-throughput drug screening. Zebrafish has recently been discovered as a promising model for drug research and discovery of therapeutic targets, however, only a few research has been done on insulin resistance models. Objective: This study aims to evaluate the putative pathways involved in insulin resistance zebrafish model and correlates it with human insulin signalling pathway by deploying Next Generation Sequencing (NGS) and bioinformatics analysis. Methodology: Insulin resistance zebrafish model have been established by immersion method with 250 nM and re-challenged with 100 nM human recombinant insulins at age of 72 hour-post-fertilization (hpf) and 96 hpf respectively. The validation of dynamic glucose study using fluorometric assay and lipid analysis using Oil Red-O (ORO) staining have shown the achievement of insulin resistance in zebrafish model. Genes of interest were analysed using qPCR and NGS have been used to obtain the differential expressed genes (DEGs). Reactome analysis which is an open pathway database was then performed to identify the functional pathways of the DEGs. Results: A total of 96 DEGs were found to be involved in insulin resistance zebrafish model, with 2 genes (SPTBN4 and PRKCA) from the 49 upregulated genes were enriched in MAPK family signalling cascades, and integration of energy metabolism pathway respectively and 1 gene (fbp2) from the 47 downregulated genes was enriched in glucose metabolism as curated and annotated in the molecular mechanism of Reactome analysis. Discussion: This study has revealed the correlation between the pathways involved in insulin resistance zebrafish model with human as most of the pathways involved are the characteristics for T2DM and had demonstrated the usefulness of zebrafish model for potential drug discovery in insulin resistance. Conclusion: Bioinformatics analysis has provided the insights on the broad molecular event pathways of insulin resistance associated between zebrafish and human which suggest for further research to be done using the animal model and utilising NGS application.

Item Type:	Project Paper
Faculty:	Faculty of Medicine and Health Science
Depositing User:	Ms. Nor Safa'aton Saidin
Date Deposited:	22 Aug 2023 07:46
Last Modified:	22 Aug 2023 07:46
URI:	http://psaspb.upm.edu.my/id/eprint/1124

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