D2.107 - Effect of n-3 long-chain polyunsaturated fatty acid and vitamin D supplementation on human lung organoids
Background
Randomized clinical trials (RCTs) suggest that n-3 long-chain polyunsaturated fatty acids (n-3 LCPUFA) and high-dose vitamin D supplementation during pregnancy may protect against childhood asthma. However, the underlying mechanisms remain unclear. In this study, we aimed to investigate the biological effects of varying doses of n-3 LCPUFA and vitamin D supplementation on the development of human lung organoids, with a focus on asthma pathophysiology.
Method
At Sanger institute (Cambridge, UK), we cultured human lung organoids derived from BILX and SEHP human-induced pluripotent stem cell lines with none, low (0.01 µl/ml) and high (0.1 µl/ml) concentrations of n-3 LCPUFA and none, low (5 pM) and high (50 pM) concentrations of vitamin D and measured the impact of the supplementations using qPCR and RNA sequencing. Raw sequencing reads were processed using FastQC, trimmed to remove adapters and low-quality bases with Trimmomatic, and aligned to the reference transcriptome using Salmon to generate normalized transcript abundance estimates. These estimates were analyzed using DESeq2 to identify differentially expressed genes. Pathway enrichment analysis was conducted using the pathfindR package in R.
Results
Our findings revealed that gene expression in lung organoids derived from human iPSC lines is more affected by increasing doses of n-3 LCPUFA than by vitamin D (differentially expressed genes: n = 907 vs. n = 23). CPT1A and ANGPTL4 genes were highly expressed in media cultured with a high dose of n-3 LCPUFA, while CYP24A1 was among the highly expressed genes in media cultured with a high dose of vitamin D. Enrichment analysis showed activation of PPAR pathways, suggesting that n-3 LCPUFA supplementation may protect against asthma by regulating lipid metabolism and inflammation.
Conclusion
We identified several genes and pathways that may provide insights into the biological effects of n-3 LCPUFA and vitamin D supplementation on asthma pathophysiology.
