PT  - JOURNAL ARTICLE
AU  - Singh, Manish Kumar
AU  - Rallabandi, Harikrishna Reddy
AU  - Zhou, Xu-Jie
AU  - Qi, Yuan-Yuan
AU  - Zhao, Zhan-Zheng
AU  - Gan, Ting
AU  - Zhang, Hong
AU  - Looger, Loren L
AU  - Nath, Swapan K
TI  - <em>KLF2</em> enhancer variant rs4808485 increases lupus risk by modulating inflammasome machinery and cellular homoeostasis
AID  - 10.1136/ard-2023-224953
DP  - 2024 Jul 01
TA  - Annals of the Rheumatic Diseases
PG  - 879--888
VI  - 83
IP  - 7
4099  - http://ard.bmj.com/content/83/7/879.short
4100  - http://ard.bmj.com/content/83/7/879.full
SO  - Ann Rheum Dis2024 Jul 01; 83
AB  - Objective A recent genome-wide association study linked KLF2 as a novel Asian-specific locus for systemic lupus erythematosus (SLE) susceptibility. However, the underlying causal functional variant(s), cognate target gene(s) and genetic mechanisms associated with SLE risk are unknown.Methods We used bioinformatics to prioritise likely functional variants and validated the best candidate with diverse experimental techniques, including genome editing. Gene expression was compared between healthy controls (HCs) and patients with SLE with or without lupus nephritis (LN+, LN−).Results Through bioinformatics and expression quantitative trait locus analyses, we prioritised rs4808485 in active chromatin, predicted to modulate KLF2 expression. Luciferase reporter assays and chromatin immunoprecipitation-qPCR demonstrated differential allele-specific enhancer activity and binding of active histone marks (H3K27ac, H3K4me3 and H3K4me1), Pol II, CTCF, P300 and the transcription factor PARP1. Chromosome conformation capture-qPCR revealed long-range chromatin interactions between rs4808485 and the KLF2 promoter. These were directly validated by CRISPR-based genetic and epigenetic editing in Jurkat and lymphoblastoid cells. Deleting the rs4808485 enhancer in Jurkat (KO) cells disrupted NLRP3 inflammasome machinery by reducing KLF2 and increasing CASPASE1, IL-1β and GSDMD levels. Knockout cells also exhibited higher proliferation and cell-cycle progression than wild type. RNA-seq validated interplay between KLF2 and inflammasome machinery in HC, LN+ and LN−.Conclusions We demonstrate how rs4808485 modulates the inflammasome and cellular homoeostasis through regulating KLF2 expression. This establishes mechanistic connections between rs4808485 and SLE susceptibility.Data are available on reasonable request.