Project description:Facial infiltrating lipomatosis is a congenital malformation characterized primarily by the excessive proliferation of adipose tissue. However, the pathogenesis of it remains unclear. To compare the gene expression differences between the adipose tissue of facial infiltrating lipomatosis and normal facial subcutaneous adipose tissue, we collected a total of 14 adipose tissue samples for high-throughput RNA sequencing, aiming to identify potential pathogenic genes.
Project description:Facial infiltrating lipomatosis (FIL) is a congenital disorder characterized by unilateral facial enlargement. Although next-generation sequencing has revealed that the pathogenesis of FIL is associated with phosphatidylinositol 3-kinase catalytic subunit alpha (PIK3CA) mutations, the underlying molecular mechanisms remain undetermined. We found that the adipose tissue in FIL patients demonstrated tissue infiltration accompanied by adipocytes hypertrophy and increased lipid accumulation. All FIL-ADSCs harboured PIK3CA mutations. Compared to ADSCs obtained from normal subcutaneous adipose tissue, FIL-ADSCs exhibited a greater capacity for adipogenesis. Suppression of PIK3CA resulted in a reduction in the adipogenic potential of FIL-ADSCs. Furthermore, WX390, a novel dual-target PI3K/mTOR inhibitor, was found to impede PIK3CA-mediated adipogenesis both in vivo and in vitro. RNA-seq revealed that the expression of transient receptor potential vanilloid subtype 1 (TRPV1) was upregulated after PI3K pathway inhibition, and overexpression and activation of TRPV1 both inhibited adipogenesis of FIL-ADSCs. Our study showed that PIK3CA mutations promoted adipogenesis in FIL-ADSCs and that this effect was achieved by suppressing the expression of TPRV1. Pathogenesis experiments suggested that WX390 may serve as an agent for the treatment of FIL.
Project description:To investigate the function of N6-methyladenosine methylome (m6A) in adipose stem and progenitor cells isolated from facial infiltrating lipomatosis (FIL-ASPCs), we analyzed m6A enrichment level in FIL-ASPCs with or without FTO inhibitor (FTO-IN-1) treatment through methylated RNA immunoprecipitation (MeRIP) sequencing.
Project description:Facial infiltrating lipomatosis is characterized by excessive growth of adipose tissue, The etiology is associated with somatic PIK3CA variant, but the specific mechanisms are not yet fully understood. In this study, we collected facial adipose tissue from both FIL patients and non-FIL individuals, isolated the stromal vascular fraction (SVF) and performed single-cell transcriptome sequencing on these samples. We mapped out the cellular landscape within the SVF and specifically focused on a deeper analysis of fibro-adipogenic precursor cells (FAPs).
Project description:In this study, we found that H3K18la level is elevated in adipose stem and progenitor cells of facial infiltrating lipomatosis (FIL-ASPCs). To further explore the potential functional significance of H3K18la in FIL, we performed genome-wide cleavage under targets and tagmentation (CUT&Tag) analysis to identify candidate genes regulated by H3K18la in FIL-ASPCs and CON-ASPCs. Following CUT&Tag, H3K18la-associated DNAs were amplified using non-biased conditions, labeled, and sequenced with Illumina NovaSeq 150PE.
Project description:Facial infiltrating lipomatosis (FIL) is a congenital disorder characterized by unilateral facial enlargement. Although next-generation sequencing has revealed that the pathogenesis of FIL is associated with phosphatidylinositol 3-kinase catalytic subunit alpha (PIK3CA) mutations, the underlying molecular mechanisms remain undetermined. We found that the adipose tissue in FIL patients demonstrated tissue infiltration accompanied by adipocytes hypertrophy and increased lipid accumulation. All FIL derived fibro-adipose progenitor cells (FIL-FAPs) harboured PIK3CA mutations. Compared to FAPs obtained from normal subcutaneous adipose tissue, FIL-FAPs exhibited a greater capacity for adipogenesis. Suppression of PIK3CA resulted in a reduction in the adipogenic potential of FIL-FAPs. Through single cell sequencing, we find that FIL-FAPs had higher expression of FKBP5. Inhibiting FKBP5 can impair the adipogenic capacity of FIL-FAPs. We also verified that PI3K-AKT pathway regulating FKBP5 expression. To find the downstream target of FKBP5, we performed RNA-seq of FIL-FAPs treated with FKBP5 inhibitor SAFit2 in DMSO or DMSO alone.
