The non-alcoholic fatty liver disease (NAFLD), increasing the global healthcare burden in the past decade, is a nutrition-triggered liver disorder associated with high heritability, genetic pleiotropy, type 2 diabetes, and obesity. NAFLD involves 20-60% cystic fibrosis (CF) cases with unknown pathogenic mechanisms. CF is the common potentially fatal inherited disease, mainly compromises pulmonary functions. The high-temperature, high-resolution gas-chromatography mass spectrometry (GC-MS) system was established in Dr. Yang’s lab, resolving the protective lipid moieties in cultivated hepatic cell line and mice model under cellular senescence. The GC-MS system can detect NAFLD/CF patients’ metabolite modification, which provides excellent metabolomic information of inherited disease. Since the primary gene of cystic fibrosis transmembrane conductance regulator (CFTR) is not expressed on differentiated hepatocytes, the etiology might be related to multiple environmental factors, including malnutrition, oxidative stress, and insulin resistance. Organoid and scRNA-seq immediately become great tools to investigate the effect of CFTR in a mini hepatocyte tissue. In the NAFLD study, we include the following aim in this sub-project to address the environmental issue well.
Aim 3-1: To establish the NAFLD model with hiPSC-derived 3D liver organoids from SF, obese-NAFLD, and non-obese-NAFLD patients.
Aim 3-2: Identification novel functional lipids in NAFLD hiPSC-derived 3D liver organoids by quantitative lipidomic analysis.
Aim 3-3: To determine the cellular components and differential effects of functional lipids in hiPSC 3D organoids by CRISPR/cas9 technology and single-cell RNA-seq.
Aim 3-4: To evaluate the efficiency delivery formulation of functional lipids for therapeutic applications with patient-specific hiPSC-derived 3D organoids.