CRISPR/Cas9-mediated deletion of adipocyte genes associated with NAFLD alters adipocyte lipid handling and reduces steatosis in hepatocytes in vitro

Lopez-Yus, Marta ; Frendo-Cumbo, Scott ; del Moral-Bergos, Raquel ; Garcia-Sobreviela, Maria Pilar ; Bernal-Monterde, Vanesa (Universidad de Zaragoza) ; Rydén, Mikael ; Lorente-Cebrian, Silvia (Universidad de Zaragoza) ; Arbones-Mainar, Jose M.

Resumen: Obesity is a major risk factor for the development of nonalcoholic fatty liver disease (NAFLD), and the subcutaneous white adi-pose tissue (scWAT) is the primary lipid storage depot and regulates lipidfluxes to other organs. Our previous work identifiedgenes upregulated in scWAT of patients with NAFLD:SOCS3,DUSP1,andSIK1. Herein, we knocked down (KD) their expres-sion in human adipose-derived mesenchymal stem cells (hADMSCs) using clustered regularly interspaced short palindromicrepeats (CRISPR)/Cas9 technology and characterized their phenotype. We found thatSOCS3,DUSP1,andSIK1expression inhADMSC-derived adipocytes was not critical for adipogenesis. However, the metabolic characterization of the cells suggestedthat the genes played important roles in lipid metabolism. Reduction ofSIK1expression significantly increased both de novolipogenesis (DNL) and palmitate-induced lipogenesis (PIL). Editing outSOCS3reduced DNL while increasing isoproterenol-induced lipolysis and insulin-induced palmitate accumulation. Conversely,DUSP1reduced PIL and DNL. Moreover, RNA-sequencing analysis of edited cells showed that these genes not only altered lipid metabolism but also other biological path-ways related to inflammatory processes, in the case ofDUSP1, extracellular matrix remodeling forSOCS3, or cellular transportforSIK1. Finally, to evaluate a possible adipocyte-hepatocyte axis, human hepatoma HepG2 cells were cocultured with editedhADMSCs-derived adipocytes in the presence of [3H]-palmitate. All HepG2 cells cultured with DUSP1-, SIK1-, or SOCS3-KD adi-pocytes decreased [3H]-palmitate accumulation compared with control adipocytes. These results support our hypotheses thatSOCS3,DUSP1,andSIK1regulate multiple aspects of adipocyte function, which may play a role in the progression of obesity-associated comorbidities, such as NAFLD.

Idioma: Inglés

DOI: 10.1152/ajpcell.00291.2023

Año: 2023

Publicado en: American Journal of Physiology - Cell Physiology 325, 5 (2023), C1178-C1189
ISSN: 0363-6143

Factor impacto JCR: 5.0 (2023)

Categ. JCR: PHYSIOLOGY rank: 8 / 85 = 0.094 (2023) - Q1 - T1
Categ. JCR: CELL BIOLOGY rank: 66 / 205 = 0.322 (2023) - Q2 - T1

Factor impacto CITESCORE: 9.1 - Physiology (Q1) - Cell Biology (Q2)

Factor impacto SCIMAGO: 1.711 - Physiology (Q1) - Cell Biology (Q1)

Financiación: info:eu-repo/grantAgreement/ES/DGA/B03-23R
Financiación: info:eu-repo/grantAgreement/ES/ISCIII/PI22-01366
Financiación: info:eu-repo/grantAgreement/ES/MICINN /BFU2015-65937-R
Financiación: info:eu-repo/grantAgreement/ES/UZ/JIUZ-2021-BIO-03

Tipo y forma: Artículo (Versión definitiva)

Área (Departamento): Área Fisiología (Dpto. Farmac.Fisiol.y Med.L.F.)
Área (Departamento): Area Medicina (Dpto. Medicina, Psiqu. y Derm.)