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FOXA1/2 depletion drives global reprogramming of differentiation state and metabolism in a human liver cell line and inhibits differentiation of human stem cell-derived hepatic progenitor cells.
Warren, Iyan; Moeller, Michael M; Guiggey, Daniel; Chiang, Alexander; Maloy, Mitchell; Ogoke, Ogechi; Groth, Theodore; Mon, Tala; Meamardoost, Saber; Liu, Xiaojun; Thompson, Sarah; Szeglowski, Antoni; Thompson, Ryan; Chen, Peter; Paulmurugan, Ramasamy; Yarmush, Martin L; Kidambi, Srivatsan; Parashurama, Natesh.
Afiliação
  • Warren I; Department of Chemical and Biological Engineering, University at Buffalo (State University of New York), Buffalo, New York, USA.
  • Moeller MM; Department of Chemical and Biomolecular Engineering, University of Nebraska- Lincoln, Lincoln, Nebraska, USA.
  • Guiggey D; Department of Chemical and Biological Engineering, University at Buffalo (State University of New York), Buffalo, New York, USA.
  • Chiang A; Department of Chemical and Biological Engineering, University at Buffalo (State University of New York), Buffalo, New York, USA.
  • Maloy M; Department of Chemical and Biological Engineering, University at Buffalo (State University of New York), Buffalo, New York, USA.
  • Ogoke O; Department of Chemical and Biological Engineering, University at Buffalo (State University of New York), Buffalo, New York, USA.
  • Groth T; Department of Chemical and Biological Engineering, University at Buffalo (State University of New York), Buffalo, New York, USA.
  • Mon T; Department of Chemical and Biological Engineering, University at Buffalo (State University of New York), Buffalo, New York, USA.
  • Meamardoost S; Department of Chemical and Biological Engineering, University at Buffalo (State University of New York), Buffalo, New York, USA.
  • Liu X; Department of Chemical and Biological Engineering, University at Buffalo (State University of New York), Buffalo, New York, USA.
  • Thompson S; Department of Chemical and Biological Engineering, University at Buffalo (State University of New York), Buffalo, New York, USA.
  • Szeglowski A; Department of Chemical and Biological Engineering, University at Buffalo (State University of New York), Buffalo, New York, USA.
  • Thompson R; Department of Chemical and Biological Engineering, University at Buffalo (State University of New York), Buffalo, New York, USA.
  • Chen P; Department of Biomedical Engineering, University at Buffalo (State University of New York), Buffalo, New York, USA.
  • Paulmurugan R; Department of Radiology, Canary Center for Early Cancer Detection and the Molecular Imaging Program at Stanford, Stanford University, Palo Alto, California, USA.
  • Yarmush ML; Center for Engineering in Medicine and Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA.
  • Kidambi S; Department of Biomedical Engineering, Rutgers University, Piscataway, New Jersey, USA.
  • Parashurama N; Department of Chemical and Biomolecular Engineering, University of Nebraska- Lincoln, Lincoln, Nebraska, USA.
FASEB J ; 37(1): e22652, 2023 01.
Article em En | MEDLINE | ID: mdl-36515690
ABSTRACT
FOXA factors are critical members of the developmental gene regulatory network (GRN) composed of master transcription factors (TF) which regulate murine cell fate and metabolism in the gut and liver. How FOXA factors dictate human liver cell fate, differentiation, and simultaneously regulate metabolic pathways is poorly understood. Here, we aimed to determine the role of FOXA2 (and FOXA1 which is believed to compensate for FOXA2) in controlling hepatic differentiation and cell metabolism in a human hepatic cell line (HepG2). siRNA mediated knockdown of FOXA1/2 in HepG2 cells significantly downregulated albumin (p < .05) and GRN TF gene expression (HNF4α, HEX, HNF1ß, TBX3) (p < .05) and significantly upregulated endoderm/gut/hepatic endoderm markers (goosecoid [GSC], FOXA3, and GATA4), gut TF (CDX2), pluripotent TF (NANOG), and neuroectodermal TF (PAX6) (p < .05), all consistent with partial/transient reprograming. shFOXA1/2 targeting resulted in similar findings and demonstrated evidence of reversibility of phenotype. RNA-seq followed by bioinformatic analysis of shFOXA1/2 knockdown HepG2 cells demonstrated 235 significant downregulated genes and 448 upregulated genes, including upregulation of markers for alternate germ layers lineages (cardiac, endothelial, muscle) and neurectoderm (eye, neural). We found widespread downregulation of glycolysis, citric acid cycle, mitochondrial genes, and alterations in lipid metabolism, pentose phosphate pathway, and ketogenesis. Functional metabolic analysis agreed with these findings, demonstrating significantly diminished glycolysis and mitochondrial respiration, with concomitant accumulation of lipid droplets. We hypothesized that FOXA1/2 inhibit the initiation of human liver differentiation in vitro. During human pluripotent stem cells (hPSC)-hepatic differentiation, siRNA knockdown demonstrated de-differentiation and unexpectedly, activation of pluripotency factors and neuroectoderm. shRNA knockdown demonstrated similar results and activation of SOX9 (hepatobiliary). These results demonstrate that FOXA1/2 controls hepatic and developmental GRN, and their knockdown leads to reprogramming of both differentiation and metabolism, with applications in studies of cancer, differentiation, and organogenesis.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Células-Tronco Pluripotentes / Fígado Limite: Animals / Humans Idioma: En Ano de publicação: 2023 Tipo de documento: Article
Texto completo
Imprimir
XML
PubMed Links
Buscar no Google

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Células-Tronco Pluripotentes / Fígado Limite: Animals / Humans Idioma: En Ano de publicação: 2023 Tipo de documento: Article