Adult and iPS-derived non-parenchymal cells regulate liver organoid development through differential modulation of Wnt and TGF-beta

Autor(es):Mostrar menos -

Goulart, Ernesto ^[1] ; de Caires-Junior, Luiz Carlos ^[1] ; Telles-Silva, Kayque Alves ^[1] ; Silva Araujo, Bruno Henrique ^[2] ; Kobayashi, Gerson S. ^[1] ; Musso, Camila Manso ^[1] ; Assoni, Amanda Faria ^[1] ; Oliveira, Danyllo ^[1] ; Caldini, Elia ^[3] ; Gerstenhaber, Jonathan A. ^[4] ; Raia, Silvano ^[5] ; Lelkes, Peter I. ^[4] ; Zatz, Mayana ^[1]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Dept Genet & Evolutionary Biol, Inst Biosci, Human Genome & Stem Cell Res Ctr HUG CEL, Sao Paulo, SP - Brazil

^[2] Brazilian Ctr Res Energy & Mat CNPEM, Brazilian Biosci Natl Lab LNBio, BR-13083970 Campinas, SP - Brazil

^[3] Univ Sao Paulo, Med Sch, Dept Pathol, Lab Cellular Biol, Sao Paulo, SP - Brazil

^[4] Temple Univ, Dept Bioengn, Philadelphia, PA 19122 - USA

^[5] Univ Sao Paulo, Med Sch, Dept Surg, Sao Paulo, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: STEM CELL RESEARCH & THERAPY; v. 10, n. 1 AUG 15 2019.
• Citações Web of Science: 1

Resumo

Background Liver organoid technology holds great promises to be used in large-scale population-based drug screening and in future regenerative medicine strategies. Recently, some studies reported robust protocols for generating isogenic liver organoids using liver parenchymal and non-parenchymal cells derived from induced pluripotent stem cells (iPS) or using isogenic adult primary non-parenchymal cells. However, the use of whole iPS-derived cells could represent great challenges for a translational perspective. Methods Here, we evaluated the influence of isogenic versus heterogenic non-parenchymal cells, using iPS-derived or adult primary cell lines, in the liver organoid development. We tested four groups comprised of all different combinations of non-parenchymal cells for the liver functionality in vitro. Gene expression and protein secretion of important hepatic function markers were evaluated. Additionally, liver development-associated signaling pathways were tested. Finally, organoid label-free proteomic analysis and non-parenchymal cell secretome were performed in all groups at day 12. Results We show that liver organoids generated using primary mesenchymal stromal cells and iPS-derived endothelial cells expressed and produced significantly more albumin and showed increased expression of CYP1A1, CYP1A2, and TDO2 while presented reduced TGF-beta and Wnt signaling activity. Proteomics analysis revealed that major shifts in protein expression induced by this specific combination of non-parenchymal cells are related to integrin profile and TGF-beta/Wnt signaling activity. Conclusion Aiming the translation of this technology bench-to-bedside, this work highlights the role of important developmental pathways that are modulated by non-parenchymal cells enhancing the liver organoid maturation. (AU)

• Processo FAPESP: 13/08028-1 - CEGH-CEL - Centro de Estudos do Genoma Humano e de Células-Tronco
• Beneficiário: Mayana Zatz
• Linha de fomento: Auxílio à Pesquisa - Centros de Pesquisa, Inovação e Difusão - CEPIDs