TY - JOUR

T1 - 3D Adipose Tissue Culture Links the Organotypic Microenvironment to Improved Adipogenesis

AU - Shen, Joanne X.

AU - Couchet, Morgane

AU - Dufau, Jérémy

AU - de Castro Barbosa, Thais

AU - Ulbrich, Maximilian H.

AU - Helmstädter, Martin

AU - Kemas, Aurino M.

AU - Zandi Shafagh, Reza

AU - Marques, Marie-Adeline

AU - Hansen, Jacob B.

AU - Mejhert, Niklas

AU - Langin, Dominique

AU - Rydén, Mikael

AU - Lauschke, Volker M.

N1 - Funding Information: The work was supported by the European Research Council (ERC) Synergy Grant SPHERES (agreement No. 856404) under the European Union's Horizon 2020 research and innovation program (erc‐spheres.univ‐tlse3.fr), the Swedish Research Council [grant agreement numbers: 2016‐01153, 2016‐01154, 2018‐02488 and 2019‐01837], the EU/EFPIA/OICR/McGill/KTH/Diamond Innovative Medicines Initiative 2 Joint Undertaking (EUbOPEN grant number 875510) and by the Strategic Research Programmes in Diabetes (SFO Diabetes) and Stem Cells and Regenerative Medicine (SFO StratRegen). Furthermore, the authors were supported by grants from the Margareta af Uggla's Foundation, the Novo Nordisk Foundation (including the Tripartite Immuno‐metabolism Consortium Grant NNF15CC0018486, the MeRIAD consortium grant number 0064142, the MSAM Consortium NNF15SA0018346 and NNF20OC0061149), Knut and Alice Wallenberg Foundation, CIMED, the Swedish Diabetes Foundation, the Stockholm County Council, Fondation pour la Recherche Médicale (DEQ20170336720), Agence Nationale de la Recherche ANR‐17‐CE14‐0015Hepadialogue and AstraZeneca France. The authors gratefully acknowledge support from the Bioinformatics and Expression Analysis Core Facility and the VirusTech Core Facility at Karolinska Institutet for integrating RNA‐Seq datasets from different sources and provision of AAV‐GFP serotypes, respectively. SVF spheroid and TERT‐hWA imaging were performed at the Biomedicum Imaging Core (BIC) facility and the LCI facility/Nikon Center of Excellence, Karolinska Institutet. The authors also acknowledge the assistance of Dr. Sonia Youhanna with spheroid culture. Publisher Copyright: © 2021 The Authors. Advanced Science published by Wiley-VCH GmbH

PY - 2021

Y1 - 2021

N2 - Obesity and type 2 diabetes are strongly associated with adipose tissue dysfunction and impaired adipogenesis. Understanding the molecular underpinnings that control adipogenesis is thus of fundamental importance for the development of novel therapeutics against metabolic disorders. However, translational approaches are hampered as current models do not accurately recapitulate adipogenesis. Here, a scaffold-free versatile 3D adipocyte culture platform with chemically defined conditions is presented in which primary human preadipocytes accurately recapitulate adipogenesis. Following differentiation, multi-omics profiling and functional tests demonstrate that 3D adipocyte cultures feature mature molecular and cellular phenotypes similar to freshly isolated mature adipocytes. Spheroids exhibit physiologically relevant gene expression signatures with 4704 differentially expressed genes compared to conventional 2D cultures (false discovery rate < 0.05), including the concerted expression of factors shaping the adipogenic niche. Furthermore, lipid profiles of >1000 lipid species closely resemble patterns of the corresponding isogenic mature adipocytes in vivo (R2 = 0.97). Integration of multi-omics signatures with analyses of the activity profiles of 503 transcription factors using global promoter motif inference reveals a complex signaling network, involving YAP, Hedgehog, and TGFβ signaling, that links the organotypic microenvironment in 3D culture to the activation and reinforcement of PPARγ and CEBP activity resulting in improved adipogenesis.

AB - Obesity and type 2 diabetes are strongly associated with adipose tissue dysfunction and impaired adipogenesis. Understanding the molecular underpinnings that control adipogenesis is thus of fundamental importance for the development of novel therapeutics against metabolic disorders. However, translational approaches are hampered as current models do not accurately recapitulate adipogenesis. Here, a scaffold-free versatile 3D adipocyte culture platform with chemically defined conditions is presented in which primary human preadipocytes accurately recapitulate adipogenesis. Following differentiation, multi-omics profiling and functional tests demonstrate that 3D adipocyte cultures feature mature molecular and cellular phenotypes similar to freshly isolated mature adipocytes. Spheroids exhibit physiologically relevant gene expression signatures with 4704 differentially expressed genes compared to conventional 2D cultures (false discovery rate < 0.05), including the concerted expression of factors shaping the adipogenic niche. Furthermore, lipid profiles of >1000 lipid species closely resemble patterns of the corresponding isogenic mature adipocytes in vivo (R2 = 0.97). Integration of multi-omics signatures with analyses of the activity profiles of 503 transcription factors using global promoter motif inference reveals a complex signaling network, involving YAP, Hedgehog, and TGFβ signaling, that links the organotypic microenvironment in 3D culture to the activation and reinforcement of PPARγ and CEBP activity resulting in improved adipogenesis.

KW - adipose stem cells

KW - fat cells

KW - lipidomics

KW - organotypic cell culture

KW - preadipocytes

KW - stromal vascular fraction

KW - transcriptomics

U2 - 10.1002/advs.202100106

DO - 10.1002/advs.202100106

M3 - Journal article

C2 - 34165908

AN - SCOPUS:85108645664

VL - 8

JO - Advanced Science

JF - Advanced Science

SN - 2198-3844

IS - 16

M1 - 2100106

ER -