Innovating a human adipocyte spheroid platform to explore the role of metabolite-sensing GPCRs in metabolic disease

Duncan, Elaine Margaret (2026) Innovating a human adipocyte spheroid platform to explore the role of metabolite-sensing GPCRs in metabolic disease. PhD thesis, University of Glasgow.

Abstract

Metabolic diseases such as obesity and type 2 diabetes are a global healthcare and economic burden affecting over 1 billion individuals worldwide. These diseases are characterised by excessive accumulation and dysfunction of adipose tissue, leading to impaired metabolic regulation and severe downstream consequences for patients. Low level chronic inflammation of adipose tissue is an important hallmark of metabolic disease, and there is growing evidence that metabolite-sensing G Protein-Coupled Receptors (m-GPCRs) can regulate metabolic function through autocrine and paracrine signalling loops. However, it has been challenging to dissect these complex signalling networks using traditional 2D cell culture or in vivo experimental models. This thesis aimed to address this gap by engineering and validating an innovative 3D in vitro model of adipose tissue suitable for investigating the role of m-GPCRs in metabolic disease.

First, a series of novel genetically encoded NanoBiT biosensors were designed and optimised to quantify real time signalling of unmodified GPCRs in live cells. Next, spheroids were generated by seeding human Simpson Golabi Behmel Syndrome (SGBS) preadipocytes in ultra-low adhesion plates and differentiating them into adipocytes. Comprehensive morphological, transcriptional and protein-level characterisation demonstrated that spheroids accumulate lipid droplets and upregulate key markers of adipogenesis during differentiation. Critically, the differentiated spheroids show characteristic adipocyte functions, including β-adrenergic-stimulated lipolysis, and insulin-stimulated glucose uptake. To mimic a metabolic disease-relevant phenotype, spheroids were treated with Tumor Necrosis Factor which induced a pro-inflammatory, insulin resistant microenvironment, and revealed upregulation of the FFA4 receptor in disease-relevant conditions. Finally, the role of m-GPCRs in adipogenesis and lipolysis could be defined by treating adipocyte spheroids with pharmacological tool compounds, and genetically encoded biosensors were incorporated to directly measure receptor signalling in adipocytes.

Overall, a novel human adipocyte spheroid platform has been developed which has allowed the investigation of m-GPCR function within a physiologically- and disease-relevant context. This therefore provides a strong foundation to interrogate the complex metabolic-immune signalling networks within adipose tissue, and may ultimately lead to new GPCR-focused therapeutic strategies for metabolic disease.

• Item Type: Thesis (PhD)
• Qualification Level: Doctoral
• Subjects: Q Science > QH Natural history > QH345 Biochemistry
• Colleges/Schools: College of Medical Veterinary and Life Sciences > School of Molecular Biosciences
• Funder's Name: Engineering and Physical Sciences Research Council (EPSRC)
• Supervisor's Name: Hudson, Dr. Brian, Dalby, Professor Matthew and Berry, Dr. Catherine
• Date of Award: 2026
• Depositing User: Theses Team
• Unique ID: glathesis:2026-85780
• Copyright: Copyright of this thesis is held by the author.
• Date Deposited: 24 Feb 2026 10:34
• Last Modified: 24 Feb 2026 11:50
• Thesis DOI: 10.5525/gla.thesis.85780
• URI: https://theses.gla.ac.uk/id/eprint/85780

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