3D Human Orbital Fibroblast Platform for Inflammatory Biomarker Discovery

Thyroid eye disease (TED) is an autoimmune inflammatory disorder and the most common cause of orbital disease in adults. Often associated with Graves’ disease, TED can lead to debilitating visual impairment due to bulging of the eyes (proptosis) from severe swelling of the extraocular muscles and orbital tissues. The pathophysiology of TED is thought to involve activation of periorbital fibroblasts, triggering an inflammatory cascade that results in local accumulation of lipids and glycosaminoglycans and, ultimately, fibrosis. Fibroblast-driven extracellular matrix remodeling and fibrosis are also hallmarks of the stromal compartment in many solid tumors, linking TED biology to broader questions in tumor microenvironment research. To better understand the mechanistic pathophysiology of TED and to support discovery of agents that target fibrotic stroma, we have developed an in vitro 3D culture model using primary human orbital fibroblasts. In this system, fibroblasts form spheroidal 3D structures that provide a more physiologically relevant context than conventional monolayer culture. Using this platform as a bridge between clinic and bench, we assay surface, intracellular, and secreted inflammatory biomarkers following IGF-1 stimulation—a signaling axis with established roles in both autoimmunity and cancer—using ELISA, MSD, JESS, and flow cytometry. Endpoint data are integrated through a bespoke AI- and machine-learning-based pipeline to identify and rank multidimensional biomarker signatures. Taken together, these data strongly validate our 3D human orbital fibroblast culture as a useful model to investigate the molecular mechanisms underlying the pathogenesis of TED and to screen potential therapeutics via assays developed in-house with clients and partners in the autoimmune space. More broadly, this IGF-1-responsive 3D human stromal platform provides a tractable model of fibrotic, inflammatory fibroblast biology that is directly relevant to the tumor microenvironment and can be extended to future co-culture studies with tumor and immune cells to support preclinical oncology drug discovery. Citation Format: Mengling Liu, Gene Lin, Noel Derecki, Lily Li, Paula Pellon, Calvin Ha, . Autoimmunity in a Dish: A 3D human orbital fibroblast platform for inflammatory biomarker discovery and fibrotic stromal biology [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 7531.