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Abstract: PO0510

Physiological Replication of the Glomerulus Using a Triple Culture Microphysiological System

Session Information

Category: Bioengineering

  • 300 Bioengineering

Authors

  • Pajoumshariati, Seyedramin, AstraZeneca PLC, Mölndal, Sweden
  • Ewart, Lorna, Emulate Inc, Cambridge, Massachusetts, United States
  • Luc, Raymond, Emulate Inc, Cambridge, Massachusetts, United States
  • Kujala, Ville J., Emulate Inc, Cambridge, Massachusetts, United States
  • Peel, Samantha, AstraZeneca PLC, Mölndal, Sweden
  • Weber, Heather, AstraZeneca PLC, Mölndal, Sweden
  • Laerkegaard Hansen, Pernille B., AstraZeneca PLC, Mölndal, Sweden
  • Williams, Julie, AstraZeneca PLC, Mölndal, Sweden
Background

The glomerulus is a complex structure highly adapted for its function. A true representation of the in vivo cell-cell/ECM interactions, which provide the semi-permeable filtration barrier, is essential to interrogate physiological and pathophysiological processes and for understanding the impact of novel therapies.

Methods

We have developed a human microphysiological system with high fidelity to glomerular physiology and structure. For the first time the three resident cell types (glomerular endothelial cells (GECs), induced pluripotent stem cell (hiPSC)-derived podocytes and mesangial cells (MCs)) reside in a relevant 3D structure under flow conditions. Analysis was performed on the individual cell types using both transcriptomics (NGS) and high content imaging to shed light on the impact of each cell type on its neighbors. Inulin and albumin permeability assays (fluorescent) were performed to evaluate the integrity of the glomerular barrier.

Results

Transcriptomic analyses demonstrated crosstalk between cells in our microfluidic tri-culture system. An influence of MCs was observed on both podocytes and GECs. For GECs, MCs increased tight junction proteins (CLDN7) and for podocytes there was modulation of cell cycle control (WDR70). The differentiation of podocytes in the chip was able to regulate matrix and cell adhesion in MC (COL6A3, ITGA2) and influence angiogenic signals in GEC (KDR, THBS1). Analysis of pathways expressed in cells in the less complex comparator systems showed that they displayed transcriptomic signals akin to human disease phenotypes (comparison with signatures found in Nephroseq).
Imaging showed increases in maturation markers such as synaptopodin in podocytes in triculture. Permeability assays revealed that as cell maturity increased barrier function improved and the passage of molecules was selectively hindered.

Conclusion

Our tri-culture model provides a highly physiologically relevant tool to study healthy glomerular function. This will enable improved understanding of the mechanisms underlying glomerulopathies and improved qualification of new therapies.

Funding

  • Commercial Support