Abstract: PO0494
Renal Proximal Tubule Chip (RPTC) for Disease Modeling and Drug Toxicity Testing
Session Information
- Bioengineering: Organoids and Organs-on-a-Chip
November 04, 2021 | Location: On-Demand, Virtual Only
Abstract Time: 10:00 AM - 12:00 PM
Category: Bioengineering
- 300 Bioengineering
Authors
- Donoghue, Leslie, The University of Alabama at Birmingham, Birmingham, Alabama, United States
- Kotru, Arushi, The University of Alabama at Birmingham, Birmingham, Alabama, United States
- Guzman, Alfredo Jose, The University of Alabama at Birmingham, Birmingham, Alabama, United States
- Sethu, Palaniappan, The University of Alabama at Birmingham, Birmingham, Alabama, United States
Background
Tissue chips are an emerging technology in disease modeling and screening therapeutics to address discrepancies between animal models and human clinical trials. They utilize tissue engineering, fluid mechanics, and biomaterials to replicate in vivo architectures and functions of complex organs and tissues. For the renal proximal tubule (PT), there are currently limited options in terms of human cell types, scalable platforms for evaluation of drug toxicity, and tissue engineered solutions where the complexity of the PT is accurately modeled.
Methods
We developed both 2D and 3D versions of the RPTC which incorporates immortalized human renal PT epithelial cells (hRPTEC-TERT1) under static and perfused conditions (i.e. physiological pressure, shear stress, and stretch). Additionally, we have begun generating peritubular vascular networks using a co-culture of human umbilical vein endothelial cells (hUVECs) and human dermal fibroblasts (hDFs) in gelatin methacryloyl (GelMA). These models were then used to investigate the effect of pressure and flow on nephrotoxicity by introducing drugs with known levels of toxicity. Our initial evaluations have been limited to non-invasive measurements such as transepithelial electrical resistance (TEER) and pro-inflammatory soluble factors, and ICC.
Results
Compared to static controls, hRPTEC-TERT1 subjected to fluid shear demonstrate that culture under physiologically relevant forces results in cytoskeletal reorganization, establishment of barrier function (adherens and tight junctions) and increased expression of transporters like aquaporin 1 and mechanosensors like α-tubulin. Additionally, noninvasive readouts such as TEER indicate the greater integrity of the renal proximal epithelium. Lastly, after 7 days, we can form dense microvascular networks to mimic the surrounding peritubular capillary networks which can actively reabsorb solutes from the glomerular filtrate. This network will enable us to test drugs in an environment where both reabsorption and secretion functions of the tubule are replicated.
Conclusion
These results provide preliminary evidence of our ability to subject hRPTEC-TERT1 to in vivo like flow conditions and demonstrate that replication of biomechanical cues from fluid flow significantly enhances the attainment of an in vivo–like phenotype which enhances the relevance of our in vitro models.
Funding
- NIDDK Support