A High-Throughput Kidney-on-a-Chip Platform for CKD Therapy Discovery
- Bioengineering: Modeling, Diagnosis, Therapy
November 04, 2023 | Location: Exhibit Hall, Pennsylvania Convention Center
Abstract Time: 10:00 AM - 12:00 PM
- 400 Bioengineering
- Chandler, Morgan, MIMETAS US, Gaithersburg, Maryland, United States
- Alsebahi, Alaa, MIMETAS US, Gaithersburg, Maryland, United States
- Zhang, Qi, Children's Hospital Los Angeles, Los Angeles, California, United States
- Da Sacco, Stefano, Children's Hospital Los Angeles, Los Angeles, California, United States
- Perin, Laura, Children's Hospital Los Angeles, Los Angeles, California, United States
- Bircsak, Kristin, MIMETAS US, Gaithersburg, Maryland, United States
Glomerular and/or proximal tubule (PT) dysfunction often culminates in the onset of chronic kidney disease (CKD). Existing therapies address symptoms rather than the underlying causes, resulting in significant side effects. More targeted therapies with increased specificity to the glomerulus or PT are needed, but their development is greatly hindered by the lack of relevant human model systems. To address this, we developed a three-dimensional in vitro kidney-on-a-chip mimicking glomerular and PT function in a high-throughput platform: the NephroPlate.
The NephroPlate was established in MIMETAS’ OrganoPlate, a 40-assay format allowing culture with inner tissue flow in a standard microtiter plate. The glomerular unit was prepared using primary human podocytes and glomerular endothelial cells. The PT unit was prepared using renal PT epithelial cells (RPTEC) and human umbilical vein endothelial cells (HUVEC). Barriers were assessed using transepithelial electric resistance via Organo(TEER). To assess glomerular filtration (GF), FITC-albumin was added to cultures mimicking either healthy or Membranous Nephropathy (MN) conditions. Expression was confirmed by immunostaining and media were collected for evaluation of secreted factors.
Cultures within the NephroPlate maintained cell phenotypes in a three-dimensional format, facilitating the de novo production of glomerular basement membrane (GBM) components and mimicking GF. While all donor-derived cultures recapitulated healthy functions, treatment with serum from patients with MN resulted in a donor-dependent loss of GF and TEER. In CKD models, treatment with α-melanocyte-stimulating hormone rescued the MN serum-mediated loss of GF as demonstrated by albumin retention, while TEER did not recover within the experimental timeframe. Upon transfer of CKD glomerular filtrate to the PT, elevated KIM-1, a marker of PT injury, was observed.
The NephroPlate captures many aspects of the kidney microenvironment, including complex composition as well as GF function and its dynamic response to mimic CKD. The high-throughput capabilities of our system allow for up to 40 conditions to be assessed simultaneously, enhancing our ability to screen existing and novel drugs as well as identifying better, more targeted therapies for CKD.
- NIDDK Support