Abstract: FR-PO690
miR-193a and Nanoparticle Technology: A Novel Therapeutic Target in Alport Syndrome
Session Information
- Glomerular Diseases: Podocyte Biology - I
November 04, 2022 | Location: Exhibit Hall, Orange County Convention Center‚ West Building
Abstract Time: 10:00 AM - 12:00 PM
Category: Glomerular Diseases
- 1304 Glomerular Diseases: Podocyte Biology
Authors
- Perin, Laura, Children's Hospital Los Angeles, Los Angeles, California, United States
- Petrosyan, Astgik, Children's Hospital Los Angeles, Los Angeles, California, United States
- Kargin, Sinem, Children's Hospital Los Angeles, Los Angeles, California, United States
- Aguiari, Paola, Children's Hospital Los Angeles, Los Angeles, California, United States
- Zhang, Qi, Children's Hospital Los Angeles, Los Angeles, California, United States
- Hou, Xiaogang, Children's Hospital Los Angeles, Los Angeles, California, United States
- De Filippo, Roger E., Children's Hospital Los Angeles, Los Angeles, California, United States
- Lemley, Kevin V., Children's Hospital Los Angeles, Los Angeles, California, United States
- Chung, Eun ji, University of Southern California, Los Angeles, California, United States
- Da Sacco, Stefano, Children's Hospital Los Angeles, Los Angeles, California, United States
Background
Significant molecular and functional changes within the glomerulus, and specifically in podocytes, the cells in charge of the ultrafiltration, are responsible for the initiation and progression of renal damage. Our data indicate that elevation of miR-193a plays a key role in regulating podocyte biology by controlling podocyte cell cycle phases. Using different tools (glomerulus-on-a-chip: GOAC, human COL4-defective podocytes, FUCCI mice, nanoparticles, and spatial transcriptomics), we have identified miR-193a as a possible specific disease target in our model of CKD, Alport Syndrome (AS).
Methods
GOAC was seeded with human glomerular endothelial cells and podocytes derived from amniotic fluid of AS and healthy patients to recapitulate the function and structure of the glomerular filtration barrier. miR-193a studies were performed using mimics and inhibitors. Nanoparticles (micelle) containing miR-193a inhibitor were designed to specifically target podocytes. AS FUCCI mouse model (cell cycle indicator mouse model) was used to track podocyte miR-193a cell cycle modulation. Biopsies of Alport patients were used to confirm miR-193a expression by in situ hybridization and to perform Digital Spatial Profiling (DSP) using Nanostring technology.
Results
Spatial transcriptomics identified in human AS glomeruli altered gene expression for podocyte structure (foot process and slit diaphragm), GEC structure (glycocalyx), matrix turnover proteins, miR-193a targets, and cell cycle. Generated AS-GOAC presented impaired permselectivity, and proteomics revealed a distinctive AS signature. In AS-GOAC, miR-193a inhibition, delivered with innovative nanoparticles designed targeting podocytes, restored to normal the altered podocyte cell cycle, and regulated downstream miR-193a targets (WT1, ItgαVβ3/osteopontin, and VEGF) necessary for podocyte homeostasis.
Conclusion
We show that upregulating of miR-193a induces changes in gene expression and alteration of the cell cycle phases specifically in AS podocytes. Inhibiting miR-193a using micelle technology may re-establish glomerular function by modulating important molecular pathways responsible for podocyte survival representing a therapeutic target in AS settings.
Funding
- NIDDK Support