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Abstract: SA-PO711

miR-193a and Nanoparticle Technology: A Novel Therapeutic Approach for Alport Syndrome

Session Information

Category: Glomerular Diseases

  • 1401 Glomerular Diseases: Mechanisms, including Podocyte Biology

Authors

  • Perin, Laura, Children's Hospital Los Angeles, Los Angeles, California, United States
  • Aguiari, Paola, Children's Hospital Los Angeles, Los Angeles, California, United States
  • Petrosyan, Astgik, Children's Hospital Los Angeles, Los Angeles, California, United States
  • Huang, Yi, University of Southern California, 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 changes in podocyte cell cycle regulation are responsible for the progression of renal damage. miR-193a has been reported to regulate podocyte phenotype and to ensure maintenance of their differentiated G0 state. We observed elevated levels of miR-193a in human podocytes from Alport Syndrome (AS) biopsies and in AS mice. Here, we study the role of miR-193a in Alport podocytes and if miR-193a inhibition by podocyte-targeting nanoparticles could restore podocyte quiescent state.

Methods

To study the effects of miR-193a, we used miR-193a mimic and antagomir on podocytes from AS FUCCI mice (cell cycle indicator mouse model) and in the glomerulus-on-a-chip system (GOAC), generated with AS patient-derived podocytes. Podocyte-targeted nanoparticles (micelles) were developed and tested for specificity in vitro and in the GOAC and for assessing the efficiency of a miR-193a inhibition therapy. In situ hybridization, Digital Spatial Profiling (DSP, Nanostring), and protein arrays were used to investigate changes in miR-193a targets in murine podocytes and human AS biopsies.

Results

We observed specific upregulation of miR-193a in podocytes vs other glomerular cells, and upregulation of miR-193a targets osteopontin, VEGFA, and ItgαV, and downregulation of WT1 in AS glomeruli. Our data shows that miR-193a regulates the cell cycle: WT podocytes re-enter the cell cycle when exposed to miR-193a (G1/S), while miR-193a inhibition rescues the AS podocyte G0 and modulates P18, cyclin A1 and cyclin C, important cell cycle regulators. We successfully generated miR-193a inhibitor-mediated micelles enhancing targeting to podocytes and preliminarily proved effective in safeguarding the glomerular filtration barrier. In human AS glomeruli, DSP revealed changes in gene expression in miR-193a targets and cell cycle regulators.

Conclusion

Our data indicate that the elevation of miR-193a plays a key role in regulating podocyte biology by controlling the podocyte cell cycle. Inhibiting miR-193a using micelle technology may re-establish glomerular function by modulating important molecular pathways responsible for podocyte survival to prevent further injury representing a therapeutic target for AS.

Funding

  • NIDDK Support