Abstract: SA-PO561
Characterization of Extracellular Vesicles Derived from Human Amniotic Fluid Stem Cells (hAFSC-EVs) and Their Therapeutic Effect in Alport Syndrome
Session Information
- Glomerular Diseases: Fibrosis, Extracellular Matrix
November 09, 2019 | Location: Exhibit Hall, Walter E. Washington Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Glomerular Diseases
- 1201 Glomerular Diseases: Fibrosis and Extracellular Matrix
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
- Dedhia, Charmi, Children's Hospital Los Angeles, Los Angeles, California, United States
- Petrosyan, Astgik, Saban Research Institute- Children's Hospital Los Angeles, Los Angeles, California, United States
- Villani, Valentina, Children's Hospital Los Angeles, Los Angeles, California, United States
- Soloyan, Hasmik, Children's Hospital Los Angeles, Los Angeles, California, United States
- Reems, Jo-Anna, University of Utah, Salt Lake City, Utah, United States
- Sedrakyan, Sargis, Children's Hospital Los Angeles, Los Angeles, California, United States
Background
Alport Syndrome (AS) is an inherited disease characterized by a loss of glomeruli cells and kidney function. Based on our previous results that EVs from AFSC modulated glomerular crosstalk, we proposed that AFSC-EVs might represent a new therapeutic approach to treat AS. In light of clinical translation, we characterized AFSC-EVs of human origin and evaluated whether they would present a therapeutic effect in an AS mouse model.
Methods
Human clonal AFSC were derived from amniotic fluid collected after volunteer donors provided consent. EVs were obtained from AFSC and characterized by RNA-seq and proteomics. hAFSC-EVs were injected into Alport mice and their therapeutic effect was studied by evaluation of renal function and life-span. RNA-seq was performed on glomeruli obtained from injected and non-injected mice.
Results
Proteomic profiling identified 675 intact proteins and RNA-seq data identified 2,535 miRs in hAFSC-EVs. hAFSC-EV “fingerprint” was assessed by performing GO analysis on the 100 most highly expressed proteins and miRs. The results identified pathways involved in tissue homeostasis, the mTOR pathway, and TGFβ and VEGF pathways. When injected in vivo into AS mice, biodistribution studies showed hAFSC-EVs localized in the kidney, corrected proteinuria and prolonged the life-span of treated mice. No side effects (including teratoma) were noted in the treated mice. RNA-seq of glomeruli obtained from treated AS mice showed similar gene expression patterns to WT, by cluster analysis. Our data indicated that hEVs highly modulated pathways involved in collagen deposition remodeling, in addition to downstream targets of VEGF, FGF, TNF, angiotensin and preserved glomerular cells structure and function.
Conclusion
Our protocol for hEVs derivation is reproducible and allows derivation of EV lots with the same identity (specific cargo of proteins and miRs), purity (absence of contaminants) and potency (present therapeutic effect in AS). hAFSC-EVs modulated signaling pathways that are central to maintaining glomerular homeostasis and preserved glomeruli structure with improved kidney function. This suggests the possibility of using hAFSC-EVs as a new therapeutic option for treating AS in humans.
Funding
- Private Foundation Support