Maggiore, Joseph
(2024)
Engineering endothelialized human kidney organoids for studying kidney injury and HDAC8 targeted therapy.
Doctoral Dissertation, University of Pittsburgh.
(Unpublished)
Abstract
There is an extreme need to develop representative in vitro models of organs to understand human disease and develop therapeutics. There is no greater need than in the setting of kidney disease, which effects ~850M people annually. Patient derived kidney organoids derived from induced pluripotent stem cells have risen as an attractive model of the kidney for their high throughput nature, patient specificity, complex morphology, and multicellular heterogeneity. Current kidney organoids, however, lack sufficient vascularization, maturity, and certain physiologically relevant cell types. Here, we advanced in vitro kidney organoid models by developing an inducible endothelial stem cell line. This inducible endothelial line generates a heterogenous population of endothelial progenitor cells. When combined to a prior developed kidney organoid protocol, the resulting kidney organoid is highly vascularized with kidney-specific endothelial cells, contains mature epithelial cell populations, and the emergence of renin+ cells. We then utilized this kidney organoid model to understand mechanisms of kidney disease and drug targeting. Based on evidence demonstrating histone deacetylase 8 (HDAC8) as a regulator of kidney injury and cancer mesenchymal transition, we aimed to understand the role HDAC8 plays in kidney disease regulating dedifferentiation. We found that in hemin injured kidney organoids HDAC8 knockout protected against DNA damage and loss of epithelial cell state globally. We then found that in a glomerular toxicity setting, doxorubicin caused dedifferentiation of podocytes and in endothelial cells, subsequent mesenchymal transition. We then found that with HDAC8 inhibitor, PCI-34051, there was preservation of epithelial cell state and prevention of downstream fibrosis and vascular rarefaction. Based on this data, we demonstrated a method for vascularizing human kidney organoids to enhance maturity, as well as a case study in understanding mechanisms of kidney disease and the role HDAC8 plays.
Share
| Citation/Export: |
|
| Social Networking: |
|
Details
| Item Type: |
University of Pittsburgh ETD
|
| Status: |
Unpublished |
| Creators/Authors: |
|
| ETD Committee: |
| Title | Member | Email Address | Pitt Username | ORCID |
|---|
| Committee Chair | Kleyman, Thomas | kleyman@pitt.edu | | | | Thesis Advisor | Hukriede, Neil A. | hukriede@pitt.edu | | | | Committee Member | Shankland, Stuart | | | | | Committee Member | Monga, Sardarshan | | | | | Committee Member | Sims-Lucas, Sunder | | | |
|
| Date: |
15 July 2024 |
| Defense Date: |
29 July 2024 |
| Approval Date: |
14 October 2024 |
| Submission Date: |
2 August 2024 |
| Access Restriction: |
No restriction; Release the ETD for access worldwide immediately. |
| Number of Pages: |
248 |
| Institution: |
University of Pittsburgh |
| Schools and Programs: |
School of Medicine > Integrative Systems Biology |
| Degree: |
PhD - Doctor of Philosophy |
| Thesis Type: |
Doctoral Dissertation |
| Refereed: |
Yes |
| Uncontrolled Keywords: |
kidney organoid, tissue engineering, kidney injury, glomerular injury, kidney disease, vascularization, genetic engineering |
| Date Deposited: |
14 Oct 2024 15:51 |
| Last Modified: |
15 Oct 2024 13:06 |
| URI: |
http://d-scholarship.pitt.edu/id/eprint/46815 |
Metrics
Monthly Views for the past 3 years
Plum Analytics
Actions (login required)
 |
View Item |
![[img]](https://d-scholarship.pitt.edu/style/images/fileicons/application_pdf.png)
![[feed]](/images/feed-icon-32x32.png){kind=icon}