GECKO · Generation of cartilage-free kidney organoids: a small molecule strategy
Horizon Europe — Marie Skłodowska-Curie Actions
- Duration
- 2022-09-01 → 2024-08-31
- EU contribution
- €203,464
- Participants
- 1
- Scheme
- HORIZON-TMA-MSCA-PF-EF
Lines connect the coordinator with its partners.
Results in brief
Generation of cartilage-free kidney organoids: a small molecule strategy
Chronic kidney disease (CKD), a progressive loss of kidney function affecting up to 17.3% of Europeans, is becoming increasingly prevalent worldwide, with an estimated 9.7 million people needing kidney replacement therapy. However, due to donor shortages, only a fraction receives treatment, underscoring the need for alternative therapies. Recent advances in generating kidney organoids from human induced pluripotent stem cells (hiPSCs) offer a promising regenerative solution, as these organoids can mimic human kidney structures and exhibit functional properties, such as glomerular filtration. Such organoids hold potential as functional kidney substitutes, providing relief from dialysis and improving patient quality of life. The GECKO project aimed to address a key challenge in organoid development: the unintended formation of cartilage within kidney organoids, which hinders their functional application. The primary objective was to create a protocol for cartilage-free kidney organoids by identifying and modulating the pathways responsible for off-target chondrocyte formation, thus optimizing the organoids for therapeutic use. GECKO followed a strategic approach to achieve these goals: Assessing Cartilage in Organoids: Cartilage presence was evaluated at key developmental stages (days 7+18 to 7+25) using molecular markers and staining techniques, identifying when off-target chondrocytes emerge and providing a foundation for improved organoid protocols. Understanding Cartilage-Related Pathways: Key pathways, including EGFR, SOX9, and Notch, were analyzed, with findings indicating that early signaling events influence cartilage development. Notch and protein kinase A (PKA) modulation proved effective in reducing off-target cell populations. Inhibiting Cartilage Formation with Small Molecules: Small molecules were screened to prevent cartilage formation, with FGF9 and time-dependent Notch modulation successfully reducing chondrocytes. These results underscore a viable approach for developing functional kidney organoids free from off-target cells. This work lays the groundwork for kidney organoid applications in regenerative medicine, ultimately aiming to improve therapy options for CKD patients and reduce dependence on donor kidneys and dialysis.
Data: CORDIS, © European Union
Project objective
The prevalence of chronic kidney disease in Europe varies from 3.3–17.3% and has risen during the last decades. In the world, 9.7 million people need kidney replacement therapy, but only 2.6 million will receive it. These numbers are expected to double within the next 10 years, increasing the pressure to find alternative solutions. Recent scientific developments to generate kidney organoids in vitro have opened the possibility for a regenerative medicine–based approach that would provide a functional substitute to the failing kidney. These kidney organoids can recapitulate renal structures as well as the cellular complexity of human kidney, and may restore glomerular filtration upon transplantation. In this project, we make use of human induced pluripotent stem cells that can be differentiated with a cocktail of biomolecules and aggregated to form of kidney organoids that are cultured at the air–liquid interface. This protocol leads to the formation of complex renal structures including glomeruli and tubules. While kidney organoids show great therapeutic potential, they also present several drawbacks, one of which is the appearance of off-target cell populations, such as neurons, myocytes and chondrocytes within the organoid. Preventing the appearance of these off-target cells will greatly improve the quality of the organoids. Preventing cartilage formation is most desirable because cartilage completely disrupts the organoids and renders them dysfunctional and unsuitable for in vivo use.The reasons for the consistent observation of chondrocytes during organoid growth and transplantation are not yet understood. The objective of this work is to understand the molecular mechanisms of cartilage formation in kidney organoids and reduce it using a small molecule strategy.
Original text from CORDIS.
Participants
- UNIVERSITEIT MAASTRICHT · MaastrichtCoordinatorNetherlands
Links
Data: CORDIS, © European Union