Developing new strategies to understand human kidney development and target human disease
- Autores: Patricia Katherine Prado Peralta
- Directores de la Tesis: Núria Montserrat Pulido (dir. tes.), Elena Garreta Bahima (codir. tes.)
- Lectura: En la Universitat Pompeu Fabra ( España ) en 2021
- Idioma: español
- Tribunal Calificador de la Tesis: Marc Guell Cargol (presid.), María Pía Cosma (secret.), Martin Johannes Hoogduijn (voc.)
- Programa de doctorado: Programa de Doctorado en Biomedicina por la Universidad Pompeu Fabra
- Materias:
- Enlaces
- Tesis en acceso abierto en: TDX
- Resumen
In recent years considerable progress has been made in the development of approaches for human pluripotent stem cells (hPSCs) differentiation. An important step in this direction has been the derivation of hPSCs derived organoids to understand early steps of human organ development and disease. Here, we have developed new techniques and fundamental knowledge to generate hPSCs-kidney organoids. Towards this aim we have explored on the possibility to emulate early steps of kidney embryogenesis forcing cell to cell and cell to extracellular matrix contact as a new approach to generate hPSCs-kidney organoids with superior treats of differentiation and function. To further assess on the impact of environmental stimuli during kidney organoid differentiation we have interrogated on the interaction of metabolic cues in this process identifying metabolic regulators, namely Esrrα, which are responsible of kidney differentiation and human chronic kidney disease. Into the light of the current COVID19 crisis we have further exploited hPSCs-kidney organoids to understand first steps of SARS-CoV-2 infection also identifying a therapeutic compound which nowadays is in a phase II b clinical trial for COVID19 patients in more than 6 countries in the world. Based on the current developments, in the present thesis we emphasize the major achievements in the field of kidney morphogenesis, including technological advances for kidney embryonic cell culture and the use of animal models to understand kidney development. We further discuss on ongoing challenges of bringing together all this knowledge to establish hPSCs-kidney organoids to study human kidney differentiation and disease.
Conclusions:
I. We have established a procedure that suffices for the generation of posterior intermediate mesoderm (PIM) committed hPSCs which upon further assembly and culture in 3D conditions (transwell system or free floating suspension) leads to the generation of kidney organoids transcriptomically matching the human kidney of the second trimester gestation upon 16 days in culture.
II. We have established a procedure for the implantation of day 16 kidney organoids into the chick developing chorioallantoic membrane (CAM) that sustains for the endogenous organization of human endothelial cells and the growth of developing glomeruli.
III. We have identified different cell culture media promoting metabolic activities in developing kidney organoids that further correlated with changes in the expression of metabolic regulators and the generation of tubular-like cells on demand.
IV. We have identified metabolic regulators of the lipid metabolism (Estrogen-related receptor alpha, namely Esrrα) involved in the differentiation of tubular-like cells in developing kidney organoids and further assessed on the impact of this factor in damage protection on these cells.
V. We have established a procedure for the study of the first steps of SARS-CoV-2 infection taking advantage of kidney organoids upon their characterization for the endogenous expression of Angiotensin Converting Enzyme 2 (ACE2) using single cell RNA seq and an exhaustive profiling for the expression of tubular markers in hPSCs-kidney organoids.
