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How to regenerate the mammalian kidney

Periodic Reporting for period 3 - REGMAMKID (How to regenerate the mammalian kidney)

Reporting period: 2018-11-01 to 2020-04-30

Kidney-related disease remains one of the major health issues in modern society. Nowadays, Chronic kidney disease (CKD) is thought to affect around 10% of the EU population. The large number of people affected by CKD is of concern, first, because some will progress to End Stage Renal Disease (ESRD), and second, because CKD increases the risk of premature death from cardiovascular diseases. In REGMAMKID our aim is to generate innovative strategies for kidney healing. Our project stands in the use of animal models with endogenous capacity to regenerate after injury (neonatal mice), together with human pluripotent stem cells. Taking advantage of these systems, we will be able to identify endogenous molecular mechanisms responsible for kidney regeneration in mammals, and to define novel culture systems for kidney cell replacement and therapy.
The activities performed in the project’s first period have mostly covered the generation of novel protocols for the derivation of three-dimensional (3D) kidney derived organoids from human pluripotent stem cells (hPSCs). Taking advantage of this in vitro system we have started to explore the epigenetic/transcriptomic landscapes during renal commitment in the human setting. Moreover, we are currently taking advantage of innovative strategies to enhance kidney differentiation from hPSCs (i.e. 3D bioprinting by combining kidney differentiated cells together with biomimetic matrices, hydrogels with tailored stiffness mimicking kidney and embryonic milieu, among others). Specifically, we have been able to generate kidney analogues by 3D bioprinting that sustain hPSCs differentiation towards renal progenitors. In parallel, we are generating a novel cellular platform by means of CRISPR/Cas9 technology for the derivation of mutant cell lines for kidney disease modeling and differentiation. Lastly, and in an effort to explore endogenous responses sustaining the de novo formation of nephrons upon injury, we have explored the capacity of WT1GFP neonatal mice to regenerate after the amputation of the 20% of the kidney mass.
Our activities have led to the generation of fundamental knowledge about kidney differentiation by the definition of novel approaches for the derivation of 3D kidney organoids from hPSCs. Such platform represents a faithful in vitro system for kidney disease modeling, offering an attractive scenario for drug screening. Furthermore, we are repurposing the use of inducible strategies for CRISPR/Cas9-based genome editing in order to engineer hPSCs for further applications related to kidney disease modeling and differentiation. The generation of CRISPR/Cas9 engineered hPSCs lines will allow not only for the generation of commercial hPSCs lines suitable for human disease modeling, but to further provide “ready-to-use” sgRNA Library cell lines (also including other relevant human cell lines-HEK293, CHO, or other cancer cell lines) in order to explore novel applications related to drug screening and human disease modeling. The potential of such strategy is therefore expected to be of considerable interest for pharmaceutical companies and represent a great advance in the field of regenerative and personalized medicine.