Generating kidney organoids and mature renal cell types via directed iPSC differentiation. Chronic kidney disease is increasing in prevalence by 6.3% per annum. Hence there is a desperate need for the development of novel treatments.
The aim of the proposal is to direct the differentiation of human pluripotent stem cells towards kidney organoids containing all four key progenitor cells capable of generating a mature organ, including the nephrogenic mesenchyme, ureteric epithelium, stroma and vascular progenitors. These organoids will serve as a source of specific kidney cell types of value for cellular therapy as well as an accurate model of the organ as a whole. The proposal draws on our published data defining the methodology for generating kidney organoids and our experience in the generation of reporter iPSC lines. Results from the studies proposed will generate tools for the research community (a suite a reporter cell lines), knowledge about the development of the kidney in humans, improved methods for differentiation and proof of concept data for the application of the resulting cells in drug/nephrotoxicity screening and cellular therapy. The validity of the kidney organoids as a model of the developing organ will be interrogated using expression profiling, including single cell RNA-seq analyses. Single, double and triple fluorescent reporter iPSC lines will be generated to facilitate the subsequent isolation of specifi cell types from the organoids post differentiation. These cell types will also be characterized, providing novel information about specific kidney cell types during human development. Validating the use of organoids as a model of the developing organ will be performed so as to develop the use of organoids for drug/nephrotoxicity screening. Reporter iPSC lines will also be generated to report defined responses to injury. Nephrotoxicity screening will be performed on organoids generated using such reporter. Specific kidney cell types will be isolated from organoids via FACS and used to evaluate cellular therapy approaches for the treatment of chronic kidney injury via transplantation into immunocompromised mice. Reporter iPSC lines will also be generated to incorporate the expression of luciferase to allow for the monitoring via bioluminescence of the location of the delivered cells in live animals across time. The efficacy of delivering kidney cells derived from pluripotent stem cells into preclinical models will therefore be determined.
Generating kidney organoids and mature renal cell types via directed iPSC differentiation. Chronic kidney disease affects up to 10% of the US population and is increasing in incidence by 6.3% per annum. This represents a health burden in the US alone of >$USD40 billion per annum. With increasing prevalence and decreasing availability of donor organs, there is a critical need for the development of alternative therapies. We have developed a method for encouraging human pluripotent stem cells to form mini-kidneys. These mini- kidneys contain all the anticipated cell types of the developing organ and may therefore represent a novel source of kidney cells for the treatment of disease. In this project, we will optimize this process, isolate out of these organs specific cell types and test their ability to improve renal function in models of kidney disease. The second most common cause for a drug to fail to reach market is toxicity to the kidney. As this is not evident until drug development ha reached pre-clinical or Phase 1 trials in humans, this represents a major challenge to the pharmaceutical industry. We will evaluate whether mini-kidneys generated from pluripotent stem cells can be used to accurately predict drug reactions early in the development phase. Use of mini-kidneys for screening may also be of value in developing models of kidney disease using patient-derived stem cells.
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| Volovelsky, Oded; Nguyen, Thi; Jarmas, Alison E et al. (2018) Hamartin regulates cessation of mouse nephrogenesis independently of Mtor. Proc Natl Acad Sci U S A 115:5998-6003 |
| Little, Melissa Helen; Kairath, Pamela (2017) Does Renal Repair Recapitulate Kidney Development? J Am Soc Nephrol 28:34-46 |
