Regenerative Medicine, Disease Modelling, and Drug Discovery in Human Pluripotent Stem Cell-Derived Kidney Tissue

Abstract

The multitude of research clarifying critical factors in embryonic organ development has been instrumental in human stem cell research. Mammalian organogenesis serves as the archetype for directed differentiation protocols, subdividing the process into a series of distinct intermediate stages that can be chemically induced and monitored for the expression of stage-specific markers. Significant advances over the past few years include established directed differentiation protocols of human embryonic stem cells and human induced pluripotent stem cells (hiPSC) into human kidney organoids in vitro. Human kidney tissue in vitro simulates the in vivo response when subjected to nephrotoxins, providing a novel screening platform during drug discovery to facilitate identification of lead candidates, reduce developmental expenditures, and reduce future rates of drug-induced acute kidney injury. Patient-derived hiPSC, which bear naturally occurring DNA mutations, may allow for modelling of human genetic diseases to enable determination of pathological mechanisms and screening for novel therapeutics. In addition, recent advances in genome editing with clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 enable the generation of specific mutations to study genetic disease, with non-mutated lines serving as an ideal isogenic control. The growing population of patients with end-stage kidney disease is a worldwide healthcare problem, with high morbidity and mortality rates, that warrants the discovery of novel forms of renal replacement therapy. Coupling the outlined advances in hiPSC research with innovative bioengineering techniques, such as decellularised kidney and three-dimensional printed scaffolds, may contribute to the development of bioengineered transplantable human kidney tissue as a means of renal replacement therapy.

Single-cell RNA sequencing and kidney organoid differentiation

“…The generating kidney organoids protocols have been established from hPSCs based on the knowledge of mammalian kidney development [ 11 , 12 ]. 3D culture technology has enhanced the ability of embryonic and adult stem cells to self-organize and form multi-cellular tissues, enabling differentiation into kidney tissue.…”

Precision nephrotoxicity testing using 3D in vitro models

“…These 3D spherical structures are generated by differentiating human pluripotent stem cells into kidney-specific cell types, closely resembling the kidney tissue. They can recapitulate various aspects of kidney composition and structure, including the formation of nephrons and interactions between different cell types [47,56,57]. They offer several advantages [45,58,59]: (a) they recapitulate renal development: Organoids mimic key features of kidney development, providing insights into developmental processes and disease mechanisms.…”

ATP/ADP biosensor organoids for drug nephrotoxicity assessment

“…Recently, several in vitro protocols involving either directed differentiation or transcription factor-based reprogramming into kidney cells and organoids have been established. These biotechnological advancements have enabled the generation of nephron progenitor cells and kidney organoids from human pluripotent stem cells (hPSCs) including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) (Mae et al, 2013;Taguchi et al, 2014;Morizane et al, 2015;Takasato et al, 2016;Morizane and Bonventre, 2017b;Gupta et al, 2017;Morizane et al, 2017). Kidney organoids contain epithelial nephron-like structures expressing markers of podocytes, proximal tubules, loops of Henle, and distal nephrons in an organized, continuous arrangement that resembles the nephron in vivo (Morizane et al, 2015).…”