Convergence of human pluripotent stem cell, organoid, and genome editing technologies

Abstract

The last decade has seen many exciting technological breakthroughs that greatly expanded the toolboxes for biological and biomedical research, yet few have had more impact than induced pluripotent stem cells and modern-day genome editing. These technologies are providing unprecedented opportunities to improve physiological relevance of experimental models, further our understanding of developmental processes, and develop novel therapies. One of the research areas that benefit greatly from these technological advances is the three-dimensional human organoid culture systems that resemble human tissues morphologically and physiologically. Here we summarize the development of human pluripotent stem cells and their differentiation through organoid formation. We further discuss how genetic modifications, genome editing in particular, were applied to answer basic biological and biomedical questions using organoid cultures of both somatic and pluripotent stem cell origins. Finally, we discuss the potential challenges of applying human pluripotent stem cell and organoid technologies for safety and efficiency evaluation of emerging genome editing tools.

Advanced Cellular Models for Rare Disease Study: Exploring Neural, Muscle and Skeletal Organoids

“…The development of iPSCs and their differentiation through the formation of organoids that morphologically and physiologically resemble human tissues already represent significant progress, as described above. However, in some experimental settings, further genetic modification of iPSCs and organoids may be required before downstream applications to address basic biological and biomedical questions [42]. Advances in genome editing technologies make it possible to precisely edit specific genes in iPSCs, modifying them to express or induce specific characteristics.…”

“…In general, mutagenesis can take place either in a tissue that serves as a source of adult stem cells or in isolated cells used to generate organoids; the second option is preferable as it is more efficient and less costly. Over the years, attempts have been made to genetically modify different types of ASC-derived organoids, as described in detail by others [13,42]. Schwank et al, Matano et al and Kavasaki et al are just some of the researchers showing that CRISPR/Cas9 could be applied in gastrointestinal tract organoids for gene knock-out or mutation repair [47][48][49].…”

Revolutionizing Disease Modeling: The Emergence of Organoids in Cellular Systems

“…Hence, lentiviral transgenesis, CRISPR/Cas9 gene editing, and single-cell readouts were established to study genes function and bring out genetic alterations in organoids. However, the dynamic environment of organoids and their level of complexity also present challenges for genetic editions [ 275 , 276 ].…”

Genome Editing in iPSC-Based Neural Systems: From Disease Models to Future Therapeutic Strategies

“…The emerging technology of personalized “organ on a chip” systems combined with high throughput transcriptomics is an exciting development that may have great relevance for testing and translation of precision therapies (Ronaldson-Bouchard and Vunjak-Novakovic, 2018 ; Ramme et al, 2019 ). Although iPSC-derived systems will not replace animal models for assessing future genetic treatments' safety, inter-species differences in sequence homology will impact targeting strategies and render testing in human-derived models an important part of the translational process (Wang et al, 2021 ).…”