Adventures and Advances in Time Travel With Induced Pluripotent Stem Cells and Automated Patch Clamp

Abstract

In the Hollywood blockbuster “The Curious Case of Benjamin Button” a fantastical fable unfolds of a man’s life that travels through time reversing the aging process; as the tale progresses, the frail old man becomes a vigorous, vivacious young man, then man becomes boy and boy becomes baby. The reality of cellular time travel, however, is far more wondrous: we now have the ability to both reverse and then forward time on mature cells. Four proteins were found to rewind the molecular clock of adult cells back to their embryonic, “blank canvas” pluripotent stem cell state, allowing these pluripotent stem cells to then be differentiated to fast forward their molecular clocks to the desired adult specialist cell types. These four proteins – the “Yamanaka factors” – form critical elements of this cellular time travel, which deservedly won Shinya Yamanaka the Nobel Prize for his lab’s work discovering them. Human induced pluripotent stem cells (hiPSCs) hold much promise in our understanding of physiology and medicine. They encapsulate the signaling pathways of the desired cell types, such as cardiomyocytes or neurons, and thus act as model cells for defining the critical ion channel activity in healthy and disease states. Since hiPSCs can be derived from any patient, highly specific, personalized (or stratified) physiology, and/or pathophysiology can be defined, leading to exciting developments in personalized medicines and interventions. As such, hiPSC married with high throughput automated patch clamp (APC) ion channel recording platforms provide a foundation for significant physiological, medical and drug discovery advances. This review aims to summarize the current state of affairs of hiPSC and APC: the background and recent advances made; and the pros, cons and challenges of these technologies. Whilst the authors have yet to finalize a fully functional time traveling machine, they will endeavor to provide plausible future projections on where hiPSC and APC are likely to carry us. One future projection the authors are confident in making is the increasing necessity and adoption of these technologies in the discovery of the next blockbuster, this time a life-enhancing ion channel drug, not a fantastical movie.

A modern automated patch-clamp approach for high throughput electrophysiology recordings in native cardiomyocytes

“…This discrepancy illustrates a major limitation for the investigation of excitable cells and their regulatory units. Recently, tremendous progress has been made in the development of high-performance automated patch-clamp (APC) systems that allow for high throughput electrophysiological measurements [6][7][8] (Supplementary Fig. 1).…”

Bridging the translational gap: what can synaptopathies tell us about autism?

“…For example, it may be possible to develop a 3D structure of microneedle electrodes within which an organoid is growing to record neuron-neuron communication throughout the organoid (Choi et al, 2021), rather than a handful of cells on the surface of a sliced organoid (Sharf et al, 2022). It is possible to reliably quantify circuitry using viral labelling and retrograde tracing (Miura et al, 2022), low-throughput methods such as patch-clamp electrophysiology (Avazzadeh et al, 2021;Rosholm et al, 2022), or high-throughput methods such as multi-electrode arrays (MEAs; Alsaqati et al, 2020). MEAs have been deployed to investigate gene mutations associated with monogenic forms of autism in hiPSC-neurons such as CNTN5, EHMT2, KCNQ2, ATRX, and SCN2A (Deneault et al, 2018;Deneault et al, 2019).…”

From squid giant axon to automated patch-clamp: electrophysiology in venom and antivenom research

“…The increased throughput of APCs has made them a valuable tool not only for drug discovery and high-throughput screening in the pharmaceutical industry, but also for basic research across several disciplines, from neurological ( Rosholm et al, 2022 ; Ghovanloo et al, 2023 ) and cardiovascular sciences ( Bell and Dallas, 2021 ) to venom ( Olamendi-Portugal et al, 2008 ) and antivenom research ( Restano-Cassulini et al, 2017 ; Ledsgaard et al, 2022 ; 2023 ; Miersch et al, 2022 ).…”