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The pain target Na V 1.7 is expressed late during human iPS cell differentiation into sensory neurons as determined in high-resolution imaging.
Pflügers Archiv : European Journal of Physiology 2024 March 28
Human-induced pluripotent stem cells (iPS cells) are efficiently differentiated into sensory neurons. These cells express the voltage-gated sodium channel NaV 1.7, which is a validated pain target. NaV 1.7 deficiency leads to pain insensitivity, whereas NaV 1.7 gain-of-function mutants are associated with chronic pain. During differentiation, the sensory neurons start spontaneous action potential firing around day 22, with increasing firing rate until day 40. Here, we used CRISPR/Cas9 genome editing to generate a HA-tag NaV 1.7 to follow its expression during differentiation. We used two protocols to generate sensory neurons: the classical small molecule approach and a directed differentiation methodology and assessed surface NaV 1.7 expression by Airyscan high-resolution microscopy. Our results show that maturation of at least 49 days is necessary to observe robust NaV 1.7 surface expression in both protocols. Electric activity of the sensory neurons precedes NaV 1.7 surface expression. A clinically effective NaV 1.7 blocker is still missing, and we expect this iPS cell model system to be useful for drug discovery and disease modeling.
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