|Title||Engineering prokaryotic channels for control of mammalian tissue excitability.|
|Publication Type||Journal Article|
|Year of Publication||2016|
|Authors||HX Nguyen, RD Kirkton, and N Bursac|
The ability to directly enhance electrical excitability of human cells is hampered by the lack of methods to efficiently overexpress large mammalian voltage-gated sodium channels (VGSC). Here we describe the use of small prokaryotic sodium channels (BacNa<sub>v</sub>) to create de novo excitable human tissues and augment impaired action potential conduction in vitro. Lentiviral co-expression of specific BacNa<sub>v</sub> orthologues, an inward-rectifying potassium channel, and connexin-43 in primary human fibroblasts from the heart, skin or brain yields actively conducting cells with customizable electrophysiological phenotypes. Engineered fibroblasts ('E-Fibs') retain stable functional properties following extensive subculture or differentiation into myofibroblasts and rescue conduction slowing in an in vitro model of cardiac interstitial fibrosis. Co-expression of engineered BacNa<sub>v</sub> with endogenous mammalian VGSCs enhances action potential conduction and prevents conduction failure during depolarization by elevated extracellular K<sup>+</sup>, decoupling or ischaemia. These studies establish the utility of engineered BacNa<sub>v</sub> channels for induction, control and recovery of mammalian tissue excitability.
|Short Title||Nature Communications|