Channels: hERG, NaV1.5, CaV1.2,Kir2.1, KV4.3, KV7.1
Cells:
HEK, CHO
Tools:
SyncroPatch 384/768PE

Application Note

Simultaneous Assessment of CiPA Stipulated
Ion Channels on the SyncroPatch 384PE
The

electrophysiology

team

atNanion

Technologies

GmbH,Munich.

Cell lines kindly provided by Charles River.

SummaryResults

The cardiac action potential is defined by multiple
voltage-dependent ion channels (see Fig. 1). A drugcandidate’s capacity tointeractwiththeionchannels involved in thedepolarization
larizationorphasesrepooftheA combined voltage step-ramp protocol (Fig. 2) was applied simultaneously to HEK or CHO cells expressing different cardiac ion channels. The first segment was used toelicit NaV1.5 or CaV1.2 (1) followed by a classical hERG-

like segment to activate KV4.3, KV11.1 and KV7.1 (2) and fi-

nally a ramp protocol to elicit inwardly rectifying Kir2.1 (3).

cardiac action potential isimportant for drug safetyassessment. Until now, safety testing has focussed oninteraction with the hERG
channel and QT prolongation which can lead topotentiallyades

defataltors-pointes(TdP).

Although this approach has been largely successful in preventing new drugs reaching the market with
unexpected potential to cause TdP, it is also possiblethat potentially valuable therapeutics have failed
due to this early screening. A new paradigm, theComprehensive In-vitro Proarrhythmia Assay (CiPA), wasintroduced in 2013 to provide a more complete assessment of proarrhythmic risk1,2. An assessment of a multitude
of cardiac ion channels, in addition to hERG, should provide a more accurate prediction of the proarrhythmic riskof a compound compared with testing on hERG alone.
Here we show recordings from HEK or CHO cells
expressing the CiPA stipulated ion channels; NaV1.5,CaV1.2, hERG, KV7.1, Kir2.1 or KV4.3, activated within one
single experiment on the SyncroPatch 384PE.

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Figure 2: A voltage protocol was used to activate 6 distinct cardiac ionchannels. The top panel shows the voltage protocol used on all wells tosimultaneously activate the 6 distinct cardiac ion channel currents shown inthe bottom panels.

Application Note

Figure 3: Six different cardiac channels recorded simultaneously on the SyncroPatch® 384PE. Shown is a screenshot of the data acquisition and analysis
software used on the SyncroPatch® 384PE showing an experiment recording six different cardiac channels at once. A representative image of the current
recording for each ion channel is shown at the top.

In conclusion, the SyncroPatch 384PE can be used toto be key targets for safety testing. In this way, a moreassess the effects of compounds simultaneously on

accurate risk assessment of compounds can be mademultiple cardiac ion channels at a high throughput. Thesetaking into account a multitude of ion channels.

ion channels have been indicated by the CiPA initiative
Cell culture & electrophysiology

References
1. https://www.ilsiextra.org/hesi/science/cardiac/cipa/SitePages/Home.
aspx
2. Sager, P.T., et al., 2014. Am. Heart J. 167(3): 292-300

Methods

Cells were cultured and harvested according toNanion’s standard cell culture protocols and perforated
patch clamp recordings were conducted according toNanion’s standard procedures for the SyncroPatch 384PE
using 20 mM Escin .

Cells

CHO cells expressing NaV1.5, CaV1.2, KV4.3 or KV7.1;
HEK293 cells expressing hERG or Kir2.1 from Charles River.
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