Ion Channels and Respiratory Diseases
Did You Know: Ion Channels and their function in respiratory disease targets
The SARS-CoV-2 pandemic which emerged in late 2019, coupled with continued seasonal and pandemic influenza infections, has caused a significant problem for global public health and health systems worldwide. Antivirals and vaccines effective against these viruses are desperately needed in order to stem the pandemic and save lives.
A number of viroporins and ion channels are involved in viral infections and respiratory disorders, a selection is provided below.
Viroporins
Viroporins are potential drug targets for block of viral replication and the spread of infection.
Useful Reviews
Scott, C., & Griffin, S. 2015. Viroporins: structure, function and potential as antiviral targets. Journal of General Virology
Nieva, J.L., Madan, V., Carrasco, L. 2012. Viroporins: structure and biological functions. Nature Reviews Microbiology.
Wang, K., Xie, S., Sun, B. 2011. Viral proteins function as ion channels. Biochim Biophys Acta Biomembr.
Viroporins of Sars-CoV
Sars-CoV 3a forms a potassium channel located on the surface of Sars-CoV infected cells and promotes virus release (Lu, W., et al., 2006). Additionally, the Sars-CoV 3a and E proteins have been shown to activate the NLRP3 inflammasome (Chen, I-Y., et al., 2019) and are involved in virus replication and pathogenesis (Castaño-Rodriguez et al, 2018).
Viroporins of Influenza virus
The M2 protein of influenza A is a proton-gated, proton selective ion channel that is essential for viral replication (Jalily, P.H., et al, 2020). Amantadine is a well-known blocker of the influenza A M2 proton channel, and has been used as an anti-influenza drug for many years. Unfortunately it is associated with a number of severe side effects and is no longer recommended for the treatment of influenza A due to resistance as a result of mutations in the M2 protein
Measuring viroporins
Viroporins can be measured using automated patch clamp when expressed in cell lines, or using the Orbit instruments when they are recombinantly expressed in bilayers.
High throughput automated patch clamp
Parallel bilayer recordings
Cytotoxicity measurements
Ca2+-release activated calcium channel (CRAC)
Useful Reviews
Staudermann, T.A. 2018. CRAC channels as targets for drug discovery and development. Cell Calcium
Measuring CRAC Channels
CRAC channels can be measured using automated patch clamp when expressed in cell lines, or using the Orbit instruments when they are recombinantly expressed in bilayers.
High throughput automated patch clamp
Parallel bilayer recordings
Cytotoxicity measurements
Volume-regulated anion channels (VRAC), LRRC8
Key Publications and Reviews
Zhou, C., Chen, X., Planells-Cases, R., et al. 2020. Transfer of cGAMP into Bystander Cells via LRRC8 Volume-Regulated Anion Channels Augments STING-Mediated Interferon Responses and Anti-viral Immunity. Immunity
Strange, K., Yamada, T., & Denton, G. 2019. A 30-year journey from volume-regulated anion currents to molecular structure of the LRRC8 channel. Journal of General Physiology
Measuring VRAC Channels
VRAC Channels can be measured using automated patch clamp when expressed in cell lines, or using the Orbit instruments when they are recombinantly expressed in bilayers.
High throughput automated patch clamp
Parallel bilayer recordings
Cytotoxicity measurements
VRAC Publications using Nanion Instruments
2016 - Human T cells in silico: Modelling their electrophysiological behaviour in health and disease
Patchliner publication in Journal of Theoretical Biology (2016)
Authors:
Ehling P., Meuth P., Eichinger P., Hermann A.M., Bittner S., Pawlowski M., Pankratz S., Herty M., Budde T., Meuth S.G.
Two pore channels (TPC)
Key Publications
Sakurai, Y., Kolokoltsov, A.A., Chen, C-C., et al. 2015. Two-pore channels control Ebola virus host cell entry and are drug targets for disease treatment. Science
Castonguay, J., Orth, J.H.C., Müller, T., et al. 2017. The two-pore channel TPC1 is required for efcient protein processing through early and recycling endosomes. Nature Scientific Reports
Measuring Two Pore Channels
Two Pore Channels can be measured using automated patch clamp when expressed in cell lines, in lysosomes, or using the Orbit instruments when they are recombinantly expressed in bilayers.
Automated patch clamp
Parallel bilayer recordings
Cytotoxicity measurements
Publications using Instruments from Nanion
2017 - Organelle membrane derived patches: reshaping classical methods for new targets
Port-a-Patch and 
Vesicle Prep Pro publication in Nature Scientific Reports (2017)
Authors:
Shapovalov G., Ritaine A., Bidaux G., Slomianny, C., Borowiec A-S., Gordienko D., Bultynck G., Skryma R., Prevarskaya N.
2014 - High susceptibility to fatty liver disease in two-pore channel 2-deficient mice
Port-a-Patch publication in Nature Communications (2014)
Authors:
Grimm C., Holdt L.M., Chen C.C., Hassan S., Müller C., Jörs S., Cuny H., Kissing S., Schröder B., Butz B., Northoff B., Castonguay J., Luber C.A., Moser M., Spahn S., Lüllmann-Rauch R., Fendel C., Klugbauer N., Griesbeck O., Haas A., Mann M., Bracher F., Teupser D., Saftig P., Biel M., Wahl-Schott C.
2010 - Planar Patch Clamp Approach to Characterize Ionic Currents from Intact Lysosomes
Port-a-Patch publication in Science Signaling (2010)
Authors:
Schieder M., Rötzer K., Brüggemann A., Biel M., Wahl-Schott C.
2010 - Characterization of two pore channel 2 (TPCN2) -mediated Ca2+ currents in isolated lysosomes
Port-a-Patch publication in The Journal of Biological Chemistry (2010)
Authors:
Schieder M., Rötzer K., Brüggemann A., Biel M., Wahl-Schott C.A.
Transporters - SLC6A19 in SARS-CoV-2
Key Publications
Yan, R., Zhang, Y., Li, Y., et al. 2020. Structural basis for the recognition of SARS-CoV-2 by full-length human ACE2. Science
Measuring SLC19A and other Transporters
SLC19A can be measured using the SSM-electrophysiology technique. It may also be measured in bilayers if reconstituted and the effect of drugs acting on transporters can be measured using impedance.
Transporter measurements
Parallel bilayer recordings
Cytotoxicity measurements
TMEM16A
Key Publication
Danahay, H.L., Lilley, S., Fox, R., et al. 2019. TMEM16A Potentiation: A Novel Therapeutic Approach for the Treatment of Cystic Fibrosis. American Journal of Respiratory and Critical Care Medicine
Measuring TMEM16A
TMEM16A Channels can be measured using automated patch clamp when expressed in cell lines, and can be activated using the internal perfusion feature of the SyncroPatch 384i or using the Orbit instruments when they are recombinantly expressed in bilayers.
High throughput automated patch clamp
Parallel bilayer recordings
Cytotoxicity measurements
Application Notes
TMEM16A (ANO1) - "Internal perfusion of Ca2+ to activate TMEM16A/ANO1 on the SyncroPatch 384PE"
SyncroPatch 384PE (a predecessor model of SyncroPatch 384) application note 
(1.1 MB)
Cells were kindly provided by SB Drug Discovery.
P2X3 in chronic cough
Key Publications
Ford, A.P. & Undem, B.J. 2013. The therapeutic promise of ATP antagonism at P2X3 receptors in respiratory and urological disorders. Frontiers in Cellular Neuroscience
Ford, A.P. 2012. In pursuit of P2X3 antagonists: novel therapeutics for chronic pain and afferent sensitization. Purinergic Signalling
Measuring P2X3 Channels
P2X3 Channels can be measured using automated patch clamp when expressed in cell lines, or using the Orbit instruments when they are recombinantly expressed in bilayers.
High throughput automated patch clamp
Parallel bilayer recordings
Cytotoxicity measurements
Application Notes
P2X3 - "Activation and inhibition of P2X3 channels recorded on the SyncroPatch 384PE"
SyncroPatch 384PE (a predecessor model of SyncroPatch 384) application note: (1.5 MB)
Cells were engineered and kindly provided by Axxam S.p.A., Milan.
P2X3 - "Activation and inhibition of P2X3 channels recorded on the Patchliner"
Patchliner application note: (0.6 MB)
Cells were engineered and kindly provided by Axxam S.p.A., Milan.
P2X2 / P2X3 - "Pharmacology of P2X2/3 channels recorded on the SyncroPatch 384PE"
SyncroPatch 384PE (a predecessor model of SyncroPatch 384) application note: (5 MB)
Cells were engineered and kindly provided by Axxam S.p.A., Milan.
P2X2 / P2X3 - "P2X2/3 receptors recorded on Nanion's Patchliner"
Patchliner application note: (0.4 MB)
Cells were kindly provided by Evotec.
Publications using Instruments from Nanion
2019 - Action of MK‐7264 (Gefapixant) at human P2X3 and P2X2/3 receptors and in vivo efficacy in models of sensitisation
Port-a-Patch publication in British Journal of Pharmacology (2019)
Authors:
Richards D., Gever J.R., Ford A.P., Fountain S.J.
2011 - State-of-the-art automated patch clamp devices: heat activation, action potentials, and high throughput in ion channel screening
Port-a-Patch, Patchliner and SyncroPatch 96 (a predecessor model of SyncroPatch 384PE) publication in Frontiers in Pharmacology (2011)
Authors:
Stoelzle S., Obergrussberger A., Brüggemann A., Haarmann C., George M., Kettenhofen R., Fertig N.
CFTR
Useful Reviews
Noone, P.G. & Knowles, M.R. 2001. 'CFTR-opathies': disease phenotypes associated with cystic fibrosis transmembrane regulator gene mutations. Respiratory Research
Mall, M.A., & Hartl, D. 2014. CFTR: cystic fibrosis and beyond. European Respiratory Journal
Measuring CFTR Channels
CFTR Channels can be measured using automated patch clamp when expressed in cell lines, or using the Orbit instruments when they are recombinantly expressed in bilayers. The CFTR channel is an ABC transporter and can also be measured using the SSM-electrophysiology technique.
High throughput automated patch clamp
Parallel bilayer recordings
Transporter measurements
Application Notes
Publications using Instruments from Nanion
2021 - The rescue of F508del-CFTR by elexacaftor/tezacaftor/ivacaftor (Trikafta) in human airway epithelial cells is underestimated due to the presence of ivacaftor
Patchliner Publication in European Respiratory Journal (2021)
Authors:
Becq F., Mirval S., Carrez T., Lévêque M., Billet A., Coraux C., Sage E., Cantereau A.
2021 - Cell engineering method using fluorogenic oligonucleotide signaling probes and flow cytometry
Patchliner publication in Biotechnology Letters (2021)
Authors:
Shekdar K., Langer J., Venkatachalan S., Schmid L., Anobile J., Shah P., Lancaster A., Babich O., Dedova O., Sawchuck D.
2020 - Targeting different binding sites in the CFTR structures allows to synergistically potentiate channel activity
Patchliner publication in European Journal of Medicinal Chemistry (2020)
Authors:
Froux L., Elbahnsi A., Boucherle B., Billet A., Baatallah N., Hoffmann B., Alliot J., Zelli R., Zeinyeh W., Haudecoeur R., Chevalier B., Fortuné A., Mirval S., Simard C., Lehn P., Mornon J-P., Hinzpeter A., Becq F., Callebaut I., Décout J-L.
2017 - Development of Automated Patch Clamp Technique to Investigate CFTR Chloride Channel Function
Patchliner publication in Frontiers in Pharmacology (2017)
Authors:
Billet A.,Froux L., Hanrahan J.W., Becq F.
2017 - Buserelin alleviates chloride transport defect in human cystic fibrosis nasal epithelial cells
Port-a-Patch publication in PLoS ONE (2017)
Authors:
Calvez M.L., Benz N., Huguet F., Saint-Pierre A., Rouillé E., Coraux C., Férec C., Kerbiriou M., Trouvé P.
2014 - Improvement of Chloride Transport Defect by Gonadotropin-Releasing Hormone (GnRH) in Cystic Fibrosis Epithelial Cells
Port-a-Patch publication in PLoS One (2014)
Authors:
Benz N., Le Hir S., Norez C., Kerbiriou M., Calvez M.-L., Becq F., Trouvé P. , Férec C.