Ria Dinsdale

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• BK channel activity in skin fibroblasts from patients with neurological disorder CHANNELS Dinsdale, R. L., Middendorf, T. R., Disilvestre, D., Adams, D., Gahl, W., Macnamara, E. F., Wolfe, L., Toro, C., Tifft, C. J., Meredith, A. L. 2025; 19 (1): 2542811

  Abstract

  Seventy-five unique variants in the KCNMA1 gene have been identified from individuals with neurological disorders. However, variant pathogenicity and evidence for disease causality are lacking in most cases. In this study, the KCNMA1 variants N999S and E656A (rs886039469 and rs149000684, respectively) were investigated from two individuals presenting with neurological disorders. N999S was previously shown to produce strong gain-of-function (GOF) changes in homomeric BK channel properties in vitro and is found as a heterozygous allele associated with epilepsy and paroxysmal dyskinesia in humans. Although its pathogenicity has been demonstrated in heterozygous animal models, the GOF classification for N999S has not been validated in a heterozygous patient-derived tissue. Conversely, the GOF pathogenicity for E656A is based solely on homomeric channels expressed in vitro and is inconclusive. For either variant, the properties of single heterozygous channels and allele expression is unknown. In this study, we profiled the wild-type and mutant KCNMA1 transcripts from primary human skin fibroblasts of heterozygous patients and unaffected controls and performed patch-clamp electrophysiology to characterize endogenous BK channel current properties. GOF gating was observed in single BK channel recordings from both channel types. Fibroblasts from the individual harboring the E656A variant showed decreases in the number of BK channels detected and E656A-containing transcripts compared to controls. These results show that single BK channels can be reliably detected in primary fibroblasts obtained from human skin biopsies, suggesting their utility for establishing variant pathogenicity, and reveal the BK channel expression and functional changes associated with two heterozygous patient genotypes.

  View details for DOI 10.1080/19336950.2025.2542811

  View details for Web of Science ID 001547963700001

  View details for PubMedID 40785052

  View details for PubMedCentralID PMC12341059

• Pharmacological profiling of small molecule modulators of the TMEM16A channel and their implications for the control of artery and capillary function BRITISH JOURNAL OF PHARMACOLOGY Al-Hosni, R., Agostinelli, E., Ilkan, Z., Scofano, L., Kaye, R., Dinsdale, R. L., Acheson, K., Macdonald, A., Rivers, D., Biosa, A., Gunthorpe, M. J., Platt, F., Tammaro, P. 2025

  Abstract

  TMEM16A chloride channels constitute a depolarising mechanism in arterial smooth muscle cells (SMCs) and contractile cerebral pericytes. TMEM16A pharmacology is incompletely defined. We elucidated the mode of action and selectivity of a recently identified positive allosteric modulator of TMEM16A (PAM_16A) and of a range of TMEM16A inhibitors. We also explore the consequences of selective modulation of TMEM16A activity on arterial and capillary function.Patch-clamp electrophysiology, isometric tension recordings, live imaging of cerebral cortical capillaries and assessment of cell death were employed to explore the effect of selective pharmacological control of TMEM16A on vascular function.In low intracellular free Ca2+ concentrations ([Ca2+]i), nanomolar concentrations of PAM_16A activated heterologous TMEM16A channels, while being almost ineffective on the closely related TMEM16B channel. In either the absence of Ca2+ or in saturating [Ca2+]i, PAM_16A had no effect on TMEM16A currents at physiological potentials. PAM_16A selectively activated TMEM16A currents in SMCs and enhanced aortic contraction caused by phenylephrine or angiotensin-II and capillary (pericyte) constriction evoked by endothelin-1 or oxygen-glucose deprivation (OGD) to simulate cerebral ischaemia. Conversely, selective TMEM16A inhibition with Ani9 facilitated aortic, mesenteric and pericyte relaxation, and protected against OGD-mediated pericyte cell death. Unlike PAM_16A and Ani9, a range of other available modulators were found to interfere with endogenous cationic currents in SMCs.Arterial tone and capillary diameter can be controlled with TMEM16A modulators, highlighting TMEM16A as a target for disorders with a vascular component, including hypertension, stroke, Alzheimer's disease and vascular dementia.

  View details for DOI 10.1111/bph.17383

  View details for Web of Science ID 001400010300001

  View details for PubMedID 39829151