The role of SLC12A family of cation-chloride cotransporters and drug discovery methodologies

Shiyao Zhang; Nur Farah Meor Azlan; Sunday Solomon Josiah; Jing Zhou; Xiaoxia Zhou; Lingjun Jie; Yanhui Zhang; Cuilian Dai; Dong Liang; Peifeng Li; Zhengqiu Li; Zhen Wang; Yun Wang; Ke Ding; Yan Wang; Jinwei Zhang

doi:10.1016/j.jpha.2023.09.002

The role of SLC12A family of cation-chloride cotransporters and drug discovery methodologies

J Pharm Anal. 2023 Dec;13(12):1471-1495. doi: 10.1016/j.jpha.2023.09.002. Epub 2023 Sep 9.

Authors

Shiyao Zhang¹, Nur Farah Meor Azlan², Sunday Solomon Josiah², Jing Zhou³, Xiaoxia Zhou¹, Lingjun Jie¹, Yanhui Zhang¹, Cuilian Dai¹, Dong Liang⁴, Peifeng Li⁵, Zhengqiu Li⁶, Zhen Wang⁷, Yun Wang³, Ke Ding⁷, Yan Wang¹, Jinwei Zhang^{1

2

7}

Affiliations

¹ Xiamen Cardiovascular Hospital of Xiamen University, School of Medicine, Xiamen University, Xiamen, Fujian, 363001, China.
² Institute of Biomedical and Clinical Sciences, Medical School, Faculty of Health and Life Sciences, University of Exeter, Exeter, EX4 4PS, UK.
³ Department of Neurology, Institutes of Brain Science, State Key Laboratory of Medical Neurobiology and MOE Frontiers Center for Brain Science, Institute of Biological Science, Zhongshan Hospital, Fudan University, Shanghai, 200032, China.
⁴ Aurora Discovery Inc., Foshan, Guangdong, 528300, China.
⁵ Institute for Translational Medicine, Qingdao University, Qingdao, Shandong, 266021, China.
⁶ School of Pharmacy, Jinan University, Guangzhou, 510632, China.
⁷ State Key Laboratory of Chemical Biology, Research Center of Chemical Kinomics, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai, 200032, China.

Abstract

The solute carrier family 12 (SLC12) of cation-chloride cotransporters (CCCs) comprises potassium chloride cotransporters (KCCs, e.g. KCC1, KCC2, KCC3, and KCC4)-mediated Cl^- extrusion, and sodium potassium chloride cotransporters (N[K]CCs, NKCC1, NKCC2, and NCC)-mediated Cl^- loading. The CCCs play vital roles in cell volume regulation and ion homeostasis. Gain-of-function or loss-of-function of these ion transporters can cause diseases in many tissues. In recent years, there have been considerable advances in our understanding of CCCs' control mechanisms in cell volume regulations, with many techniques developed in studying the functions and activities of CCCs. Classic approaches to directly measure CCC activity involve assays that measure the transport of potassium substitutes through the CCCs. These techniques include the ammonium pulse technique, radioactive or nonradioactive rubidium ion uptake-assay, and thallium ion-uptake assay. CCCs' activity can also be indirectly observed by measuring γ-aminobutyric acid (GABA) activity with patch-clamp electrophysiology and intracellular chloride concentration with sensitive microelectrodes, radiotracer ³⁶Cl^-, and fluorescent dyes. Other techniques include directly looking at kinase regulatory sites phosphorylation, flame photometry, ²²Na⁺ uptake assay, structural biology, molecular modeling, and high-throughput drug screening. This review summarizes the role of CCCs in genetic disorders and cell volume regulation, current methods applied in studying CCCs biology, and compounds developed that directly or indirectly target the CCCs for disease treatments.

Keywords: Cation-chloride cotransporters; Chloride volume regulation; Cotransporter assays; Drug discovery.

Publication types

Review