A debilitating illness, pulmonary arterial hypertension (PAH) arises from deleterious remodeling of pulmonary arterioles, leading to increased pulmonary artery pressure, a rise in pulmonary vascular resistance, right sided heart failure and death. The pathogenesis of the disease is incompletely understood; however, certain established pathological features have guided medical treatments to improve mortality rates. For instance, an imbalance of vasoconstrictor molecules, such as endothelin-1, to vasodilator compounds, such as nitric oxide, contributes to excessive pulmonary arterial constriction, and a propensity for pulmonary arterial smooth muscle and endothelial cell proliferation. Therapeutic strategies may aim to restore this imbalance with the use of endothelin receptor antagonists, prostacyclin analogs, and other vasodilating agents.
Mutations in the BMPR2 gene, the most common genetic cause of PAH, leads to aberrant TGF-ß signaling, which promotes uncontrollable cell proliferation and pathological changes in pulmonary arterioles. Genetic studies have revealed PAH-associated mutations in several other genes within the TGF-ß signaling pathway. More recently, our research group discovered loss-of-function mutations in the KCNK3 gene encoding the KCNK3 two-pore domain potassium channel in patients with idiopathic and familial PAH.
KCNK3 (also referred to as TASK-1, or K2P3.1) represents the first ion channelopathy as a cause of PAH. KCNK3 is expressed in human pulmonary artery smooth muscle and endothelial cells. Loss of KCNK3 channel currents leads to membrane depolarization and predisposes to deleterious pulmonary arterial remodeling. Chapter 1 of my thesis explores the impact of KCNK3 mutations on potassium channel function in cellular models of heterozygous conditions, as all patients with PAH-associated KCNK3 mutations in our study were heterozygous at the KCNK3 gene locus.
Furthermore, we explored function of mutant and non-mutant KCNK3 channels in cultured human pulmonary artery smooth muscle cells to better define the electrophysiological consequence of KCNK3 dysfunction, and used a KCNK3-activating pharmacological agent, ONO-RS-082, to gauge the therapeutic potential of KCNK3 as a pharmacological target in PAH. Moreover, the study of KCNK3 channel activity when assembled with the closely related KCNK9 channel provided a platform for exploring the lung-specific phenotype in patients with heterozygous KCNK3 mutations, despite widespread tissue expression KCNK3 in the body.
In Chapter 2 of my thesis work, the discovery of a second potassium channelopathy in PAH is characterized. Heterozygous mutations in the ABCC8 gene, encoding the sulfonylurea receptor 1 (SUR1) protein, were found in pediatric and adult patients with idiopathic and familial PAH. SUR1, a beta subunit of the ATP-sensitive potassium channel (KATP), assembles with the pore-forming Kir6.2 alpha subunit to form KATP, a channel sensitive to inhibition by intracellular ATP. At the plasma membrane, KATP inwardly rectifying potassium currents contribute to the resting potential, and may play a pathophysiological role in PAH via dysfunction in pulmonary artery smooth muscle and/or endothelial cells. In this chapter, eight ABCC8 mutations associated with PAH were functionally characterized, and pharmacological agents were employed to examine the therapeutic potential in targeting SUR1-containing KATP channels in PAH.
Altogether, the research presented in this dissertation identifies and explores potassium channel dysfunction as a pathogenic mechanism in PAH, due to heterozygous genetic mutations in KCNK3 and ABCC8. Evidence of restoration of mutant KCNK3 and KATP channel function by pharmacological agents suggests that targeting potassium channels as a therapeutic strategy may alleviate the severe morbidity and mortality burden in patients with PAH.
| Identifer | oai:union.ndltd.org:columbia.edu/oai:academiccommons.columbia.edu:10.7916/D89K4J52 |
| Date | January 2017 |
| Creators | Bohnen, Michael S. |
| Source Sets | Columbia University |
| Language | English |
| Detected Language | English |
| Type | Theses |
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