Background: Resolution of inflammation is orchestrated by specialized proresolving lipid mediators (SPMs), and this would be impaired in some cardiovascular diseases. Among SPMs, resolvins (Rv) have beneficial effects in cardiovascular pathologies, but little is known about their effect on cardiovascular damage in hypertension. Methods: Aorta, small mesenteric arteries, heart, and peritoneal macrophages were taken from C57BL/6J mice, infused or not with angiotensin II (AngII; 1.44 mg/kg/day, 14 days) in presence or absence of resolvin D2 (RvD2) (100 ng/mice, every second day) starting 1 day before or 7 days after AngII infusion. Results: Enzymes and receptors involved in SPMs biosynthesis and signaling were increased in aorta or heart from AngII-infused mice. We also observed a differential regulation of SPMs in heart from these mice. Preventive treatment with RvD2 partially avoided AngII-induced hypertension and protected the heart and large and small vessels against functional and structural alterations induced by AngII. RvD2 increased the availability of vasoprotective factors, modified SPMs profile, decreased cardiovascular fibrosis, and increased the infiltration of pro-resolving macrophages. When administered in hypertensive animals with established cardiovascular damage, RvD2 partially improved cardiovascular function and structure, decreased fibrosis, reduced the infiltration of neutrophils, and shifted macrophage phenotype toward a pro-resolving phenotype. Conclusions: There is a disbalance between proinflammatory and resolution mediators in hypertension. RvD2 protects cardiovascular function and structure when administered before and after the development of hypertension by modulating vascular factors, fibrosis and inflammation. Activating resolution mechanisms by treatment with RvD2 may represent a novel therapeutic strategy for the treatment of hypertensive cardiovascular disease.

Ion channels are macromolecular complexes present in the plasma membrane and intracellular organelles of cells. Dysfunction of ion channels results in a group of disorders named channelopathies, which represent an extraordinary challenge for study and treatment. In this review, we will focus on voltage-gated potassium channels (K_V), specifically on the K_V4-family. The activation of these channels generates outward currents operating at subthreshold membrane potentials as recorded from myocardial cells (I_TO, transient outward current) and from the somata of hippocampal neurons (I_SA). In the heart, K_V4 dysfunctions are related to Brugada syndrome, atrial fibrillation, hypertrophy, and heart failure. In hippocampus, K_V4.x channelopathies are linked to schizophrenia, epilepsy, and Alzheimer’s disease. K_V4.x channels need to assemble with other accessory subunits (β) to fully reproduce the I_TO and I_SA currents. β Subunits affect channel gating and/or the traffic to the plasma membrane, and their dysfunctions may influence channel pharmacology. Among K_V4 regulatory subunits, this review aims to analyze the K_V4/KChIPs interaction and the effect of small molecule KChIP ligands in the A-type currents generated by the modulation of the K_V4/KChIP channel complex. Knowledge gained from structural and functional studies using activators or inhibitors of the potassium current mediated by K_V4/KChIPs will better help understand the underlying mechanism involving K_V4-mediated-channelopathies, establishing the foundations for drug discovery, and hence their treatments.

Immune cells have an important role in the tumor-microenvironment. Macrophages may tune the immune response toward inflammatory or tolerance pathways. Tumor-associated macrophages (TAM) have a string of immunosuppressive functions and they are considered a therapeutic target in cancer. This study aimed to analyze the effects of trabectedin, an antitumor agent, on the tumor-microenvironment through the characterization of the electrophysiological and molecular phenotype of macrophages. Experiments were performed using the whole-cell configuration of the patch-clamp technique in resident peritoneal mouse macrophages. Trabectedin does not directly interact with K_V1.5 and K_V1.3 channels, but their treatment (16 h) with sub-cytotoxic concentrations of trabectedin increased their K_V current due to an upregulation of K_V1.3 channels. In vitro generated TAM (TAM_iv) exhibited an M2-like phenotype. TAM_iv generated a small K_V current and express high levels of M2 markers. K⁺ current from TAMs isolated from tumors generated in mice is a mixture of K_V and K_Ca, and in TAM isolated from tumors generated in trabectedin-treated mice, the current is mostly driven by K_Ca. We conclude that the antitumor capacity of trabectedin is not only due to its effects on tumor cells, but also to the modulation of the tumor microenvironment, due, at least in part, to the modulation of the expression of different macrophage ion channels.

Background and Purpose: Local anaesthetics block sodium and a variety of potassium channels. Although previous studies identified a residue in the pore signature sequence together with three residues in the S6 segment as a putative binding site, the precise molecular basis of inhibition of K_v channels by local anaesthetics remained unknown. Crystal structures of K_v channels predict that some of these residues point away from the central cavity and face into a drug binding site called side pockets. Thus, the question arises whether the binding site of local anaesthetics is exclusively located in the central cavity or also involves the side pockets.Experimental Approach: A systematic functional alanine mutagenesis approach, scanning 58 mutants, together with in silico docking experiments and molecular dynamics simulations was utilized to elucidate the binding site of bupivacaine and ropivacaine.Key Results: Inhibition of K_v1.5 channels by local anaesthetics requires binding to the central cavity and the side pockets, and the latter requires interactions with residues of the S5 and the back of the S6 segments. Mutations in the side pockets remove stereoselectivity of inhibition of K_v1.5 channels by bupivacaine. Although binding to the side pockets is conserved for different local anaesthetics, the binding mode in the central cavity and the side pockets shows considerable variations.Conclusion and Implications: Local anaesthetics bind to the central cavity and the side pockets, which provide a crucial key to the molecular understanding of their K_v channel affinity and stereoselectivity, as well as their spectrum of side effects.

Background and Purpose: TheK_vβ1.3 subunit modifies the gating and pharmacology ofK_v1.5 channels in aPKC‐dependent manner, decreasing channel sensitivity to bupivacaine‐ and quinidine‐mediated blockade. CardiacK_v1.5 channels associate with receptor for activated C kinase 1 (RACK1), theK_vβ1.3 subunit and differentPKCisoforms, resulting in the formation of a functional channelosome. The aim of the present study was to investigate the effects ofPKCinhibition on bupivacaine and quinidine block ofK_v1.5 +K_vβ1.3 channels.Experimental Approach: HEK293 cells were transfected withK_v1.5 +K_vβ1.3 channels, and currents were recorded using the whole‐cell configuration of the patch‐clamp technique.PKCinhibition was achieved by incubating the cells with either calphostinCor bisindolylmaleimideIIand the effects of bupivacaine and quinidine were analysed.Key Results: The voltage‐dependent inactivation ofK_v1.5 +K_vβ1.3 channels and their pharmacological behaviour afterPKCinhibition with calphostinCwere similar to those displayed byK_v1.5 channels alone. Indeed, theIC₅₀values for bupivacaine were similar in cells whosePKCwas inhibited with calphostinCor bisindolylmaleimideII. Similar results were also observed in the presence of quinidine.Conclusions and Implications: The finding that the voltage‐dependence of inactivation and the pharmacology ofK_v1.5 +K_vβ1.3 channels afterPKCinhibition resembled that observed inK_v1.5 channels suggests that both processes are dependent onPKC‐mediated phosphorylation. These results may have clinical relevance in diseases that are characterized by alterations in kinase activity.