News and Highlights
On this page we present recently published research carried out by SSEP members
Tamoxifen in Duchenne Muscular Dystrophy – promising first results
(2021) Neuromusc Disorders 31, 9, 801-802
Tamoxifen is one of the most often prescribed drugs to treat estrogen-dependent breast cancer. It is orally active, has a good safety profile and is available at low cost. Our earlier preclinical investigations focusing on an approach to treat Duchenne Muscular Dystrophy (DMD) via molecular targets in the pathogenesis of the disease led to tamoxifen. Thus, in 2017, two clinical trials were initiated. In this Editorial, results of the first trial for repurposing tamoxifen in DMD for are discussed. Even though only few DMD patients were treated, results are encouraging as numerous tests showed a stabilization of the progress of the disease. Once results from the larger placebo-controlled trial ongoing in eleven European Centers and coordinated by the University Hospital Basel are known, these findings might lead to an improved treatment of DMD patients.
Historical account and current research of Pharmacology in Switzerland
Urs Rüegg and Gabriele Weitz-Schmidt
(2020) Pharmacological Research 154, 1-12
The path of the Swiss Society of Pharmacology & Toxicology from its foundation in 1965 is described together with the past and the current state of research in the swiss academic and industrial centers of pharmacology and toxicology. Measures to promote the role of pharmacology in Switzerland are proposed, such as Personalized and Precision Medicine. An improved integration of academic and industrial pharmacological research will be required to master the challenges and realize the opportunities ahead.
Mesoporous Silica Nanoparticles as pH-Responsive Carrier for the Immune-Activating Drug Resiquimod Enhance the Local Immune Response in Mice
Julia Wagner, Dorothée Gößl, Natasha Ustyanovska, Mengyao Xiong, Daniel Hauser, Olga Zhuzhgova, Sandra Hočevar, Betül Taskoparan,
Laura Poller, Stefan Datz, Hanna Engelke, Youssef Daali, Thomas Bein, and Carole Bourquin
(2021) ACS Nano, 15, 3, 4450–4466
Immune-activating drugs, such as the TLR7 agonist resiquimod, are known to potently activate dendritic cells (DCs) and have the potential to boost anti-cancer immune responses. Indeed, the synthetic TLR7 ligand imiquimod is already in clinical use for the topical treatment of basal cell carcinoma. Yet the systemic administration of these small-molecule drugs for cancer immunotherapy has thus far been limited by their quick distribution and elimination from the body before they reach the DCs in high enough doses. We therefore developed stimuli-responsive mesoporous silica nanoparticles (MSNs) as carriers for resiquimod to improve the drug’s pharmacokinetics and enhance the activation of DCs. The MSNs, equipped with a pH-responsive gatekeeper, showed efficient release of their cargo in an environmental pH of 5.5 or below. We furthermore demonstrated that the resiquimod-loaded MSNs were rapidly taken up by DCs into the acidic environment of the lysosomes, which resulted in the potent activation of these immune cells. Upon subcutaneous injection into mice, the loaded particles strongly enhanced the activation of migratory DCs in the draining lymph nodes. Moreover, the pharmacokinetic profile of resiquimod significantly improved while the systemic exposure of the drug was reduced. When we simultaneously delivered the model antigen OVA and the adjuvant resiquimod through MSNs, the antigen-specific T-cell response increased, suggesting a potential use for MSNs as carriers for cancer vaccines.
Physiological concentrations of cyanide stimulate mitochondrial Complex IV and enhance cellular bioenergetics
Elisa B Randi, Karim Zuhra, Laszlo Pecze, Theodora Panagaki, Csaba Szabo
(2021) Proc Natl Acad Sci U S A, 18;118(20):e2026245118.
Cyanide is generally viewed as a cytotoxic agent to mammalian cells, because it inhibits mitochondrial Complex IV. However, this report demonstrates that the effect of cyanide is biphasic, and at low (nM - µM) concentrations cyanide stimulates CCOx, while the classical inhibitory effects only occur at higher concentrations. These studies, coupled with studies demonstrating that cyanide can be produced by various enzymes in mammalian cells raise the possibility that – in addition to NO, CO and H2S – cyanide may also serve regulatory roles as the fourth gasotransmitter in mammalian cells and tissues.
PLCγ1 suppression promotes the adaptation of KRAS-mutant lung adenocarcinomas to hypoxia
Maria Saliakoura, Matteo Rossi Sebastiano, Chiara Pozzato, Florian H Heidel, Tina M Schnöder, Spasenija Savic Prince, Lukas Bubendorf, Paolo Pinton, Ralph A Schmid, Johanna Baumgartner, Stefan Freigang, Sabina A Berezowska, Alessandro Rimessi, Georgia Konstantinidou
(2020) Nat Cell Biol, 22(11):1382-1395
Hypoxia is a powerful driver of aggressive behaviour in solid cancers and is linked to metabolic reprogramming, with suppression of mitochondrial respiration in favour of glycolysis. In KRAS-driven lung cancer, this metabolic adaptation to hypoxia has been attributed to the effect of hypoxia-inducible factor-1α (HIF-1α)-dependent signalling and transcriptional regulation. Hypoxia can lead to profound rearrangements in cancer lipid metabolism, including inhibition of fatty acid oxidation and accumulation of triglycerides in lipid droplets. Nevertheless, the identity and contribution of lipid species that are important for cancer cell adaptation to hypoxia are still mostly unknown. We found that hypoxia alters the saturation level and number of carbons of phosphoinositide (PI) lipids with vital role in membrane signalling propagation, namely PI, PI4P and PI(4,5)P2. Mechanistically, this was due to the suppression of phospholipase Cγ1 (PLCγ1) levels in hypoxia, through a mechanism that is, at least in part, HIF-1α-dependent. The hypoxia-induced PLCγ1 suppression triggered a sharp functional effect, driving proliferation and allowing lung cancer cells to override apoptosis. The survival advantage conferred by PLCγ1 downregulation could be attributed to the suppression of mitochondrial ROS and further amplification of the glycolytic profile of cancer cells, two key events that allow them to efficiently adapt to hypoxia. By assessing the protein levels of PLCγ1 in a lung tumour tissue microarray, we also provided evidence that the level of PLCγ1 has a predictive value for overall tumour resilience and lung cancer patient survival. Given the fact that hypoxia is a future of virtually all solid tumors, our results are expected to make a major contribution to the field of KRAS-driven lung cancer.
Para-Halogenation of Amphetamine and Methcathinone Increases the Mitochondrial Toxicity in Undifferentiated and Differentiated SH-SY5Y Cells
Zhou X, Bouitbir J, Liechti ME, Krähenbühl S, and Mancuso RV
(2020) Int J Mol Sci, 21(8), 2841
New psychoactive substances (NPS) are a wide variety of psychoactive compounds, structurally and pharmacologically similar to traditional drugs of abuse, not yet defined as illegal due to their novelty. A common method to obtain NPS is the halogenation of amphetamines and methcathinones. The para-halogenated derivatives of amphetamine and methcathinone are available over the internet and have entered the illicit drug market. The present study explores the neurotoxicity of amphetamine, methcathinone and their para-halogenated derivatives 4-fluoroamphetamine (4-FA), 4-chloroamphetamine (PCA), 4-fluoromethcathinone (4-FMC), and 4-chloromethcathinone (4-CMC) in undifferentiated and differentiated neuronal SH-SY5Y cells. Para-halogenation of amphetamine and methcathinone increases their neurotoxic properties in undifferentiated and differentiated SH-SY5Y cells, due to the impairment of mitochondrial function and induction of apoptosis. The toxicity rank order of the substituents in the p-position for disturbing mitochondrial function was for both amphetamines and methcathinones (Cl > F > H), which was also the case for cytotoxicity. Moreover, epidemiological studies have reported an increased risk to develop Parkinson’s disease (PD) in amphetamine consumers. The current study suggests that para-halogenation of amphetamines and methcathinones may increase such risk, and explain the loss of dopaminergic neurons in the nigrostriatal system due to mitochondrial damage.
Hyperthermia Increases Neurotoxicity Associated with Novel Methcathinones
Zhou X, Bouitbir J, Liechti ME, Krähenbühl S, and Mancuso RV
(2020) Cells, 9(4), 965
Hyperthermia, also known as “overheating”, is the one of the prominent acute manifestations of stimulant drug abuse, and it is often associated to life-threatening conditions. The abuse of new psychoactive substances (NPS), a broad group of drugs that are not controlled by classic international drug laws, is a major problem worldwide. Synthesis cathinones, a new class of NPS, have emerged in recent years on the black market, and their use as recreational drugs has grown rapidly. Structurally, synthetic cathinones are b-keto-amphetamine derivatives, with pharmacological and toxicological properties similar to amphetamines. The present study investigates in vitro the role of hyperthermia on methcathinone-induced neurotoxicity using the well-established SH-SY5Y neuronal cell model. The investigated synthetic cathinones result to be mitochondrial toxicants whose toxicity is increased by shifting the temperature from 37 to 40.5 °C. SH-SY5Y cells exposed to 40.5 °C activate cellular defense mechanisms, such as the expression of Hsp70 proteins, which can partially prevent early apoptosis and necrosis. With time, the activation of additional defense mechanisms, such as autophagy, is necessary to prevent cell dysfunction and cell death. It is well known that the aggregation of misfolded proteins, which may result from the production of defective proteins and/or impaired function of the protein quality control systems, is a common pathological feature of many neurodegenerative diseases, such as Alzheimer’s disease (AD), Parkinson’s disease (PD), and Huntington disease (HD). Many studies have shown a link between autophagy and neurodegenerative diseases. With this study, we suggest that repetitive ingestion of neurotoxic drugs, such as methcathinones, may aggravate or even provoke such conditions.
Anti-alphaLbeta2 antibodies reveal novel endocytotic crossmodulatory functionality
Mancuso RV, Casper J, Schmidt AG, Krahenbuhl S, Weitz-Schmidt G
(2020) Br J Pharmacol, 10.1111/bph.14996
Antibodies targeting cell surface receptors are considered to deliver highly selective therapeutic interventions for immune disorders and cancer. Their biologic profiles are found, generally, to represent the net effects of the antibody-target interaction. The former therapeutic anti-integrin aLb2 (LFA-1) antibody efalizumab seems to defeat this paradigm by eliciting, via mechanisms currently unknown, much broader effects than would be predicted based on its target specificity. The present study identifies a hitherto unknown functionality of inhibitory anti-integrin aLb2 antibodies (including efalizumab) which clarifies, at least in part, their perplexingly broad and profound biologic effects. The newly described phenomenon, referred to as endocytotic crossmodulation, accounts for the antibodies’ unexpected downmodulation of integrin immune receptors which are unrelated to the antibodies’ target integrin aLb2. In other words, endocytotic crossmodulation renders specifically acting antibodies biologically non-selective. The phenomenon is of high therapeutic relevance because it has the potential of fundamentally altering an antibody’s benefit risk profile, as evident with efalizumab which was withdrawn from markets in 2009. Intriguingly, the crossmodulation observed with antibodies does not occur when integrin aLb2 is inhibited by small molecule-based pharmacologies. The newly described phenomenon of endocytotic crossmodulation may be of relevance to other therapeutic antibodies, particularly to those targeting cluster-forming receptors. It should be on the radar of pharmacologists investigating such antibodies, in consequence.
Overproduction of H2S, generated by CBS, inhibits mitochondrial Complex IV and suppresses oxidative phosphorylation in Down syndrome
Panagaki T, Randi EB, Augsburger F, and Szabo C
(2019) Proc Natl Acad Sci U S A, 116(38):18769-18771
In Down Syndrome there is an extra 21st chromosome in the cells, which produces many extra proteins, leading to severe biochemical problems. One of these extra proteins is an enzyme called CBS. This enzyme has important physiological roles: it gets rid off some potentially toxic metabolites (e.g. homocysteine), but it also makes a biological gas called H2S. In the Down Syndrome cells, too much H2S produced, and at this level, H2S is toxic. The current paper shows that this excess H2S inhibits the ability of the mitochondria (a cell organelle that specializes in making ATP, the 'currency' of energy in our cells) to move electrons. When the H2S homeostasis is normalized (by deleting CBS or pharmacologically inhibiting its ability to make H2S), the "toxic H2S gas cloud" can be lifted and the cells regained their ability to produce energy. The findings give us hope that that future, therapeutic inhibition of CBS may improve metabolic, neuronal, or possibly cognitive function in Down Syndrome.
The architecture of the IgG anti-carbohydrate repertoire in primary antibody deficiencies (PADs)
Jandus P, Frias Boligan K, Smith DF, de Graauw E, Grimbacher B, Jandus C, Abdelhafez MM, Despont A, Bovin N, Simon D, Rieben R, Simon HU, Cummings RD, von Gunten S.
(2019) Blood.2019 Sep 19. pii: blood.2019001705
The surface of each living cell, including microbial pathogens and tumor cells,is coated with a variety of often characteristic glycans, which can be recognized by glycan-specific antibodies. Given the importance of antibodies in immune defence, an atlas of the glycan-specific IgG repertoire in primary antibody deficiency (PAD) was established. Patients with certain types of PAD exhibited a significant lack of IgG antibodies to bacterial antigens. Furthermore, a lack of naturally-occurring tumour-specific antibodies was found in patient subsets known to be at increased risk to develop malignancies. On the other hand, the observed lack of Gal-alpha IgG antibodies in PAD patients suggested tolerance for xenotransplants, which was confirmed in functional experiments. Taken together, we identified significant differences in the architecture of the glycan-specific IgG antibody repertoire, which may have important implications for diagnostics, preventive and therapeutic avenues and the clinical course in immunodeficiencies.
Siglec-9 regulates an effector memory CD8+T-cell subset that congregates in the melanoma tumor microenvironment
Haas Q, Boligan KF, Jandus C, Schneider C, Simillion C, Stanczak MA, Haubitz M, Seyed Jafari SM, Zippelius A, Baerlocher GM, Läubli H, Hunger RE, Romero P, Simon HU, von Gunten S.
(2019) Cancer Immunol Res 7(5):707-718
Tumors are known to exhibit altered surface glycosylation patterns, which have functional implications. In this study, it wasdemonstrated that hypersialylation,one of the most common tumor glycosylation changes, preventsefficient immune attack by cytotoxic CD8+T cells. This attenuation of T cell anti-tumor responses was shown to involve the interaction of tumor-associated carbohydrate antigens (TACA) with the inhibitory Siglec-9 receptor, which in melanoma was up-regulated on tumor-infiltrating effector memory T cells. This work highlights the role of tumor glycosylation interaction with glycan-binding proteins (GBP) as a further immune checkpoint that might be targeted for improved immunotherapeutic interventionsfor the treatment of cancer.
2018 and older
Identification of potent and selective small molecule inhibitors of the cation channel TRPM4
Ozhathil LC, Delalande C, Bianchi B, Nemeth G, Kappel S, Thomet U, Ross-Kaschitza D, Simonin C, Rubin M, Gertsch J, Lochner M, Peinelt C, Reymond JL, Abriel H
(2018) Br J Pharmacol, 10.1111/bph.14220
Ion channels are membrane proteins found in virtually every cell allowing the passage of different ions such as Na+, K+ and Ca2+ and Cl-. Many drugs are modulating their function. In this study, we looked for new chemical compounds able to block the ion channel called TRPM4 found at the membrane of many cell types such as cardiac and cancer cells. In particular, we developed a novel fluorescence-based cellular assay and screened a library of ~800 compounds to identify a potent and selective blocker of TRPM4 channel. Our data also demonstrate that the identified compound in addition to being a blocker of TRPM4 also rescues the membrane trafficking of TRPM4 loss-of-expression mutants found in cardiac patients. The new chemical structure identified in our study will be useful to understand the role of TRPM4 in disease and possibly develop new clinical drug candidates.
ROS and glutathionylation balance cytoskeletal dynamics in neutrophil extracellular trap formation
Stojkov D, Amini P, Oberson K, Sokollik C, Duppenthaler A, Simon HU, Yousefi S
(2017) J Cell Biol 216: 4073-4090
Neutrophils represent the most prominent cells in the blood circulation and their antimicrobial defense activity has been defined by their ability to phagocytose microbes or to form Neutrophil Extracellular Traps (NETs) that can entrap and kill bacteria in the extracellular space. In our present manuscript, we report novel findings which indicate that a dynamic intracellular transport by the cytoskeleton plays a role in bringing granule proteins and mitochondrial DNA together to form NETs. Our data also demonstrate that reactive oxygen species (ROS), generated by the enzyme NADPH oxidase, act as signaling molecules involved in the regulation of the cytoskeleton. Overall, our work suggests that the activation of actin polymerization might be a new strategy for improving neutrophil function in NADPH oxidase deficiency.
BH3 mimetics efficiently induce apoptosis in mouse basophils and mast cells
Reinhart R, Rohner L, Wicki S, Fux M, Kaufmann T
(2018) Cell Death Differ 25: 204-216
Basophils and mast cells are known to play critical roles in the pathogenesis of diverse allergic diseases. The survival of both cell types seems to strongly depend on distinct pro-survival proteins of the Bcl-2 family. We thus hypothesized that a novel class of small molecule inhibitors specifically targeting individual Bcl-2 family members, co-called BH3 mimetics, may be useful to induce apoptosis in naïve or activated basophils and mast cells. Indeed, our work revealed the crucial importance of BCL-2 and BCL-XL for survival of in vitro differentiated mouse basophils, whereas mouse mast cells and human basophils highly depended on BCL-2 and MCL-1, respectively. Cell survival was strongly increased by the key cytokine IL-3, but this effect could still be counteracted by the right combination of BH3 mimetic compounds. In conclusion, our results indicate that BH3 mimetics, besides their application in anti-cancer therapy, may have a potential worthwhile exploring in the treatment of basophil and mast cell mediated allergic disorders.
Evidence of an abnormal epithelial barrier in active, untreated and corticosteroid-treated eosinophilic esophagitis
Simon D, Page B, Vogel M, Bussmann C, Blanchard C, Straumann A, Simon HU
(2017) Allergy, 10.1111/all.13244
Eosinophilic esophagitis (EoE) is a chronic, immune-mediated disease characterized by symptoms related to esophageal dysfunction and an eosinophil-predominant inflammation. Here, we asked the question whether Candida albicans colonization and sensitization observed in pediatric and adult EoE patients is the consequence of an epithelial dysfunction associated with the disease and/or is a consequence of the treatment with corticosteroids (CS) that is the first line therapy for EoE. Using immunofluorescence techniques on esophageal tissue sections, we observed increased numbers of eosinophils and mast cells, higher expression levels of alarmins, antimicrobial peptides, and proteases in EoE as compared with controls, while reduced expression of a protease inhibitor and barrier proteins, i.e. filaggrin, E-cadherin, claudin, occludin, demoglein-1 was found, independent of CS therapy. CS seemed to have selective effects on improving the epithelial barrier. Instead, CS reduced the expression of cathelicidin as well as the numbers of LHC and eosinophils, thereby possibly even further promoting Candida albicans colonization and invasion.
IVIG regulates the survival of human but not mouse neutrophils
Schneider C, Wicki S, Graeter S, Timcheva TM, Keller CW, Quast I, Leontyev D, Djoumerska-Alexieva IK, Kasermann F, Jakob SM, Dimitrova PA, Branch DR, Cummings RD, Lunemann JD, Kaufmann T, Simon HU, von Gunten S
(2017) Sci Rep 7: 1296
Intravenous immunoglobulin (IVIG) has anti-inflammatory effects when administered at high concentrations to patients with chronic inflammatory and autoimmune disorders. IVIG is a pluripotent drug and a number of immunoregulatory mechanisms have been described including induced cell death of activated neutrophils. Schneider C et al. found that latter mechanism is specific, Fab but not Fc-mediated, and occurs in human, but not mouse neutrophils. In addition to xenogeneic effects of human IVIG if used in mice, this finding highlights potential species-differences in the mechanisms of IVIG action depending on the experimental disease model in vivo.
Conformational dynamics and role of the acidic pocket in ASIC pH-dependent gating
Vullo S, Bonifacio G, Roy S, Johner N, Berneche S, Kellenberger S
(2017) Proc Natl Acad Sci U S A 114: 3768-3773
Acid-sensing ion channels (ASICs) are neuronal Na channels that are activated by a drop in extracellular pH. They contribute to physiological and pathological processes such as learning, fear and pain sensation, and neuronal death after ischemic stroke. Although ASICs are potentially interesting drug targets, their activation mechanism is only poorly understood. This study shows that pH sensing in a channel domain that had been proposed as the ligand binding site of ASICs, the “acidic pocket”, has only a modulatory, but not an essential role for the activation of these channels. The study further describes the mechanisms of the ASIC regulation by the acidic pocket, which remains an interesting target site for drug candidates acting on ASICs.
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