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1.  The Role of HIF1α in Renal Cell Carcinoma Tumorigenesis 
The transcription factor HIF1α is implicated in the development of clear cell renal cell carcinoma (ccRCC). Although HIF1α was initially believed to be essential for ccRCC development, recent studies hypothesize an oncogenic role for HIF2α in ccRCC, but a tumor suppressor role for HIF1α [1], leading to uncertainty as to the precise roles of the different HIF transcription factors in this disease.
Using evidence available from studies with human ccRCC cell lines, mouse xenografts, murine models of ccRCC, and human ccRCC specimens, we evaluate the roles of HIF1α and HIF2α in the pathogenesis of ccRCC. We present a convergence of clinical and mechanistic data supporting an important role for HIF1α in promoting tumorigenesis in a clinically important and large subset of ccRCC. This indicates that current understanding of the exact roles of HIF1α and HIF2α is incomplete and that further research is required to determine the diverse roles of HIF1α and HIF2α in ccRCC.
doi:10.1007/s00109-014-1180-z
PMCID: PMC4119538  PMID: 24916472
Clear cell renal cell carcinoma; Cancer stem cells; Hypoxia inducible factor 1α; Kidney cancer; Carcinogenesis; Review
2.  Molecules in medicine mini review: the αβ T cell receptor 
As an integral part of the mammalian immune system, a distributed network of tissues, cells, and extra-cellular factors, T lymphocytes perform and control a multitude of activities that collectively contribute to the effective establishment, maintenance, and restoration of tissue and organismal integrity. Development and function of T cells is controlled by the T cell receptor (TCR), a heterodimeric cell surface protein uniquely expressed on T cells. During T cell development, the TCR undergoes extensive somatic diversification that generates a diverse T cell repertoire capable of recognizing an extraordinary range of protein and nonprotein antigens presented in the context of major histocompatibility complex molecules (MHC). In this review, we provide an introduction to the TCR, describing underlying principles that position this molecule as a central regulator of the adaptive immune system involved in responses ranging from tissue protection and preservation to pathology and autoimmunity.
doi:10.1007/s00109-014-1145-2
PMCID: PMC4269364  PMID: 24848996
Tcell receptor; Major histocompatibility complex; Tcell repertoire
3.  The pneumococcus: why a commensal misbehaves 
Several characteristics of Streptococcus pneumoniae (pneumococcus) combine to make it a particularly problematic pathogen. Firstly, the pneumococcus has the capacity to cause disease through the expression of virulence factors such as its polysaccharide capsule and pore-forming toxin. In addition, the pneumococcus is highly adaptable as demonstrated by its ability to acquire and disseminate resistance to multiple antibiotics. Although the pneumococcus is a major cause of disease, the organism is most commonly an “asymptomatic” colonizer of its human host (the carrier state), with transmission occurring exclusively from this reservoir of commensal organisms. Thus, it is unclear how the organism’s virulence and adaptability promote its persistence or host to host spread during its carrier state. This review summarizes current understanding of how these characteristics may contribute to the commensal lifestyle of the pneumococcus.
doi:10.1007/s00109-009-0557-x
PMCID: PMC4487619  PMID: 19898768
Colonization; Virulence factor; Neutrophil; Pneumococcus
4.  Ubiquitous Points of Control over Regulatory T cells 
Posttranslational modification by ubiquitin tagging is crucial for regulating the stability, activity and cellular localization of many target proteins and processes including DNA repair, cell cycle progression, protein quality control and signal transduction. It has long been appreciated that ubiquitin-mediated events are important for certain signaling pathways leading to leukocyte activation and the stimulation of effector function. It is now clear that the activities of molecules and pathways central to immune regulation are also modified and regulated through ubiquitin. Among the mechanisms of immune control, regulatory T cells (or Tregs) are themselves particularly sensitive to such regulation. E3 ligases and deubiquitinases both influence Tregs through their effects on signaling pathways pertinent for these cells or through the direct, posttranslational regulation of Foxp3. In this review we will summarize and discuss several examples of ubiquitin-mediated control over multiple aspects of biology of Tregs including their generation, function and phenotypic fidelity. Fully explored and exploited, these potential opportunities for Treg modulation may lead to novel immunotherapies for both positive and negative fine-tuning of immune restraint.
doi:10.1007/s00109-014-1156-z
PMCID: PMC4083097  PMID: 24777637
E3 ligase; Foxp3; Treg; Ubiquitin; Deubiquitinase
5.  Control of Autophagy Maturation by Acid Sphingomyelinase in Mouse Coronary Arterial Smooth Muscle Cells: Protective Role in Atherosclerosis 
Recent studies have indicated a protective role of autophagy in regulating vascular smooth muscle cells homeostasis in atherogenesis, but the mechanisms controlling autophagy, particularly autophagy maturation, are poorly understood. Here, we investigated whether acid sphingomyelinase (ASM)-regulated lysosome function is involved in autophagy maturation in coronary arterial smooth muscle cells (CASMCs) in the pathogenesis of atherosclerosis. In coronary arterial wall of ASM-deficient (Smpd1−/−) mice on Western diet, there were high expression levels of both LC3B, a robust marker of autophagosomes (APs), and p62, a selective autophagy substrate, compared to those in wild type (Smpd1+/+) mice. By Western blotting and flow cytometry, atherogenic stimulation of Smpd1+/+ CASMCs with 7-ketocholesterol was found to significantly enhance LC3B expression and increase the content of both APs and autophagolysosomes (APLs). In Smpd1−/− CASMCs, such 7-ketocholesterol-induced increases in LC3B and p62 expression and APs were further augmented, but APLs formation was abolished. Analysis of fluorescence resonance energy transfer (FRET) between fluorescence-labeled LC3B and Lamp1 (lysosome marker) showed that 7-ketocholesterol markedly induced fusion of APs with lysosomes in Smpd1+/+ CASMCs, which was abolished in Smpd1−/− CASMCs. Moreover, 7-ketocholesterol-induced expression of cell dedifferentiation marker vimentin and proliferation was enhanced in Smpd1−/− CASMCs compared to those in Smpd1+/+ CASMCs. Lastly, overexpression of ASM further increased APLs formation in Smpd1+/+ CASMCs and restored APLs formation in Smpd1−/− CASMCs indicating that increased ASM expression is highly correlated with enhanced APLs formation. Taken together, our data suggest that the control of lysosome trafficking and fusion by ASM is essential to a normal autophagic flux in CASMCs, which implicates that the deficiency of ASM-mediated regulation of autophagy maturation may result in imbalance of arterial smooth muscle cell homeostasis and thus serve as an important atherogenic mechanism in coronary arteries.
doi:10.1007/s00109-014-1120-y
PMCID: PMC4211081  PMID: 24463558
Acid sphingomyelinase; autophagic flux; lysosome; coronary arterial smooth muscle cell; coronary artery; atherosclerosis
6.  EVOLUTIONARY FOUNDATIONS FOR MOLECULAR MEDICINE 
Evolution has long provided a foundation for population genetics, but many major advances in evolutionary biology from the 20th century are only now being applied in molecular medicine. They include the distinction between proximate and evolutionary explanations, kin selection, evolutionary models for cooperation, and new strategies for tracing phylogenies and identifying signals of selection. Recent advances in genomics are further transforming evolutionary biology and creating yet more opportunities for progress at the interface of evolution with genetics, medicine, and public health. This article reviews 15 evolutionary principles and their applications in molecular medicine in hopes that readers will use them and others to speed the development of evolutionary molecular medicine.
doi:10.1007/s00109-012-0889-9
PMCID: PMC4416654  PMID: 22544168
Evolution; biology; genetics; Darwinian medicine; evolutionary medicine
7.  Nilotinib-induced autophagic changes increase endogenous parkin level and ubiquitination, leading to amyloid clearance 
Alzheimer’s disease (AD) is a neurodegenerative disorder associated with amyloid accumulation and autophagic changes. Parkin is an E3 ubiquitin ligase involved in proteasomal and autophagic clearance. We previously demonstrated decreased parkin solubility and interaction with the key autophagy enzyme Beclin-1 in AD, but tyrosine kinase inhibition restored parkin-Beclin-1 interaction. In the current studies we determined the mechanisms of Nilotinib-induced parkin-Beclin-1 interaction, which leads to amyloid clearance. Nilotinib increased endogenous parkin levels and ubiquitination, which may enhance parkin recycling via the proteasome, leading to increased activity and interaction with Beclin-1. Parkin solubility was decreased and autophagy was altered in amyloid expressing mice, suggesting that amyloid stress affects parkin stability, leading to failure of protein clearance via the lysosome. Isolation of autophagic vacuoles revealed amyloid and parkin accumulation in autophagic compartments but Nilotinib decreased insoluble parkin levels and facilitated amyloid deposition into lysosomes in wild type, but not parkin−/− mice, further underscoring an essential role for endogenous parkin in amyloid clearance. These results suggest that Nilotinib boosts the autophagic machinery, leading to increased level of endogenous parkin that undergoes ubiquitination and interacts with Beclin-1 to facilitate amyloid clearance. These data suggest that Nilotinib-mediated autophagic changes may trigger parkin response via increased protein levels, providing a therapeutic strategy to reduce Aβ and Tau in AD.
doi:10.1007/s00109-013-1112-3
PMCID: PMC3975659  PMID: 24337465
Ubiquitination; parkin; autophagy; Tau; amyloid; Alzheimer’s
8.  Light induces NLRP3 inflammasome activation in retinal pigment epithelial cells via lipofuscin-mediated photooxidative damage 
Abstract
Photooxidative damage and chronic innate immune activation have been implicated in retinal pigment epithelium (RPE) dysfunction, a process that underlies blinding diseases such as age-related macular degeneration (AMD). To identify a potential molecular link between these mechanisms, we investigated whether lipofuscin-mediated phototoxicity activates the NLRP3 inflammasome in RPE cells in vitro. We found that blue light irradiation (dominant wavelength 448 nm, irradiance 0.8 mW/cm2, duration 6 h) of lipofuscin-loaded primary human RPE cells and ARPE-19 cells induced photooxidative damage, lysosomal membrane permeabilization (79.5 % of cells vs. 3.8 % in nonirradiated controls), and cytosolic leakage of lysosomal enzymes. This resulted in activation of the inflammasome with activation of caspase-1 and secretion of interleukin-1β (14.6 vs. 0.9 pg/ml in nonirradiated controls) and interleukin-18 (87.7 vs. 0.2 pg/ml in nonirradiated controls). Interleukin secretion was dependent on the activity of NLRP3, caspase-1, and lysosomal proteases cathepsin B and L. These results demonstrate that accumulation of lipofuscin-like material in vitro renders RPE cells susceptible to phototoxic destabilization of lysosomes, resulting in NLRP3 inflammasome activation and secretion of inflammatory cytokines. This new mechanism of inflammasome activation links photooxidative damage and innate immune activation in RPE pathology and may provide novel targets for therapeutic intervention in retinal diseases such as AMD.
Key message
• Visible light irradiation of lipofuscin-loaded RPE cells activates inflammasome.
• Inflammasome activation results from lysosomal permeabilization and enzyme leakage.
• Inflammasome activation induces secretion of inflammatory cytokines by RPE cells.
• Photooxidative damage by visible light as new mechanism of inflammasome activation.
• Novel link between hallmark pathogenetic features of retinal degenerative diseases.
doi:10.1007/s00109-015-1275-1
PMCID: PMC4510924  PMID: 25783493
Age-related macular degeneration; Retinal pigment epithelium; Interleukin-1β; Lysosomal membrane permeabilization; Lipid peroxidation
9.  A novel COCH mutation associated with autosomal dominant nonsyndromic hearing loss disrupts the structural stability of the vWFA2 domain 
Mutations in COCH have been associated with autosomal dominant nonsyndromic hearing loss (DFNA9) and are frequently accompanied by vestibular hypofunction. Here, we report identification of a novel missense mutation, p.F527C, located in the vWFA2 domain in members of a Korean family with late-onset and progressive hearing loss. To assess the molecular characteristics of this cochlin mutant, we constructed both wild-type and mutant cochlin constructs and transfected these into mammalian cell lines. Results of immunocytochemistry analysis demonstrated localization of the cochlin mutant in the ER/Golgi complex, whereas western blot analyses of cell lysates revealed that the mutant cochlin tends to form covalent complexes that are retained in the cell. Biochemical analyses of recombinant vWFA2 domain of cochlin carrying the p.F527C mutation revealed that the mutation increases propensity of the protein to form covalent disulfide-bonded dimers and affects the structural stability but not the collagen-affinity of the vWFA2 domain. We suggest that the instability of mutant cochlin is the major driving force for cochlin aggregation in the inner ear in DFNA9 patients carrying the COCH p.F527C mutation.
doi:10.1007/s00109-012-0911-2
PMCID: PMC4361775  PMID: 22610276
nonsyndromic hearing loss; DFNA9; cochlin; mutation; protein stability
10.  Metal-deficient SOD1 in amyotrophic lateral sclerosis 
Mutations to the ubiquitous antioxidant enzyme Cu/Zn superoxide dismutase (SOD1) were the first established genetic cause of the fatal, adult-onset neurodegenerative disease amyotrophic lateral sclerosis (ALS). It is widely accepted that these mutations do not cause ALS via a loss of antioxidant function, but elucidating the alternate toxic gain of function has proven to be elusive. Under physiological conditions, SOD1 binds one copper ion and one zinc ion per monomer to form a highly stable and functional homodimer, but there is now ample evidence to indicate aberrant persistence of SOD1 in an intermediate metal-deficient state may contribute to the protein’s involvement in ALS. This review briefly discusses some of the data to support a role for metal-deficient SOD1 in the development of ALS and some of the outcomes from drug development studies that have aimed to modify the symptoms of ALS by targeting the metal state of SOD1. The implications for the metal state of SOD1 in cases of sporadic ALS that do not involve mutant SOD1 are also discussed.
doi:10.1007/s00109-015-1273-3
PMCID: PMC4408375  PMID: 25754173
Amyotrophic lateral sclerosis (ALS); Motor neuron disease (MND); Copper (Cu); Zinc (Zn); Cu/Zn superoxide dismutase (SOD1); Protein misfolding; Diacetylbis(4-methylthiosemicarbazonato)copperII
11.  NLRP3 inflammasome activation is required for fibrosis development in NAFLD 
NLR inflammasomes, caspase 1 activation platforms critical for processing key pro-inflammatory cytokines, have been implicated in the development of nonalcoholic fatty liver disease (NAFLD). As the direct role of the NLRP3 inflammasome remains unclear, we tested effects of persistent NLRP3 activation as a contributor to NAFLD development and, in particular, as a modulator of progression from benign hepatic steatosis to steatohepatitis during diet-induced NAFLD. Gain of function tamoxifen-inducible Nlrp3 knock-in mice allowing for in vivo temporal control of NLRP3 activation and loss of function Nlrp3 knockout mice were placed on short-term choline-deficient amino acid-defined (CDAA) diet, to induce isolated hepatic steatosis or long-term CDAA exposure, to induce severe steatohepatitis and fibrosis, respectively. Expression of NLRP3 associated proteins was assessed in liver biopsies of a well-characterized group of patients with the full spectrum of NAFLD. Nlrp3−/− mice were protected from long-term feeding CDAA-induced hepatomegaly, liver injury, and infiltration of activated macrophages. More importantly, Nlrp3−/−mice showed marked protection from CDAA-induced liver fibrosis. After 4 weeks on CDAA diet, wild-type (WT) animals showed isolated hepatic steatosis while Nlrp3 knock-in mice showed severe liver inflammation, with increased infiltration of activated macrophages and early signs of liver fibrosis. In the liver samples of patients with NAFLD, inflammasome components were significantly increased in those patients with nonalcoholic steatohepatitis (NASH) when compared to those with non-NASH NAFLD with mRNA levels of pro-IL1 beta correlated to levels of COL1A1. Our study uncovers a crucial role for the NLRP3 inflammasome in the development of NAFLD. These findings may lead to novel therapeutic strategies aimed at halting the progression of hepatic steatosis to the more severe forms of this disease.
doi:10.1007/s00109-014-1170-1
PMCID: PMC4349416  PMID: 24861026
NLRP3; Inflammation; Liver fibrosis; NASH; Steatoheptatitis
12.  MitoTimer: A Novel Protein for Monitoring Mitochondrial Turnover 
Mitochondrial quality control refers to the coordinated cellular systems involved in maintaining a population of healthy mitochondria. In addition to mitochondrial protein chaperones (Hsp10, Hsp60 and others) and proteases (Lon, AAA proteases) needed for refolding or degrading individual proteins, mitochondrial integrity is maintained through regulation of protein import via the TOM/TIM complex and protein redistribution across the network via fusion and fission; and through mitophagy and biogenesis, key determinants of mitochondrial turnover. A growing number of studies point to the importance of mitochondrial dynamics (fusion/fission) and mitochondrial autophagy in the heart. Mitochondrial biogenesis must keep pace with mitophagy in order to maintain a stable number of mitochondria. In this review we will discuss the use of MitoTimer as a tool to monitor mitochondrial turnover.
doi:10.1007/s00109-014-1230-6
PMCID: PMC4333239  PMID: 25479961
13.  Complement evasion by Bordetella pertussis: implications for improving current vaccines 
Bordetella pertussis causes whooping cough or pertussis, a highly contagious disease of the respiratory tract. Despite high vaccination coverage, reported cases of pertussis are rising worldwide and it has become clear that the current vaccines must be improved. In addition to the well-known protective role of antibodies and T cells during B. pertussis infection, innate immune responses such as the complement system play an essential role in B. pertussis killing. In order to evade this complement activation and colonize the human host, B. pertussis expresses several molecules that inhibit complement activation. Interestingly, one of the known complement evasion proteins, autotransporter Vag8, is highly expressed in the recently emerged B. pertussis isolates. Here, we describe the current knowledge on how B. pertussis evades complement-mediated killing. In addition, we compare this to complement evasion strategies used by other bacterial species. Finally, we discuss the consequences of complement evasion by B. pertussis on adaptive immunity and how identification of the bacterial molecules and the mechanisms involved in complement evasion might help improve pertussis vaccines.
doi:10.1007/s00109-015-1259-1
PMCID: PMC4366546  PMID: 25686752
Complement; Bordetella pertussis; Innate immunity; Evasion; Vaccine
14.  Both GLS silencing and GLS2 overexpression synergize with oxidative stress against proliferation of glioma cells 
Mitochondrial glutaminase (GA) plays an essential role in cancer cell metabolism, contributing to biosynthesis, bioenergetics and redox balance. Humans contain several GA isozymes encoded by the GLS and GLS2 genes, but the specific roles of each in cancer metabolism are still unclear. In this study, glioma SFxL and LN229 cells with silenced isoenzyme glutaminase KGA (encoded by GLS) showed lower survival ratios and a reduced GSH-dependent antioxidant capacity. These GLS-silenced cells also demonstrated induction of apoptosis indicated by enhanced annexin V binding capacity and caspase 3 activity. GLS silencing was associated with decreased mitochondrial membrane potential (ΔΨm) (JC-1 dye test), indicating that apoptosis was mediated by mitochondrial dysfunction. Similar observations were made in T98 glioma cells overexpressing glutaminase isoenzyme GAB, encoded by GLS2, though some characteristics (GSH/GSSG ratio) were different in the differently treated cell lines. Thus, control of GA isoenzyme expression may prove to be a key tool to alter both metabolic and oxidative stress in cancer therapy. Interestingly, reactive oxygen species (ROS) generation by treatment with oxidizing agents: arsenic trioxide or hydrogen peroxide, synergizes with either KGA silencing or GAB overexpression to suppress malignant properties of glioma cells, including the reduction of cellular motility. Of note, negative modulation of GLS isoforms or GAB overexpression evoked lower c-myc and bcl-2 expression, as well as higher pro-apoptotic bid expression. Combination of modulation of GA expression and treatment with oxidizing agents may become a therapeutic strategy for intractable cancers and provides a multi-angle evaluation system for anti-glioma pre-clinical investigations.
doi:10.1007/s00109-013-1105-2
PMCID: PMC4327995  PMID: 24276018
Apoptosis; Cancer; Glioma; Glutaminase; Glutathione; ROS
15.  Cancer genomics: why rare is valuable 
Rare conditions are sometimes ignored in biomedical research because of difficulties in obtaining specimens and limited interest from fund raisers. However, the study of rare diseases such as unusual cancers has again and again led to breakthroughs in our understanding of more common diseases. It is therefore unsurprising that with the development and accessibility of next-generation sequencing, much has been learnt from studying cancers that are rare and in particular those with uniform biological and clinical behavior. Herein, we describe how shotgun sequencing of cancers such as granulosa cell tumor, endometrial stromal sarcoma, epithelioid hemangioendothelioma, ameloblastoma, small-cell carcinoma of the ovary, clear-cell carcinoma of the ovary, nonepithelial ovarian tumors, chondroblastoma, and giant cell tumor of the bone has led to rapidly translatable discoveries in diagnostics and tumor taxonomies, as well as providing insights into cancer biology.
doi:10.1007/s00109-015-1260-8
PMCID: PMC4366545  PMID: 25676695
Next-generation sequencing; Formefruste; Rare tumors; Genomics
16.  Mitochondrial biogenesis in the metabolic syndrome and cardiovascular disease 
The metabolic syndrome is a constellation of metabolic disorders including obesity, hypertension, and insulin resistance, components which are risk factors for the development of diabetes, hypertension, cardiovascular, and renal disease. Pathophysiological abnormalities that contribute to the development of the metabolic syndrome include impaired mitochondrial oxidative phosphorylation and mitochondrial biogenesis, dampened insulin metabolic signaling, endothelial dysfunction, and associated myocardial functional abnormalities. Recent evidence suggests that impaired myocardial mitochondrial biogenesis, fatty acid metabolism, and antioxidant defense mechanisms lead to diminished cardiac substrate flexibility, decreased cardiac energetic efficiency, and diastolic dysfunction. In addition, enhanced activation of the renin–angiotensin–aldosterone system and associated increases in oxidative stress can lead to mitochondrial apoptosis and degradation, altered bioenergetics, and accumulation of lipids in the heart. In addition to impairments in metabolic signaling and oxidative stress, genetic and environmental factors, aging, and hyperglycemia all contribute to reduced mitochondrial biogenesis and mitochondrial dysfunction. These mitochondrial abnormalities can predispose a metabolic cardiomyopathy characterized by diastolic dysfunction. Mitochondrial dysfunction and resulting lipid accumulation in skeletal muscle, liver, and pancreas also impede insulin metabolic signaling and glucose metabolism, ultimately leading to a further increase in mitochondrial dysfunction. Interventions to improve mitochondrial function have been shown to correct insulin metabolic signaling and other metabolic and cardiovascular abnormalities. This review explores mechanisms of mitochondrial dysfunction with a focus on impaired oxidative phosphorylation and mitochondrial biogenesis in the pathophysiology of metabolic heart disease.
doi:10.1007/s00109-010-0663-9
PMCID: PMC4319704  PMID: 20725711
Metabolic impairment; Oxidative phosphorylation; Mitochondrial biogenesis
17.  New therapeutics to modulate mitochondrial dynamics and mitophagy in cardiac diseases 
The processes that control the number and shape of the mitochondria (mitochondrial dynamics) and the removal of damaged mitochondria (mitophagy) have been the subject of intense research. Recent work indicates that these processes may contribute to the pathology associated with cardiac diseases. This review describes some of the key proteins that regulate these processes and their potential as therapeutic targets for cardiac diseases.
doi:10.1007/s00109-015-1256-4
PMCID: PMC4333238  PMID: 25652199
Cardiac disease; Mitochondria; Mitophagy; Autophagy; Fission; Fusion; Mitochondrial dynamics; Heart
18.  [No title available] 
PMCID: PMC3926703  PMID: 24297496
19.  [No title available] 
PMCID: PMC3946681  PMID: 24248265
20.  [No title available] 
PMCID: PMC3946714  PMID: 24068256
21.  Acid sphingomyelinase inhibition protects mice from lung edema and lethal Staphylococcus aureus sepsis 
Abstract
Pulmonary edema associated with increased vascular permeability is a severe complication of Staphylococcus aureus–induced sepsis and an important cause of human pathology and death. We investigated the role of the mammalian acid sphingomyelinase (Asm)/ceramide system in the development of lung edema caused by S. aureus. Our findings demonstrate that genetic deficiency or pharmacologic inhibition of Asm reduced lung edema in mice infected with S. aureus. The Asm/ceramide system triggered the formation of superoxide, resulting in degradation of tight junction proteins followed by lung edema. Treatment of infected mice with amitriptyline, a potent inhibitor of Asm, protected mice from lung edema caused by S. aureus, but did not reduce systemic bacterial numbers. In turn, treatment with antibiotics reduced bacterial numbers but did not protect mice from lung edema. In contrast, only the combination of antibiotics and amitriptyline inhibited both pulmonary edema and bacteremia protecting mice from lethal sepsis and lung dysfunction suggesting the combination of both drugs as novel treatment option for sepsis.
Key messages
Antibiotics are often insufficient to cure S. aureus–induced sepsis.S. aureus induces lung edema via the Asm/ceramide system.Genetic deficiency of Asm inhibits lung dysfunction upon infection with S. aureus.Pharmacologic inhibition of Asm reduces lung edema induced by S. aureus.Antibiotics plus amitriptyline protect mice from lung edema and lethal S. aureus sepsis.
Electronic supplementary material
The online version of this article (doi:10.1007/s00109-014-1246-y) contains supplementary material, which is available to authorized users.
doi:10.1007/s00109-014-1246-y
PMCID: PMC4432103  PMID: 25616357
Acid sphingomyelinase/ceramide system; Lung edema; Superoxide; Staphylococcus aureus
22.  Metastasis Suppressors in Breast Cancers: Mechanistic Insights and Clinical Potential 
doi:10.1007/s00109-013-1109-y
PMCID: PMC3923422  PMID: 24311119
metastasis suppressor; Nm23; RECK; BRMS1; KISS1; TIMP; E-cadherin; MKK4; KAI1; adhesion; invasion; MMP; intravasation; cohesion; motility; endothelium; angiogenesis; review
23.  Biological functions of ecto-enzymes in regulating extracellular adenosine levels in neoplastic and inflammatory disease states 
When present in the extracellular environment, the nucleoside adenosine protects cells and tissues from excessive inflammation and immune-mediated damage while promoting healing processes. This role has been highlighted experimentally using distinct disease models, including those of colitis, diabetes, asthma, sepsis, and ischemic injury. Adenosine also suppresses immune responses, as in the tumor microenvironment, assisting immune evasion while promoting angiogenesis. The mechanisms involved in adenosine signaling are addressed elsewhere in this issue. Here, the authors specifically address the generation of adenosine from extracellular nucleotides. This process is catalyzed by a series of plasma membrane ectonucleotidases, with the focus in this article on members of the CD39, CD73, and CD38 families and on their role in inflammatory and neoplastic hematological diseases. Pharmacological modulation of adenosine generation by drugs that either have or modulate ectonucleotidase function might be exploited to treat these diverse conditions.
doi:10.1007/s00109-012-0991-z
PMCID: PMC4266600  PMID: 23292173
Adenosine; Cancer; Colitis; Ectonucleotidase; Inflammation; Ischemic injury
24.  Mutation of EpCAM leads to intestinal barrier and ion transport dysfunction 
Abstract
Congenital tufting enteropathy (CTE) is a devastating diarrheal disease seen in infancy that is typically associated with villous changes and the appearance of epithelial tufts. We previously found mutations in epithelial cell adhesion molecule (EpCAM) to be causative in CTE. We developed a knock-down cell model of CTE through transfection of an EpCAM shRNA construct into T84 colonic epithelial cells to elucidate the in vitro role of EpCAM in barrier function and ion transport. Cells with EpCAM deficiency exhibited decreased electrical resistance, increased permeability, and decreased ion transport. Based on mutations in CTE patients, an in vivo mouse model was developed, with tamoxifen-inducible deletion of exon 4 in Epcam resulting in mutant protein with decreased expression. Tamoxifen treatment of EpcamΔ4/Δ4 mice resulted in pathological features of villous atrophy and epithelial tufts, similar to those in human CTE patients, within 4 days post induction. EpcamΔ4/Δ4 mice also showed decreased expression of tight junctional proteins, increased permeability, and decreased ion transport in the intestines. Taken together, these findings reveal mechanisms that may underlie disease in CTE.
Key messages
Knock-down EpCAM cell model of congenital tufting enteropathy was developed.In vivo inducible mouse model was developed resulting in mutant EpCAM protein.Cells with EpCAM deficiency demonstrated barrier and ion transport dysfunction.Tamoxifen-treated EpcamΔ4/Δ4 mice demonstrated pathological features.EpcamΔ4/Δ4 mice showed improper barrier function and ion transport.
Electronic supplementary material
The online version of this article (doi:10.1007/s00109-014-1239-x) contains supplementary material, which is available to authorized users.
doi:10.1007/s00109-014-1239-x
PMCID: PMC4408367  PMID: 25482158
Congenital tufting enteropathy; EpCAM; Ion transport; Barrier function
25.  MitoTimer: a novel protein for monitoring mitochondrial turnover in the heart 
Mitochondrial quality control refers to the coordinated cellular systems involved in maintaining a population of healthy mitochondria. In addition to mitochondrial protein chaperones (Hsp10, Hsp60, and others) and proteases (Lon, AAA proteases) needed for refolding or degrading individual proteins, mitochondrial integrity is maintained through the regulation of protein import via the TOM/TIM complex and protein redistribution across the network via fusion and fission and through mitophagy and biogenesis, key determinants of mitochondrial turnover. A growing number of studies point to the importance of mitochondrial dynamics (fusion/fission) and mitochondrial autophagy in the heart. Mitochondrial biogenesis must keep pace with mitophagy in order to maintain a stable number of mitochondria. In this review, we will discuss the use of MitoTimer as a tool to monitor mitochondrial turnover.
doi:10.1007/s00109-014-1230-6
PMCID: PMC4333239  PMID: 25479961
MitoTimer; Mitochondrial biogenesis; Mitophagy; Fluorescence microscopy

Results 1-25 (257)