Explore the vast range of titles available.

ABPP combined with quantitative MS enabled identification of specific on- and off-targets of covalent kinase inhibitors. Modifications to inhibitors that alter specificity beyond a defined window can promote kinase-independent toxicity. Kinases are principal components of signal transduction pathways and the focus of intense basic and drug discovery research. Irreversible inhibitors that covalently modify non-catalytic cysteines in kinase active sites have emerged as valuable probes and approved drugs. Many protein classes, however, have functional cysteines, and therefore understanding the proteome-wide selectivity of covalent kinase inhibitors is imperative. Here, we accomplish this objective using activity-based protein profiling coupled with quantitative MS to globally map the targets, both specific and nonspecific, of covalent kinase inhibitors in human cells. Many of the specific off-targets represent nonkinase proteins that, notably, have conserved active site cysteines. We define windows of selectivity for covalent kinase inhibitors and show that, when these windows are exceeded, rampant proteome-wide reactivity and kinase target–independent cell death conjointly occur. Our findings, taken together, provide an experimental road map to illuminate opportunities and surmount challenges for the development of covalent kinase inhibitors.

Antibody-based therapeutics have experienced a rapid growth in recent years and are now utilized in various modalities spanning from conventional antibodies, antibody-drug conjugates, bispecific antibodies to chimeric antigen receptor (CAR) T cells. Many next generation antibody therapeutics achieve enhanced potency but often increase the risk of adverse events. Antibody scaffolds capable of exhibiting inducible affinities could reduce the risk of adverse events by enabling a transient suspension of antibody activity. To demonstrate this, we develop conditionally activated, single-module CARs, in which tumor antigen recognition is directly modulated by an FDA-approved small molecule drug. The resulting CAR T cells demonstrate specific cytotoxicity of tumor cells comparable to that of traditional CARs, but the cytotoxicity is reversibly attenuated by the addition of the small molecule. The exogenous control of conditional CAR T cell activity allows continual modulation of therapeutic activity to improve the safety profile of CAR T cells across all disease indications. Many next-generation antibody therapeutics have enhanced potency but the risk of adverse events. Here the authors develop a conditionally activated, single-module CAR.

Key Points Obesity and type 2 diabetes have become epidemic medical problems and will cause significant morbidity and mortality if new therapeutic agents cannot be brought forward. Protein tyrosine phosphatase 1B (PTP1B) has been shown to be involved in the negative regulation of both insulin and leptin action at the in vitro, ex vivo and in vivo levels. A growing body of human genetic data also support the hypothesis that PTP1B has an important role in insulin signalling and possibly in obesity in humans. PTP1B has been implicated in diabetes and obesity in knockout-mouse studies on a normal background. When crossed onto ob/ob mice, improvement in blood glucose and obesity is seen. When Ptp1b antisense oligonucleotides are administered in vivo to ob/ob mice, normalized blood glucose levels are obtained. Insulin sensitivity is improved at the cellular level. PTP1B is tractable to structure-based drug design, and the crystal structure is well known. Recently, the crystal structure of the closely related T-cell protein tyrosine phosphatase (TCPTP) has become available, raising the possibility for designing selective inhibitors. Potent ( K i values in the nM range) inhibitors of PTP1B are known, and oxamic acids are a common structural motif found in these compounds. The high polar surface area (PSA) and charge of this functionality make further optimization difficult, and an intense search to uncover phosphotyrosine mimetics with improved physico-chemical properties is in progress. Pro-drugs of the oxamic acids and replacement of the acid functionality with isosteric functional groups might provide compounds with improved physico-chemical properties. Lipophilic compounds have been reported — one of the two reported structures seems to be a competitive inhibitor of PTP1B and the other does not. This highlights the need for a clear understanding of the mechanism of inhibition and an understanding of the kinetics of compounds. Increased incidence of type 2 diabetes mellitus and obesity has elevated the medical need for new agents to treat these disease states. Resistance to the hormones insulin and leptin are hallmarks of both type 2 diabetes and obesity. Drugs that can ameliorate this resistance should be effective in treating type 2 diabetes and possibly obesity. Protein tyrosine phosphatase 1B (PTP1B) is thought to function as a negative regulator of insulin and leptin signal transduction. This article reviews PTP1B as a novel target for type 2 diabetes, and looks at the challenges in developing small-molecule inhibitors of this phosphatase.

Kinases are principal components of signal transduction pathways and the focus of intense basic and drug discovery research. Irreversible inhibitors that covalently modify non-catalytic cysteines in kinase active-sites have emerged as valuable probes and approved drugs. Many protein classes, however, possess functional cysteines and therefore understanding the proteome-wide selectivity of covalent kinase inhibitors is imperative. Here, we accomplish this objective using activity-based protein profiling coupled with quantitative mass spectrometry to globally map the targets, both specific and non-specific, of covalent kinase inhibitors in human cells. Many of the specific off-targets represent non-kinase proteins that, interestingly, possess conserved, active-site cysteines. We define windows of selectivity for covalent kinase inhibitors and show that, when these windows are exceeded, rampant proteome-wide reactivity and kinase target-independent cell death conjointly occur. Our findings, taken together, provide an experimental roadmap to illuminate opportunities and surmount challenges for the development of covalent kinase inhibitors.

A dominant histopathological feature in neuromuscular diseases, including amyotrophic lateral sclerosis and inclusion body myopathy, is cytoplasmic aggregation of the RNA-binding protein TDP-43. Although rare mutations in TARDBP —the gene that encodes TDP-43—that lead to protein misfolding often cause protein aggregation, most patients do not have any mutations in TARDBP . Therefore, aggregates of wild-type TDP-43 arise in most patients by an unknown mechanism. Here we show that TDP-43 is an essential protein for normal skeletal muscle formation that unexpectedly forms cytoplasmic, amyloid-like oligomeric assemblies, which we call myo-granules, during regeneration of skeletal muscle in mice and humans. Myo-granules bind to mRNAs that encode sarcomeric proteins and are cleared as myofibres mature. Although myo-granules occur during normal skeletal-muscle regeneration, myo-granules can seed TDP-43 amyloid fibrils in vitro and are increased in a mouse model of inclusion body myopathy. Therefore, increased assembly or decreased clearance of functionally normal myo-granules could be the source of cytoplasmic TDP-43 aggregates that commonly occur in neuromuscular disease. Cytoplasmic, amyloid-like oligomeric assemblies that contain TDP-43 are increased in damaged tissues with elevated regeneration, thereby enhancing the possibility of amyloid fibre formation and/or aggregation of TDP-43 in disease.

Nocturia is associated with poor sleep quality; however, little is known about the relationship between nocturia and sleep quality across different workforce-relevant age groups of adults. This has implications for developing new treatment strategies that are well tolerated across populations. We conducted a cross-sectional study involving merged data from the 2005–2006 and 2007–2008 waves of the National Health and Nutrition Examination Survey. Participants responded to validated questions on nocturia frequency and sleep from the Functional Outcomes of Sleep Questionnaire General Productivity subscale (FOSQ-gp, range 1–4). Analyses included multivariable linear regression with stratification by gender to examine associations between nocturia frequency (higher worse) and the FOSQ-gp scores (lower scores indicating worse daytime function related to sleep disturbance). Of 10,512 adults aged ≥20 years who completed the survey, 9148 (87%) had complete nocturia and FOSQ-gp data. The population age-adjusted prevalence of nocturia at least twice nightly was 21.1% among men and 26.6% among women (P < 0.001), and nocturia increased with age (P < 0.001). Compared with those with no or 1 episode of nocturia, those with nocturia at least twice nightly reported lower mean FOSQ-gp scores (3.65; 95% CI, 3.61–3.69 vs 3.19; 95% CI, 3.09–3.31 for men and 3.52; 95% CI, 3.48–3.56 vs 3.09; 95% CI, 3.02–3.16 for women). Older adults (aged >65 years) with greater nocturia frequency reported worse FOSQ-gp scores compared with younger adults with similar nocturia frequency (P < 0.001 among men and women). In a population-based sample of community-dwelling men and women, the association between nocturia and worsened functional outcomes of sleep was greater among adults older than 65 years—a group more vulnerable to drug side effects, and in whom nocturia is typically multifactorial. Additionally, these analyses found that the association between nocturia and functional outcomes of sleep is stronger with increasing age among men. Effective treatment strategies that are well tolerated by older adults, such as multicomponent treatments that simultaneously address the combined effects of lower urinary tract and sleep dysfunction, are needed.

IntroductionVaping and vape shops pose risk for COVID-19 and its transmission.ObjectivesWe examined vape shop non-compliance with state-ordered business closures during COVID-19, changes in their marketing and experiences among consumers.MethodsAs part of a longitudinal study of vape retail in six metropolitan statistical areas (MSAs; Atlanta, Boston, Minneapolis, Oklahoma City, San Diego and Seattle), we conducted: (1) legal research to determine whether statewide COVID-19 orders required vape shops to close; (2) phone-based and web-based surveillance to assess vape shop activity in March–June 2020 during shelter-in-place periods; and (3) a concurrent online survey of e-cigarette users about their experiences with vape retail.ResultsNon-essential business closure varied in timing/duration across states and applied to vape shops in California, Massachusetts, Minnesota, Oklahoma (for a brief period) and Washington (Georgia’s orders were ambiguous). Surveillance analysis focused on the five MSAs in these states. Of 156 vape shops, 53.2% were open as usual, 11.5% permanently closed and 3.8% temporarily closed; 31.4% offered pick-up/delivery services. Among survey respondents (n=354, M age =23.9±4.6; 46.9% male, 71.8% white, 13.0% Hispanic), 27.4% worried their vape shop would close/go out of business during COVID-19; 7.3% said their vape shop did so. Few noticed increases in vape product delivery options (7.3%), discounts/price promotions (9.9%) and/or prices (9.3%). While 20.3% stockpiled vape products, 20.3% tried to reduce use and 15.8% tried to quit.ConclusionsMany vape shops were non-compliant with state COVID-19 orders. E-cigarette users were as likely to stockpile vape products as to attempt to reduce or quit using e-cigarettes.

Human cytomegalovirus (HCMV) causes substantial disease in transplant patients and harms the development of the nervous system in babies infected in utero. Thus, there is a major focus on developing safe and effective HCMV vaccines. Evidence has been presented that a major target of neutralizing antibodies (NAbs) is the HCMV pentamer glycoprotein gH/gL/UL128-131. In some studies, most of the NAbs in animal or human sera were found to recognize the pentamer,which mediates HCMV entry into endothelial and epithelial cells. It was also reported that pentamer-specific antibodies correlate with protection against transmission from mothers to babies. One problem with the studies on pentamer-specific NAbs to date has been that the studies did not compare the pentamer to the other major form of gH/gL, the gH/gL/gO trimer, which is essential for entry into all cell types. Here, we demonstrate that both trimer and pentamer NAbs are frequently found in human transplant patients’ and pregnant mothers’ sera. Depletion of human sera with trimer caused reductions in NAbs similar to that observed following depletion with the pentamer. The trimer- and pentamer-specific antibodies acted in a synergistic fashion to neutralize HCMV and also to prevent virus cell-to-cell spread. Importantly, there was no correlation between the titers of trimer- and pentamer-specific NAbs and transmission of HCMV from mothers to babies. Therefore, both the trimer and pentamer are important targets of NAbs. Nevertheless, these antibodies do not protect against transmission of HCMV from mothers to babies.

Abstract Purpose Nocturia and sleep problems are common in older adults. We developed and tested a novel intervention, multicomponent behavioral treatment and exercise therapy (M-BET), that may reduce nocturia and improve sleep in men. We compared reductions in nocturia and improvement in sleep in men with M-BET versus an active drug comparator (α-blocker) used alone or in combination (M-BET + α-blocker) Methods This randomized, controlled trial was conducted in the ambulatory setting in 2 US Department of Veterans Affairs medical centers in men at least 40 years of age with nocturia (defined as ≥2 nightly episodes). Participants were randomized to receive either M-BET, including pelvic floor muscle training, urge-suppression techniques, delayed voiding, fluid management, sleep hygiene, and peripheral edema management; an active comparator of known efficacy (the α-blocker tamsulosin, one 0.4-mg tablet nightly); or both therapies combined. Participants received interventions over 12 weeks. Outcomes were assessed via voiding diaries, wrist actigraphy, and validated questionnaires. The primary outcome was change in diary-recorded nocturia, assessed using ANCOVA for the between-group changes and paired t tests for within-group changes. Findings A total of 72 men with a mean age of 65.8 years participated. At 12 weeks, mean diary-recorded nocturia changed with M-BET by –1.39 episodes/night ( P < 0.001), with α-blocker therapy by –0.59 episodes/night ( P < 0.01), and with combination therapy by –1.03 episodes/night ( P < 0.01). Reductions were not statistically different across treatment groups ( P = 0.41). M-BET also showed statistically significant improvements in sleep quality, bother from nocturia, and nocturia-specific quality of life. All treatment groups indicated global satisfaction with treatment. Implications Behavioral therapy in men, alone or combined with α-blocker therapy, consistently showed large and statistically significant nocturia reductions and favorable effects on sleep and quality of life. Based on these findings, behavioral therapy, while not statistically superior to α-blocker therapy, may provide a meaningful treatment option for men with nocturia. Future research should include the development of behavioral treatment and exercise therapy interventions that could be more easily deployed. ClinicalTrials.gov identifier: NCT00824200.

The goal of this study was to determine if the US adult population with nocturia (waking from sleep at night to void) can easily take medications (desmopressin acetate) approved by the US Food and Drug Administration for nocturia. The study examined: (1) the prevalence of comorbid conditions, laboratory abnormalities, and concomitant medications that increase risk of desmopressin use; and (2) whether these factors are associated with age or nocturia frequency. Using a cross-sectional analysis of four US National Health and Nutrition Examination Survey (NHANES) waves (2005–2012), a total of 4111 participants aged ≥50 years who reported ≥2 nightly episodes of nocturia were identified. The main outcome was frequency of contraindications and drug interactions as described in US Food and Drug Administration–approved prescribing information. These prescribing concerns were matched to examination findings, medical conditions, concomitant medications, and laboratory results of NHANES participants. The associations between prescribing concerns and nocturia severity and age groups were examined. The mean participant age was 65.7 years (95% CI, 65.3–66.1), and 45.5% were male. Desmopressin prescribing concerns were present in 80.5% (95% CI, 78.0–82.9) of those ≥50 years of age with nocturia; 50.0% (95% CI, 47.0–53.0) had contraindications, and 41.6% (95% CI, 39.3–44.0) took a concomitant drug that could increase risk of low serum sodium. Desmopressin contraindications were higher with older age (P < 0.001) and present in 73.2% (95% CI, 69.3–77.1) of those ≥80 years of age. Using NHANES data, this study showed that older US adults with nocturia have a high prevalence of medical conditions, concomitant medications, and baseline laboratory abnormalities that likely increase the risk of potentially severe adverse side effects from desmopressin use. A medication designed and approved for a clinical symptom that is most common in older adults could not be taken by most of the older adults with the symptom. •US FDA-approved desmopressin drugs for nocturia have boxed warning for hyponatremia risk.•Older adults are the most likely population to have nocturia, more frequent nocturia.•Most US older adults with nocturia have comorbidities, take other meds that increase risk.•The older the age group, the higher the contraindications, contraindications + interacting meds.•Desmopressin trial participants were not representative of the US older adult population with nocturia.