miRNA analysis reveals novel dysregulated pathways in amyotrophic lateral sclerosis.

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease. Its complex pathogenesis and phenotypic heterogeneity hinder therapeutic development and early diagnosis. Altered RNA metabolism is a recurrent pathophysiologic theme, including distinct microRNA (miRNA) profiles in ALS tissues. We profiled miRNAs in accessible biosamples, including skin fibroblasts and whole blood and compared them in age- and sex-matched healthy controls versus ALS participants with and without repeat expansions to chromosome 9 open reading frame 72 (C9orf72; C9-ALS and nonC9-ALS), the most frequent ALS mutation. We identified unique and shared profiles of differential miRNA (DmiRNA) levels in each C9-ALS and nonC9-ALS tissues versus controls. Fibroblast DmiRNAs were validated by quantitative real-time PCR and their target mRNAs by 5-bromouridine and 5-bromouridine-chase sequencing. We also performed pathway analysis to infer biological meaning, revealing anticipated, tissue-specific pathways and pathways previously linked to ALS, as well as novel pathways that could inform future research directions. Overall, we report a comprehensive study of a miRNA profile dataset from C9-ALS and nonC9-ALS participants across two accessible biosamples, providing evidence of dysregulated miRNAs in ALS and possible targets of interest. Distinct miRNA patterns in accessible tissues may also be leveraged to distinguish ALS participants from healthy controls for earlier diagnosis. Future directions may look at potential correlations of miRNA profiles with clinical parameters.

Gut microbiome correlates with plasma lipids in amyotrophic lateral sclerosis.

Amyotrophic lateral sclerosis (ALS) is a complex, fatal neurodegenerative disease. Disease pathophysiology is incompletely understood but evidence suggests gut dysbiosis occurs in ALS, linked to impaired gastrointestinal integrity, immune system dysregulation and altered metabolism. Gut microbiome and plasma metabolome have been separately investigated in ALS, but little is known about gut microbe-plasma metabolite correlations, which could identify robust disease biomarkers and potentially shed mechanistic insight. Here, gut microbiome changes were longitudinally profiled in ALS and correlated to plasma metabolome. Gut microbial structure at the phylum level differed in ALS versus control participants, with differential abundance of several distinct genera. Unsupervised clustering of microbe and metabolite levels identified modules, which differed significantly in ALS versus control participants. Network analysis found several prominent amplicon sequence variants strongly linked to a group of metabolites, primarily lipids. Similarly, identifying the features that contributed most to case versus control separation pinpointed several bacteria correlated to metabolites, predominantly lipids. Mendelian randomization indicated possible causality from specific lipids related to fatty acid and acylcarnitine metabolism. Overall, the results suggest ALS cases and controls differ in their gut microbiome, which correlates with plasma metabolites, particularly lipids, through specific genera. These findings have the potential to identify robust disease biomarkers and shed mechanistic insight into ALS.

Dysregulated biodynamics in metabolic attractor systems precede the emergence of amyotrophic lateral sclerosis.

Evolutionarily conserved mechanisms maintain homeostasis of essential elements, and are believed to be highly time-variant. However, current approaches measure elemental biomarkers at a few discrete time-points, ignoring complex higher-order dynamical features. To study dynamical properties of elemental homeostasis, we apply laser ablation inductively-coupled plasma mass spectrometry (LA-ICP-MS) to tooth samples to generate 500 temporally sequential measurements of elemental concentrations from birth to 10 years. We applied dynamical system and Information Theory-based analyses to reveal the longest-known attractor system in mammalian biology underlying the metabolism of nutrient elements, and identify distinct and consistent transitions between stable and unstable states throughout development. Extending these dynamical features to disease prediction, we find that attractor topography of nutrient metabolism is altered in amyotrophic lateral sclerosis (ALS), as early as childhood, suggesting these pathways are involved in disease risk. Mechanistic analysis was undertaken in a transgenic mouse model of ALS, where we find similar marked disruptions in elemental attractor systems as in humans. Our results demonstrate the application of a phenomological analysis of dynamical systems underlying elemental metabolism, and emphasize the utility of these measures in characterizing risk of disease.

Cutting Edge: Transcriptional Profiling Reveals Multifunctional and Cytotoxic Antiviral Responses of Zika Virus-Specific CD8+ T Cells.

Zika virus (ZIKV) constitutes an increasing public health problem. Previous studies have shown that CD8+ T cells play an important role in ZIKV-specific protective immunity. We have previously defined antigenic targets of the ZIKV-specific CD8+ T cell response in humans. In this study, we characterized the quality and phenotypes of these responses by a combined use of flow cytometry and transcriptomic methods, using PBMCs from donors deriving from different geographical locations collected in the convalescent phase of infection. We show that ZIKV-specific CD8+ T cells are characterized by a polyfunctional IFN-γ signature with upregulation of TNF-α, TNF receptors, and related activation markers, such as CD69, as well as a cytotoxic signature characterized by strong upregulation of GZMB and CRTAM. The signature is stable and not influenced by previous dengue virus exposure, geographical location, or time of sample collection postinfection. To our knowledge, this work elucidates the first in-depth characterization of human CD8+ T cells responding to ZIKV infection.

Differences in substance use, psychiatric disorders and social factors between Mexican adolescents and young adults.

BACKGROUND AND OBJECTIVES: Substance use disorders (SUDs) have high comorbidities with psychiatric disorders. Childhood and adolescence are particularly vulnerable developmental periods for the onset of SUDs. The objective of this study was to explore the differences, if any, between Mexican adolescents and young adults with respect to the prevalences of groups of psychiatric disorders, the types of substances used and the social factors involved. METHODS: This cross-sectional study included 781 patients evaluated at the Youth Integration Center in Mexico City. The diagnostic criteria for SUDs and psychiatric disorders were defined according to the DSM-IV and ICD-10. Associations between SUDs and psychiatric disorders were evaluated via multivariate analysis using logistic regression models. RESULTS: The adolescents were more frequently substance abusers, whereas the adults had legal problems more often than the adolescents. We showed that adolescents using inhalants or cocaine were 1.62 more likely to have attention deficit hyperactivity disorder (ADHD). Moreover, adults using inhalants were 3.33 times more likely to meet the criteria for a psychotic disorder. DISCUSSION AND CONCLUSIONS: We found that adolescents diagnosed with ADHD were more likely to have problems with use or abuse of or dependence on inhalants, and an elevated prevalence of parental SUDs was found in both the adolescent and adult groups. SCIENTIFIC SIGNIFICANCE: Our findings indicate that earlier diagnosis and intervention are necessary in adolescents with ADHD and/or parental SUDs to prevent more advanced psychiatric diseases and adverse social consequences during adulthood. (Am J Addict 2018;XX:1-7).

Expression of microRNAs in human post-mortem amyotrophic lateral sclerosis spinal cords provides insight into disease mechanisms.

Amyotrophic lateral sclerosis is a late-onset and terminal neurodegenerative disease. The majority of cases are sporadic with unknown causes and only a small number of cases are genetically linked. Recent evidence suggests that post-transcriptional regulation and epigenetic mechanisms, such as microRNAs, underlie the onset and progression of neurodegenerative disorders; therefore, altered microRNA expression may result in the dysregulation of key genes and biological pathways that contribute to the development of sporadic amyotrophic lateral sclerosis. Using systems biology analyses on postmortem human spinal cord tissue, we identified dysregulated mature microRNAs and their potential targets previously implicated in functional process and pathways associated with the pathogenesis of ALS. Furthermore, we report a global reduction of mature microRNAs, alterations in microRNA processing, and support for a role of the nucleotide binding protein, TAR DNA binding protein 43, in regulating sporadic amyotrophic lateral sclerosis-associated microRNAs, thereby offering a potential underlying mechanism for sporadic amyotrophic lateral sclerosis.

Insulin Resistance Prevents AMPK-induced Tau Dephosphorylation through Akt-mediated Increase in AMPKSer-485 Phosphorylation.

Metabolic syndrome (MetS) is a cluster of cardiovascular risk factors including obesity, diabetes, and dyslipidemia, and insulin resistance (IR) is the central feature of MetS. Recent studies suggest that MetS is a risk factor for Alzheimer disease (AD). AMP-activated kinase (AMPK) is an evolutionarily conserved fuel-sensing enzyme and a key player in regulating energy metabolism. In this report, we examined the role of IR on the regulation of AMPK phosphorylation and AMPK-mediated Tau phosphorylation. We found that AMPK(Ser-485), but not AMPK(Thr-172), phosphorylation is increased in the cortex of db/db and high fat diet-fed obese mice, two mouse models of IR. In vitro, treatment of human cortical stem cell line (HK-5320) and primary mouse embryonic cortical neurons with the AMPK activator, 5-aminoimidazole-4-carboxamide 1-ß-D-ribofuranoside (AICAR), induced AMPK phosphorylation at both Thr-172 and Ser-485. AMPK activation also triggered Tau dephosphorylation. When IR was mimicked in vitro by chronically treating the cells with insulin, AICAR specifically induced AMPK(Ser-485), but not AMPK(Thr-172), hyperphosphorylation whereas AICAR-induced Tau dephosphorylation was inhibited. IR also resulted in the overactivation of Akt by AICAR treatment; however, preventing Akt overactivation during IR prevented AMPK(Ser-485) hyperphosphorylation and restored AMPK-mediated Tau dephosphorylation. Transfection of AMPK(S485A) mutant caused similar results. Therefore, our results suggest the following mechanism for the adverse effect of IR on AD pathology: IR → chronic overactivation of Akt → AMPK(Ser-485) hyperphosphorylation → inhibition of AMPK-mediated Tau dephosphorylation. Together, our results show for the first time a possible contribution of IR-induced AMPK(Ser-485) phosphorylation to the increased risk of AD in obesity and diabetes.

Improving credibility and transparency of conservation impact evaluations through the partial identification approach.

The fundamental challenge of evaluating the impact of conservation interventions is that researchers must estimate the difference between the outcome after an intervention occurred and what the outcome would have been without it (counterfactual). Because the counterfactual is unobservable, researchers must make an untestable assumption that some units (e.g., organisms or sites) that were not exposed to the intervention can be used as a surrogate for the counterfactual (control). The conventional approach is to make a point estimate (i.e., single number along with a confidence interval) of impact, using, for example, regression. Point estimates provide powerful conclusions, but in nonexperimental contexts they depend on strong assumptions about the counterfactual that often lack transparency and credibility. An alternative approach, called partial identification (PI), is to first estimate what the counterfactual bounds would be if the weakest possible assumptions were made. Then, one narrows the bounds by using stronger but credible assumptions based on an understanding of why units were selected for the intervention and how they might respond to it. We applied this approach and compared it with conventional approaches by estimating the impact of a conservation program that removed invasive trees in part of the Cape Floristic Region. Even when we used our largest PI impact estimate, the program's control costs were 1.4 times higher than previously estimated. PI holds promise for applications in conservation science because it encourages researchers to better understand and account for treatment selection biases; can offer insights into the plausibility of conventional point-estimate approaches; could reduce the problem of advocacy in science; might be easier for stakeholders to agree on a bounded estimate than a point estimate where impacts are contentious; and requires only basic arithmetic skills.

Ecto-nucleotidases activities in the contents of ovarian endometriomas: potential biomarkers of endometriosis.

Endometriosis, defined as the growth of endometrial tissue outside the uterus, is a common gynecologic condition affecting millions of women worldwide. It is an inflammatory, estrogen-dependent complex disorder, with broad symptomatic variability, pelvic pain, and infertility being the main characteristics. Ovarian endometriomas are frequently developed in women with endometriosis. Late diagnosis is one of the main problems of endometriosis; thus, it is important to identify biomarkers for early diagnosis. The aim of the present work is to evaluate the ecto-nucleotidases activities in the contents of endometriomas. These enzymes, through the regulation of extracellular ATP and adenosine levels, are key enzymes in inflammatory processes, and their expression has been previously characterized in human endometrium. To achieve our objective, the echo-guided aspirated fluids of endometriomas were analyzed by evaluating the ecto-nucleotidases activities and compared with simple cysts. Our results show that enzyme activities are quantifiable in the ovarian cysts aspirates and that endometriomas show significantly higher ecto-nucleotidases activities than simple cysts (5.5-fold increase for ATPase and 20-fold for ADPase), thus being possible candidates for new endometriosis biomarkers. Moreover, we demonstrate the presence of ecto-nucleotidases bearing exosomes in these fluids. These results add up to the knowledge of the physiopathologic mechanisms underlying endometriosis and, open up a promising new field of study.

Refractory pseudoseptic arthritis in Behçet's disease successfully treated with infliximab: a case report and literature review.

Arthritis associated with Behçet's disease is typically nonerosive and nondeforming, and most patients respond to colchicine treatment. However, destructive arthritis and refractory arthritis have also been reported on occasion. Elevated white blood cell counts may occur in synovial fluid in BD, but pseudoseptic arthritis is a very rare event in this disease. We report a patient with BD and rapidly progressive deforming pseudoseptic arthritis refractory to colchicine, corticosteroids, and methotrexate, who entered remission after infliximab treatment.

Certified and uncertified logging concessions compared in Gabon: changes in stand structure, tree species, and biomass.

Forest management certification is assumed to promote sustainable forest management, but there is little field-based evidence to support this claim. To help fill this gap, we compared a Forest Stewardship Council (FSC)-certified with an adjacent uncertified, conventionally logged concession (CL) in Gabon on the basis of logging damage, above-ground biomass (AGB), and tree species diversity and composition. Before logging, we marked, mapped, and measured all trees >10 cm dbh in 20 and twelve 1-ha permanent plots in the FSC and CL areas, respectively. Soil and tree damage due to felling, skidding, and road-related activities was then assessed 2-3 months after the 508 ha FSC study area and the 200 ha CL study area were selectively logged at respective intensities of 5.7 m(3)/ha (0.39 trees/ha) and 11.4 m(3)/ha (0.76 trees/ha). For each tree felled, averages of 9.1 and 20.9 other trees were damaged in the FSC and CL plots, respectively; when expressed as the impacts per timber volume extracted, the values did not differ between the two treatments. Skid trails covered 2.9 % more of the CL surface, but skid trail length per unit timber volume extracted was not greater. Logging roads were wider in the CL than FSC site and disturbed 4.7 % more of the surface. Overall, logging caused declines in AGB of 7.1 and 13.4 % at the FSC and CL sites, respectively. Changes in tree species composition were small but greater for the CL site. Based on these findings and in light of the pseudoreplicated study design with less-than perfect counterfactual, we cautiously conclude that certification yields environmental benefits even after accounting for differences in logging intensities.

The community capacity curve applied to reforestation: a framework to support success.

Community involvement is critical for the success of many interventions designed to promote reforestation. To secure this involvement, it helps to recognize that communities are heterogenous both within and among themselves and possess diverse mixes of livelihood assets required to implement reforestation. We explore the relationship between livelihood assets and reforestation success and outline a conceptual model that we call the community capacity curve (CCC) applied to reforestation. We argue that the shape of the CCC is sigmoidal. Importantly, communities at the lower end of the CCC have limited capacity to implement reforestation projects without substantial and ongoing capacity building and other sorts of support, including through livelihood projects that improve food security and provide cash benefits. Communities at the higher part of the CCC have greater capacity to implement reforestation projects, especially projects focused on biodiversity and environmental services. The CCC can help design, implement, monitor and assess reforestation projects, select appropriate livelihood activities and types of reforestation, select communities suited to a reforestation project, guide implementation and understand projects' successes and failure. The CCC also provides a framework to engage with policy makers and funding bodies to explore the types of support for communities to reforest successfully. This article is part of the theme issue 'Understanding forest landscape restoration: reinforcing scientific foundations for the UN Decade on Ecosystem Restoration'.

Attitudes Toward Lesbians, Gay Men, and Their Rights in a Sample of Ecuadorian Cisgender Men and Women.

Since 1997, Ecuador has undergone a series of changes to ensure family rights to sexual minorities. However, there is still limited research regarding attitudes toward them. This study focused on the attitudes toward lesbians (L), gay men (G), and their rights. A sample of 318 cisgender Ecuadorians who responded to an online survey was recruited. Analyses indicated that men, heterosexuals, who practice their religion, attend more frequently to religious services, and identify as conservative showed higher levels of prejudice against LG as well as less support toward their rights. Further, participants who did not have LG acquaintances, friends, family members, and those who did not know any LG parented family showed less support toward these populations. Multiple regression analyses indicated that believing that a person's sexual orientation is learned significantly predicted the attitudes measured in our study. Implications of these findings to help reduce prejudice against LG individuals are discussed.

Gut microbiota in a mouse model of obesity and peripheral neuropathy associated with plasma and nerve lipidomics and nerve transcriptomics.

BACKGROUND: Peripheral neuropathy (PN) is a common complication in obesity, prediabetes, and type 2 diabetes, though its pathogenesis remains incompletely understood. In a murine high-fat diet (HFD) obesity model of PN, dietary reversal (HFD-R) to a low-fat standard diet (SD) restores nerve function and the nerve lipidome to normal. As the gut microbiome represents a potential link between dietary fat intake and nerve health, the current study assessed shifts in microbiome community structure by 16S rRNA profiling during the paradigm of dietary reversal (HFD-R) in various gut niches. Dietary fat content (HFD versus SD) was also correlated to gut flora and metabolic and PN phenotypes. Finally, PN-associated microbial taxa that correlated with the plasma and sciatic nerve lipidome and nerve transcriptome were used to identify lipid species and genes intimately related to PN phenotypes. RESULTS: Microbiome structure was altered in HFD relative to SD but rapidly reversed with HFD-R. Specific taxa variants correlating positively with metabolic health associated inversely with PN, while specific taxa negatively linked to metabolic health positively associated with PN. In HFD, PN-associated taxa variants, including Lactobacillus, Lachnoclostridium, and Anaerotruncus, also positively correlated with several lipid species, especially elevated plasma sphingomyelins and sciatic nerve triglycerides. Negative correlations were additionally present with other taxa variants. Moreover, relationships that emerged between specific PN-associated taxa variants and the sciatic nerve transcriptome were related to inflammation, lipid metabolism, and antioxidant defense pathways, which are all established in PN pathogenesis. CONCLUSIONS: The current results indicate that microbiome structure is altered with HFD, and that certain taxa variants correlate with metabolic health and PN. Apparent links between PN-associated taxa and certain lipid species and nerve transcriptome-related pathways additionally provide insight into new targets for microbiota and the associated underlying mechanisms of action in PN. Thus, these findings strengthen the possibility of a gut-microbiome-peripheral nervous system signature in PN and support continuing studies focused on defining the connection between the gut microbiome and nerve health to inform mechanistic insight and therapeutic opportunities. Video Abstract.

Cross-reactive humoral and CD4+ T cell responses to Mu and Gamma SARS-CoV-2 variants in a Colombian population.

The SARS CoV-2 antibody and CD4+ T cell responses induced by natural infection and/or vaccination decline over time and cross-recognize other viral variants at different levels. However, there are few studies evaluating the levels and durability of the SARS CoV-2-specific antibody and CD4+ T cell response against the Mu, Gamma, and Delta variants. Here, we examined, in two ambispective cohorts of naturally-infected and/or vaccinated individuals, the titers of anti-RBD antibodies and the frequency of SARS-CoV-2-specific CD4+ T cells up to 6 months after the last antigen exposure. In naturally-infected individuals, the SARS-CoV-2 antibody response declined 6 months post-symptoms onset. However, the kinetic observed depended on the severity of the disease, since individuals who developed severe COVID-19 maintained the binding antibody titers. Also, there was detectable binding antibody cross-recognition for the Gamma, Mu, and Delta variants, but antibodies poorly neutralized Mu. COVID-19 vaccines induced an increase in antibody titers 15-30 days after receiving the second dose, but these levels decreased at 6 months. However, as expected, a third dose of the vaccine caused a rise in antibody titers. The dynamics of the antibody response upon vaccination depended on the previous SARS-CoV-2 exposure. Lower levels of vaccine-induced antibodies were associated with the development of breakthrough infections. Vaccination resulted in central memory spike-specific CD4+ T cell responses that cross-recognized peptides from the Gamma and Mu variants, and their duration also depended on previous SARS-CoV-2 exposure. In addition, we found cross-reactive CD4+ T cell responses in unexposed and unvaccinated individuals. These results have important implications for vaccine design for new SARS-CoV-2 variants of interest and concern.

Corrigendum: Cross-reactive humoral and CD4+ T cell responses to Mu and Gamma SARS-CoV-2 variants in a Colombian population.

[This corrects the article DOI: 10.3389/fimmu.2023.1241038.].

A 50-Year-Old Man With a History of Recurrent Exudative Right-Sided Pleural Effusion.

In this case report, we describe a 50-year-old man who presented to our facility for a second opinion after a year-long history of recurrent and now persistent right-sided exudative pleural effusion. On review of previous records, negative findings were seen in microbiological studies, including acid-fast bacilli, cytology, flow cytometry, and pleural biopsy using video-assisted thoracoscopy. On transthoracic echocardiography performed during our evaluation, the expected respiratory variations across the mitral and tricuspid valves were not appreciated. This necessitated subsequent cardiac workup via magnetic resonance imaging, which showed a small pericardial fluid, thickened pericardium, and a septal bounce. The patient was surgically treated using a phrenic-to-phrenic pericardiectomy, following which his symptoms resolved completely. Pleural effusions occur in approximately 40-60% of patients with constrictive pericarditis, and despite the known association of pleural effusions with constrictive pericarditis, the diagnosis of constrictive pericarditis is not readily entertained in patients with undiagnosed pleural effusions.

Sustained timber yield claims, considerations, and tradeoffs for selectively logged forests.

What is meant by sustainability depends on what is sustained and at what level. Sustainable forest management, for example, requires maintenance of a variety of values not the least of which is sustained timber yields (STYs). For the 1 Bha of the world's forests subjected to selective or partial logging, failure to maintain yields can be hidden by regulatory requirements and questionable auditing practices such as increasing the number of commercial species with each harvest, reducing the minimum size at which trees can be harvested and accepting logs of lower quality. For assertions of STY to be credible, clarity is needed about all these issues, as well as about the associated ecological and economic tradeoffs. Lack of clarity about sustainability heightens risks of unsubstantiated claims and unseen losses. STY is possible but often requires cutting cycles that are longer and logging intensities that are lower than prescribed by law, as well as effective use of low-impact logging practices and application of silvicultural treatments to promote timber stock recovery. These departures from business-as-usual practices will lower profit margins but generally benefit biodiversity and ecosystem services.

Evaluation of Zika rapid tests as aids for clinical diagnosis and epidemic preparedness.

Background: Development and evaluation of diagnostics for diseases of epidemic potential are often funded during epidemics, but not afterwards, leaving countries unprepared for the next epidemic. United Nations Children's Emergency Fund (UNICEF) partnered with the United States Agency for International Development (USAID) to address this important gap by investing in an advance purchase commitment (APC) mechanism to accelerate the development and evaluation of Zika rapid diagnostic tests (RDTs) for case detection and surveillance. This paper describes the performance evaluation of five Zika RDTs eligible for procurement. Methods: A network of European Union-funded ZikaPLAN sites in Africa, Asia, Latin America with access to relevant serum specimens were selected to evaluate RDTs developed for the UNICEF APC mechanism. A standardised protocol and evaluation panels were developed and a call for specimens for the evaluation panels issued to different sites. Each site contributed specimens to the evaluation from their biobank. Data were collated, analysed and presented to the UNICEF Procurement Review Group for review. Findings: Three RDTs met the criteria for UNICEF procurement of sensitivity and specificity of 85% against a refence standard. The sensitivity/specificity of the ChemBio anti-Zika Virus (ZIKV) immunoglobulin M (IgM) test was 86.4 %/86.7% and the ChemBio ZCD system for anti-ZIKV IgM was 79.0%/97.1%, anti-dengue virus (DENV) IgM 90.0%/89.2%, anti-Chikungunya virus (CHIKV) IgM 90.6%/97.2%. The sensitivity/specificity of the SD Biosensor anti-ZIKV IgM was 96.8 %/90.8%, anti-DENV IgM 71.8%/83.5%, the DENV nonstructural protein 1 (NS1) glycoprotein 90.0%/90.2%, anti- yellow fever virus (YFV) IgM 84.6%/92.4%, anti-CHIKV IgM 86.3%/97.5%. Interpretation: Three RDTs fulfilled the performance thresholds set by WHO and were eligible for UNICEF procurement. These tests will improve the diagnosis of ZIKV and other arboviral infections as well as providing countries with better tools for surveillance and response to future epidemics. Funding: This work was supported by the USAID grant GHA-G-00-07-00007 and ZikaPLAN (European Union's Horizon 2020 Research and Innovation Programme under Grant Agreement No. 734584).

Tofacitinib Suppresses Natural Killer Cells In Vitro and In Vivo: Implications for Amyotrophic Lateral Sclerosis.

Amyotrophic lateral sclerosis (ALS) is a fatal and incurable neurodegenerative disease with few therapeutic options. However, the immune system, including natural killer (NK) cells, is linked to ALS progression and may constitute a viable therapeutic ALS target. Tofacitinib is an FDA-approved immunomodulating small molecule which suppresses immune cell function by blocking proinflammatory cytokine signaling. This includes the cytokine IL-15 which is the primary cytokine associated with NK cell function and proliferation. However, the impact of tofacitinib on NK activation and cytotoxicity has not been thoroughly investigated, particularly in ALS. We therefore tested the ability of tofacitinib to suppress cytotoxicity and cytokine production in an NK cell line and in primary NK cells derived from control and ALS participants. We also investigated whether tofacitinib protected ALS neurons from NK cell cytotoxicity. Finally, we conducted a comprehensive pharmacokinetic study of tofacitinib in mice and tested the feasibility of administration formulated in chow. Success was assessed through the impact of tofacitinib on peripheral NK cell levels in mice. We found tofacitinib suppressed IL-15-induced activation as measured by STAT1 phosphorylation, cytotoxicity, pro-inflammatory gene expression, and pro-inflammatory cytokine secretion in both an NK cell line and primary NK cells. Furthermore, tofacitinib protected ALS neurons from NK cell-mediated cytotoxicity. In mice, we found tofacitinib bioavailability was 37% in both male and female mice; using these data we formulated mouse containing low and high doses of tofacitinib and found that the drug suppressed peripheral NK cell levels in a dose-dependent manner. These results demonstrate that tofacitinib can suppress NK cell function and may be a viable therapeutic strategy for ALS.