A cross-disease, pleiotropy-driven approach for therapeutic target prioritization and evaluation.

Cross-disease genome-wide association studies (GWASs) unveil pleiotropic loci, mostly situated within the non-coding genome, each of which exerts pleiotropic effects across multiple diseases. However, the challenge "W-H-W" (namely, whether, how, and in which specific diseases pleiotropy can inform clinical therapeutics) calls for effective and integrative approaches and tools. We here introduce a pleiotropy-driven approach specifically designed for therapeutic target prioritization and evaluation from cross-disease GWAS summary data, with its validity demonstrated through applications to two systems of disorders (neuropsychiatric and inflammatory). We illustrate its improved performance in recovering clinical proof-of-concept therapeutic targets. Importantly, it identifies specific diseases where pleiotropy informs clinical therapeutics. Furthermore, we illustrate its versatility in accomplishing advanced tasks, including pathway crosstalk identification and downstream crosstalk-based analyses. To conclude, our integrated solution helps bridge the gap between pleiotropy studies and therapeutics discovery.

Priority index for critical Covid-19 identifies clinically actionable targets and drugs.

While genome-wide studies have identified genomic loci in hosts associated with life-threatening Covid-19 (critical Covid-19), the challenge of resolving these loci hinders further identification of clinically actionable targets and drugs. Building upon our previous success, we here present a priority index solution designed to address this challenge, generating the target and drug resource that consists of two indexes: the target index and the drug index. The primary purpose of the target index is to identify clinically actionable targets by prioritising genes associated with Covid-19. We illustrate the validity of the target index by demonstrating its ability to identify pre-existing Covid-19 phase-III drug targets, with the majority of these targets being found at the leading prioritisation (leading targets). These leading targets have their evolutionary origins in Amniota ('four-leg vertebrates') and are predominantly involved in cytokine-cytokine receptor interactions and JAK-STAT signaling. The drug index highlights opportunities for repurposing clinically approved JAK-STAT inhibitors, either individually or in combination. This proposed strategic focus on the JAK-STAT pathway is supported by the active pursuit of therapeutic agents targeting this pathway in ongoing phase-II/III clinical trials for Covid-19.

Priority index for asthma (PIA): In silico discovery of shared and distinct drug targets for adult- and childhood-onset disease.

Asthma is a chronic disease that is caused by a combination of genetic risks and environmental triggers and can affect both adults and children. Genome-wide association studies have revealed partly distinct genetic architectures for its two age-of-onset subtypes (namely, adult-onset and childhood-onset). We reason that identifying shared and distinct drug targets between these subtypes may inform the development of subtype-specific therapeutic strategies. In attempting this, we here introduce Priority Index for Asthma or PIA, a genetics-led and network-driven drug target prioritisation tool for asthma. We demonstrate the validity of the tool in improving drug target prioritisation for asthma compared to the status quo methods, as well as in capturing the underlying etiology and existing therapeutics for the disease. We also illustrate how PIA can be used to prioritise drug targets for adult- and childhood-onset asthma, as well as to identify shared and distinct pathway crosstalk genes. Shared crosstalk genes are mostly involved in JAK-STAT signaling, with clinical evidence supporting that targeting this pathway may be a promising drug repurposing opportunity for both subtypes. Crosstalk genes specific to childhood-onset asthma are enriched for PI3K-AKT-mTOR signaling, and we identify genes that are already targeted by licensed medications as repurposed drug candidates for this subtype. We make all our results accessible and reproducible at http://www.genetictargets.com/PIA. Collectively, our study has significant implications for asthma computational medicine research and can guide the future development of subtype-specific therapeutic strategies for the disease.

Donnan Effect-Engineered Covalent Organic Framework Membranes toward Size- and Charge-Based Precise Molecular Sieving.

Covalent organic frameworks (COFs), with ordered pores and well-defined topology, are ideal materials for nanofiltration (NF) membranes because of their capacity of transcending the permeance/selectivity trade-off predicament. However, most reported COF-based membranes are focused on separating molecules with different sizes, resulting in low selectivity to similar molecules with different charges. Here, the negatively charged COF layer was fabricated in situ on a microporous support for the separation of molecules with different sizes and charges. Ultrahigh water permeance (216.56 L m-2 h-1 bar-1) was obtained because of the ordered pores and excellent hydrophilicity, which exceeds that of most membranes with similar rejections. For the first time, we used multifarious dyes with different sizes and charges, for the investigation of the selectivity behavior caused by the Donnan effect and size exclusion. The obtained membranes represent superior rejections to negatively and neutrally charged dyes larger than 1.3 nm, while positively charged dyes with a size of 1.6 nm can pass through the membrane, resulting in the separation of negative/positive mixed dyes with similar molecular sizes. This strategy of combining the Donnan effect and size exclusion in nanoporous materials may evolve into a generic platform for sophisticated separation.

Various Mobile Genetic Elements Involved in the Dissemination of the Phenicol-Oxazolidinone Resistance Gene optrA in the Zoonotic Pathogen Streptococcus suis: a Nonignorable Risk to Public Health.

The rapid increase of phenicol-oxazolidinone (PhO) resistance in Streptococcus suis due to transferable resistance gene optrA is a matter of concern. However, genetic mechanisms for the dissemination of the optrA gene remain to be discovered. Here, we selected 33 optrA-positive S. suis isolates for whole-genome sequencing and analysis. The IS1216E element was present in 85% of the optrA-carrying contigs despite genetic variation observed in the flanking region. IS1216E-optrA-carrying segments could be inserted into larger mobile genetic elements (MGEs), including integrative and conjugative elements, plasmids, prophages, and antibiotic resistance-associated genomic islands. IS1216E-mediated circularization occurred to form the IS1216E-optrA-carrying translocatable units, suggesting a crucial role of IS1216E in optrA spreading. Three optrA-carrying MGEs (ICESsuAKJ47_SSU1797, plasmid pSH0918, and prophage ΦSsuFJSM5_rum) were successfully transferred via conjugation at different transfer frequencies. Interestingly, two types of transconjugants were observed due to the multilocus integration of ICESsuAKJ47 into an alternative SSU1943 attachment site along with the primary SSU1797 attachment site (type 1) or into the single SSU1797 attachment site (type 2). In addition, conjugative transfer of an optrA-carrying plasmid and prophage in streptococci was validated for the first time. Considering the abundance of MGEs in S. suis and the mobility of IS1216E-optrA-carrying translocatable units, attention should be paid to the potential risks to public health from the emergence and spread of PhO-resistant S. suis. IMPORTANCE Antimicrobial resistance to phenicols and oxazolidinones by the dissemination of the optrA gene leads to treatment failure in both veterinary and human medicine. However, information about the profile of these MGEs (mobilome) that carry optrA and their transferability in streptococci was limited, especially for the zoonotic pathogen S. suis. This study showed that the optrA-carrying mobilome in S. suis includes integrative and conjugative elements (ICEs), plasmids, prophages, and antibiotic resistance-associated genomic islands. IS1216E-mediated formation of optrA-carrying translocatable units played important roles in optrA spreading between types of MGEs, and conjugative transfer of various optrA-carrying MGEs (ICEs, plasmids, and prophages) further facilitated the transfer of optrA across strains, highlighting a nonignorable risk to public health of optrA dissemination to other streptococci and even to bacteria of other genera.

Identification of N- and C-3-Modified Laudanosoline Derivatives as Novel Influenza PAN Endonuclease Inhibitors.

Influenza PAN inhibitors are of particular importance in current efforts to develop a new generation of antiviral drugs due to the growing emergence of highly pathogenic influenza viruses and the resistance to existing antiviral inhibitors. Herein, we design and synthesize a set of 1,3-cis-N-substituted-1,2,3,4-tetrahydroisoquinoline derivatives to enhance their potency by further exploiting the pockets 3 and 4 in the PAN endonuclease based on the hit d,l-laudanosoline. Particularly, the lead compound 35 exhibited potent and broad anti-influenza virus effects with EC50 values ranging from 0.43 to 1.12 µM in vitro and good inhibitory activity in a mouse model. Mechanistic studies demonstrated that 35 could bind tightly to the PAN endonuclease of RNA-dependent RNA polymerase, thus blocking the viral replication to exert antiviral activity. Overall, our study might establish the importance of 1,2,3,4-tetrahydroisoquinoline-6,7-diol-based derivatives for the development of novel PAN inhibitors of influenza viruses.

Comparison of an online versus conventional multidisciplinary collaborative weight loss programme in type 2 diabetes mellitus: A randomized controlled trial.

AIM: The aim of this study was to examine the effect of an online multidisciplinary weight loss management programme. METHODS: Between July 2016 and July 2017 this randomized controlled trial recruited patients in Nanjing, China who were living with type 2 diabetes mellitus and who were obese or overweight and randomized them to online versus conventional groups. All participants were managed by a multidisciplinary team. The experimental group was managed using the Why Wait WeChat Platform for Weight Reduction Management. RESULTS: There were 55 and 52 participants in the online and conventional groups, respectively. The decreases in fasting blood glucose (-4.26 vs. -2.99 mmol/L), 2-h postprandial blood glucose (-4.48 vs. -2.68 mmol/L) and glycated haemoglobin (-22.11 vs. -6.21 mmol/mol) were more pronounced in the online compared to conventional group (all P < 0.05). After the intervention, self-management ability parameters, including diet control, foot care and total score, were improved in the online group compared with the conventional group, as well as all indexes of quality of life (all P < 0.05). CONCLUSION: The online multidisciplinary weight loss management programme improved blood glucose in obese or overweight patients living with type 2 diabetes mellitus. Self-management ability parameters (including diet control, foot care and total score) and quality of life were improved in the online group compared with the conventional group.

Targeting the PI3K/AKT/mTOR Signaling Pathway in the Treatment of Human Diseases: Current Status, Trends, and Solutions.

The phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) pathway is one of the most important intracellular pathways involved in cell proliferation, growth, differentiation, and survival. Therefore, this route is a prospective biological target for treating various human diseases, such as tumors, neurodegenerative diseases, pulmonary fibrosis, and diabetes. An increasing number of clinical studies emphasize the necessity of developing novel molecules targeting the PI3K/AKT/mTOR pathway. This review focuses on recent advances in ATP-competitive inhibitors, allosteric inhibitors, covalent inhibitors, and proteolysis-targeting chimeras against the PI3K/AKT/mTOR pathway, and highlights possible solutions for overcoming the toxicities and acquired drug resistance of currently available drugs. We also provide recommendations for the future design and development of promising drugs targeting this pathway.

Effects of slice thickness on CT radiomics features and models for staging liver fibrosis caused by chronic liver disease.

OBJECTIVES: To investigate the effects of slice thickness on CT radiomics features and models for staging liver fibrosis. METHODS: A total of 108 pathologically confirmed liver fibrosis patients from a single center were retrospectively collected and divided into different groups. Both thick (5- or 7-mm) and thin slices (1.3- or 2-mm) were analyzed. A fivefold cross-validation with 100 repeats was conducted. The minimum redundancy-maximum relevance algorithm was used to reduce the radiomics features, and the top 10 ranking features were included for further analysis for each loop. The random forest was used for model establishment. The models with median AUC were selected for the assessment of the discriminative performance for both datasets. Mutual features selected by the models with AUC > 0.8 were searched and considered as the most predictive ones. RESULTS: A total of 162 and 643 radiomics features with excellent reliability were selected from thick- and thin-slice datasets, respectively. The overall discriminative performance of the 500 AUCs from the thin-slice dataset was better than the thick slice. The median AUC values of the thick-sliced datasets were significantly lower than those of the thin-sliced datasets (0.78 and 0.90 for differentiating F1 vs. F2-4, 0.72 and 0.85 for differentiating F1-2 vs. F3-4, both P = 0.03). For differentiating F1-3 vs. F4, no significant difference was found (0.85 vs 0.94, P = 0.15). Six mutual predictive features across all the datasets were found. CONCLUSIONS: The radiomics features extracted from thin-slice images and their corresponding models were better and more stable for staging liver fibrosis.

Eco-friendly chitosan@silver/plant fiber membranes for masks with thermal comfortability and self-sterilization.

The surgical masks have been essential consumables for public in the COVID-19 pandemic. However, long-time wearing masks will make wearers feel uncomfortable and massive discarded non-biodegradable masks lead to a heavy burden on our environment. In this paper, we adopt degradable chitosan@silver (CS@Ag) core-shell fibers and plant fibers to prepare an eco-friendly mask with excellent thermal comfort, self-sterilization, and antiviral effects. The thermal network of CS@Ag core-shell fibers highly improves the in-plane thermal conductivity of masks, which is 4.45 times higher than that of commercial masks. Because of the electrical conductivity of Ag, the fabricated mask can be electrically heated to warm the wearer in a cold environment and disinfect COVID-19 facilely at room temperature. Meanwhile, the in-situ reduced silver nanoparticles (AgNPs) endow the mask with superior antibacterial properties. Therefore, this mask shows a great potential to address the urgent need for a thermally comfortable, antibacterial, antiviral, and eco-friendly mask. Supplementary Information: The online version contains supplementary material available at 10.1007/s10570-022-04582-x.

Genetic Susceptibility to Enteric Fever in Experimentally Challenged Human Volunteers.

Infections with Salmonella enterica serovars Typhi and Paratyphi A cause an estimated 14 million cases of enteric fever annually. Here, the controlled nature of challenge studies is exploited to identify genetic variants associated with enteric fever susceptibility. Human challenge participants were genotyped by Illumina OmniExpress-24 BeadChip array (n = 176) and/or transcriptionally profiled by RNA sequencing (n = 174). While the study was underpowered to detect any single nucleotide polymorphisms (SNPs) significant at the whole-genome level, two SNPs within CAPN14 and MIATNB were identified with P < 10-5 for association with development of symptoms or bacteremia following oral S. Typhi or S. Paratyphi A challenge. Imputation of classical human leukocyte antigen (HLA) types from genomic and transcriptomic data identified HLA-B*27:05, previously associated with nontyphoidal Salmonella-induced reactive arthritis, as the HLA type most strongly associated with enteric fever susceptibility (P = 0.011). Gene sets relating to the unfolded protein response/heat shock and endoplasmic reticulum-associated protein degradation were overrepresented in HLA-B*27:05+ participants following challenge. Furthermore, intracellular replication of S. Typhi is higher in C1R cells transfected with HLA-B*27:05 (P = 0.02). These data suggest that activation of the unfolded protein response by HLA-B*27:05 misfolding may create an intracellular environment conducive to S. Typhi replication, increasing susceptibility to enteric fever.

Highly thermally conductive and eco-friendly OH-h-BN/chitosan nanocomposites by constructing a honeycomb thermal network.

More than 110,000,000 tons of mismanaged plastics were to be produced in 2020. Polymers are favored in the preparation of thermally conductive materials due to their excellent comprehensive properties. However, most polymers fabricated for thermally conductive materials are difficult to degrade in the natural environment. To alleviate the increasingly severe environmental problems, we reported a novel eco-friendly material with high thermal conductivity, which was composited of chitosan microspheres (CSM) and hydroxyl-functionalized hexagonal boron nitride (OH-h-BN) nanoplatelets. Utilizing their significant difference in scales, the OH-h-BN nanoplatelets were arranged between each CSM. Their overall structure was similar to the honeycomb: CSM were honeycomb cores, and OH-h-BN nanoplatelets were honeycomb network. The routine-structure OH-h-BN/CS nanocomposites were only 0.94 ± 0.02 W·m-1·K-1 at 50 wt% in thermal conductivity. However, the OH-h-BN/CSM nanocomposites with honeycomb structure can reach 5.66 ± 0.32 W·m-1·K-1 in the same loading, for enhancement of 502% and 1914% than OH-h-BN/CS nanocomposites and pure CS, respectively.

Ruthenium-Catalyzed, Chemoselective and Regioselective Oxidation of Polyisobutene.

Polyolefins are important commodity plastics, yet their lack of functional groups limits their applications. The functionalization of C-H bonds holds promise for incorporating functionalities into polymers of ethylene and linear α-olefins. However, the selective functionalization of polyolefins derived from branched alkenes, even monobranched, 1,1-substituted alkenes, has not been achieved. These polymers are less reactive, due to steric effects, and they are prone to chain scission that degrades the polymer. We report the chemoselective and regioselective oxidation of a commercially important polymer of a branched olefin, polyisobutene. A polyfluorinated ruthenium-porphyrin catalyst incorporates ketone units into polyisobutene at methylene positions without chain cleavage. The oxidized polymer is thermally stable, yet it undergoes programmed reactions and possesses enhanced wetting properties.

Heat Conduction and Antibacterial Hexagonal Boron Nitride/Polypropylene Nanocomposite Fibrous Membranes for Face Masks with Long-Time Wearing Performance.

Wearing surgical masks is one of the best protective measures to protect humans from viral invasion during the 2019 coronavirus (COVID-19) outbreak. However, wearing surgical masks for extended periods will cause uncomfortable sweltering sense to users and are easy to breed bacteria. Here, we reported a novel fibrous membrane with outstanding comfortability and antibacterial activity prepared by PP ultrafine fiber nonwovens and antibacterial functionalized h-BN nanoparticles (QAC/h-BN). The thermal conductivity of commercial PP nonwovens was only 0.13 W m-1 K-1, but that of the QAC/h-BN/PP nanocomposite fibrous membranes can reach 0.88 W m-1 K-1, an enhancement of 706.5% than commercial PP nonwovens. The surface temperature of commercial PP surgical masks was 31.8 °C when the wearing time was 60 min. In contrast, QAC/h-BN/PP surgical masks can reach 33.6 °C at the same tested time, exhibiting stronger heat dissipation than commercial PP surgical masks. Besides, the antibacterial rates of QAC/h-BN/PP nanocomposite fibrous membranes were 99.3% for E. coli and 96.1% for S. aureus, and their antibacterial mechanism was based on "contact killing" without the release of unfavorable biocides. We think that the QAC/h-BN/PP nanocomposite fibrous membranes could provide better protection to people.

Functional Genomic Analysis of a RUNX3 Polymorphism Associated With Ankylosing Spondylitis.

OBJECTIVE: To investigate the functional consequences of the single-nucleotide polymorphism rs4648889 in a putative enhancer upstream of the RUNX3 promoter associated with susceptibility to ankylosing spondylitis (AS). METHODS: Using nuclear extracts from Jurkat cells and primary human CD8+ T cells, the effects of rs4648889 on allele-specific transcription factor (TF) binding were investigated by DNA pull-down assay and quantitative mass spectrometry (qMS), with validation by electrophoretic mobility shift assay (EMSA), Western blotting of the pulled-down eluates, and chromatin immunoprecipitation (ChIP)-quantitative polymerase chain reaction (qPCR) analysis. Further functional effects were tested by small interfering RNA knockdown of the gene for interferon regulatory factor 5 (IRF5), followed by reverse transcription-qPCR (RT-qPCR) and enzyme-linked immunosorbent assay (ELISA) to measure the levels of IFNγ messenger RNA (mRNA) and protein, respectively. RESULTS: In nuclear extracts from CD8+ T cells, results of qMS showed that relative TF binding to the AS-risk A allele of rs4648889 was increased 3.7-fold (P < 0.03) for Ikaros family zinc-finger protein 3 (IKZF3; Aiolos) and components of the NuRD complex, including chromodomain helicase DNA binding protein 4 (CHD4) (3.6-fold increase; P < 0.05) and retinoblastoma binding protein 4 (RBBP4) (4.1-fold increase; P < 0.03). In contrast, IRF5 bound significantly more to the AS-protective G allele compared to the AS-risk A allele (fold change 8.2; P = 0.003). Validation with Western blotting, EMSA, and ChIP-qPCR confirmed the differential allelic binding of IKZF3, CHD4, RBBP4, and IRF5. Silencing of IRF5 in CD8+ T cells increased the levels of IFNγ mRNA as measured by RT-qPCR (P = 0.03) and IFNγ protein as measured by ELISA (P = 0.02). CONCLUSION: These findings suggest that the association of rs4648889 with AS reflects allele-specific binding of this enhancer-like region to certain TFs, including IRF5, IKZF3, and members of the NuRD complex. IRF5 may have crucial influences on the functions of CD8+ lymphocytes, a finding that could reveal new therapeutic targets for the management of AS.

The effect of platelet-rich plasma on the fusion rate and clinical outcome of spinal fusion surgery: A systematic review and meta-analysis.

BACKGROUND: Platelet-rich plasma (PRP) is widely used in many orthopedic surgeries and spinal disease treatments; however, the effect of PRP on spinal fusion remains controversial. QUESTIONS/PURPOSES: To assess the fusion rate and clinical results of PRP compared with non-PRP administration in the treatment of spinal fusion with regard to decreasing pain and improving healing and function. PATIENTS AND METHODS: Studies comparing PRP to non-PRP treatment with respect to the fusion rate and clinical outcome in patients who underwent spinal fusion surgery were included. RESULT: Three randomized controlled trials (RCTs) and 7 prospective cohort studies were identified. The spinal fusion rate was not significantly different between the groups in all RCTs or cohort studies at the final follow-up. In comparison, PRP significantly reduced pain after surgery as evaluated in the RCT analysis and the complication rate did not differ significantly between the two groups. CONCLUSION: According to the available studies, PRP does not contribute to the union rate, relieve pain or increase the complication rate in spinal fusion surgery. As clinical heterogeneity exists in these studies, further large, well-designed RCTs that focus on the standard assessment of PRP are needed.

Aryl Amination Using Soluble Weak Base Enabled by a Water-Assisted Mechanism.

The amination of aryl halides has become one of the most commonly practiced C-N bond-forming reactions in pharmaceutical and laboratory syntheses. The widespread use of strong or poorly soluble inorganic bases for amine activation nevertheless complicates the compatibility of this important reaction class with sensitive substrates as well as applications in flow and automated synthesis, to name a few. We report a palladium-catalyzed C-N coupling using Et3N as a weak, soluble base, which allows a broad substrate scope that includes bromo- and chloro(hetero)arenes, primary anilines, secondary amines, and amide type nucleophiles together with tolerance for a range of base-sensitive functional groups. Mechanistic data have established a unique pathway for these reactions in which water serves multiple beneficial roles. In particular, ionization of a neutral catalytic intermediate via halide displacement by H2O generates, after proton loss, a coordinatively unsaturated Pd-OH species that can bind amine substrate triggering intramolecular N-H heterolysis. This water-assisted pathway operates efficiently with even weak terminal bases, such as Et3N. The use of a simple, commercially available ligand, PAd3, is key to this water-assisted mechanism by promoting coordinative unsaturation in catalytic intermediates responsible for the heterolytic activation of strong element-hydrogen bonds, which enables broad compatibility of carbon-heteroatom cross-coupling reactions with sensitive substrates and functionality.

Sacroiliac joint fusion VS conservative management for chronic low back pain attributed to the sacroiliac joint: A protocol for systematic review and meta analysis.

INTRODUCTION: Low back pain (LBP) is high prevalent and it is the leading cause of years lived with disability in both developed and developing countries. The sacroiliac joint (SIJ) is a common reason that caused LBP. At present, the treatment of chronic LBP attributed to SIJ is mainly conservative treatment and surgical treatment. However, there are still controversies between the 2 treating methods, and there is no recognized standard of treatment or surgical indications. Recent publications indicated that minimally invasive sacroiliac joint arthrodesis was safe and more effective improving pain, disability, and quality of life compared with conservative management in 2 years follow-up, which re-raise the focus of sacroiliac joints fusion. This paper will systematically review the available evidence, comparing the effectiveness of sacroiliac joint fusion and conservative therapy for the treatment of gait retraining for patients suffered from LBP attributed to the sacroiliac joint. METHOD AND ANALYSIS: A systematic review and meta-analysis of relevant studies in Pubmed, Embase, SCOPUS, and Cochrane Library will be synthesized. Inclusion criteria will be studies evaluating clinical outcomes (i.e., changes to pain and/or function) comparing sacroiliac joint fusion and conservative therapy in populations sacroiliac join related LBP; studies with less than 10 participants in total will be excluded. The primary outcomes measured will be pain score, Oswestry Disability Index (ODI), and adverse events during treatment. Review Manager (Revman; Version 5.3) software will be used for data synthesis, sensitivity analysis, meta-regression, subgroup analysis, and risk of bias assessment. A funnel plot will be developed to evaluate reporting bias and Begg and Egger tests will be used to assess funnel plot symmetries. We will use the Grading of Recommendations Assessment, Development and Evaluation system to assess the quality of evidence. ETHICS AND DISSEMINATION: Our aim is to publish this systematic review and meta-analysis in a peer-reviewed journal. Our findings will provide information comparing the efficacy and safety comparing sacroiliac joint fusion and non-surgical treatment for patients with LBP attributed to the sacroiliac joint. This review will not require ethical approval as there are no issues about participant privacy.

GM-CSF Primes Proinflammatory Monocyte Responses in Ankylosing Spondylitis.

Objectives: GM-CSF is a pro-inflammatory cytokine with multiple actions predominantly on myeloid cells. Enhanced GM-CSF expression by lymphocytes from patients with Ankylosing Spondylitis (AS) has recently been described, however, its potential pathogenic role(s) in AS are unknown. Methods: The effects of GM-CSF on TNF, IL-23, and CCL17 production by blood, PBMCs and isolated CD14+ monocytes from AS patients and healthy controls (HCs) were studied using ELISA. Serum CCL17 and GM-CSF and T cell GM-CSF production were studied in AS patients including pre-and on TNFi therapy. Results: GM-CSF markedly increased TNF production by LPS-stimulated whole blood, peripheral blood mononuclear cells (PBMC) and purified monocytes from AS patients, with 2 h GM-CSF exposure sufficient for monocyte "priming." Blocking of GM-CSF significantly reduced the production of TNF by whole blood from AS patients but not HCs. GM-CSF priming increased IL-23 production from LPS-stimulated AS and HC whole blood 5-fold, with baseline and stimulated IL-23 levels being significantly higher in AS whole blood. GM-CSF also stimulated CCL17 production from AS and HC blood and CCL17 levels were elevated in AS plasma. GM-CSF could be detected in plasma from 14/46 (30%) AS patients compared to 3/18 (17%) HC. Conclusion: We provide evidence that GM-CSF primes TNF and IL-23 responses in myeloid cells from AS patients and HC. We also show CCL17 levels, downstream of GM-CSF, were elevated in plasma samples of AS patients. Taken together these observations are supportive of GM-CSF neutralization as a potential novel therapeutic approach for the treatment of AS.