Background: Hidradenitis suppurativa (HS) is a chronic inflammatory disease characterized by painful inflamed nodules, abscesses, and pus-draining tunnels appearing in axillary, inguinal, and perianal skin areas. HS lesions contain various types of immigrated immune cells. Objective: This study aimed to characterize mediators that support lesional B/plasma cell persistence in HS. Methods: Skin samples from several cohorts of HS patients and control cohorts were assessed by mRNA sequencing, quantitative PCR on reverse-transcribed RNA, flow cytometry, and immunohistofluorescence. Blood plasma and cultured skin biopsy samples, keratinocytes, dermal fibroblasts, neutrophilic granulocytes (neutrophils), monocytes, and B cells were analyzed. Complex systems biology approaches were used to evaluate bulk and single-cell RNA sequencing data. Results: Proportions of B/plasma cells, neutrophils, CD81 T cells, and M0 and M1 macrophages were elevated in HS lesions compared to skin of healthy and perilesional intertriginous areas. There was an association between B/plasma cells, neutrophils, and B-cell activating factor (BAFF, aka TNFSF13B). BAFF was abundant in HS lesions, particularly in nodules and abscesses. Among the cell types present in HS lesions, myeloid cells were the main BAFF producers. Mechanistically, granulocyte colony-stimulating factor in the presence of bacterial products was the major stimulus for neutrophils' BAFF secretion. Lesional upregulation of BAFF receptors was attributed to B cells (TNFRSF13C/BAFFR and TNFRSF13B/TACI) and plasma cells (TNFRSF17/BCMA). Characterization of the lesional BAFF pathway revealed molecules involved in migration/adhesion (eg, CXCR4, CD37, CD53, SELL), proliferation/survival (eg, BST2), activation (eg, KLF2, PRKCB), and reactive oxygen species production (eg, NCF1, CYBC1) of B/plasma cells. Conclusion: Neutrophil-derived BAFF supports B/plasma cell persistence and function in HS lesions. (J Allergy Clin Immunol 2023;151:1015-26.)

ABSTRACTObjectivesTo investigate the role of endogenous TSG-6 in human osteoarthritis (OA) and assess the disease-modifying potential of a TSG-6-based biological treatment in cell, explant and animal models of OA.MethodsKnee articular cartilages from OA patients were analysed for TSG-6 protein and mRNA expression using immunohistochemistry and RNAscope, respectively. The inhibitory activities of TSG-6 and its isolated Link module domain (Link_TSG6) on cytokine-induced glycosaminoglycan loss in OA cartilage explants were compared. Mesenchymal stem/stromal cell (MSC)-derived chondrocyte pellet cultures were used to determine the effects of Link_TSG6 and full-length TSG-6 on IL-1α-, IL-1β- or TNF-stimulated ADAMTS4, ADAMTS5 and MMP13 mRNA expression. Link_TSG6 was administered i.a. to the rat ACLTpMMx model and cartilage damage and tactile allodynia were assayed.ResultsTSG-6 is predominantly associated with chondrocytes in regions of cartilage damage and its expression is negatively correlated with MMP13, the major collagenase implicated in OA progression. Link_TSG6 is more potent than full-length TSG-6 at dose-dependently inhibiting cytokine-mediated matrix breakdown in human OA cartilage explants; about 50% of donor cartilages, from 59 tested, were responsive to Link_TSG6 treatment. Similarly, Link_TSG6 displayed more potent effects in 3D pellet cultures, suppressing aggrecanase and collagenase gene expression. Link_TSG6 treatment reduced touch-evoked pain and dose-dependently inhibited cartilage damage in a rodent model of surgically-induced OA.ConclusionsNative TSG-6 is associated with a low catabolic chondrocyte phenotype in OA cartilage. Link_TSG6, which has enhanced chondroprotective activity compared to the full-length TSG-6 protein, demonstrates potential as a disease modifying OA drug (DMOAD) and warrants further investigation and development.KEY MESSAGESWhat is already known about this subject?TSG-6, a protein with anti-inflammatory and protective effects in other tissues, is expressed in joints affected by osteoarthritis – a condition for which there are no disease-modifying drugs.What does this study add?A novel protective mechanism has been identified, whereby TSG-6 inhibits inflammatory cytokine-induced catabolic pathways in cartilage.The Link module of TSG-6 (Link_TSG6) has been shown to have greater potency than TSG-6 as an inhibitor of cartilage damage and is efficacious in a rat model of osteoarthritis.Data from cartilage explants indicate that OA patients can be stratified for Link_TSG6 responsiveness.How might this impact on clinical practice or future developments?Link_TSG6 has been identified as potential disease-modifying OA drug that mimics an intrinsic protective process.

Hidradenitis suppurativa (HS) is a chronic inflammatory disease, characterized by painful, purulent and destructive skin alterations in intertriginous areas.

This study's aim was to demonstrate that the combination of patient immune profiling and testing in a humanized mouse model of ulcerative colitis (UC) might lead to patient stratification for treatment with oxelumab. First, immunological profiles of UC patients and non-UC donors were analyzed for CD4+ T cells expressing OX40 (CD134; also known as TNFRSF4) and CD14+ monocytes expressing OX40L (CD252; also known as TNFSF4) by flow cytometric analysis. A significant difference was observed between the groups for CD14+ OX40L+ (UC: n=11, 85.44±21.17, mean±s.d.; non-UC: n=5, 30.7±34.92; P=0.02), whereas no significant difference was detected for CD4+ OX40+. CD14+ OX40L+ monocytes were correlated significantly with T helper 1 and 2 cells. Second, NOD/Scid IL2Rγ null mice were reconstituted with peripheral blood mononuclear cells from UC donors exhibiting elevated levels of OX40L, and the efficacy of oxelumab was compared with that of adalimumab. The clinical, colon and histological scores and the serum concentrations of IL-6, IL-1β and glutamic acid were assessed. Treatment with oxelumab or adalimumab resulted in significantly reduced clinical, colon and histological scores, reduced serum concentrations of IL-6 and reduced frequencies of splenic human effector memory T cells and switched B cells. Comparison of the efficacy of adalimumab and oxelumab by orthogonal partial least squares discrimination analysis revealed that oxelumab was slightly superior to adalimumab; however, elevated serum concentrations of glutamic acid suggested ongoing inflammation. These results suggest that oxelumab addresses the pro-inflammatory arm of inflammation while promoting the remodeling arm and that patients exhibiting elevated levels of OX40L might benefit from treatment with oxelumab.

Due to the clinical development of drugs such as secukinumab, ustekinumab and dupilumab, major changes have been achieved in the treatment of patients diagnosed with psoriasis and atopic dermatitis. In academia and the pharmaceutical industry, research is increasingly moving towards the development of bispecific antibodies and multi‐specific nanobodies, as there is a compelling need for new treatment modalities for patients suffering from autoimmune or malignant disease. The purpose of this review is to discuss aspects of translational drug development with a particular emphasis on indications such as psoriasis and atopic dermatitis. The identification of biomarkers, the assessment of target organ pharmacokinetic and pharmacodynamics interactions and a wide range of in vitro, ex vivo and in vivo models should contribute to an appropriate prediction of a biological effect in the clinical setting. As human biology may not be perfectly reflected by approaches such as skin equivalents or animal models, novel approaches such as the use of human skin and dermal microperfusion assays in healthy volunteers and patients appear both reasonable and mandatory. These models may indeed generate highly translationally relevant data that have the potential to reduce the failure rate of drugs currently undergoing clinical development.

Genoskin recently developed InflammaSkin®, a T cell-driven skin inflammation model for psoriasis based on the activation and differentiation into a Th17/Th1 phenotype of skin resident T cells. This ex vivo human skin model is composed of epidermis and dermis with or without the underlying subcutaneous tissue. In the presence of the adipose tissue, it is designed to support subcutaneous delivery of therapeutic compounds. T-cell activation and differentiation in InflammaSkin® with or without adipose tissue, led to a strong upregulation of the release of IL-17A, IL-22, IL-27 and IFNγ pro-inflammatory cytokines after 3- and 7-days culture, as well as an important loss of tissue integrity and cell viability at the histological level. Release of T cell-derived cytokines also led to overexpression of S100A7 and Keratin 16 epidermal activation marker. In order to assess pharmacological response of the model to subcutaneous administration of biologics, adalimumab was injected in the fat tissue layer of the model after 3 days of in situ activation and polarization of resident T-cells. Histological analysis showed improvement of skin cells viability and partial restoration of skin structure 4 days after adalimumab delivery, as well as a strong decrease in S100A7 expression in the epidermis suprabasal layers. Furthermore, analysis of cytokine levels in culture media showed dose-dependent downregulation of IL-17A, IL-22 and IFNγ but not IL-27 release, indicating anti-inflammatory effect of therapeutic treatment with adalimumab, a well-known anti-TNF immunosuppressive monoclonal antibody. In conclusion, we demonstrated that the InflammaSkin® model with adipose tissue successfully responds to subcutaneous injection of biologics and could become key to assess the efficacy of subcutaneously-delivered therapeutic compounds.

Background: To date, responsiveness to tumor necrosis factor alpha inhibitors in ulcerative colitis (UC) patients is not predictable. This is partially due to a lack of understanding of the underlying inflammatory processes. The aim of this study was to identify immunological subgroups of patients with UC and to test responsiveness to adalimumab in these subgroups in the mouse model of ulcerative colitis (UC), which is based on NOD/scid IL-2R gamma(null) (NSG) mice reconstituted with peripheral blood mononuclear cells (PBMCs; NSG-UC). Methods: The immunological profiles of 40 UC patients and 16 non-UC donors were determined by flow cytometric analysis of PBMCs in a snapshot and longitudinal study and analyzed by principal component, orthogonal partial least square discrimination (oPLS-DA), and hierarchical clustering analysis. NSG mice were reconstituted 5 times at consecutive time points with PBMCs from a single donor and were analyzed for frequencies of human leukocytes and histological phenotype. The response to adalimumab of 2 identified subgroups was tested in the NSG-UC model. We used the clinical, colon, and histological score, serum levels of glutamic and aspartic acid, and IL-6 and IL-1 beta. Response was analyzed by oPLS-DA. Results: Analysis revealed a distinction between UC and non-UC donors. Hierarchical clustering identified 2 major subgroups in UC patients. Group I was characterized by TH17 and M1 monocytes, group II by TH2/TH1, and switched B cells. These subgroups reflect the dynamics of inflammation as patients. NSG-UC mice achieved an immunological phenotype reflecting the patient's immunological phenotype. oPLS-DA revealed that NSG-UC mice reconstituted with PBMCs from group II responded better to adalimumab. Conclusions: The combination of profiling and testing of therapeutics in the NSG-UC model may lead to individualized and phase-dependent therapies.

To evaluate in an interventional trial on knee osteoarthritis (OA) the level and change of two serum biomarkers and their correlation with imaging parameters. The previously reported interventional OA study (ClinicalTrials.gov: NCT00536302) identified a positive effect of collagen hydrolysate (CH) on cartilage morphology in patients with knee OA using delayed gadolinium enhanced magnetic resonance imaging (dGEMRIC). It was the objective in this research project to evaluate in an interventional clinical trial on knee OA the level and change of two serum biomarkers and their correlation with imaging parameters. In blood samples of study participants, we determined the concentration of procollagen type II N-terminal propeptide (PIIANP) and aggrecan chondroitin sulfate 846 epitope (CS846) at baseline (BL) and at the follow-up (FU) visits at 24 and 48 weeks. We measured the level and change of biomarker concentrations in both study groups, and the correlation of those changes with changes in dGEMRIC. For the biomarker PIIANP, we observed a significantly greater increase in the CH group (29.9% vs. 1.2% at week 24, P= 0.001). For CS846, the mean concentration was lower among the CH treated participants at 24 weeks (78% vs. 96%, P= 0.045). Consistent correlations of changes in biomarkers PIIANP and CS846 with changes of the dGEMRIC score could not be observed. In this study, different changes per treatment group, CH and placebo were seen for dGEMRIC and PIIANP BL to 24 weeks FU, but only weak correlations between changes in dGEMRIC and biochemical markers.

Purpose: TSG-6 is a protein that is not constitutively expressed in most adult tissues, but is secreted (by stromal and immune cells) in response to inflammatory mediators and is believed to play a role in protecting tissues from the damaging effects of inflammation. For example, many of the anti-inflammatory and tissue reparative properties of mesenchymal stromal cells (MSCs) have recently been attributed to TSG-6 production. Several mechanisms for this have been proposed/identified, including TSG-6's suppression of neutrophil migration; in this context the isolated recombinant Link module of human TSG-6 (Link_TSG6) is as potent as the full-length recombinant protein (rhTSG-6). Previously, TSG-6 treatment/overexpression has been found to be chondroprotective, reducing cartilage breakdown in mouse models of inflammatory arthritis, where this has been attributed, in large part, to its inhibitory effect on neutrophils. However, the effects of TSG-6 on chondrocyte function have not been studied. The aim here was to investigate whether TSG-6 can suppress the effects of pro-inflammatory cytokines on chondrocytes and test the activities of TSG-6 in models of osteoarthritis (OA). Methods: Chondrocytes in 3D pellets were treated with IL-1 or TNF with/without recombinant TSG-6 proteins (Link_TSG6 or rhTSG-6); expression of ADAMTS4, ADAMTS5 and MMP13 were quantified by qPCR. Human OA cartilage explants were cultured with IL-1 and oncostatin M with/without TSG-6 for 7 or 14 days and proteoglycan release measured using the dimethylene blue assay. A rat surgically-induced model of OA (ACLTpMx) was treated with intra-articular injections of Link_TSG6 protein at 7-, 14- and 21-days after surgery; joint damage was evaluated at the 4-week endpoint by macroscopy and histology, and pain was assessed by tactile allodynia with von Frey hairs. TSG-6 protein was immunolocalised in cartilage obtained from OA patients. Results: Treatment with Link_TSG6 and rhTSG-6 proteins suppressed the response of MSC-derived chondrocytes (in 3D pellet cultures) to inflammatory cytokines (IL-1 and TNF), inhibiting their expression of ADAMTS4, ADAMTS5 and MMP13; i.e. the aggrecanse and collagenase enzymes responsible for cartilage degradation in OA. Link_TSG6 was generally more potent than rhTSG-6 and, for example, was able to completely abolish the expression of MMP13 in response to IL-1β. Link_TSG6 significantly reduced proteoglycan loss from cytokine-stimulated human cartilage explants obtained from OA patients (e.g. a reduction of 31.0±6.3% over 7 days with a 1 μM dose when data are pooled for 7 donors); rhTSG-6 had a small (but not significant) inhibitory effect. Intra-articular administration of Link_TSG6 in a rat model of OA resulted in a dose-dependent (and significant) reduction in cartilage damage (e.g. fibrillation and ulceration). For example, at the highest dose tested only 1 animal (out of 18) had exposed bone and 5 had intact cartilage as compared to 5 and 1 animals, respectively, in the vehicle control group of 20 rats. Link_TSG6 treatment also significantly reduced pain. High levels of TSG-6 staining in OA cartilage were associated with the cells and matrix in the most damaged margins of the tissue, but staining was also detectable in a low percentage of cells within the deep and mid zones of macroscopically intact regions; this localisation pattern is consistent with TSG-6 expression being a response to tissue disruption/disease, which might serve to limit excessive matrix turnover. Conclusions: These results indicate that Link_TSG6 has the potential to be developed as a biological therapy for osteoarthritis. The chondroprotective activity of TSG-6 combined with its anti-inflammatory and anti-bone-resorptive properties make it a unique therapeutic target.

OBJECTIVE:To evaluate the extent to which the current designs of clinical trials in knee osteoarthritis (OA) permit detection of a therapeutic effect of disease-modifying OA drugs (DMOADs) on the incidence of knee replacement, and to provide estimates of the required sample sizes. METHODS:We selected distinct subcohorts of the Osteoarthritis Initiative (OAI), based on available information on eligibility criteria for clinical knee OA trials (ClinicalTrials.gov) and additional subcohorts stratified for age, sex, and the severity of radiographic OA. The observed incidence of knee replacement in these OAI subcohorts was used to estimate the expected incidence of knee replacement in the control group of a clinical trial. Based on this estimate, the sample sizes required to detect hypothetical treatment effects on the incidence of knee replacement were calculated, assuming observation periods of 2, 5, or 7 years. RESULTS:The cumulative knee replacement incidence rates in the OAI subcohorts ranged from 0.9% to 12.9%. The corresponding sample sizes required to detect 50% improvement by the DMOAD, with a power of 80% and 95% confidence, were 5,459 and 362, respectively. Including only women with advanced age and radiographic OA increased the incidence of knee replacement and decreased the required sample size. CONCLUSION:The sample sizes that are commonly used in clinical trials do not enable the effects of a DMOAD on incident knee replacement to be detected with sufficient power and confidence. The estimated incidence rates of knee replacement and the corresponding sample sizes are important for informing the design of trials for disease course-modifying effects as well as for socioeconomic evaluation of a DMOAD in terms of preventing knee replacement.

Near-infrared fluorophore (NIRF)-labeled imaging probes are becoming increasingly important in bio-molecular imaging applications, that is, in animal models for tumor imaging or inflammation studies. In this study we showed that the previously introduced chemical concept of ‘Reverse Design’ represents an efficient strategy for the generation of selective probes for cysteine proteases from chemically optimized protease inhibitors for investigations in proteomic lysates as well as for in vivo molecular imaging studies. The newly developed activity-based probe AW-091 was demonstrated to be highly selective for cathepsin S in vitro and proved useful in monitoring cysteine cathepsin activity in vivo, that is, in zymosan-induced mouse model of inflammation. AW-091 showed higher signal-to-background ratios at earlier time points than the commercially available polymer-based ProSense680 (VisEn Medical) and thus represents an efficient new tool for studying early proteolytic processes leading to various diseases, including inflammation, cancer, and rheumatoid arthritis. In addition, the fluorescent signal originating from the cleaved AW-091 was shown to be reduced by the administration of an anti-inflammatory drug, dexamethasone and by the cathepsin inhibitor E-64, providing a valuable system for the evaluation of small-molecule inhibitors of cathepsins.

European Journal of PainVolume 10, Issue S1 p. S29c-S30 98 MICROARRAY-BASED GENE EXPRESSION PROFILING OF DRG SAMPLES FROM TWO MOUSE STRAINS WITH DIFFERENT SUSCEPTIBILITY TO NEUROPATHIC PAIN M. Gebauer, M. Gebauer Genomic Sciences, Sanofi-Aventis Deutschland GmbH, Frankfurt am Main, GermanySearch for more papers by this authorS. Jordan, S. Jordan Genomic Sciences, Sanofi-Aventis Deutschland GmbH, Frankfurt am Main, GermanySearch for more papers by this authorC. Metz-Weidmann, C. Metz-Weidmann Genomic Sciences, Sanofi-Aventis Deutschland GmbH, Frankfurt am Main, GermanySearch for more papers by this authorA.M. Schulte, A.M. Schulte Genomic Sciences, Sanofi-Aventis Deutschland GmbH, Frankfurt am Main, GermanySearch for more papers by this authorD. Ding-Pfennigdorff, D. Ding-Pfennigdorff Therapeutic Department Thrombosis and Angiogenesis, Sanofi-Aventis Deutschland GmbH, Frankfurt am Main, GermanySearch for more papers by this authorE. Bartnik, E. Bartnik Therapeutic Department Thrombosis and Angiogenesis, Sanofi-Aventis Deutschland GmbH, Frankfurt am Main, GermanySearch for more papers by this authorM. Michaelis, M. Michaelis Therapeutic Department Thrombosis and Angiogenesis, Sanofi-Aventis Deutschland GmbH, Frankfurt am Main, GermanySearch for more papers by this author M. Gebauer, M. Gebauer Genomic Sciences, Sanofi-Aventis Deutschland GmbH, Frankfurt am Main, GermanySearch for more papers by this authorS. Jordan, S. Jordan Genomic Sciences, Sanofi-Aventis Deutschland GmbH, Frankfurt am Main, GermanySearch for more papers by this authorC. Metz-Weidmann, C. Metz-Weidmann Genomic Sciences, Sanofi-Aventis Deutschland GmbH, Frankfurt am Main, GermanySearch for more papers by this authorA.M. Schulte, A.M. Schulte Genomic Sciences, Sanofi-Aventis Deutschland GmbH, Frankfurt am Main, GermanySearch for more papers by this authorD. Ding-Pfennigdorff, D. Ding-Pfennigdorff Therapeutic Department Thrombosis and Angiogenesis, Sanofi-Aventis Deutschland GmbH, Frankfurt am Main, GermanySearch for more papers by this authorE. Bartnik, E. Bartnik Therapeutic Department Thrombosis and Angiogenesis, Sanofi-Aventis Deutschland GmbH, Frankfurt am Main, GermanySearch for more papers by this authorM. Michaelis, M. Michaelis Therapeutic Department Thrombosis and Angiogenesis, Sanofi-Aventis Deutschland GmbH, Frankfurt am Main, GermanySearch for more papers by this author First published: 13 January 2012 https://doi.org/10.1016/S1090-3801(06)60101-8Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat No abstract is available for this article. Volume10, IssueS1September 2006Pages S29c-S30 RelatedInformation

The paper provides evidence that transforming growth factor-beta activated kinase 1 (TAK1, MEKK7), a downstream mediator of IL-1β signal transduction, plays an important role in the regulation of catabolic events and inflammatory processes in the context of degenerative joint diseases. We investigated the expression of TAK1 in human articular chondrocytes and in the murine growth plate by cDNA array, quantitative RT-PCR and immunohistochemistry, respectively. The human chondrosarcoma cell line SW1353 was stimulated with the proinflammatory cytokine IL-1β. The subsequent expression of proteolytic enzymes and proinflammatory cytokines was quantified. TAK1 specific siRNA was used to study the influence of TAK1 downregulation on the expression of MMP-13, MMP1 and TNF-α. As a result we demonstrated the expression of TAK1 in normal and osteoarthritic human articular cartilage. Expression of TAK1 in the hypertrophic zone of the growth plate gave us a first evidence for a catabolic function of TAK1 concerning cartilage metabolism. By gene suppression with RNAi technology we could show that TAK1 downregulation leads to a 60–70% reduced release of TNF-α, a 40–50% reduced release of MMP13, and a 20–30% reduction of MMP1 release. As TNF-α is a main player in inflammatory processes, and MMP13 is one of the major proteases involved in cartilage degradation, our results suggests that TAK1 has an important regulatory role in the context of degenerative joint diseases and thus is an attractive drug target in attempts to reduce inflammation and suppress structural changes in OA induced by IL-1β.