Mast Cell Stabilizer Research Articles

Non-Nitrogen-Containing Bisphosphonates Prevent Pyrophosphorylation of Exocytosis Proteins.

Clodronate, a non-nitrogen-containing bisphosphonate (non-NBP), is intracellularly converted into non-hydrolyzable ATP analogs. Clodronate and its analogs impair normal cell functions, including the exocytosis process. However, how this occurs in mast cells is still not well characterized. To summarize the possible mechanisms of clodronate-mediated exocytosis inhibition in mast cells. Non-NBPs display several possible mechanisms of exocytosis inhibition in various cell types, including vesicular nucleotide transporter (VNUT) and purinergic receptor inhibition. Inhibition of purinergic receptors has been shown in mast cells, but VNUT inhibition remains to be confirmed. Inhibition of protein prenylation by non-NBPs has also been shown; however, direct evidence of non-NBPs in prenylated exocytosis proteins is still contradictory. Finally, non-NBPs may inhibit mast cell exocytosis via impairment of protein pyrophosphorylation. This mechanism is less studied, and direct evidence of the involvement of pyrophosphorylated proteins in exocytosis is still lacking. Non-NBPs may affect mast cell exocytosis by interacting with purinergic receptors or VNUT or by preventing post-translational modifications of exocytosis protein(s), i.e., prenylation and pyrophosphorylation. The latter needs further investigation to provide direct evidence of a role for non- NBPs.

Mast cells promote viral entry of SARS-CoV-2 via formation of chymase/spike protein complex

The pulmonary pathological findings associated with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) result from the release of multiple proinflammatory cytokines, which causes the subsequential damage of the lungs. The current study was undertaken to investigate the responses of mast cells to viral inoculation and their contribution to host defenses from the point of view of viral entry. Pseudovirions, in which the spike glycoprotein of SARS-CoV-2 was incorporated, triggered activation of mast cells, and a mast cell-derived chymase, MCP2, formed a complex with spike protein, which promoted protease-dependent viral entry. According to the quantification results of viral entry, 10 μM quercetin, a mast cell stabilizer, potentially potently inhibited 41.3% of viral entry, while 100 μM chymostatin, which served as a chymase inhibitor, suppressed 52.1% of viral entry, compared to non-treated cells. Study using mast cell-deficient mice showed that the absence of mast cells may influence early viral loading in the upper respiratory tract, which consequently increases the risk of viral invasion into the lower respiratory system. Furthermore, mast cell-deficient mice exhibited ongoing infection in the late phase post-viral inoculation, while clearance of virus-positive cells was observed in wild-type mice. In conclusion, mast cells act as a multifaceted immune modulator that is equipped with both protective effects and pathogenic influences on viral entry of SARS-CoV-2. The utility of mast cell stabilizers and chymase inhibitors in the treatment of SARS-CoV-2-induced acute respiratory syndrome should be optimized regarding the infection stage and the risk of cytokine storm.

Evaluation of Toxicity and Efficacy of Inotodiol as an Anti-Inflammatory Agent Using Animal Model.

Chaga mushroom (Inonotus obliquus) comprises polyphenolic compounds, triterpenoids, polysaccharides, and sterols. Among the triterpenoid components, inotodiol has been broadly examined because of its various biological activities. The purpose of this study is to examine inotodiol from a safety point of view and to present the potential possibilities of inotodiol for medical usage. From chaga mushroom extract, crude inotodiol (INO20) and pure inotodiol (INO95) were produced. Mice were treated with either INO20 or INO95 once daily using oral administration for repeated dose toxicity evaluation. Serum biochemistry parameters were analyzed, and the level of pro-inflammatory cytokines in the serum was quantified. In parallel, the effect of inotodiol on food allergic symptoms was investigated. Repeated administration of inotodiol did not show any mortality or abnormalities in organs. In food allergy studies, the symptoms of diarrhea were ameliorated by administration with INO95 and INO20. Furthermore, the level of MCPT-1 decreased by treatment with inotodiol. In this study, we demonstrated for the first time that inotodiol does not cause any detrimental effect by showing anti-allergic activities in vivo by inhibiting mast cell function. Our data highlight the potential to use inotodiol as an immune modulator for diseases related to inflammation.

α-Asarone alleviates allergic asthma by stabilizing mast cells through inhibition of ERK/JAK2-STAT3 pathway.

Asthma is a heterogeneous disease related to numerous inflammatory cells, among which mast cells play an important role in the early stages of asthma. Therefore, treatment of asthma targeting mast cells is of great research value. α-Asarone is an important anti-inflammatory component of the traditional Chinese medicine Acorus calamus L, which has a variety of medicinal values. To investigate whether α-asarone can alleviate asthma symptoms and its mechanism. In this study, we investigated the effect of α-asarone on mast cell activation in vivo and in vitro. The release of chemokines or cytokines, AHR (airway hyperresponsiveness), and mast cell activation were examined in a mast cell-dependent asthma model. Western blot was performed to determine the underlying pathway. α-Asarone inhibited the degranulation of LAD2 (laboratory allergic disease 2) cells and decreased IL-8, MCP-1, histamine, and TNF-α in vitro. α-Asarone reduced paw swelling and leakage of Evans blue, as well as serum histamine, CCL2, and TNF-α in vivo. In the asthma model, α-asarone showed an inhibitory effect on AHR, inflammation, mast cells activation, infiltration of inflammatory cells, and the release of IL-5 and IL-13 in lung tissue. α-Asarone decreased the levels of phosphorylated JAK2, phosphorylated ERK, and phosphorylated STAT3 induced by C48/80. Our findings suggest that α-asarone alleviates allergic asthma by inhibiting mast cell activation through the ERK/JAK2-STAT3 pathway.

LB975 The role of gut flora metabolite butyrate in inhibiting mast cell activation via deacetylase in chronic spontaneous urticaria

Chronic spontaneous urticaria (CSU) is an immune-related skin disease characterized by abnormal activation of mast cells (MCs). With the advanced research on the gut flora in immune-related diseases, it is suggested that dysbiosis of the gut flora may contribute to the pathogenesis of CSU, but the mechanisms involved are unknown. Butyrate, a metabolite of the gut flora, is involved in a variety of cellular physiological processes as a histone deacetylase inhibitor (HDACi) and exerts beneficial effects on the intestinal tract and other organs.

Mast cell activation syndrome and the link with long COVID

Mast cells are innate immune cells found in connective tissues throughout the body, most prevalent at tissue-environment interfaces. They possess multiple cell-surface receptors which react to various stimuli and, after activation, release many mediators including histamine, heparin, cytokines, prostaglandins, leukotrienes and proteases. In mast cell activation syndrome, excessive amounts of inflammatory mediators are released in response to triggers such as foods, fragrances, stress, exercise, medications or temperature changes. Diagnostic markers may be difficult to assess because of their rapid degradation; these include urinary N-methyl histamine, urinary prostaglandins D2, DM and F2α and serum tryptase (which is stable) in the UK. Self-management techniques, medications and avoiding triggers may improve quality of life. Treatments include mast cell mediator blockers, mast cell stabilisers and anti-inflammatory agents. ‘Long COVID’ describes post-COVID-19 syndrome when symptoms persist for more than 12 weeks after initial infection with no alternative diagnosis. Both mast cell activation syndrome and long COVID cause multiple symptoms. It is theorised that COVID-19 infection could lead to exaggeration of existing undiagnosed mast cell activation syndrome, or could activate normal mast cells owing to the persistence of viral particles. Other similarities include the relapse-remission cycle and improvements with similar treatments. Importantly, however, aside from mast cell disorders, long COVID could potentially be attributed to several other conditions.

Pyridoxine Synergistically Potentiates Mast Cell-Stabilizing Property of Ascorbic Acid.

Besides their physiological properties, vitamins, such as vitamin C (ascorbic acid) and B6 (pyridoxine), ameliorate the symptoms of allergic disorders. Because exocytosis in mast cells can be detected electrophysiologically by the changes in the membrane capacitance (Cm), its continuous monitoring in the presence of these vitamins would determine their mast cell-stabilizing, anti-allergic properties. Employing the whole-cell patch-clamp technique in rat peritoneal mast cells, we examined the effects of ascorbic acid and pyridoxine on the degranulation of mast cells and the increase in the Cm during exocytosis. Both ascorbic acid and pyridoxine dose-dependently suppressed the GTP-γ-S-induced increase in the Cm and inhibited the degranulation from mast cells. Surprisingly enough, relatively low concentrations of pyridoxine (1, 2 mM) synergistically enhanced the suppressive effect of 2 mM ascorbic acid on mast cell degranulation. These results provided electrophysiological evidence for the first time that ascorbic acid and pyridoxine inhibited the process of exocytosis in a dose-dependent manner. At relatively lower concentrations, these vitamins were not enough to stabilize mast cells. However, such concentrations of pyridoxine synergistically potentiated the mast cell-stabilizing property of ascorbic acid.

Evaluation of Mast Cell Stabilizing, Anti-Inflammatory, and Anti-Oxidant Activity of Seed Extracts of Saraca Asoka (Roxb.), De. Wild

Saraca Asoka (Caesalpiniaceae) is a medicinal plant used traditionally for the treatment of various diseases. There is no scientific evidence for the anti-asthmatic activity of Saraca Asoka. Preliminary phytochemical screening of extract has revealed the presence various phytochemical components such as flavonoids, tannins, saponins, carbohydrates, phenols, glycosides, and fixed oils and fats. However, no alkaloids, proteins and amino acids were found in the extracts. The present study was evaluated for phytochemical screening, mast cell stabilizing, anti-inflammatory, and antioxidant activity of the methanolic extract of Saraca Asoka. In the present study, DPPH radical scavenging activity was highest in methanol extract (94.5 % ± 1.8%) of Saraca Asoka seeds. The in vivo anti-inflammatory activity was evaluated in rats using carrageenan-induced paw edema and in vitro antioxidant activity was performed by 1, 1-diphenyl-2-picryl-hydrazyl (DPPH) and ABTS. Quantitative estimation of total polyphenolic content of the (SA-ME) was estimated by Folin-Ciocalteu method. SA-ME (200 mg/kg body wt.) significantly decreased paw volume, after oral administration of SA-ME in carrageenan and formaldehyde injection. SA-ME also exhibit significant antioxidant activity. Total polyphenolic content was found to be (179 ± 0.27 mg/ml) and exhibited highest flavonoid content (8.42 ± 0.25 mg/ml). These results show that the methanol extract of Saraca Asoka seeds shows potential mast cell stabilizing, anti-inflammatory and anti-oxidant activity.

Abstract 6382: Targeting mast cells restores T cell infiltration and sensitizes sarcomas to PD-L1 inhibition

Abstract Despite recent advances in cancer research, developments in sarcoma treatment have been slow. The efficacy of chemotherapy in sarcoma is limited by the pathophysiology of surrounding tumor stroma. Abnormalities in the tumor microenvironment (TME) result in stiffening of the tumor and accumulation of mechanical stresses which in turn cause the extreme deformation and collapse of intratumoral vessels. Strategies to induce TME reprogramming with the aim to restore mechanical properties of the tissue as well as tumor blood vessel functionality involve the use of “mechanotherapeutics”. Herein, we exploited the potential of the antihistamine drug and mast cell stabilizer, ketotifen, to induce mechanotherapeutic effects in murine models of soft tissue and bone sarcoma. Mast cells are the first population of immune cells drawn to the tumorigenic nidus where they interact with multiple components of the TME. Yet, their role in tumor stroma remains enigmatic as they either promote or inhibit tumor development depending on the conditions. In agreement with other mechanotherapeutics, we found that ketotifen induces vascular normalization in a dose dependent manner as indicated by the increase in perfusion, higher open lumen fraction and pericyte coverage of the vessels. Moreover, in vivo ultrasound shear wave elastography confirmed the mechanotherapeutic properties of ketotifen in reducing tissue stiffness, while further ex vivo validation indicated that TME reprogramming is mediated by a significant reduction in cancer associated fibroblast and inhibition of collagen synthesis as well as of that of other extracellular matrix components. Furthermore, we found that ketotifen modulates the immune microenvironment to promote immunostimulation. Immune stimulation was mediated by both intratumoral T cell accumulation and higher cytotoxic to regulatory T cell ratio concomitantly with a pronounced reduction in tissue hypoxia and upregulation of leukocyte adhesion molecules on endothelium. As a result of ketotifen-induced TME reprogramming, the addition of ketotifen to doxorubicin-aPDL1 combination treatment exhibited therapeutic superiority as opposed to doxorubicin-aPDL1, offering a durable tumor remission and immunological memory following tumor rechallenge experiments. Taken together, our findings demonstrate that ketotifen has a dual role in shaping the TME: i) reducing intratumoral physical forces to improve perfusion and ii) establishing favorable immunogenic conditions. Therefore, incorporating such agents in clinical practice holds great promise in the treatment of refractory, advanced sarcomas. Citation Format: Myrofora Panagi, Fotios Mpekris, Chrysovalantis Voutouri, Christina Michael, Anastasia Constantinidou, John D. Martin, Triantafyllos Stylianopoulos. Targeting mast cells restores T cell infiltration and sensitizes sarcomas to PD-L1 inhibition [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6382.

Green Liquid Chromatography Method for the Determination of Related Substances Present in Olopatadine HCl Nasal Spray Formulation, Robustness by Design Expert.

Dual therapeutic nature drug mast cell stabilizer and histamine receptor antagonist olopatadine hydrochloride (OPT) nasal spray does not have an official monograph, and no literature is available. Eye drops formulation had the official monograph for impurities, but the determination was done in two methods. A simple and effective green liquid chromatography method to develop and validate for the related substances of OPT nasal spray formulation. A 25 min gradient method was employed to separate impurities and OPT with a 1.0 mL/min flow rate using a Boston green C8 (150 mm × 4.6 mm, 5 µm) HPLC column. The set wavelength and column oven temperatures were 299 nm and 30°C, respectively. pH 3.5 phosphate buffer-acetonitrile in the ratio of (70:30, v/v) as mobile phase A and (50:50, v/v) ratio as mobile phase B. A Quality by Design (QbD) based Design of Experiments (DoE) was employed to evaluate the robustness characteristics of the analytical method validation. The obtained RSD from the precision and intermediate precision was 0.4 to 4.1%. The % recovery of the impurities from LOQ to 150% of specification level was 87.5 to 110.3%. The linear regression curves for the impurities with a correlation coefficient of >0.999 indicate that all peak responses are linear with the concentration. The sample and standard solutions were stable for 24 h at benchtop and refrigerator conditions. All the critical peaks were well separated from the forced degradation studies' diluent, placebo, and generated degradation peaks. The method validation data and QbD based robustness study results indicate that the developed impurities method fits the routine quality control laboratory use. National Environmental Index (NMEI), Green Analytical Procedure Index (GAPI), Analytical Eco-scale and Analytical Greenness (AGREE) tools expressed the method's greenness. The proposed method is QbD utilized and green chemistry assessed impurities determination method for OPT in nasal spray formulation.

Contact Allergens in Over-the-Counter Topical Antihistamine and Mast Cell Stabilizer Ophthalmic Agents.

Contact Allergens in Over-the-Counter Topical Antihistamine and Mast Cell Stabilizer Ophthalmic Agents.

WITHDRAWN: Elevated histamine etiology model for most major vaccine associated adverse events including SARS-CoV-2 spike vaccines

Vaccinees experience no adverse events, mild adverse events, multiple adverse events, or serious adverse events post vaccination. Many of these vaccine adverse events occur with different vaccines with different occurrence frequencies. Many of these adverse events are generally considered as associated with immune responses to the active vaccine components (antigens) and/or to possibly one or more of the vaccine excipients. Most of these vaccine adverse events are self-limiting and resolve within days. Many of these adverse events symptoms overlap symptoms associated with elevated histamine levels. Based on these observations, the hypothesis that the majority of vaccine associated reactogenicity adverse events are caused by temporal histamine intolerance in vaccinees is proposed. This hypothesis is based on a model of innate immune responses releasing a surge of inflammatory molecules including histamine; this surge is hypothesized to exceed the normal histamine tolerance level for vaccinees with reactogenicity adverse events. Further, these symptoms resolve as histamine levels fall below the vaccinee's tolerance threshold. This model can be evaluated by the detection of elevated histamine levels in vaccinees corresponding to timing of symptoms onset. If confirmed, a direct consequence of this model predicts that some antihistamine treatments, mast cell stabilizers, and possibly diamine oxidase enzyme may reduce the incidence or severity of adverse events experienced by vaccinees post vaccinations for most or all high reactogenicity vaccines including coronavirus disease 2019 (COVID-19) Spike vaccines.

Anti-pseudo-allergic components in licorice extract inhibit mast cell degranulation and calcium influx.

Pseudo-allergic reactions (PARs) widely occur upon application of drugs or functional foods. Anti-pseudo-allergic ingredients from natural products have attracted much attention. This study aimed to investigate anti-pseudo-allergic compounds in licorice. The anti-pseudo-allergic effect of licorice extract was evaluated in rat basophilic leukemia 2H3 (RBL-2H3) cells. Anti-pseudo-allergic compounds were screened by using RBL-2H3 cell extraction and the effects of target components were verified further in RBL-2H3 cells, mouse peritoneal mast cells (MPMCs) and mice. Molecular docking and human MRGPRX2-expressing HEK293T cells (MRGPRX2-HEK293T cells) extraction were performed to determine the potential ligands of MAS-related G protein-coupled receptor-X2 (MRGPRX2), a pivotal target for PARs. Glycyrrhizic acid (GA) and licorice chalcone A (LA) were screened and shown to inhibit Compound48/80-induced degranulation and calcium influx in RBL-2H3 cells. GA and LA also inhibited degranulation in MPMCs and increase of histamine and TNF-α in mice. LA could bind to MRGPRX2, as determined by molecular docking and MRGPRX2-HEK293T cell extraction. Our study provides a strong rationale for using GA and LA as novel treatment options for PARs. LA is a potential ligand of MRGPRX2.

Inhibition effect of PPAR-γ signaling on mast cell-mediated allergic inflammation through down-regulation of PAK1/ NF-κB activation

As a ligand-activated transcription factor, peroxisome proliferator-activated receptor gamma (PPAR-γ) plays a crucial role in allergic inflammation. Recently, the nuclear factor kappa B (NF-κB) pathway and PAK1 have been indicated to be associated with allergic diseases. However, the effect of PPAR-γ on NF-κB and PAK1 production in food allergies is not known. This study aims to 1) systematically validate that the activation of PPAR-γ attenuates allergic reactions and 2) elucidate the mechanism by which PPAR-γ regulates mast cell degranulation. Brown Norway rats were separated into control, ovalbumin, ovalbumin + rosiglitazone, and ovalbumin + GW9662 groups. In vivo experiments demonstrated that rosiglitazone administration markedly inhibited the clinical symptoms and the serum levels of immunoglobulins E and G1. In addition, cytokine release was regulated by activated PPAR-γ and characterized by increased levels of IFN-γ and decreased levels of IL-4, IL-5, and TNF-α. Our data showed that activated PPAR-γ has the potential to alleviate food allergies by enhancing intestinal mucosal integrity and tight junctions. Moreover, we found that PPAR-γ activation inhibited mast cell degranulation both in vivo and in vitro. Our in vitro findings also showed that the activated PPAR-γ signal could inhibit PAK1 phosphorylation and the expression of p65. Furthermore, the interaction between p65 and p-PAK1 during ovalbumin treatment was attenuated after PPAR-γ activation. Collectively, these results demonstrate that PPAR-γ is an important regulator of mast cell degranulation and the Th2-type response, which sheds new light on the importance of PPAR-γ in food allergies.

A mast cell stabilizer attenuates pathological angiogenesis in a model of choroidal neovascularization

Age‐related macular degeneration (AMD) is one of the leading causes of irreversible blindness in the elderly worldwide. Wet AMD is characterized by pathologic choroidal neovascularization that invades the RPE and compromises retinal function. Although anti‐vascular endothelial growth factor (anti‐VEGF) therapy has been shown to improve symptoms of wet AMD in a large number of people, long‐term treatment is associated with the development of geographic atrophy. The development of new therapeutic alternatives to prevent the progression of exudative AMD is still necessary. Inflammation and deterioration of the RPE are considered to be the main sources of the pathogenesis of AMD. Recent studies have reported accumulation and degranulation of mast cells in the compromised choroid of patients with AMD. When activated, mast cells can secrete a variety of biologically active mediators, including inflammatory cytokines and proteolytic enzymes such as tryptase. However, the precise role of mast cells in the pathogenesis of AMD remains largely unknown. We are interested in how mast cells participate in the initiation of inflammation in the choroid which results in pathological choroidal neovascularization. We investigated the effect of a mast cell stabilizer on angiogenic response in the choroid. The angiogenic effect of a mast cell stabilizer, ketotifen fumarate, was evaluated in laser‐induced CNV model. Mice received intraperitoneal injections of ketotifen fumarate (5 and 25 mg/kg/d) or vehicle. CNV area and mononuclear phagocytes were determined respectively by using lectin staining and an anti–IBA‐1 antibody on RPE/choroid flat mounts. CNV was attenuated by ketotifen fumarate with no effect on inflammatory cell recruitment. Conditioned media from activated peritoneal mast cells exerted proangiogenic effects on vascular sprouting. Tryptase increased choroidal endothelial cell migration and choroidal vascular sprouting. The study supports possible modulation of mast cells as a therapeutic modality for CNV.

Ligation of Siglec-9 inhibits FcɛRI-dependent mediator release from human mast cells

Abstract Mast cell activation is critical for the development of allergic diseases. Ligation of Sialic acid-binding immunoglobin-like lectins (Siglecs) such as Siglec-7 and 8 has been shown to inhibit mast cell activation and is considered one of the main approaches to reduce mast cell contribution to allergic disease. Recent high throughput studies provided evidence that human mast cells express Siglec-9, an inhibitory receptor that is mainly expressed by innate immune cells such as neutrophils, monocytes and eosinophils. Based on this evidence, we aimed to examine Siglec-9 expression and function in human mast cells. Flow cytometry analysis showed that Siglec-9 is expressed in the human mast cell lines LAD2, LUVA, and HMC-1, and in peripheral blood-derived cultured human mast cells (PBCMCs). The expression of Siglec-9 in PBCMCs peaks at week 5 of culture and correlates positively with the expression of the high affinity receptor for IgE (FcɛRI). Pre-treatment of human mast cells with the Siglec-9 agonists glycophorin A or high molecular weight hyaluronic acid (HMW-HA) followed by FcɛRI-dependent stimulation has an inhibitory effect on mast cell degranulation. SIGLEC9 gene disruption by CRISPR/Cas9 editing resulted in a significant reduction in Siglec-9 expression in LAD2 cells that also became impervious to inhibition by Siglec-9 agonists. Together, our data shows that human mast cells express Siglec-9 and that engaging this inhibitory receptor can reduce mast cell degranulation.

Role of Mast Cells in HIV-associated Chronic Widespread Pain

Chronic widespread pain (CWP) in people with HIV-1 (PWH) is associated with a high rate of disability and low quality of life, with prevalence estimates ranging from 25%-85%. However, the specific mechanisms that contribute to CWP in HIV are not understood. Previously we demonstrated that PWH with CWP have increased hemolysis, elevated plasma levels of cell-free heme, and attenuated levels of the heme degrading enzyme, heme oxygenase 1 (HO-1). We also found that high cell-free heme and low HO-1 were associated with hypersensitivity in animals. In this study, we hypothesized that high heme and low HO-1 activates mast cells leading to degranulation and release of pain mediators like, histamine and bradykinin in HIV. PWH who self-report CWP were recruited from the University of Alabama at Birmingham HIV clinic. Animal models used in the study included, C57BL/6 mice subjected to hemolysis by intraperitoneal injection of phenylhydrazine hydrochloride (PHZ), HO-1 knockout mice, and HIV-1 transgenic rats overexpressing HIV-1 proteins. Cellular studies were performed in MC/9 mast cells. Human findings show that PWH with CWP have elevated plasma levels of histamine and bradykinin. HIV-1 transgenic rats also have elevated plasma levels of histamine and bradykinin and are hypersensitive as estimated by a decrease in withdrawal threshold to von Frey stimulation. In addition, data in mice with elevated plasma levels of free-heme, or decreased expression of HO-1, show significantly elevated levels of the pain mediators, histamine and bradykinin. Cellular data shows that high heme or low HO-1 induces mast cell degranulation and the release of histamine and bradykinin. Together, these data suggest that heme-dependent mast cell degranulation and the release of pain mediators may be contributing CWP in HIV. Future studies will focus on determining strategies to stabilize mast cell membrane in HIV and reduce hypersensitivity. Grant Number: RO1DA049657 Grant Name: Chronic Widespread Pain in HIV: Novel Mechanisms and Therapeutics.

Ocular surface mast cells promote inflammatory lymphangiogenesis.

Mast cells, sentinel immune cells, are most abundantly expressed in vascularized tissues that interface the external environment, such as the skin and ocular surface. Our previous reports have shown mast cells reside closely with vascular endothelial cells and mediate the pathogenic angiogenic response. However, the contribution of mast cells and their underlying mechanisms on lymphangiogenesis have not been fully deciphered. Using a murine model of inflammatory corneal angiogenesis, we observed adjacent migration of activated mast cells with new lymph vessel growth. Our in vitro co-culture assays demonstrate that mast cells express high levels of of VEGF-D and directly promote lymphatic endothelial cell tube formation and proliferation. Moreover, our loss-of-function approaches, using mast cell knockout mice and cromolyn-mediated mast cell inhibition, showed mast cell deficiency suppresses the induction of inflammatory lymphangiogenesis and VEGF-D expression at the ocular surface following corneal tissue insult. Our findings suggest blockade of mast cells as a potential therapeutic strategy to inhibit pathological lymphangiogenesis.

Mast Cells Released Inflammatory Markers Play a Vital Role in Mandibular Remodeling of Odontogenic Keratocyst Patients

Background: Odontogenic Keratocyst (OKC) falls among the distinctive and rare odontogenic developmental cyst which is derivative of the dental lamina and contains clear fluid along with a cheesy material which resembles keratin debris. Treatment strategies for OKC are for the most part classified as conservative or aggressive. Mast Cells play an important role in the pathogenesis of inflammatory conditions. Objective: The goal of our study was to observe the participation of inflammatory mediators (RANKL, TNF-α, TGF-β and MMP-9) in the pathogenesis as well as the recovery process of OKC. Methods: Enrolled patients were divided into three groups while a fourth group was formed of the control individuals. Sampling of all groups was done once after required data collection. Blood samples were taken, mRNA extraction and cDNA was then subjected to qRT-PCR. IHC for mast cells was done on patient samples. Results: qRT-PCR showed higher expressions of inflammatory markers in patients undergoing treatment as compared to individuals with completed treatment procedure. Levels of RANKL were high in patients that were currently undergoing bone development as compared to other patients. The levels of TNF-α were comparatively higher in patients who showed visible signs of inflammation like swelling and pain. TGF-β and MMP-9 expression levels showed correlation with each other in all three groups. Both these cytokines were also found to be in association with mast cells. The results of histochemical staining of mast cells suggested the involvement of these cells in the pathogenesis of the cystic lesion via inflammation. Conclusions: The findings of the study suggest that the expressions of inflammatory mediators correlate with the presence of mast cells, therefore, application of mast cells stabilizers in addition to marsupialization in the treatment of OKC can be a prospective treatment strategy in future clinical settings.

Tyrosine kinase inhibitors for the treatment of indolent systemic mastocytosis: Are we there yet?

Indolent systemic mastocytosis (ISM) is the most prevalent form of systemic mastocytosis. Many patients with ISM suffer from mast cell (MC) mediator-related symptoms. In a small number of patients, hematologic progression is seen in the follow-up. In some patients with ISM, symptoms arising from MC-derived mediators including gastrointestinal symptoms, anaphylaxis, and neuropsychiatric symptoms are kept under control with conventional mediator-targeting drugs or MC-stabilizing agents. However, in a substantial number of patients, the symptoms are refractory to such conventional therapy. For these patients, novel drugs and targeted approaches are considered. One reasonable approach may be to apply tyrosine kinase inhibitors directed against KIT and other key kinase targets expressed in neoplastic MCs in ISM. Because MCs in more than 90% of all patients with typical ISM display the KIT D816V mutant receptor, clinically effective KIT-targeting drugs have to be active against this mutant form of KIT. The 2 such most effective and well-studied agents currently available are midostaurin and avapritinib. Other KIT-targeting drugs, such as imatinib or masitinib, are less effective or even noneffective against KIT D816V and are thus recommended for use only in patients with other KIT mutant forms (noncodon 816 mutations) or with wild-type KIT. In the present article, we review the current state in the treatment of ISM with tyrosine kinase inhibitors, with special emphasis on treatment responses and potential adverse effects. In fact, all of these agents also have unique and common adverse effects, and their use to treat patients with ISM should be balanced against their toxicity and short- and long-term safety.