Biology subjects

Rothenberg, M.

Publications and source records attributed to Rothenberg, M..

6 recordsLinked to original sources

bioRxiv2025

Effects of C3aR activated mast cells in eosinophilic esophagitis

BackgroundEosinophilic esophagitis is a chronic food induced allergic inflammatory disease of the esophagus. Eosinophil-depleting antibodies have not shown significant improvement in clinical symptoms, which has turned attention to mast cells. These cells also accumulate in the esophagus of EoE at levels that correlate with symptoms, demonstrate extensive activation including degranulation, correlate and remain elevated and poised for reactivation even in EoE patients in remission as defined by reduced eosinophil levels. Herein, we examine a mechanism of mast cell activation in EoE. MethodsEsophageal mast cell degranulation was quantitated by CD63 expression by flow cytometry. Mast cell degranulation activity of esophageal biopsies was examined employing a bioassay with human CD34 progenitor cell derived primary human mast cells. Bulk and single cell RNA sequencing data of the esophagus were analyzed for complement dysregulation and mast cell properties as a function of disease state. Human esophageal biopsies were stained for C3 and analyzed for C3a and C3 protein content. Primary human mast cells and fibroblasts were further interrogated by flow cytometry, immunostaining, and RNA sequencing. ResultsEsophageal mast cells demonstrated increased CD63 expression, a measure of degranulation, in subjects with active compared with remission EoE. Esophageal biopsy lysates induced mast cell degranulation. Analysis of esophageal bulk RNA sequencing data demonstrated that the mast cell marker CPA3 strongly correlated with C3AR1. Accordingly, the level of C3a receptor on esophageal mast cells was proportional with CD63 expression. C3AR1 expression was increased in the fibrostenotic endoscopic phenotype compared to normal appearing active EoE patients. Esophageal biopsies were notably enriched in the expression of multiple complement genes (including C3AR1, C3, CFB, C1QA, C1QB, and C1QC) some of which were increased in active vs remission EoE (C3AR1, CFB, C1QA, C1QB, and C1QC). Single cell RNA (scRNA) sequencing data revealed enrichment of C3 in fibroblasts at levels higher in EoE compared with non-EoE controls. The ratio of C3a/C3 protein levels in the esophagus was increased in EoE compared to control. Mast cells and fibroblasts have increased proximity in EoE compared with non-EoE control subjects. Additionally, C3a stimulated mast cells to express increased CD117 and release multiple EoE-relevant cytokines including fibroblast active mediators including granzyme B, which is enriched in esophageal mast cells. ConclusionTaken together, our findings present evidence for the interplay of mast cells and fibroblasts via C3a in the pathogenesis of EoE.

immunology↗

bioRxiv2025

Distinct roles for thymic stromal lymphopoietin (TSLP) and IL-33 in experimental eosinophilic esophagitis

RationaleThymic stromal lymphopoietin (TSLP) and IL-33 are alarmins implicated in EoE pathogenesis by activating multiple cells including mast cells (MCs). Whether TSLP or IL-33 have a role in EoE and whether their activities are distinct requires further investigation. MethodsExperimental EoE was induced in wild type (WT) Il33-/-and Crlf2-/- mice. TSLP or IL-5 were neutralized using antibodies. Esophageal histopathology was determined by H&E, anti-Ki67, anti-CD31 and anti-MBP staining. Esophageal RNA was subjected to RNA sequencing. Bone marrow-derived MCs were activated with TSLP and IL-13 was determined (ELISA) ResultsTSLP and IL-33 were overexpressed in human and experimental EoE. Human and mouse esophageal MCs displayed the highest level of Crlf2 (TSLPR) compared to other immune cells. Crlf2-/- mice were nearly-completely protected from EoE, and TSLP neutralization resulted in decreased basal cell proliferation, eosinophilia, lamina propria thickening and vascularization. Induction of experimental EoE in Il33-/- mice resulted in reduced eosinophilia but no alterations in tissue remodeling were observed compared to WT mice. RNA sequencing revealed that TSLP regulates the expression of key genes associated with human EoE (e.g. eotaxins, Il19, Klk5, Flg, Il36rn, Il1r2) and suggest a role for TSLP in regulating IL-1 signaling, barrier integrity and epithelial cell differentiation. Experimental EoE was characterized by a MC-associated gene signature and elevated MCs. Activation of MCs with TSLP resulted in secretion of IL-13. ConclusionTSLP and IL-33 have non-redundant functions in experimental EoE. This study highlights TSLP as an upstream regulator of IL-13 and a potential therapeutic target for EoE.

immunology↗

bioRxiv2024

Identification of a Neuroimmune Circuit that Regulates Allergic Inflammation in the Esophagus

Eosinophilic esophagitis is a chronic food antigen-driven allergic inflammatory disease associated with symptoms involving the nervous system such as refractory pain. Yet, the role of the nervous system in disease pathogenesis has not received much attention. Herein, we demonstrate that allergen exposure evokes pain-like behavior in association with increased nociceptor signaling and transcriptional responses in dorsal root ganglia. NaV1.8+ sensory nerves were found traveling along the length of the esophagus, organized in distinct bundles adjacent to the basal epithelium, with beta III-tubulin+ sensory nerves distributed more distal to the lumen. Targeted deletion of Il4ra in NaV1.8+ neurons impeded allergen-induced increases in nerve innervation density. Furthermore, Il4ra-/-NaV1.8 mice had diminished allergen-induced allergic inflammation in the esophagus including eosinophilia and transcription of pro-inflammatory genes. Translational studies revealed extensive myelinated nerve innervation in the human esophagus, which was increased in patients with eosinophilic esophagitis. Taken together, these data indicate that allergic inflammation is associated with an increase in non-evoked pain, esophageal nerve density, altered sensitivity of sensory neurons, and transcriptional changes in dorsal root ganglia. These finding identify a type 2 neuroimmune circuit that involves the interplay of allergen-induced IL-4 receptor-dependent DRG responses that modify esophageal end-organ inflammatory responses.

immunology↗

bioRxiv2024

Macrophage-derived CCL24 promotes fibrosis and worsens cardiac dysfunction during heart failure

Inflammation is a significant risk factor and contributes to cardiovascular disease by driving both adaptive and maladaptive processes. Macrophages are the most abundant immune cells in the heart and play an important role in the remodeling of cardiac tissue. We have previously shown an overall protective function of resident cardiac macrophages after pressure-overloaded injury. However, a subpopulation of resident macrophages also expresses high levels of the profibrotic CC motif chemokine ligand 24 (CCL24), suggesting a dichotomous role in pressure overload-induced cardiac remodeling. Here, we report that following transverse aortic constriction CCL24 knockout (CCL24 KO) mice have improved systolic function, cardiac wall enlargement, as well as increased myocyte surface area and hypertrophy, suggesting that CCL24 disrupts compensatory hypertrophy. TAC-operated CCL24 KO mice also displayed reduced fibrosis and diminished expression of fibrotic genes, implying a pro-fibrotic role for CCL24. Indeed, CCL24 induced the proliferation and activation of primary mouse fibroblasts in a process that required CCR3, the sole G protein-coupled receptor for CCL24. Correspondingly, selective ablation of CCR3 in fibroblasts improved cardiac function and ameliorated fibrosis following pressure overload. Administration of a CCL24 blocking antibody or a CCR3 antagonist both improved cardiac function in pressure-overloaded mice, highlighting the CCL24-CCR3 axis as a potential therapeutic target for heart failure. Finally, CCL24 deficiency improved cardiac function and ameliorated fibrosis during physiological aging. Overall, these results show that macrophage-derived CCL24 aggravates fibrosis via the CCR3 receptor, leading to impaired cardiac function in acute and chronic heart failure.

immunology↗

bioRxiv2018

Overestimation of the prevalence of eosinophilic colitis with reliance on a single billing code

Eosinophilic colitis (EC) is a rare disorder characterized by eosinophilic inflammation of the colon causing diarrhea, bloody stool, nausea, constipation and abdominal pain. The Consortium for Eosinophilic Gastrointestinal Disease Research sought to undertake the first multi-center study of the EC population, but faced challenges with meeting enrollment goals that were based on initial estimates. To understand the reason for this, we performed chart review of patients with ICD codes for EC at 8 tertiary care centers. Chart review revealed that the isolated use of ICD codes overestimated EC rates in the pediatric population.

epidemiology↗

bioRxiv2018

Genetic, inflammatory, and tissue-specific factors control expression of human calpain-14

Eosinophilic esophagitis (EoE) is a chronic, food-driven allergic disease resulting in eosinophilic esophageal inflammation. We recently found that EoE susceptibility is associated with genetic variants in the promoter of CAPN14, a gene with reported esophagus-specific expression. CAPN14 is dynamically up-regulated as a function of EoE disease activity and after exposure of epithelial cells to interleukin-13 (IL-13). Herein, we aimed to explore molecular modulation of CAPN14 expression. We identified three putative binding sites for the IL-13-activated transcription factor STAT6 in the promoter and first intron of CAPN14. Luciferase reporter assays revealed that the two most distal STAT6 elements were required for the ~10-fold increase in promoter activity subsequent to stimulation with IL-13 or IL-4, and also for the genotype-dependent reduction in IL-13-induced promoter activity. One of the STAT6 elements in the promoter was necessary for IL-13-mediated induction of CAPN14 promoter activity while the other STAT6 promoter element was necessary for full induction. Chromatin immunoprecipitation in IL-13 stimulated esophageal epithelial cells was used to further support STAT6 binding to the promoter of CAPN14 at these STAT6 binding sites. The highest CAPN14 and calpain-14 expression occurred with IL-13 or IL-4 stimulation of esophageal epithelial cells under culture conditions that allow the cells to differentiate into a stratified epithelium. This work corroborates a candidate molecular mechanism for EoE disease etiology in which the risk variant at 2p23 dampens mediated CAPN14 expression in differentiated esophageal epithelial cells following IL-13/STAT6 induction of CAPN14 promoter activity.

genomics↗