DIA/NORD 2012 US Conference on Rare Diseases and Orphan Products: Poster Abstracts

Abstract

DIA/NORD 2012 US Conference on Rare Diseases and Orphan Products: Poster Abstracts The accepted Poster Abstracts for the DIA/NORD 2012 US Conference on Rare Diseases and Orphan Products: Shaping the Future Now are printed below. The posters present scientific developments related to research or studies on rare diseases or public health projects. The meeting was held October 22-24, 2012 in Washington, DC. Elucidating the Mechanism of the Allergic Inflammation in Eosinophilic Esophagitis Mari Kent, Ozlem Goker-Alpan, MD, and Oral Alpan, MD 1 Laboratory of Immunopathogenesis, O&O Alpan LLC, Fairfax, VA, USA 2 James Madison University, Harrisonburg, VA, USA 3 CFCT, O&O Alpan LLC, Fairfax, VA, USA Objective: The rationale for the study is to understand the role of IgE in eosinophilic esophagitis, a rare disorder. Method: Patients with eosinophilic esophagitis (EE) were treated with a drug (omalizumab) that blocks IgE in the body. This therapy is approved for asthma but had not been used in eosinophilic esophagitis. We obtained IRB and FDA approval to test this drug to understand disease mechanism in EE. Results: This study defines improvement in patient condition with 2 variables: histology and clinical symptoms. The study defines complete histological improvement as less than 5 eosinophils/high power field at 3 months, with partial improvement being a 50% reduction in eosinophils/high power field. Total absence of vomiting, heartburn, dysphagia, nausea, and abdominal pain will be defined as complete clinical improvement, with partial improvement being a 50% reduction of symptoms or less than one symptom per week. Of the 17 participants in the study with anti-IgE therapy, 5 showed at least partial clinical improvement, indicated by self-reported symptom survey results. Symptom surveys included assessments of severity (on a 1-5 scale) and frequency of vomiting, nausea, heartburn, dysphagia, and abdominal pain. Five of the 17 participants had no noteworthy change in symptoms, though 2 of these showed partial histological improvement. Another 5 participants displayed slight clinical improvement, meaning not a 50% reduction in overall symptom occurrence but a reduction in severity or occurrence of their most severe symptom. Of these 5 participants, only 1 showed partial histological improvement; the others had no improvement. Of the 5 participants with significant clinical improvement, 3 reported partial clinical improvement and 2 reported near-complete or complete symptom resolution. Those with partial clinical improvement had histological results as follows: 1 showed complete histological improvement, and 2 had partial improvement. One of the patients showing complete clinical improvement also had complete histological improvement, and the other showed partial histological improvement. Conclusion: The mechanism by which treatment methods are evaluated for medical conditions has to vary by the prevalence of the disease. Generally, when prevalent diseases are treated according to the standard of care, laboratory documented evidence of disease and patient symptoms cease together. With a rare disease such as eosinophilic esophagitis, the process becomes more complex. Treatments falling under the standard of care for this disease often fail to produce a remarkable change in quality of life for its patients in addition to a decrease in eosinophil counts on endoscopy. Also, because there are many different ways in which eosinophilic esophagitis may present, patients’ response to the therapies traditionally applied (food avoidance, swallowed steroids) tends to be less than predictable. The task at hand, then, is finding a treatment that produces significant lessening of histological evidence of eosinophils and symptoms in any number of patients. Simultaneous resolution of eosinophil counts and symptoms is so rare that when such occurs, even in the small numbers seen with the anti-IgE therapy used in this study, the treatment warrants further investigation, as do the patients who responded to the drug. The subgroup of patients who responded to the anti-IgE therapy indicates. The evidence gathered in this study will provide a direction for future drug development or repurposing of other medications to treat eosinophilic esophagitis. In addition to developing new treatments for EE, this research will give some indication of which therapies will actually be beneficial to certain groups of patients, hopefully increasing clinical efficiency and patient satisfaction. Pyrin Localization During Integrin-Mediated Neutrophil Migration in Familial Mediterranean Fever Michelle B. Atallah, Kostas Konstantopoulos, MD, and Anjelica L. Gonzalez, PhD 1 Yale University, New Haven, CT, USA 2 Athens University Medical School, Athens, Greece Drug Information Journal 46(6) 748-758 a The Author(s) 2012 Reprints and permission: sagepub.com/journalsPermissions.nav DOI: 10.1177/0092861512461897 http://dij.sagepub.com Objective: Familial Mediterranean fever (FMF) is an autoinflammatory disease caused by mutations in the pyrin protein of neutrophils. This study aimed to compare the localization of pyrin within the neutrophils of FMF patients and healthy volunteers and to investigate its effects on the inflammatory profile. Method: Neutrophils were isolated from blood (Yale University, University of Athens) and allowed to migrate across tissue matrix proteins in cell adhesion chambers. Pyrin localization and neutrophil adhesion, polarization, and migration were analyzed, comparing FMF patient neutrophils to those of healthy volunteers. Results: Blood samples from 3 healthy human volunteers were collected at Yale University. In collaboration with the University of Athens Medical School in Athens, Greece, blood samples were also collected from 2 unrelated FMF patients with confirmed FMF-associated genetic mutations. The neutrophils from both FMF patients adhered to the glass at a rate of at least 50% less than neutrophils from healthy volunteers. A higher percentage of FMF neutrophils also became dark-phase, polarized, and activated on all 3 substrates (uncoated glass slides, and slides coated with fibrinogen or fibronectin) as compared to neutrophils from healthy volunteers. These highly activated neutrophils from FMF patients displayed a greater number of actin-rich processes, indicating more active migration. Qualitatively, time-lapse photography also showed that neutrophils from FMF patients migrated at a faster velocity than those from healthy volunteers. Fluorescent confocal microscopy using anti-pyrin antibodies revealed that FMF-associated neutrophils displayed pyrin localization in patterns distinct from those of unaffected volunteers. On uncoated glass slides, pyrin in FMF neutrophils was diffuse across the cytoplasm. On fibrinogen and fibronectin, which activate amb2 and a4b1 integrin surface receptors, respectively, pyrin colocalized with cytoplasmic F-actin. In neutrophils from unaffected individuals, pyrin localized to the microtubule organizing center on uncoated glass slides but was diffuse across the cytoplasm on fibrinogen and nuclear on fibronectin. These unaffected neutrophils did not show pyrin:actin colocalization on any substrates. Conclusion: This project is the first to examine the localization of native pyrin in human neutrophils, the effectors of familial Mediterranean fever. This physiologically relevant system offers valuable insight into the function of pyrin and possible interactions with integrin-mediated pathways. There are marked differences in the subcellular localization of pyrin between neutrophils on fibrinogenand fibronectin-coated slides. Localization of pyrin is associated with integrin signaling because pyrin colocalizes with actin only when the pyrin is mutated, and where integrin-mediated cell motility occurs. Mutations in pyrin appear to be pro-migration; compromising adhesion but significantly enhancing the ability of neutrophils to migrate towards sites of inflammation as seen during inflammatory crises of FMF. We observed that neutrophils from FMF patients adhere only weakly to slides, even when slides were coated with proteins that are present in disease states and should result in heightened adhesion. Poor adhesion contributes to actin turnover and thus facilitates cell motility. The differences in adhesion between neutrophils from patients and healthy volunteers, when combined with the observed pyrin:actin colocalization, suggest that pyrin might be involved in actin turnover. Because pyrin colocalizes with actin only on fibrinogen and fibronectin, this further indicates that integrin-specific binding to these matrix proteins elicits distinct functional neutrophil responses and has an effect on cellular adhesion and polarization. One practical application of these findings is the potential for a diagnostic assay. While analysis of genetic mutations and clinical presentation are the only current methods of diagnosis, the differences seen here at the cellular level between patients and nonpatients offer the potential to develop a cell-based assay that can more accurately predict a patient’s disease profile. Changing the Natural History of Cross-Reactive Immunologic Material (CRIM)-Negative Infantile Pompe Patients Treated With ERT Suhrad G. Banugaria, MD Duke University, Durham, NC, USA Objective: The main objective was to study the safety, efficacy, and feasibility of a clinical algorithm developed at our institution to identify CRIM-negative (CN) status in patients with infantile Pompe disease (IPD) to allow timely initiation of enzyme replacement therapy (ERT) and immune tolerance induction (ITI) in this vulnerable population. Method: CN patients were identified using mutation analysis and western blot on skin fibroblast through an IRB-approved study. These patients were treated with an ITI regimen including rituximab, methotrexate, and intravenous immunoglobulin (IVIG) in an outpatient setting at their