Progress 04/01/24 to 03/31/25

Outputs
Target Audience:During this reporting period, the target audience includes eosinophilic patients enrolled in the study, eosinophilic gastrointestinal disease researchers, andgastroenterologistclinical practitioners. Changes/Problems:We requested a no-cost extension for the grant.Enrollingthe 20 patients proposed tocomplete Goal 2'sIgA-directed food elimination diet took longer than initially anticipated. The diet is a minimum of 8 weeks and requires a patient to do afollow-upendoscopy to ensure the diet is controlling the eosinophilia. Enrolled patients had reported wanting to start a diet after certainholidays or closer to when their insurance would allow for another endoscopy, which resulted in delays in patient recruitment for dietary management. We completed the 20 patients follow-up endoscopy at the end of March this year. What opportunities for training and professional development has the project provided?Training in eosinophilic disease pathophysiology continues to be facilitated by a primary mentor and through conference attendance (Digestive Disease Week May 2025 and Food the Main Course August 2025). In addition, further training has occurred on developing the food-specific antibody assay, including training with the bead-based coupling protocol and analysis of results. To continue training in statistics, I completed all coursework required to receive a biostatistics certificate from the Department of Family Medicine at the University of Utah (completed Spring 2025). Related to professional development, I participated in a 2-day grant writing workshop through Columbia University, SHAPR NIH grant writing boot camp, andCorporate and Foundation Relations Summer Grant Funding Workshopto improve my grant searching skills, research writing, and communication. To continue teaching development, I conducted guest lectures to dietitian Master students in the nutrition and integrative physiology department (master students) on medical nutrition therapy on lower GI disorders, irritable bowel disease, and microbiome and use of probiotics in GI disorders.? How have the results been disseminated to communities of interest?The final results from the project goals have not yet been disseminated. An oral presentation on thefood-specific antibody repertoire in esophageal secretions and serum was presented at a gastrointestinal diseases research conference. Additionally, the progress of the project and preliminary results were presented at the University of Utah GI department conference. What do you plan to do during the next reporting period to accomplish the goals?For Goal 1: Objective 4: Comparison analysis needs to be completed in this set of individuals with active EoE. Objective 5: We have begun analysis to compare food-specific antibody signals among the different sample types. Objective 6: During the next review period, co-authors will provide a critical evaluation and edits before manuscript submission of both manuscripts. For Goal 2: Objective 5: Additional analyses on IgA, IgG1, and IgG4 need to be completed prior to drafting the manuscript. Data analyses, draft of the results, and discussion sections of the manuscript are underway. During the next review period, co-authors will provide a critical evaluation and edits before manuscript submission of both manuscripts.

Impacts
What was accomplished under these goals? Goal 1: Determine FSA responses in patients with EoE to top trigger whole-food proteins in a customized bead-based assay. Objective 1: Using MagPlex microspheres 'beads' we coupled component food proteins of dairy (casein and whey), wheat (gluten and alpha amylase inhibitor), and egg (ovalbumin and ovomucoid). In addition, purchasable whole food proteins (Greer) were coupled at the same time for comparison to the component food beads. Objective 2: Sample collection of esophageal secretions, saliva, and serum is ongoing through an IRB at the University of Utah. During this reporting period, we have continued to process thesesamples for use with the custom Luminex MagPix food-specific assay. We have collected anadditional 185 individuals. Objective 4: # individuals have paired whole food and component protein food-specific antibodies measured in their esophageal secretions. We measured food reactivity with IgA, IgG1, IgG4, and IgM in both bead types to determine the breadth of antibodyresponse seen in the EoE. Objective 5: An Additional # of individuals had food-specific antibodies measured in serum. As partof this cohort, a series of individuals have paired samples of serum pre- and post-diet for comparison of food-specific antibodies with the removal of food antigen. We have done the analyses of the serum and saliva food-specific antibodies. Objective 6: With the addition of individuals in the overall cohort, we have re-drafted manuscripts planned under goal 1. We have re-drafted a manuscript describing food-specific antibodies in the three different sample types, esophageal secretions, serum and saliva, within active EoE. A separate manuscript will focus on the paired analyses of esophageal secretions and serum among patients pre- and post-diet to highlight the change in food-specific antibodies with the removal of a food antigen. Data analysis, drafting of the results, and discussion are underway for this manuscript. Goal 2: Demonstrate the efficacy of a food-specific IgA elimination diet in EoE to induce resolution and determine test thresholds for FSAs in patients with active EoE. Objective 1, 3 and 4: We have recruited and followed 21 patients through an IgA-directed elimination diet. Collecting esophageal secretionsand serum when possible, at their standard-of-care endoscopies pre- and post-diet. For this aim, we successfully met our proposed sample size. We have followed a small set of patients through additional eliminations and/or reintroductions (n=5). Objective 2: We have measured and analyzed food-specific antibodies in the esophageal secretions in the patients who completed elimination diets. Reactivity to IgA was used to dictate the patient's elimination diet. However, we also have data on food-specific IgG1 and IgG4 for the manuscript. Objective 5: Analyses on the pre- and post-food-specific IgA data is completed.

Publications

Progress 04/01/23 to 03/31/24

Outputs
Target Audience: During this reporting period, the target audience includes eosinophilic patients enrolled in the study, eosinophilic gastrointestinal disease researchers, and eosinophilic gastrointestinal disease clinical practitioners. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Training in eosinophilic disease pathophysiology is facilitated by a primary mentor and through conference attendance (Digestive Disease Week Conference). In addition, further training has occurred on developing the food-specific antibody assay, including training with the bead-based coupling protocol and analysis of results. To continue training in statistics, I took coursework required to complete the biostatistics certificate (courses: Epidemiology I and Survey of Statistical Packages) andparticipated in an R programming workshop. Related to professional development, I have participated in a 3-day K grant writing workshop through the University of Michigan, Michigan Institute for Clinical & Health Research, and the University of Utah post-doctoral career workshops, such as a cover letter workshop, to improve my research writing and communication. To enhance my mentorship skills, I participated in the fundamentals of the professional mentoring workshop through the University of Utah, Utah Coaching Advancement Network (U-CAN). To continue teaching development, I attended the University of Utah Health Educators symposium, conducted guest lectures to dietitian Master students in the nutrition and integrative physiology department (master students) on medical nutrition therapy on lower GI disorders and irritable bowel disease, and presented ongastrointestinal elimination diets at the University of Utah GI departmentconference. How have the results been disseminated to communities of interest?The final results from the projectgoals have not yet been disseminated. An abstract of the food-specific antibodies found inthe serum of EoE patients has been accepted as a posterat adisease-focused conference. What do you plan to do during the next reporting period to accomplish the goals?To complete the objectives under goal 1, I will do the following in the next reporting period: We are poised to couple component proteins of beta-lactoglobulin, casein, gluten, alpha-amylase inhibitor, ovalbumin, and ovomucoid in the next month. MagPlex microspheres "beads", coupling kit reagents, and proteins are purchased. Once the coupling is complete in the spring of 2024, we will measure the component protein-specific antibody responses in the saliva, serum, and esophageal secretions already processed and aliquoted. We willcontinueto run a pooled patient-samplestandard curvewith the food-componentproteinantibody assays. To complete the objectives under goal 2, I will do the following in the next reporting period: We will continue to recruit patients with active EoE willing to undergo dietary therapy and enroll them in the food-specific IgA-directed elimination diet. We have currently collected data on five patients.We are poised todetermine the diet for twopatients with recent collections.We expect to enroll two to three patients monthly, enabling our goal of 20 patients to undergo a directed diet and follow-upby the fall of 2024. We will continue to monitorany patients who elect to do further elimination diets or reintroductions and analyze the associated food-specific signals. We expect to have data on all FSA-directed diets as of Fall 2024; once completed, amanuscript will be draftedby the end of the following review process. We will use the pooled standard curve to interpret antibody signals, which will be included in themanuscript result section.

Impacts
What was accomplished under these goals? For Goal 1, determine FSA responses in patients with EoE to top trigger whole-food proteins in a customized bead-based assay, the following objectives have been met during this review process: Whole food proteins ofcow milk, whole wheat, whole egg, textured soy protein, corn flour, rice flour, potato flour, and ground flaxseed have been coupled to MagPlex microspheres "beads" for the use in Luminex MagPix assay. Sample collection of esophageal secretions, saliva, and serum is ongoing through an approved IRB through the University of Utah. Thus far, 100 patient samples have been collected during standard-of-care endoscopies. We have processed and aliquoted 26 saliva, 75 serums, and80 esophageal secretions that can be utilized with Luminex MagPix assays. Apooled-patient sample standard curve was run with each whole food bead-basedassay. We have measured food-specific antibodies for whole food beads for 62 patient samples of esophageal secretions. Of these samples, we have data on 33 patients with active EoE, 21 remission EoE, and eight controls. We have whole food-specific antibody results for 60 serum samples and 26 saliva samples. We have begun analysis to compare food-specific antibody signals among the different sample types. For the 3 sample type manuscript will focus on the whole food reactivity, the introduction and methodsare drafted; the final sample size for comparison of saliva to esophageal secretions will be 24 patients with active EoE,and for comparison of serum to esophageal secretions, 42 patients with active EoE. Data analysis, drafting of the results, and discussionare underway. During the next review period, co-authors will provide acritical evaluationand editsbefore manuscript submission. For goal 2, demonstratethe efficacy of a food-specific IgA elimination diet in EoE to induce resolution and determine test thresholds for FSAs in patients with active EoE, the following objectives have been metduring this review process: Recruitment for an IgA-directed elimination diet is ongoing during standard-of-care endoscopies. Thus far, five patients have undergone the diet, and thefollow-up endoscopy indicated that two patients' IgA-directed elimination diet resulted in disease remission.

Publications