Diagnostic innovation for inflammatory bowel diseases in children and adults: non invasive stool and salivary testing with ecologic new methods and biomaterials

Inflammatory bowel diseases (IBD) are increasing worldwide, onset at young age and in one fourth of the cases in children, persist long-life with relapses alternating to remissions, and complications often occur. For the society IBD have a disproportionally high cost related to the need of frequent monitoring (endoscopy, radiology, histology) and repeated hospitalizations. No specific serum biomarkers are actually available for IBD diagnosis. Fecal calprotectin (fCal), marker of neutrophilic intestinal inflammation, is widely used to rule-out functional intestinal disorders and to rule-in intestinal inflammation, but its sensitivity and specificity for IBD do not exceed 70%. We patented a diagnostic system for IBD based on MALDI-TOF/MS analysis of the fecal peptidome, which showed in adult patients high sensitivity (80%) and specificity (about 100% ). Our first aim is to verify whether our patented MALDI-TOF/MS-based fecal peptidomic assay allows to diagnose and to monitor IBD in the pediatric age, distinguishing also the two main IBD forms, ulcerative colitis and Crohn's disease, and whether it is applicable in a real world scenario. To do this pediatric IBD patients will be enrolled by the pediatric gastroenterologist of the operative unit 1 while validation in a real world scenario will be performed by a prospective enrollment of outpatients with gastrointestinal symptoms at both the units 1 and 2. For this last cohorts of patients clinical data will be retrieved by the general practitioners for two years to identify any incident IBD case diagnosed in this time frame after enrollment. Our aim 2 is to improve the bioinformatic analysis of our patented MALDI-TOF/MS assay by artificial intelligence and to improve sustainability by developing "green" collection devices. The prospective data obtained in real world scenario from both units will be analyzed by machine learning using diagnoses as model outcome. Moreover, through already established collaborations with the industry new ecologic biomaterials will be tested for collection devices investigating their compatibility with biochemical assays including MALDI-TOF/MS. Our third aim derives from preliminary results from proteomic analyses of stool of IBD patients that evidenced the presence of salivary proteins. The finding of these proteins only in the stools of patients with Crohn's disease suggests the hypothesis that an altered salivary protein pattern associated with the delivery in the intestine of potentially pathogenetic proteins might underly the development of the chronic intestinal inflammation. This process might involve immune cells but also stems cells. Based on this premise we plan to establish an in vitro model based on the isolation and subsequent culture of intestinal mesenchymal stem cells to verify whether the identified salivary proteins exert any effect on their proliferation, apoptosis and differentiation. Moreover, the proteomic profile of patients' saliva will be analyzed to discover whether it carries diagnostic biomarkers.
The results of this project will improve patients' management, and this will impact mainly, but not only, in the pediatric setting. It is expected a reduction in the frequency of invasive diagnostic procedures which in turn will enhance patients' compliance and reduced health care costs. New pathogenetic insights are also expected and these might support the development of new therapies.