Objectives

1. To establish a cohort of patients with immune-mediated disease (SS) or cancer (LC, BC) in order to cultivate organoids starting from the biopsies.

2. To establish a cohort of dogs with MEL or AD to cultivate organoids starting from biopsies.

3. To standardize protocols for organoid cultures in human and dogs’ biopsies

4. To establish a ‘mini-lab organomicrolife’ to better standardize 3D cultures 5. To increase the awareness of researchers and professionals about the scientific research on ethical issues in processing personal and non-personal data in ORGANOMICROLIFE.

1. To characterize the organoids generated in this project using light and electron microscopy to determine various parameters, including immunophenotypic features of the whole structure as well as to investigate cell-related features.
2. To analyze the established 3D cultured organoids for histological, immunophenotypic and genomic concordance with parental biopsies.
3. To analyze the epigenetic state of organoids derived from biopsies.

1. To characterize the biophysical (i.e. morphology and quantity) and biochemical (i.e. lipid profiles) aspects of EVs released from 3D cell models.
2. To determine the oncometabolites derived from the amino acids L-tryptophan, L-arginine and L-phenylalanine in the secretome of organoids, as these have been shown to activate immunoregulatory functions. At the same time, we will also determine the presence of the enzymes responsible for the first limiting step of metabolism of these amino acids (IDO1, arginase 1, and IL4i1, respectively) in the EVs of the organoid secretome.
3. To determine whether Glucocorticoid hormones (GC), Glucocorticoids receptors (GR) and Glucocorticoid-Induced Leucine Zipper (GILZ) are present in and act as immunomodulatory components of EV.
4. To evaluate the expression of RAGE in organoids and the levels of RAGE ligands throughout the secretome and in EVs derived from organoids.
5. To validate functional properties of secretome/EVs from 3D organoids through peripheral blood mononuclear cells (PBMCs) co-cultures.
6. To investigate the pro-cachectic potential of the organoid secretome/EVs we will evaluate the activation state of RAGE signaling and several molecular markers of muscle wasting in human myotubes.

1. Multidisciplinary scientific insights: Through advanced statistical analyses and machine learning techniques, we aim to integrate diverse clinical data, including demographic, pathological, and lifestyle information. By correlating these data points, the project seeks to identify possible patterns and associations that contribute to dysregulated immune responses. In particular, statistics will be generated, where possible in parallel for human and canine patients, on:
– master data (sex, age, ethnicity/race)
– pathology (subtype, staging, genetic signature, family history, autoimmune response, post-operative health status, remission)
– lifestyle (negative prognostic factors such as BMI, smoking)
– psychological aspects.
2. Narrative analysis and psychological insights: Psychological insights and narrative analysis will be applied to results from questionnaires, on which to apply data analysis and narrative medicine techniques. This approach aims to investigate the psychological aspects of perceptions of both patients and public, regarding understanding, and trust in organoid technology.
3. Comprehensive Data Set Creation: Through the combination of medical records and surveys, a comprehensive and valuable set of data will be created, to provide significant insights for future research in the scientific community with potential novel associations combining clinical, psychological, and demographic data, potentially opening the path to the development of personalized treatments and interventions.