TOLerogenic Potential of Hematopoietic Stem and Progenitor Cells and Inflammatory Bowel Disease (TOL-IBD)

Exploring and Exploiting the TOLerogenic Potential of Hematopoietic Stem and Progenitor Cells to Cure Pediatric Inflammatory Bowel Disease

Pediatric refractory Inflammatory Bowel Disease (IBD) is a chronic inflammatory condition of the gastrointestinal tract, not responsive to current treatments. Since hematopoietic stem and progenitor cells (HSPCs) in the bone marrow display immunomodulatory functions and IL-10-producing regulatory cells regulate gut homeostasis, by combining state-of-the-art strategies for the ex-vivo manipulation and expansion of HSPCs and gene delivery systems to drive HLA-class II-restricted antigen presentation and expression of tolerogenic molecules, the investigators propose to dissect the antigen- (Ag-) presenting capacity of HSPCs and to exploit their tolerogenic potential to induce IL-10-mediated tolerance in the intestinal mucosa of IBD patients. The investigators hypothesize that HSPCs can be engineered using commensal-derived Ags w/wo IL-10 to drive the differentiation of Tr1 cells with the desired Ag-specificity to control intestinal inflammation in IBD. The results of this study will pave the way for defining innovative cell-based approaches for treating refractory pediatric IBD.

Study Overview

• Status: Not yet recruiting
• Conditions: Inflammatory Bowel Disease (Crohn's Disease; Ulcerative Colitis)
• Intervention / Treatment: Procedure: biological sample collection: an additional volume of peripheral blood (3-10 ml); Procedure: biological sample collection: small fragment (1-5 mm) of intestinal tissue - residual or leftover material; Procedure: biological sample collection: leftover peripheral blood samples from healthy subjects

Detailed Description

Inflammatory Bowel Disease (IBD) is a chronic inflammatory condition affecting the gastrointestinal tract, with Crohn's Disease (CD) and Ulcerative Colitis (UC) being the main types. The causes of IBD are complex and include immune dysregulation with activation of immune cells, release of inflammatory cytokines, and intestinal tissue damage. Despite significant advances in the treatment of pediatric IBD, a large unmet need for a definitive cure remains, and cell immunotherapy is under investigation. The investigators are specifically interested in refractory IBD, a chronic active condition requiring continuous treatment for symptom relief, with detrimental side effects. Despite the several treatment options currently available, 30% of pediatric IBD patients are refractory, and none of the existing treatments results in complete remission.

Interleukin 10 (IL-10) is an immunoregulatory cytokine associated with IBD pathogenesis and regulating gut homeostasis. Type 1 regulatory T (Tr1) cells produce IL-10 and their function is crucial for suppression of inflammation in IBD. The investigators recently published that antigen (Ag)-specific immune responses in Celiac Disease can be controlled via IL-10-producing Ag-presenting cells engineered to express a gliadin epitope, demonstrating that Ag-presenting cells can be manipulated to promote Tr1 cell differentiation.

Significant advances in the field of hematopoietic stem and progenitor cells (HSPCs) engineering and biology have been achieved. The development of protocols for ex-vivo manipulation and expansion of circulating (c)HSPCs and of mobilization-based chemotherapy-free approaches broadens the applicability of HSPC-based therapies to diseases for which HSPC transplantation is not the standard of care. A recent report showed that Tr1 cells can be promoted by immunogenic HSPC, which present Ags via HLA class II to CD4+ T cells in the bone marrow. This discovery opens new avenues for the development of approaches exploiting the Ag-presenting capacity and the tolerogenic potential of HSPCs to counteract inflammation in target tissues.

Based on these premises, with the final goal of developing a procedure for the induction of IL-10-mediated tolerance to control intestinal inflammation in refractory IBD pediatric patients, the investigators will:

• Investigate the type and frequency of inflammatory and regulatory immune cells, including IL-10-related regulatory cells, infiltrating the gut mucosa of IBD and non-IBD control patients. Results will indicate whether enforcement of the regulatory arm would benefit IBD patients. To this aim, it is necessary to collect and analyze intestinal tissue fragments from IBD and non-IBD control patients.
• Investigate the type and frequency of immune cells, including IL-10-related regulatory cells, circulating in the peripheral blood of IBD patients and non-IBD control patients. Results will indicate whether enforcement of the regulatory arm would benefit IBD patients. To this aim it is necessary to collect PB from IBD and non-IBD control patients.
• Characterize the presence, phenotype, and expansion potential of CD34+ HSPCs circulating in the peripheral blood of IBD patients. To this end, the investigators will collect peripheral blood from IBD patients, and i) assess the frequency, composition, and Ag-presenting potential of and ii) apply ex-vivo expansion and engineering protocols to CD34+ HSPC circulating in the peripheral blood of IBD patients. These results will be pivotal for assessing the feasibility of HSPC-based approaches to restore homeostasis in the intestinal tissue in refractory pediatric IBD.
• Evaluate the response to commensal-derived Ags (e.g., Bacteroides-derived peptides) of CD4+ T lymphocytes circulating by collecting the peripheral blood of IBD patients, non-IBD control patients, and healthy subjects as control. These results will indicate which Ag could be used for HSPC engineering and for inducing IL-10-producing regulatory T cells.
• Assess the ability of human HSPCs to promote Ag-specific Tr1 cells in vitro. To this end, selected IBD patients known to be responders commensal -derived Ags (as above) and peripheral blood will be collected at 1 year (6-18 months) follow-up. The investigators will engineer circulating HSPCs ex-vivo expanded from the peripheral blood of selected IBD patients and test for their ability to promote Ag-specific Tr1 cell upon in vitro culture with autologous CD4+ T cells.

• Study Type: Observational
• Enrollment (Estimated): 80

Contacts and Locations

• Study Contact: Name: Silvia Gregori, PhD; Phone Number: +390226434894; Email: gregori.silvia@hsr.it
• Study Contact Backup: Name: Laura Passerini, PhD; Phone Number: +390226439303; Email: passerini.laura@hsr.it

Study Locations

• Italy
  • Milan, Italy, 20132
    • Pediatric Immunohematology Unit, IRCCS Ospedale San Raffaele
    • Contact: Federica Barzaghi, MD; Phone Number: +390226432979; Email: barzaghi.federica@hsr.it
  • Rome, Italy, 00189
    • UOC Pediatria, Azienda Ospedaliero-Universitaria Sant'Andrea
    • Contact: Giovanni Di Nardo, MD; Phone Number: +390633775971; Email: giovanni.dinardo@uniroma1.it

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Child; Adult
• Accepts Healthy Volunteers: Yes

• Sampling Method: Non-Probability Sample

Study Population

Eligible participants will be consecutively enrolled as they present to the clinical centers until the planned sample size is achieved.

The Study will include pediatric subjects, both males and females belonging to the following study populations:

1. IBD patients: pediatric patients with chronic intestinal inflammation (i.e., inflammatory bowel disease (IBD), including Crohn Disease, Ulcerative Colitis) at diagnosis or follow-up (including responders to standard of care and refractory to treatment);
2. Non-IBD patients: patients with rectal bleeding with a negative colonoscopy and the exclusion of any inflammatory disorders of the gastrointestinal tract;
3. Healthy controls: healthy volunteers participating in the TIGET09 study protocol ("Collection of biological samples for the study of blood cells and their microenvironment, and for the development of novel therapeutic approaches for genetic diseases and cancer."), as control population.

Description

Inclusion Criteria:

For all groups:

• Written informed consent from parent(s)/legal guardian(s);
• Sex: Males and Females;
• Age: ≥2 years and <18 years.

For study group 1:

- Subjects with suspected or confirmed IBD diagnosis.

For study group 2:

- Subjects with rectal bleeding w/o inflammatory disorders of the gastrointestinal tract.

For study group 3:

• Written consent for participation to TIGET09 study protocol;
• healthy subjects, without known immunodeficiencies, autoimmune, inflammatory or genetic diseases, undergoing genetic, hematological, hematochemical, or HLA compatibility screenings and participating in the TIGET09 study protocol (Title: "Collection of biological samples for the study of blood cells and their microenvironment, and for the development of novel therapeutic approaches for genetic diseases and cancer").

Exclusion Criteria:

For all groups:

• Refusal or inability of the parent(s) or legal guardian(s) to provide written informed consent;
• Age: <2 years and ≥18 years;
• Presence of any medical, psychiatric, or clinical condition that, in the opinion of the clinician, may interfere with participation in the study or interpretation of the study results;

For study group 1:

- patients without IBD diagnosis or suspect;

For study group 2:

- Subjects without rectal bleeding or with known inflammatory/autoimmune disorders of the gastrointestinal tract.

For all groups:

• Refusal or inability of the parent(s) or legal guardian(s) to provide written informed consent;
• Age: <2 years and ≥18 years;
• Presence of any medical, psychiatric, or clinical condition that, in the opinion of the clinician, may interfere with participation in the study or interpretation of the study results;

For study group 1:

- patients without IBD diagnosis or suspect;

For study group 2:

- Subjects without rectal bleeding or with known inflammatory/autoimmune disorders of the gastrointestinal tract.

For study group 3:

• Lack of written consent for participation to TIGET09 study protocol;
• patients belonging to study groups 1 and 2; patients with immunodeficiencies, autoimmune, inflammatory or genetic diseases;
• subjects with signs of systemic inflammation.

Study Plan

How is the study designed?

Number of groups / cohorts

3

Cohorts and Interventions

Group / Cohort	Intervention / Treatment
Study Group 1; IBD patients: pediatric patients with chronic intestinal inflammation (i.e., inflammatory bowel disease (IBD), including Crohn Disease, Ulcerative Colitis) at diagnosis or follow-up (including responders to standard of care and refractory to treatment)	Procedure: biological sample collection: an additional volume of peripheral blood (3-10 ml); An additional volume of peripheral blood (3-10 ml) will be obtained in concomitance with clinically indicated procedures; Procedure: biological sample collection: small fragment (1-5 mm) of intestinal tissue - residual or leftover material; A small fragment (1-5 mm) of intestinal tissue - residual or leftover material -will be obtained from patients undergoing diagnostic or follow-up endoscopy
Study Group 2; Non-IBD patients: patients with rectal bleeding with a negative colonoscopy and the exclusion of any inflammatory disorders of the gastrointestinal tract	Procedure: biological sample collection: an additional volume of peripheral blood (3-10 ml); An additional volume of peripheral blood (3-10 ml) will be obtained in concomitance with clinically indicated procedures; Procedure: biological sample collection: small fragment (1-5 mm) of intestinal tissue - residual or leftover material; A small fragment (1-5 mm) of intestinal tissue - residual or leftover material -will be obtained from patients undergoing diagnostic or follow-up endoscopy
Study Group 3; Healthy volunteers (controls)	Procedure: biological sample collection: leftover peripheral blood samples from healthy subjects; Peripheral blood samples from healthy subjects, leftover from TIGET09 protocol analysis will be collected

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
To characterize pediatric IBD patients' peripheral blood and intestinal mucosa immune cell composition	Frequency (%) of predefined regulatory and inflammatory immune-cell subsets in peripheral blood and gut mucosa, including regulatory T cells (FOXP3+ Tregs and IL-10 producing Tr1 cells), T naïve/memory and effector cells, regulatory myeloid cells (DC-10), and inflammatory myeloid cells (cDC1 and cDC2), measured by flow cytometry. The primary objective will be considered met if patients with IBD show a lower frequency of IL-10-producing cells than controls, with the estimated between-group difference supporting a defect in IL-10-producing cell responses.	Baseline timepoint

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
To explore the ability of HSPCs to induce in vitro the differentiation of Ag-specific Tr1 cells.	Induction of Tr1-like cells following T-cell/HSPC co-culture, assessed by: frequency (%) of cells expressing the Tr1-associated phenotype and expression of Tr1-associated genes. Success criterion (exploratory): Generation of a T-cell population displaying Tr1-associated features.	Baseline and 1 year follow up

Collaborators and Investigators

• Sponsor: IRCCS San Raffaele
• Investigators: Principal Investigator: Silvia Gregori, PhD, IRCCS Ospedale San Raffaele; Principal Investigator: Alessandro Aiuti, MD, IRCCS Ospedale San Raffaele

Publications and helpful links

General Publications

• Parigi TL, D'Amico F, Abreu MT, Dignass A, Dotan I, Magro F, Griffiths AM, Jairath V, Iacucci M, Mantzaris GJ, O'Morain C, Reinisch W, Sachar DB, Turner D, Yamamoto T, Rubin DT, Peyrin-Biroulet L, Ghosh S, Danese S. Difficult-to-treat inflammatory bowel disease: results from an international consensus meeting. Lancet Gastroenterol Hepatol. 2023 Sep;8(9):853-859. doi: 10.1016/S2468-1253(23)00154-1. Epub 2023 Jul 6.
• Hernandez-Malmierca P, Vonficht D, Schnell A, Uckelmann HJ, Bollhagen A, Mahmoud MAA, Landua SL, van der Salm E, Trautmann CL, Raffel S, Grunschlager F, Lutz R, Ghosh M, Renders S, Correia N, Donato E, Dixon KO, Hirche C, Andresen C, Robens C, Werner PS, Boch T, Eisel D, Osen W, Pilz F, Przybylla A, Klein C, Buchholz F, Milsom MD, Essers MAG, Eichmuller SB, Hofmann WK, Nowak D, Hubschmann D, Hundemer M, Thiede C, Bullinger L, Muller-Tidow C, Armstrong SA, Trumpp A, Kuchroo VK, Haas S. Antigen presentation safeguards the integrity of the hematopoietic stem cell pool. Cell Stem Cell. 2022 May 5;29(5):760-775.e10. doi: 10.1016/j.stem.2022.04.007.
• Omer-Javed A, Pedrazzani G, Albano L, Ghaus S, Latroche C, Manzi M, Ferrari S, Fiumara M, Jacob A, Vavassori V, Nonis A, Canarutto D, Naldini L. Mobilization-based chemotherapy-free engraftment of gene-edited human hematopoietic stem cells. Cell. 2022 Jun 23;185(13):2248-2264.e21. doi: 10.1016/j.cell.2022.04.039. Epub 2022 May 25.
• Capo V, Penna S, Merelli I, Barcella M, Scala S, Basso-Ricci L, Draghici E, Palagano E, Zonari E, Desantis G, Uva P, Cusano R, Sergi Sergi L, Crisafulli L, Moshous D, Stepensky P, Drabko K, Kaya Z, Unal E, Gezdirici A, Menna G, Serafini M, Aiuti A, Locatelli SL, Carlo-Stella C, Schulz AS, Ficara F, Sobacchi C, Gentner B, Villa A. Expanded circulating hematopoietic stem/progenitor cells as novel cell source for the treatment of TCIRG1 osteopetrosis. Haematologica. 2021 Jan 1;106(1):74-86. doi: 10.3324/haematol.2019.238261.
• Passeri L, Andolfi G, Bassi V, Russo F, Giacomini G, Laudisa C, Marrocco I, Cesana L, Di Stefano M, Fanti L, Sgaramella P, Vitale S, Ziparo C, Auricchio R, Barera G, Di Nardo G, Troncone R, Gianfrani C, Annoni A, Passerini L, Gregori S. Tolerogenic IL-10-engineered dendritic cell-based therapy to restore antigen-specific tolerance in T cell mediated diseases. J Autoimmun. 2023 Jul;138:103051. doi: 10.1016/j.jaut.2023.103051. Epub 2023 May 22.
• Cook L, Stahl M, Han X, Nazli A, MacDonald KN, Wong MQ, Tsai K, Dizzell S, Jacobson K, Bressler B, Kaushic C, Vallance BA, Steiner TS, Levings MK. Suppressive and Gut-Reparative Functions of Human Type 1 T Regulatory Cells. Gastroenterology. 2019 Dec;157(6):1584-1598. doi: 10.1053/j.gastro.2019.09.002. Epub 2019 Sep 10.
• Krawiec P, Pawlowska-Kamieniak A, Pac-Kozuchowska E. Interleukin 10 and interleukin 10 receptor in paediatric inflammatory bowel disease: from bench to bedside lesson. J Inflamm (Lond). 2021 Mar 10;18(1):13. doi: 10.1186/s12950-021-00279-3.
• Neurath MF, Sands BE, Rieder F. Cellular immunotherapies and immune cell depleting therapies in inflammatory bowel diseases: the next magic bullet? Gut. 2024 Dec 10;74(1):9-14. doi: 10.1136/gutjnl-2024-332919.

Study record dates

Study Major Dates

• Study Start (Estimated): October 1, 2026
• Primary Completion (Estimated): October 1, 2029
• Study Completion (Estimated): November 1, 2029

Study Registration Dates

• First Submitted: July 23, 2026
• First Submitted That Met QC Criteria: July 28, 2026
• First Posted (Actual): July 31, 2026

Study Record Updates

• Last Update Posted (Actual): July 31, 2026
• Last Update Submitted That Met QC Criteria: July 28, 2026
• Last Verified: July 1, 2026

More Information

Terms related to this study

• Keywords: regulatory T cells; immune tolerance; hematopoietic stem and progenitor cells (HSPCs); IL-10 producing cells; commensal-derived epitopes
• Additional Relevant MeSH Terms: Intestinal Diseases; Digestive System Diseases; Gastrointestinal Diseases; Colonic Diseases; Gastroenteritis; Colitis; Colitis, Ulcerative; Crohn Disease; Inflammatory Bowel Diseases
• Other Study ID Numbers: RF-2024-12378919 (Other Grant/Funding Number: Ministero della Salute - Bando Ricerca finalizzata 2024)

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: UNDECIDED

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No