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Report

Teaser, summary, work performed and final results

Periodic Reporting for period 2 - INTENS (INtestinal Tissue ENgineering Solution)

Teaser

Summary

What is the problem/issue being addressed?
Short Bowel Syndrome (SBS) is a condition that occurs when part or the entire small intestine is missing or has been removed during surgery. This condition renders the bowel incapable of fulfilling its nutritional function (intestinal failure). There is no cure for SBS.

Why is it important for society?
Children with SBS and their families often have a very different and difficult life compared to that of normal families such as:
1. Frequent hospital visits.
2. Difficulty attending nursery and school.
3. Movement and activities of the child are impaired.
4. Difficult for parents to take care of a normal job.
5. Intensive night care and sleep deprivation.

SBS is a chronically debilitating disorder without a cure. Intravenous feeding offers a survival rate of 70% at 5 years in newborn infants. However, in the most severe cases, when only 10% of expected intestinal length is present, 5-year survival is reduced to around 20%. In particular we will focus on developing an INtestinal Tissue ENgineering Solution (INTENS) for children with SBS.

What are the overall objectives?
The objective of this programme is to deliver a functional bowel reconstruction to patients with SBS through an autologous tissue engineering strategy, overcoming the shortage of organs, and avoiding the need for immunosuppression. It will be achieved by identifying the best autologous cell source; providing the ideal scaffold; engineering functional intestine for transplantation and engaging with patients, scientists and public. The work is designed to lead directly to a clinical trial for the application of the optimal protocol for tissue-engineered intestine.

Work performed

WP 1 â€“ Autologous cell derivation
The second period (18-36 months) of WP1 in the INTENS consortium has followed the projected developments and we are currently at a stage where we can expand all the cell types from both primary tissue and pluripotent stem cells necessary to add to the scaffold that will eventually be utilised for transplantation experiments. These are now ready for expansion and implementation into the scaffold and the technology is being transferred into the teams responsible for seeding and maintaining the artificial intestines. The focus in WP1 is now to consolidate methodology for functional assessment of epithelial as well as other cells that will go into the scaffold.

WP 2 â€“ Scaffold preparation
We successfully modified the chemistry of previously developed PEG hydrogels to replace the transglutaminase Factor XIII (adding additional complexity for downstream clinical applications) with much simpler â€˜clickâ€™ chemistry. We successfully expanded mouse and human (both from iPSC and somatic stem cells) intestinal stem cells and derived organoids in these defined matrices. We successfully grew human colon organoids in fully defined synthetic hydrogels without the use of any animal-derived components such as Laminin-1. We also developed novel methodologies to generate organoids that for the first time develop (i.e. undergo symmetry breaking and patterning) in a predicable manner.

WP 3 â€“ In vitro engineering and functional studies
To optimise the repopulation of the decellularised scaffolds in vitro, we first tested seeding conditions of intestinal epithelial cells (organoids) on scaffolds under static culture using RAFT system as a surrogate step to identify the optimal seeding density. So far, we managed to successfully repopulate both RAFT, human and porcine decellularised scaffolds with epithelial cells, fibroblasts and endothelial cells under static culture. The repopulated epithelial cells were able to form polarised monolayer with sucrose digestive enzyme expression, suggesting the presence of enterocyte differentiation. More importantly, we have optimised a protocol to turn epithelial layer to form crypt-villus structures. We will continue to explore the optimal conditions for co-culturing all cell types, both static and in bioreactor, and to test the functions of the recellularised intestinal graft.

WP 4 â€“ Good Manufacturing Practice (GMP) processing and scaling up
The generation of GMP grade growth factors has been initiated and these growth factors can now be tested in cultures of human intestinal cells. GMP grade hydrogels to support the growth of organoids are being tested.

WP 5 â€“ Transplantation of engineered intestine
We have identified acceptable conditions and polymer specifications for small and large animal models of implanted donor cells and now seek to optimize these conditions with additional donor cell types.

WP 6 â€“ Dissemination, patient engagement and ethics
Dissemination and Communication:
Created a dynamic, public facing website updated with news. Built capacity within the INTENS consortia for public engagement through establishing a network of communicators, the production of tools to communicate digitally and through face to face engagement with a range of audiences including teachers, school pupils, general public, and families. Significant development and production of high profile public engagement activity, particularly in collaboration with Great Ormond Street Hospital.

Patient engagement:
Established engagement with patient networks to pave the way for a co-developed patient engagement event.

Training:
Four INTENS researchers received 7 days training at the Hydra Summer School on Stem Cell Biology.

WP 7 â€“ Management structure and procedures
WP7 activities covered all aspects of project monitoring, reporting, financial and contractual administration in accordance with the Commissionâ€™s rules, ensuring proper communication within the consorti

Final results

The potential impact is the development of a solution (medical intervention) for the treatment of Short Bowel Syndrome, a condition which can only be managed but not cured. The tools to tissue engineer sections of small bowel for implantation into patients will revolutionise patient care, increase survival and dramatically improve quality of life for patients and their families. The creation of a tissue engineered small bowel is in itself beyond the state-of-the-art as no one has ever created, through tissue engineering, such a complex structure.