Production and characterization of endocrine cells derived from human pancreas organoids for the cell-based therapy of type 1 diabetes

Informations projet

LSFM4LIFE

N° de convention de subvention: 668350

DOI

10.3030/668350

Projet clôturé

Date de signature de la CE 3 Decembre 2015

Date de début 1 Janvier 2016

Date de fin 31 Decembre 2019

Financé au titre de

• SOCIETAL CHALLENGES - Health, demographic change and well-being

Coût total

€ 5 120 275,00

Contribution de l’UE

€ 4 666 400,00

Coordonné par

JOHANN WOLFGANG GOETHE-UNIVERSITAET FRANKFURT AM MAIN
Germany

Periodic Reporting for period 3 - LSFM4LIFE (Production and characterization of endocrine cells derived from human pancreas organoids for the cell-based therapy of type 1 diabetes)

Période du rapport: 2019-01-01 au 2019-12-31

Résumé du contexte et des objectifs généraux du projet

The worldwide incidence of type 1 diabetes (T1D) is increasing at a rate of 4% yearly. This alarming epidemiological data strongly motivates the development of a cellular treatment of T1D, in which the missing beta-cells are replenished with exogenous cells. Despite many advancements, the allogeneic transplantation of Langerhans islets is only recommended for a small group of patients. The suitability of islets transplantation is limited both by donor availability and by the need for immunosuppressive treatment.
In contrast, cellular therapy with beta-cells obtained by differentiating adult stem cells can drastically revolutionize this picture and realize the dream of a curative therapy of T1D. Academic and industrial partners from France, Germany, Great Britain, Italy, Netherlands, and Switzerland joined the European consortium LSFM4LIFE to develop the hPO technology. LSFM4LIFE is supported for the period 2016-2020 by a Horizon 2020 grant.

LSFM4LIFEhas achieved its major objectives:
1) the groups the University of Cambridge has developed the technology to long-term expand hPOs, from both fresh and cryopreserved human pancreas tissue in a chemically defined, serum-free medium with no detectable tumorigenicity in vivo. hPOs are now an abundant source of pancreas ductal cells that retain the characteristics of the original tissue. This allows the modeling of diseases of the ductal epithelium, an increasing understanding of the human pancreas exocrine biology, as well as for producing insulin-secreting cells for the treatment of diabetes.
2) the Goethe University Frankfurt has developed the technology (hardware, workflow, and protocols) for performing high-throughput analysis of three-dimensional hPOs with Light Sheet Fluorescence Microscopy (LSFM). With the new imaging technology, a fast and very detailed characterization and quality control of organoids are possible.
3) the company Cellendes, has synthesized a dextran-based hydrogel that efficiently supports the growth of hPOs. The new hydrogel, fully defined and GMP compliant, allows the expansion of hPO both from already established frozen stocks as well as from tissue biopsies.
4) the Cell Factory at Policlinico hospital in Milan has accomplished a further major goal of LSFM4LIFE: a GMP workflow for the clinical-grade manufacture of hPO. This is fundamental for the translation in the clinics and the start of clinical trials. Lonza Netherlands has elaborated a full roadmap leading to the commercial production of hPO and their use as a cellular medicinal product.

Travail effectué depuis le début du projet jusqu’à la fin de la période considérée dans le rapport et principaux résultats atteints jusqu’à présent

The LSFM4LIFE consortium realized the major goals envisioned and planned in the project.

Objective 1: the groups at the University of Cambridge have shown that hPOs can be expanded long-term, from both fresh and cryopreserved human pancreas tissue in a chemically defined, serum-free medium with no detectable tumorigenicity.
hPOs can be clonally expanded, genetically manipulated and are amenable to culture in a chemically defined hydrogel. Therefore, the LSFM4LIFE consortium has successfully established hPOs as an abundant source of pancreas ductal cells that retain the characteristics of the original tissue. These results have been published in BMC Developmental Biology (https://doi.org/10.17863/CAM.48839).

Objective 2: Goethe University Frankfurt realized the hardware, workflow and the corresponding protocols for performing high-throughput analysis of hPOs with Light Sheet Fluorescence Microscopy (LSFM). The results of this work have been recently published in Nature Scientific Reports (https://doi.org/10.1038/s41598-019-53380-2). Moreover, a monolithic version of the high-throughput LSFM (mHT-LSFM) has been realized. The mHT-LSFM incorporates the operating principles of the HT-LSFM prototype in a compact and user-friendly device that can be operated by non-specialists in microscopy, thus greatly expanding the possibility of adoption of light-sheet microscopy technology by other laboratories working with organoids and further types of 3D cultures for regenerative medicine and cell-based therapy.

Objective 3: a primary goal of LSFM4LIFE, which has been achieved by Cellendes. A novel dextran-based hydrogel efficiently supporting the growth of hPO has been synthesized and tested. The new hydrogel, fully defined and GMP compliant, allows the expansion of hPO from frozen stocks as well as from tissue biopsies. These results have been published in BMC Developmental Biology (https://doi.org/10.17863/CAM.48839).

Objective 4: Cell Factory at Policlinico Hospital in Milan has accomplished a further major goal of LSFM4LIFE, namely developing the GMP workflow for the clinical-grade manufacture of hPO. These results have been published in the journal Stem Cell Research & Therapy (“Standardized GMP-compliant scalable production of human pancreas organoids”, doi: 10.1186/s13287-020-1585-2) and featured in the editorial https://doi.org/10.1186/s13287-020-1586-1).
Lonza Netherlands has accomplished all the planned tasks, elaborating in total four comprehensive dossiers, which outline a full roadmap leading to the commercial production of hPO and their use as a cellular medicinal product.

During P3, ProSciencia joined the consortium for the dissemination activities. ProSciencia has further outstandingly carried on the dissemination work of LSFM4LIFE, by ensuring a day-by-day updating of the LSFM4LIFE website (www.lsfm4life.eu) as well as daily coverage of the advancements in the research and therapy of Diabetes Type 1 on Twitter, presenting both the results of the consortium and the ones of other researchers external to LSFM4LIFE. In addition, 5 new videos describing the work of LSFM4LIFE and including interviews with the scientists working on the project have been produced in P3 (https://lsfm4life.eu/2019/new-project-videos/). Finally, during P3 the LSFM4LIFE consortium has organized and supported two international conferences: the European Organoids Symposium - EOS2019 (www.eos2019.eu) and the Light Sheet Microscopy Conference - LSFM2019 (www.lsfm2019.eu).

Progrès au-delà de l’état des connaissances et impact potentiel prévu (y compris l’impact socio-économique et les conséquences sociétales plus larges du projet jusqu’à présent)

The ambition of LSFM4LIFE is advancing the state-of-the-art in several biotechnological areas on the way towards the production of human pancreas organoids at GMP-level. The biotechnological areas of innovation pursued by LSFM4LIFE cluster into the following main fields:

• Cell-based therapy.
• Biomimetic hydrogel scaffolds.
• Therapeutic-scale production of cells at GMP standard.
• Optical technology for 3D tissues.
• Cell-based assay.

hPOs produced at GMP-level as a long-term source of pancreas stem cells, represent a breakthrough in the cellular therapy of Type 1 Diabetes. A further groundbreaking and ambitious goal is the production of a cost-effective surrogate for Matrigel, which is a hydrogel of animal origin unsuitable for clinical application. LSFM4LIFE also advances the field of cell-based assay by developing standardized assays on pancreas organoids in a multi-well format for drug and toxicity screening. Moreover, with the high-throughput light sheet microscope (HT-LSFM) LSFM4LIFE realizes an innovative optical technology for the imaging of 3D tissues, which shows much better performance than state-of-the-art confocal fluorescence microscopes in terms of high speed, high resolution and low photo-bleaching.

Pancreas organoids cultured in Matrigel. Cyan: microtubules, red: cell nuclei.