LSFM4LIFE in-depth

LSFM4LIFE ground


Type 1 Diabetes (T1D) in numbers


  • 20.7 million people are affected by T1D worldwide

  • 10 year reduction of life expectancy for people with T1D

  • 2.3 times higher medical expenses than average

  • 4% growth rate of young adults


The Global Burden of Diabetes


Amongst the 415 million people who are affected by diabetes 5% are concerned by T1D. This incurable disease is caused by the autoimmune destruction of the insulin-producing pancreas cells.

Advances in healthcare and increased public awareness resulted in improved management of this disease. However, T1D still requires a daily demanding monitoring and treatment.

A group of European researchers has gathered with the goal of sparing affected people from lifelong insulin therapy, by developing clinical-grade Human Pancreas Organoids (hPOs) for a cell-based therapy.

The LSM4LIFE project is dedicated to develop a process for the mass production of three-dimensional cellular structures of insulin-producing cells (organoids) in the laboratory.

LSFM4LIFE approach


Amongst all the advances in healthcare to overcome the deficient pancreas of patients with T1D, the LSFM4LIFE project is driven by one of the most promising alternatives: stem cell-derived beta cells. The idea is to isolate and to expand adult human pancreas stem cells and to differentiate them into insulin-producing cells.

The stem cell-derived beta cells approach has been made possible by the discovery of the group of Hans Clevers in Utrecht, which succeeded in activating the pancreas ductal cells to express the protein Lgr5, a marker of adult stem cells.

Once isolated and cultured in a 3D gel, the Lgr5- expressing cells form pancreas organoids that can be grown almost limitlessly in culture.

Pancreas organoid grown for 3 months in culture from mouse pancreas ducts

 

LSFM4LIFE challenges


Dr. Meritxell Huch from the Gurdon Institute, a partner of LSFM4LIFE, carried out a study showing that an organoid transplanted into mice differentiates in insulin-producing cells. The final goal of the LSFM4LIFE project is to transfer this technology from mouse to human.

Producing clinical-grade (GMP-compliant) hPOs as a long-term source of pancreas stem cells represent a breakthrough for cellular therapy of T1D. In order to achieve it, LSFM4LIFE partners have several challenges to undertake into different fields: cell-based therapy; 3D scaffolds; mass production of cells; optical technology ; and cell-based assay.

Here are some specific innovation challenges addressed by the LSFM4LIFE project:

One major bottleneck of current cell-based therapies based on differentiated human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) is the risk of tumor formations after transplantation. A further difficulty is the inherent genetic instability of both hESCs and iPSCs, which consists in a hallmark of cancer. Similarly to these cell-based therapeutics, immunogenicity and immunotoxicity are a possible issue of hPOs.
Matrigel is the state-of-the-art product in this field. It is composed of a complex protein hydrogel extracted from a mouse sarcoma tissue. Although it is used for many types of three-dimensional cell cultures, its batch-dependent variable composition prevents the controlled and safe production of hPOs. Moreover, the animal origin of Matrigel might be a source of pathogens and is thus unsuitable for the GMP production of cell-based therapeutics.
At the moment, exogenous dyes are used to isolate primary cells from donors’ pancreas islets, wich requires tedious optimization.
There is no strategy for the scaling-up of cells for the therapy of diabetes in Europe.
The consistency of the manufacturing process at GMP standards is critically important for the production of cellular products that can be transplanted into the patient. To date, there are no GMP procedures for the production of cell-based therapeutics for T1D in Europe.

LSFM4LIFE work packages


The project starts with an R&D protocol and finishes with a clear strategy for the necessary steps to advance hPO towards clinical trials. During the project, the hPO generation protocols are refined with respect to translation to GMP and in vitro differentiation in two parallel threads that are
closely linked.

Translation of the process of hPO expansion from the R&D level to GMP-level

WP03 & WP07: The generation of undifferentiated hPO from cells extracted from human pancreatic glands provides the starting point for all subsequent activities.

WP04:
Based on the initial R&D with undifferentiated organoids, quality criteria for organoid expansion are determined.

WP04, WP05 & WP06: Biomimetic hydrogel and the protocols for expanding the organoids in this hydrogel are optimised until all components are GMP-compliant and the organoids satisfy the quality criteria.


Establishing the differentiation of hPOs in vitro

WP04: Based on comparisons with pancreas islets phenotypic and functional quality criteria for hPO differentiation are established.

WP03 & WP04: The differentiation protocol is optimized until the quality criteria are fulfilled.

WP07: The GMP-compliant undifferentiated organoids as well as the R&D level differentiated organoids are tested in vivo with respect to their safety, long-term survival and functional efficacy.

WP06: All results of the project flow into a strategy for the further development of hPO towards later Clinical Investigations.

LSFM4LIFE outcomes


The main motivation of LSFM4LIFE is a response to the societal needs for advanced cell-based therapies for the treatment of T1D, improving the quality of life, the health and the life expectancy of the patients affected by T1D.

By intending to produce GMP-level organoids ready for the implementation of the clinical phases, the project is expected to provide an impact on several technological aspects, leading to:

LSFM4LIFE develops R&D, GMP manufacturing tools, and technologies that pave the way to clinical studies with the hPO as a news cell-based therapy for T1D. The tools include:
– Expansion of the organoids in chemically well-defined biomimetic hydrogels
– Standardized characterization of the organoids at cellular and molecular level
– The capacity to manufacture the organoids at a therapeutic-relevant quantity under GMP conditions
Since cellular organoids, devices and assays follow different regulations, LSFM4LIFE develops a quality management system that addresses all regulations. Safety and regulatory criteria are determined in the perspective of clinical implementations and up-scaled production of hPO.
The innovations within LSFM4LIFE leads to a new reproducible islet model for drug screening. Given the market and the clear feasibility, LSFM4LIFE breakthroughs generate a highly innovative cell-based medical treatment in the next decade, fostering the competitiveness of the EU over the US and Asia.
LSFM4LIFE combines the innovations produced by groups experts in different R&D fields, including stem-cell biology, clinics, optical technology and screening, material sciences and manufacturing of mammalian cells according to GMP.
LSFM4LIFE increases the attractiveness of Europe as a hub for innovative medical technologies by bringing
together a highly competitive group of academic and clinical partners with dynamic high-technology SMEs.