Genetically engineered human pluripotent stem cells, functionalized silk-fibroin platforms and bio-inks: a novel solution for large-scale ex-vivo platelet production, transfusion and drug research

Informations projet

SilkFUSION

N° de convention de subvention: 767309

DOI

10.3030/767309

Projet clôturé

Date de signature de la CE 12 Septembre 2017

Date de début 1 Novembre 2017

Date de fin 31 Octobre 2022

Financé au titre de

• EXCELLENT SCIENCE - Future and Emerging Technologies (FET)

Coût total

€ 2 998 500,01

Contribution de l’UE

€ 2 998 500,00

Coordonné par

UNIVERSITA DEGLI STUDI DI PAVIA
Italy

Periodic Reporting for period 3 - SilkFUSION (Genetically engineered human pluripotent stem cells, functionalized silk-fibroin platforms and bio-inks: a novel solution for large-scale ex-vivo platelet production, transfusion and drug research)

Période du rapport: 2020-05-01 au 2022-10-31

Résumé du contexte et des objectifs généraux du projet

The limited supply of human platelets and the rapidly growing demand for medical research and clinical applications including studies of platelet-related disease mechanisms and platelet transfusions for patients are manifest. Therefore, in transfusion medicine, there is an urgent need for new modes to generate functional platelets ex-vivo to address clinical needs as well as for insight into fundamental studies of mechanisms of physiologic and pathological platelet production. The main objective of the SilkFusion project is to be able to develop technology and methodology that allows the manufacture of "a la carte" platelets, without the need to depend on donations. Unlike blood, platelets do not have a long shelf life and that makes stock management difficult. Unlike blood, which has a half-life of storage of 30-40 days, platelets can currently only be stored for about 5 days. It has been seen that their demand is going in crescendo and this problem in the future will get worse if a solution is not found.

Our hypothesis is that platelet production ex-vivo can be optimized by providing megakaryocytes (the platelet progenitors) with the correct physical and biochemical environment. To prove this, we will develop a unique technological platform by engineering non-thrombogenic silk-fibroin biomaterial with proteins that were proven to promote a gain in platelet production of at least one order of magnitude, through the creation of a three-dimensional ex-vivo bone marrow model that will enhance platelet release. Our long-term vision is to foster the production of platelets in-vitro for clinical transfusions in humans at a scale and cost that will address current supply challenges as immunologically matched products to alloimmunised patients. The successful development and distribution of the SilkFusion platforms will also offer researchers and clinicians specialized precision instruments for determining the safety and efficacy of drugs, reducing the costs of ineffective therapies while promoting affordable functional strategies for the development of novel molecules.

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

Key developments were achieved in the third part of the project for the development of a 3D silk biomaterial platform for the large-scale generation of human platelets. The platform addresses therapeutic areas with unmet critical needs such as personalized transfusions, patients with acute hemorrhage due to trauma, obstetrics, surgery or wounded in conflict zones. Additional applications include developing personalized drug therapies and as a screening technology in the development of new compounds in thrombocytopenic and oncological patients.

The consortium completed the fabrication of 3D engineered silk models to mimic human bone marrow for platelet production. As part of the activities necessary to render the 3D environment human-like, the team focused on the functionalization of the silk scaffolds with five peptides (short chains of amino acid monomers). We proved that the scaffolds functionalized with positive peptides were able to induce an increase of platelet production from megakaryocytes derived from iPS cell lines. This represents an important result towards the scale-up of platelet production for application in transfusion medicine.

The consortium worked on the design and fabrication of a miniaturized version of the silk bioreactor to culture cells derived from a small amount of patient peripheral blood. This allowed us to study the pathogenic mechanisms of some inherited thrombocytopenia characterized by decreased peripheral blood platelet numbers. The consortium also developed hPSC lines from patients affected by different forms of inherited thrombocytopenia. The goal of this activity is to perfect the nanotechnology platform as a novel instrument to study disease mechanisms and novel therapeutic targets. This will help with the development of personalized treatments for these rare diseases.

The consortium defined the methodology for printing megakaryocytes using specialized 3D bio-printers and engineered inks. The team succeeded in printing functional megakaryocytes in silk-based bio-inks for 3D printing of the silk bone marrow platform. This constitutes one of the building blocks towards the industrialization of the nanotechnology platform for large-scale generation of human platelets, disease mechanism study, and drug testing.

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)

Throughout Europe, the demand for platelets averages 5 units per 1,000 population per annum, i.e. 2.5 million units per year costing over €375 million. Due to the ageing population and ever more aggressive chemotherapies platelet usage is increasing 10% every year while the supply of platelets is entirely dependent on allogenic donations.

SilkFusion is developing novel technologies that do not currently exist for producing human platelet ex-vivo. The potential applications are very far-reaching and could lead to the introduction of this technology in other fields, such as the production of other human stem cells or more mature cells. These technologies will have additional applications as novel tools for studying the pathogenetic mechanisms in patients of platelet-related disorders to design more effective and personalized treatments and as a screening technology for the pharmaceutical industry to predict the therapeutic effect of novel compounds in thrombocytopenic and oncological patients. The successful development and distribution of our platform will offer researchers and clinicians a specialized precision instrument for establishing the safety and efficacy of drugs, reducing the current limitations of animal models in the study of thrombocytopenia.

Our project integrates bioengineering, cell and molecular biology, and clinical expertise. The combination of the complementary expertise of the consortium will foster insight into fundamental models to mimic bone marrow physiology for platelet production with breakthrough applications for transfusions, personalized medicine, and drug testing.

Finally, in our program, we are also enrolling and supporting the research community by training young actors towards future technological leadership.

Group photo - Last Governance Meeting

Group photo - Kick-Off Meeting