We developed tools and technologies to enable the establishment of a combined ATMP, composed of therapeutic cells with the implantable medical device. Our first-time right concept based on the quality-by-design approach enabled us to identify sources of variability affecting our processes, setup measurements to control them enabling us to generate a streamlined, safe and cost-effective GMP conform process that will be translatable into the clinical setting.
Several manufacturing protocols were generated by UKW to establish a GMP compliant method for cell isolation and cultivation of BOECs using animal component free coated vessels and defined medium for cell growth and expansion. The production and purification of recombinant lentiviruses carrying the genetic code for the human FVIII gene was established. Lentiviruses are capable of effectively introducing functional copies of FVIII into the BOECs enabling them to produce functional clotting factor VIII. These steps were implemented into GMP conditions by UKW and IMS/GRU.
During and after expansion of the corrected BOECs at UNILO,the cells were analysed for their safety profile, functionality and senescence state to evaluate and define release criteria for the therapeutic cell product for future human clinical studies. As a first in vivo preliminary test, UPO conducted an initial test on BOECs in the haemophilia A mouse model showing that corrected BOECs injected interperitoneally released therapeutic levels of FVIII into the blood stream. After conducting surgical implantation, dosing and additional supporting proof of concept studies, SERC demonstrated that the cells met the defined release criteria, transplanted the corrected BOECs into the Cell Pouch™, demonstrating survival of cells for the three-month period. Cell PouchTM transplanted cells were analysed at various time points for cell survival, safety and clotting factor VIII production. For corrected BOECs, it was indicated that the cells were able to produce FVIII within the Cell Pouch™. However dosing optimization with larger sample size is required, but importantly, FVIII released from BOECs within the Cell PouchTM could be detected in blood and an improvement in clotting was shown possible.The implantation and transplantation studies of the scalable, pre vascularized, removable Cell Pouch™ will be used in developing an Instructions For Use document suitable for use in future human clinical studies.
Different sets of design and manufacturing protocols were generated, based on current European GMP regulations leading to the preparation of a common technical document (CTD) for an ATMP, composed of therapeutic cells and an implantable medical device, a combined Advanced or Gene Therapeutic Medicinal Product. With this document the first step is achieved to develop an ATMP medical product, which can be evaluated in a clinical phase I/II study.
Within the project duration, the team was present on various international Haemophilia conferences and attended Venture Capitalist events. Independent of the occasion the concept of HemAcure met with great interest. Currently we are evaluating the best way presenting the results to the scientific community.
