Hollow fiber system for tuberculosis setting up at university of Zaragoza

Ramón García, Santiago
TUBERCULOSIS 2022 - EMBO Workshop on Tuberculosis 2022 - From innovation to intervention
Participation type: 
Other authors: 
D. Aguilar Ayala, M.S. Rabodoarivelo, F. Sanz-García, R. Bailo, A. Picó Marco, B. Gracia-Diaz, A. Muzylo, A. Lucia-Quintana, S. Ramón García
Paris, France. Sep 12-16 2022

The Hollow Fiber System for tuberculosis (HFS-TB) is a preclinical tool for drug evaluation qualified by the European Medicines Agency in 2015. The system allows the reproduction of selected drug exposures to infer PK/PD parameters, which will feed in silico models able to inform the design of Phase 2/3 clinical trials. Although methodological guidelines for HFS-TB exist, there are no standard procedures for its implementation nor mandatory quality control steps for intra-laboratory reproducibility. In this context, UNIZAR was tasked with the implementation of the HFS-TB in BSL-2 and BSL-3 laboratories within the ERA4TB consortium.
The setting up was divided into two main phases: 1) assessment of experimental needs and internal capacity; and 2) standardization of operational procedures.
First, we tested and selected the more suitable equipment and materials in terms of compatibility, size, and ease to handle, to perform mono and combinatorial assays in BSL-2 and BSL-3 facilities. Capacity assessments were performed including the understanding of economical and personnel cost. Timeframe estimates revealed “protocol design” and “troubleshooting anticipation” as key steps for a successful experimental outcome.
The next phase involved the optimization of different procedures to ensure optimal experimental designs. This included bacterial growth dynamics assessment in several media, different types of cartridges and different inoculum sizes. We also evaluated sampling methods for optimizing PD measurement as well as PK profile verification.
With the info obtained, we stablished the work-flow for HFS studies: an initial study of the fiber compatibility with the drug, confirmation of PK profiles achievement, drug testing of broad conditions in the M. tuberculosis H37Ra strain (BSL2 facilities) and finally drug testing of selected conditions in the M. tuberculosis H37Rv strain (BSL-3).
In summary, we have successfully implemented HFS-TB at UNIZAR in BSL2 and BSL3 facilities. Our findings contribute to the standardization of the HFS-TB and should be considered when designing protocols to allow reproducibility between laboratories.
This work has received support from the Innovative Medicines Initiatives 2 Joint Undertaking (grant No 853989) and reflects only the author's views. The JU is not responsible for any use that may be made of the information it contains.