Nano-antibiotherapy screening in adult zebrafish for tuberculosis treatment
The purpose of the experiments is to use a well-established tuberculosis (TB) model in adult zebrafish involving Mycobacterium marinum to test different nanotherapeutic strategies to deliver encapsulated anti-TB antibiotics. The application requires the use of adult zebrafish and is subject to regulation.
The level of distress for the fish is expected to be moderate.
Technical pilot experiments will be conducted to master methods allowing to reduce the number of fish used and to minimize negative effects on the fish well-being. A requirement of 15 fish per experimental condition (e.g. NP with drug versus non-NP (free) drug) has been calculated to be needed to obtain statistically significant results. For screening and testing of the different nanoparticle formulation strategies, a maximum of 3004 adult zebrafish are planned to be used (2390 wild-type - 234 lyve1:DsRed/kdrl:GFP – 380 fli:GFP/mpeg1:mCherry).
The rationale for using adult zebrafish (wild type and GMO) are that these relatively transparent animals are small enough to perform detailed whole-organism imaging of fluorescent NP and bacteria, as well as live-imaging of (selectively labeled) host cells. These are technical limitations of mouse models, the preferred preclinical model. Compared to zebrafish larvae, the adult zebrafish possess a fully functional immune system, allowing the induction of both latent and progressive forms of infection, as well as the monitoring of NP over long period. The use of the adult zebrafish is an extension of the rapid screenings of NP in zebrafish larvae previously made. In addition to providing further insight into off-target interactions, this facilitates the selection and characterization of the most promising nanotherapeutic strategies to be further tested in a mouse model of TB. This will be a large improvement in the development and evaluation of new delivery systems for anti-tuberculosis drugs, which are both able to enhance therapeutic efficacy and reduce toxic side effects.
Collectively, these experiments will contribute greatly to reducing both the number of mice needed in the next step (in support of the 3R principle) and in volume of drugs and NP needed for testing. In addition, a power analysis has been conducted to determine the minimal number of required animal to perform reliable experiments. A special care has been taken in order to refine experimental procedures, such as the reduction of phototoxicity risks during live-imaging and enriched housing conditions..
The level of distress for the fish is expected to be moderate.
Technical pilot experiments will be conducted to master methods allowing to reduce the number of fish used and to minimize negative effects on the fish well-being. A requirement of 15 fish per experimental condition (e.g. NP with drug versus non-NP (free) drug) has been calculated to be needed to obtain statistically significant results. For screening and testing of the different nanoparticle formulation strategies, a maximum of 3004 adult zebrafish are planned to be used (2390 wild-type - 234 lyve1:DsRed/kdrl:GFP – 380 fli:GFP/mpeg1:mCherry).
The rationale for using adult zebrafish (wild type and GMO) are that these relatively transparent animals are small enough to perform detailed whole-organism imaging of fluorescent NP and bacteria, as well as live-imaging of (selectively labeled) host cells. These are technical limitations of mouse models, the preferred preclinical model. Compared to zebrafish larvae, the adult zebrafish possess a fully functional immune system, allowing the induction of both latent and progressive forms of infection, as well as the monitoring of NP over long period. The use of the adult zebrafish is an extension of the rapid screenings of NP in zebrafish larvae previously made. In addition to providing further insight into off-target interactions, this facilitates the selection and characterization of the most promising nanotherapeutic strategies to be further tested in a mouse model of TB. This will be a large improvement in the development and evaluation of new delivery systems for anti-tuberculosis drugs, which are both able to enhance therapeutic efficacy and reduce toxic side effects.
Collectively, these experiments will contribute greatly to reducing both the number of mice needed in the next step (in support of the 3R principle) and in volume of drugs and NP needed for testing. In addition, a power analysis has been conducted to determine the minimal number of required animal to perform reliable experiments. A special care has been taken in order to refine experimental procedures, such as the reduction of phototoxicity risks during live-imaging and enriched housing conditions..