#	Pagina
attuale pagina	/open-h2020/projects/200813/results.html

Report

Teaser, summary, work performed and final results

Periodic Reporting for period 1 - HISENTS (High level Integrated SEnsor for NanoToxicity Screening)

Teaser

Summary

\"The goal of HISENTS is to develop a smart multimodule screening technology for nanosafety assessment with mechanistic functionality. This will provide \"\"how\"\" and \"\"why\"\" answers to the fundamental processes of human response to nanomaterial (NM) toxicity using toxicity pathway analysis, toxicogenomics, and novel endpoints. For this purpose an innovative miniaturised high throughput screening (HTS) tool will be developed with the capability of reliably analysing bio-nano-interactions within a platform of increasing complexity of biological organisation. HISENTS will also align this experimental platform with a comprehensive physiologically based pharmacokinetic (PBPK) type model to aid in the platform design and to contribute to the pathway and mechanistic analysis. The PBPK model together with the instrumentation development is integrated into one multimodular platform screen.

HISENTS is important to society as:
1. It leads to a step increase in the speed of screening nanomaterials;
2. The HISENTS platform will have widespread applications in addition to nanosafety;
3. It enables screening systems to be fully computer controlled increasing their reliability and reproducibility;
4. The platform is very amenable to PBPK modelling in effect developing the animal-on-a-chip toxicity testing technology backed up by computer prediction;
5. The approach will derive quantitative structure activity relationship ((Q)SAR) properties of NMs so that very general principles of bio-nano-interaction will be derived;
6. It creates a technology with high translational potential to the pharma, water, health and safety and security industries.

HISENTS overall objectives are:
1. Synthesise and characterise NM for use as standard test analytes;
2. Design and test effective screening devices, each representing a particular physiological function which can be incorporated into a multimodular platform for sensing nanotoxicity;
3. Configure robust electrochemical and optical techniques to interrogate the individual devices. Develop and innovate instrumentation for interfacing to sensor chips;
4. Incorporate and integrate the screening devices into miniaturised and effective flow systems for HTS of NM along a directional pathway;
5. Develop smart automated signal processing data recognition techniques;
6. Carry out comprehensive performance evaluation of platform with respect to each individual device and the whole platform.\"

Work performed

The work performed for Period 1 (1st April 2016 to 30th September 2017) is given under individual work packages. WP1 has synthesised 4 groups of NM: Au, SiO2, FexOy and TiO2. These NM were extensively characterised and sent out to all partners in the consortium. In addition Au and Ag NM of varying size, shape and functionality have been synthesised and characterised in particular for understanding their interaction with the. biomembrane module sensor element. WP2 has developed microfluidic networks and electronic systems for interfacing to the sensor element modules developed by the individual partners. In particular a signal processing and data unscrambling algorithm has been successfully developed for analysing data generated by the biomembrane module. The biomembrane module has also been incorporated into an automated fluidic system which together with the electronic interrogation is entirely computer controlled. WP3 has identified strains of miRNA which are sensitive to NM damage. At the same time DNA-lipid conjugates can be incorporated into biomembrane sensing element so that DNA damage can be electronically detected on-chip. WP4 has carried out a comprehensive study on the interactions of novel NM with the biomembrane sensor element in the flow system. The lung-on-chip sensing device has been developed and the single cell, liver-, kidney-, intestine- and placenta-on-chip devices are in various stages of development together with their associated chip-based supports, flow systems and electronic and optical interrogation. WP5 has developed a nine compartment PBPK model to mirror the final experimental platform. The model has been tested with published results on NM accumulation in in-vivo tissues and the fit to experimental data is good. Finally WP6 has carried out two intercalibration exercises. In the first, three soluble toxicants were sent out to six partners in the consortium. The results were extremely encouraging and showed that all sensor elements responded identically to the toxicity ranking of the toxicants. This can be explained by the fact that the cell/tissues sensor elements were recording membrane damage in their toxicity assay. In the second exercise four groups of NM were sent out to the same partners. The results were not so straightforward due probably to the aggregation of the NM with time but they did show some consistency when this was taken into account.

Final results

Advances and expected results in HISENTS are as follows - HISENTS:

1. Pioneers synthesis of new types of NM. The novel NMs will be used to understand more carefully structure-activity relationships and individual pathway analyses of each specific material in the multimodular system;
2. The programme includes miniaturisation, automation and integration of in-vitro devices into a chip-based multimodular smart screening platform;
3. Develops several novel endpoints. These include: (a) membrane modification (b) cell deformability change and (c) DNA structural damage;
4. Takes miRNA and DNA transforming their interrogation to high throughput multimodular devices;
5. Substitutes the well plate for a flow module interconnected by a microfluidic network that provides each compartment with necessary reagents and media components;
6. Modifies the chip-based transducer design miniaturising it from macro to micro/nano dimensions and reconfiguring the interrogation system to allow for the highest sensitivity;
7. Integrates a series of high level toxicity assays into a multimodular platform within a microfluidic system where each module contains a sensor element at successively higher level of biological organisation;
8. Interfaces the multimodule flow-through system with smart customised multichannel electronic potentiostatic instrumentation which will synchronise flow system manipulation with sensor element handling and allow several modules to be cross-examined simultaneously;
9. Develops the conceptual framework of the PBPK model approach side by side with an experimental system.

Potential impact:
The global HISENTS technology rationalises current toxicity testing allowing it to be carried out by a single instrument rather than by a whole series of â€œwetâ€ tests. HISENTS technology will therefore revolutionise the way toxicity testing is carried out.
Wider socioeconomic impacts of HISENTS arise from its capacity as a generic automated toxicity screener for appropriate organisations e.g.:
Water industry: toxins/NM in potable water;
Environmental and marine agencies of toxins/NMs in the river, estuarine and marine environments;
Government agencies and statutory bodies: toxicants/NMs in aqueous environments in particular to develop risk assessments for the NMs\' use;
Commercial and academic NM producers for formulation of risk assessment procedures and disposal methods for the materials;
Defence and security industry throughout the EU including NATO, border control and civil defence for the toxicity of NMs where these are manufactured with terrorist ambitions.