Periodic Reporting for period 1 - PORTable STaNDOuTs (Perfused Organs-on-a-Chip integrated with TEER measurement technique: A novel approach towards studying non-targeted drug induced organ toxicity) Reporting period: 2021-04-14 to 2023-04-13 Summary of the context and overall objectives of the project The growing global concern over the outbreak of new diseases and disease conditions makes it pertinent to synthesise new drug molecules to protect the society from any undesirable health conditions. The process of drug development is time–consuming, taking about 10 to 15 years and cost-ineffective, involving more than a billion Euros. To consider a newly synthesised drug molecule safe for human administration, the drug must undergo two stages, namely, preclinical and clinical trials in the drug developmental process. During the preclinical stages, the efficacy, toxicity and pharmacokinetics-pharmacodynamics (PK-PD) based data of the drugs are collected. It has been found that about one out of ten drugs tested fail in the clinical trials owing to inefficient and unsuitable models available for the preclinical trial stage of the drug development process.In the light of this, the EU-funded MSCA-IF project, ‘PORTable STaNDOuTs’, was aimed to develop a novel sensor-based electronic device that can be integrated on Organs-on-a-chip microfluidic platform to measure trans-endo(epi)thelial electrical resistance (TEER) measurements to replace the conventional time-consuming and cost-ineffective preclinical procedures in the drug development process. TEER measurement is rapid and non-invasive, and helps in assessing the integrity of the tissue layer barrier of different organs necessary for homeostasis in the human body. The developed novel device demonstrated the promising capability to measure TEER/impedance of the tissue layer of any organ system grown on a microfluidic platform. Thus, the developed device capable of supporting growth and monitoring of the 3D biological constructs within the microfluidic platform can be appropriately termed a microphysiological system (MPS). Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far A thorough literature review was carried out at the beginning of the project to understand the ongoing works related to the TEER sensor development and its applications in different Organs-on-a-Chip microfluidic platforms. Commercially available TEER based devices and patents were also thoroughly studied. An initial write-up about the project was also published on the Cherry Biotech website as an act towards dissemination of the relevance of the project and also realising the positive impact it will create on providing an alternate reliable in vitro model as compared to the animal models for the pharmaceutical drug developmental process. The project, ‘PORTable STaNDOuTs’, served to develop a novel 4-electrodes 24 multiwell TEER measuring device aimed to overcome identified not-addressed needs as already mentioned earlier. It is to be mentioned that the device was co-developed with the biosensors team at the BioSense Institute, Serbia. As such, in course of development of the device, other outcomes related to the subject of heart cells electrophysiological stimulation and recording had also been incorporated by Cherry Biotech and BioSense Institute based on other going projects (CISTEM, Grant Agreement 778354). The TEER device was also developed in such a way that it is compatible with previously developed patented microfluidic disposables (Cherry Biotech SAS, FR3094012B1) to incorporate cells maintenance and microfluidics control in a widely used standard for biological cell culturing. Additionally, as part of the project, the ER also worked with other commercially available optical and electrochemical sensors with a vision towards the importance of integrating these sensors along with that for TEER measurements. This will help in bringing about a robust and reliable microfluidic platform capable of monitoring important parameters/metabolites such as TEER, dissolved oxygen, pH, glucose and lactate, thus leading to an all-in-one in vitro device as an alternative to animal model for the drug developmental process.The outcome of the project, ‘PORTable STaNDOuTs’, is a working prototype that is capable for making reliable and repetitive TEER or impedance-based measurements. The prototype is a novel design containing 2 pairs of electrodes in each well of a 24 multi-well plate (dimensions matching commercially available 24MW plates) that can carry out 4 terminal-based impedance measurements. The device was tested for making impedance measurements on endothelial tissue layer and is found to indicate promising results, thereby implicating the extrapolation of using this device for other cell types as well. In addition to the TEER-based experimentation, the ER was also able to optimise important parameters such as dissolved oxygen and pH by carrying out several experiments using commercially available optical flowthrough sensors when endothelial cells were subjected to perfused condition using the MPS controlling prototype developed at Cherry Biotech (CB). The outcome from these optical sensors-based experimentation will be used in the near future for other cell types when different 3D biological models will be developed at CB. Considering the novelty and the potential of the developed TEER measuring device, a patent preparation is also in progress. The impedance-based results obtained by carrying out the tasks of this project will also be published in a peer-reviewed international journal of good impact factor, that will comply to the Horizon Europe mandate for publishing scientific articles. Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far) The results of the work carried out as part of this project once commercialised can have an impact on the evaluation of therapeutic compounds during drug development research phases. The combination of the developed device with other technologies of Cherry Biotech will bring new value to them and pave the way towards the commercialisation of animal-free drug testing by means of human cells-based in vitro microfluidic devices. The devices developed also have the potential to impact positively jobs and value creation in the EU space. As an additional positive impact, this Marie Curie action has allowed the ER to gain experience and insight with the EU product-oriented research and development space, and to experiment it with the facilities and training provided at Cherry Biotech, the host institution. TEER/Impedance-based results derived from carrying out tasks involved in the PORTable STaNDOuTs proj
