Periodic Reporting for period 2 - BrainMatTrain (Development of Biomaterial-based Delivery Systems for Parkinson’s disease - an Integrated Pan-European Approach) Reporting period: 2018-01-01 to 2019-12-31 Summary of the context and overall objectives of the project The problem being addressedParkinson’s disease (PD) is a chronic neuro-degenerative disorder characterized by the progressive loss of dopamine-producing neurons in a particular part of the brain resulting in various motor and non-motor symptoms. The loss of these cells is caused by chronic neuro-inflammation and results in a feed forward cycle promoting further inflammation and neuro-degeneration within the brain which characterizes the pathophysiology of PD. Current treatments for PD are symptomatic pharmacotherapies that neither address the underlying disease pathogenesis nor prevent, slow or halt the chronic, progressive neuro-degeneration. The BrainMatTrain research program aims to develop novel biomaterial-based scaffolds, which facilitate functional regeneration in the central nervous system (CNS). Our project addresses both the inflammation and neuro-degenerative phases of the underlying pathology by using collagen reservoir scaffolds incorporating drugs and cells that are targeting these phases of the disease.Overall ObjectivesThe main scientific and technology objectives of the research program are to:• Design collagen-based scaffolds for the delivery of therapeutic moieties• Investigate their efficacy in in vitro models of Parkinson’s disease• Conduct pre-clinical studies in in vivo models of Parkinson’s disease• Scale up collagen scaffolds• Develop a device for delivery of functionalised scaffoldsImportance for societyParkinson’s disease is the second most common neuro-degenerative disease affecting more than ten million people worldwide. Many neuro-degenerative diseases are incurable due to the progressive degeneration and/or death of neuron cells. As research advances, many similarities appear that relate these diseases to one another on a sub-cellular level. Discovering these similarities offers hope for therapeutic progress that could ameliorate many diseases simultaneously. Therefore, this project supports the EU objectives on research on neuro-degenerative disease diagnosis and treatment as the European Parliament has repeatedly emphasized the importance of addressing the challenges of neuro-degenerative disease in research as well as in social and health policies. Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far Work performed and main resultsVarious collagen hydrogels have been successfully designed, optimized and fabricated which resulted in final products that proved to be non-toxic and highly stable both in vitro and in vivo. Secondly, these hydrogels have further successfully been used as a delivery system by encapsulating cells and drugs to target neuroprotection and neuro-inflammation. These so-called functionalised hydrogels did indeed decrease the inflammatory processes in the brain showing not only their biocompatibility and bio-protective qualities but also their substantial abilities to reduce the underlying inflammation that characterises PD.Additionally, new in vitro next generation PD model systems have being developed within the project. A robust protocol for re-programming adult human fibroblasts (abundantly present in the body) from patients with PD into dopaminergic neurons has been designed. This is a crucial step for future treatments as these cells derived from the patients themselves and would obviate the use of stem cells. These cells resemble the physiological profile of healthy dopaminergic neurons and they withstand encapsulation within BMT scaffolds, which is a fantastic proof of applicability for future treatments.Another model of Parkinson’s disease is the lab-on-a-chip model. A novel technique for the fabrication of microfluidic chips, termed 3D-printed soft lithography, was developed within BrainMatTrain that enables 3D printing of cells in liquid collagen as well as in hydrogels. Moreover, to refine this technique, we additionally developed 2D neurite guidance chips. Both chips will help researchers to create more in vitro models of neurological disorders as 3D-printed soft lithography allows the fabrication of microfluidic chips for the creation of interconnected neural networks in 2D.To characterize the in vivo responses of the implanted biomaterials, a robust animal model of pure neuro-inflammation was successfully developed to be analysed by the non-invasive in vivo neuro imaging system positron emission tomography (PET). This animal model is a valuable tool which will enable the evaluation of the inflammatory aspect only of Parkinson’s disease. Additionally, the long-term survival and the functionality of the implanted dopaminergic neurons, within the hydrogel scaffolds was successfully tested in a relevant animal model of PD. Therefore, the degree of the pathophysiological effects can now be measured before and after implantation of different hydrogel variations by analysing certain inflammatory markers, the changes of the protein patterns (proteomics) and the in vivo characterization via PET.Furthermore, our industry partners developed protocols and strategies for the scale-up of the biomaterial for later commercialization.For designing the device that delivers the biomaterials including cells and drugs into the brain, several physical stress conditions of the material have been tested. This enhanced the material stability when injected through the device on a larger scale.Additionally, to ensure that sufficient quantities of mesenchymal stromal cells (MSCs) will be available for use in the clinic, transformative technologies that deliver highly purified MSCs, called ORBCEL-CTM, have been successfully developed. These cells were modified and optimized to be used in a closed automated cell expansion system to scale-up the manufacture process. 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) Progress beyond the state-of-the-artBrainMatTrain trained 17 Early Stage Researchers (ESRs) in multidisciplinary fields necessary to fully engage in the tools of tissue engineering and regenerative medicine. A further cornerstone of their education within BrainMatTrain was the acquisition of many transferable skills which will enhance their future career paths in either industry or academia or elsewhere.One key aspect within the project’s lifetime was the emphasis on educational and public engagement (EPE) activities, which resulted in a vast participation of all researchers. The fellows engaged in many meaningful interactions with the public, for example a 30-minute science documentary. It has been screened 59 times covering an audience of over 314,040 members of the public, academic staff and students. The film was scheduled for festival screenings in the USA, Germany, Sweden and The Netherlands and was also broadcast in Ireland for one screening per year over a period of three years.Additionally, the newly established EPE program ‘Strength in Science’ will have an impact on society as it addresses one of the worldwide problems of obesity and lack of physical activity. BrainMatTrain researchers convincingly explained the secondary effect on brain health to teachers and 3,032 actively involved pupils. Similarly acknowledged was the newly developed stem cell game ‘STEMinator’. This card game contains scientific data emphasized by very appealing drawings. This combination of easy utilization, attractive images and fun led to its successful dissemination to the public via multiple channels including https://www.eurostemcell.org/. Simultaneously, we have fulfilled one of the missions in life science, namely elucidate the facts of stem cells in research to the public. Figure 5: Documentary Film Poster Figure 4: The new stem cell card game ‘STEMinator’ Figure 1: Lab-on-a-chip device Figure 2: Collagen particles in oil at high magnification Figure 3: BrainMatTrain fellows engaging with the public at a science fair
