Periodic Reporting for period 1 - BrainChip4MED (Brain-on-a-chip as a preclinical model tool for the screening of theranostic nanoformulations for neurodegenerative diseases)
Reporting period: 2022-03-01 to 2023-02-28
Dementia, which includes a broad category of brain disorders that decline brain function, has for a long time been a global challenge that was dismissed by pharmaceutical companies due to the low penetration efficiency of drugs that are able to cross the blood-brain barrier (BBB), as well as to the limited understanding of the underlying biophysical mechanism, which is still mostly unknown.
Recently, brain-on-a-chip (BoC) has emerged as an advanced microfluidic platform combined with 3D tissue culture techniques, with the potential to create an accurate and simple-to-use preclinical model tool, by decoupling a complex organ, such as the brain, into different cellular structures, while maintaining their interconnections. This approach allows for the precise assessment of drug molecules and/or drug nanocarriers along the different tissues, unveiling new interactions between them, essential for the development of new therapeutic strategies for neurological diseases. Also, the possibility of integrating biosensors on it could extend its monitoring and workability for longer periods of time.
Motivated by the lack of an appropriate in-vitro model to study brain-targeting drug nanocarriers, and the potentiality of BoC technology to recapitulate human biology and predict in-vivo response, the overreaching aim of this action is to develop an advanced microfluidic preclinical device with the structural and functional aspects of the brain tissue and BBB, using BoC as a preclinical model tool to assess and study possible alternatives for the diagnosis and treatment (i.e. theranostic) of neurodegenerative disorders, and to develop multifunctional novel stimuli-responsive drug nanocarrier systems (Figure 1).
By so, BrainChip4MED prototype (mimicking the neural tissue and BBB integrated with multiplexed biosensors) proposed in this engineered platform is a cutting-edge technology that allows for the study of new drug delivery strategies and treatments of neurological diseases, such as Alzheimer’s disease (AD).
Therefore, the present project goes beyond the current state-of-the-art, by addressing the main two challenges in the neuroscience field, (1) the inefficiency of FDA-approved drugs for targeting and bypassing the BBB, by developing and studying new added-value theranostic NFs, and (2) the lack of robust preclinical tools for screening and monitoring the efficacy of those NFs to cross the BBB and target diseased neurological cells. The envisioned technology has a high potential to impact the society by delivering an advanced microfluidic device capable to reduce the use of animal tests, increase the successful rates in the translation of novel nanoformulations from laboratory to clinical use, and find a successful strategy that allows a better understanding and treatment of AD and dementia, which is significantly growing worldwide among the aging population.
Recently, brain-on-a-chip (BoC) has emerged as an advanced microfluidic platform combined with 3D tissue culture techniques, with the potential to create an accurate and simple-to-use preclinical model tool, by decoupling a complex organ, such as the brain, into different cellular structures, while maintaining their interconnections. This approach allows for the precise assessment of drug molecules and/or drug nanocarriers along the different tissues, unveiling new interactions between them, essential for the development of new therapeutic strategies for neurological diseases. Also, the possibility of integrating biosensors on it could extend its monitoring and workability for longer periods of time.
Motivated by the lack of an appropriate in-vitro model to study brain-targeting drug nanocarriers, and the potentiality of BoC technology to recapitulate human biology and predict in-vivo response, the overreaching aim of this action is to develop an advanced microfluidic preclinical device with the structural and functional aspects of the brain tissue and BBB, using BoC as a preclinical model tool to assess and study possible alternatives for the diagnosis and treatment (i.e. theranostic) of neurodegenerative disorders, and to develop multifunctional novel stimuli-responsive drug nanocarrier systems (Figure 1).
By so, BrainChip4MED prototype (mimicking the neural tissue and BBB integrated with multiplexed biosensors) proposed in this engineered platform is a cutting-edge technology that allows for the study of new drug delivery strategies and treatments of neurological diseases, such as Alzheimer’s disease (AD).
Therefore, the present project goes beyond the current state-of-the-art, by addressing the main two challenges in the neuroscience field, (1) the inefficiency of FDA-approved drugs for targeting and bypassing the BBB, by developing and studying new added-value theranostic NFs, and (2) the lack of robust preclinical tools for screening and monitoring the efficacy of those NFs to cross the BBB and target diseased neurological cells. The envisioned technology has a high potential to impact the society by delivering an advanced microfluidic device capable to reduce the use of animal tests, increase the successful rates in the translation of novel nanoformulations from laboratory to clinical use, and find a successful strategy that allows a better understanding and treatment of AD and dementia, which is significantly growing worldwide among the aging population.
The main scientific and technological achievement of WP1, performed during the secondment phase at Harvard Medical School/Brigham Women’s Hospital (HMS/BWH), was the biofabrication of the BoC platform comprising both brain and BBB models. With this aim, a novel soft-hydrogel was designed to mimic the BBB ECM and act as a bio-membrane that separates the vascularized cells from the brain. This BBB model consisted of a gelatine-based hydrogel incorporating heparin, as a pro-angiogenic growth factor immobilization molecule, and hyaluronic acid (HA) as an enhancer of the mechanical strength and regulator of the physiological processes related to BBB cells. This biochemically engineered approach created a simple, rapid and non-toxic method to produce the BBB ECM. After the biofabrication, heparin served as a molecular platform to non-covalently bind the growth factors FGF-2 and VEGF. All sets of gelatine-based hydrogels were fully characterized in terms of mechanical properties, swelling, biodegradation, cell adhesion and proliferation, as well as gene expression. Overall, the results shown that the developed fabrication strategy created an innovative bio-membrane able to sustain and generate ideal conditions for the spread, adhesion and close junction of endothelial cells for several days, which represents a major advance in the current state-of-the-art to develop mimicking BBB models. In particular, our results showing that the vascularized endothelial cell layer (BBB) could be cultured in the bio-engineered membranes for seven days, were presented in a Poster communication at the 2022 Discovery Brigham (Boston, USA, November 2022).
The development of this innovative bio-membrane contributed for the improvement of the BoCs state-of-the-art, by delivering a novel platform, where the BBB is closely represented with the adhesion and close junction of endothelial cells for several days. In parallel, the brain model was also developed and optimized using commercial cell lines, particularly human iPS cell-derived astrocytes and neurons cultured in Matrigel.
The validation of the BrainChip4MED prototype in dynamic flow was conducted during at least 5 days, in continuous flow, and key-biomarkers were used to monitor the cell activity of the BBB and brain model. Additionally, immunostaining and live/dead analysis techniques were performed to screen the viability of the biomodels along the dynamic in-vitro culturing. By achieving this main research objective (RO1) planned for the outgoing phase (WP1), this bioengineered platform creates the cutting-edge technology that will allow the study of new drug delivery strategies and treatments of neurological diseases, such as Alzheimer’s disease, planned for the second year in the returning phase (WP2 and WP3).
The development of this innovative bio-membrane contributed for the improvement of the BoCs state-of-the-art, by delivering a novel platform, where the BBB is closely represented with the adhesion and close junction of endothelial cells for several days. In parallel, the brain model was also developed and optimized using commercial cell lines, particularly human iPS cell-derived astrocytes and neurons cultured in Matrigel.
The validation of the BrainChip4MED prototype in dynamic flow was conducted during at least 5 days, in continuous flow, and key-biomarkers were used to monitor the cell activity of the BBB and brain model. Additionally, immunostaining and live/dead analysis techniques were performed to screen the viability of the biomodels along the dynamic in-vitro culturing. By achieving this main research objective (RO1) planned for the outgoing phase (WP1), this bioengineered platform creates the cutting-edge technology that will allow the study of new drug delivery strategies and treatments of neurological diseases, such as Alzheimer’s disease, planned for the second year in the returning phase (WP2 and WP3).
The expected outcomes and impacts of this research project are relevant at technological, human health and economic levels, and are fully aligned with the UN Sustainable Development Goal (SDG 3) – Good health and well-being. Specifically, the analytical BoC tool and NFs for AD, in development in this action, are essential to understand the progress and prognosis of neurodegenerative diseases, and to support therapeutic decisions of drug therapy (currently ineffective). The proposed BoC is expected to deliver an innovative advanced preclinical platform that mimics the human brain physiology and surpass the use of animal models (3R’s), which besides the ethical issues, is the main responsible for the bias results and failure of new medicines in clinical trials. Thus, this project expects to deliver for the first time, a prototype that integrates in the same platform, BBB and brain models in situ monitored by biosensor’s analysis. This achievement represents a step further in the automatization and robustness of these advanced preclinical platforms, with the potential to reduce the typical extended costs and time consumed in the development/screening of new medicines. Thus, possibly instigating the research of new strategies to treat dementia at lower market prices. Also, nanomedicines in study can increase the efficiency of drugs for AD, decreasing their side effects and ensuring better population ageing condition. Moreover, the analytical tool/prototype that will be obtained has great potential as a commercial product. At the same time, the proposed application has great outreach for academia and pharmaceutical research, contributing to the advancement of nanomedicine and neurosciences in Europe and beyond.