Periodic Reporting for period 4 - MINERVA (MIcrobiota-Gut-BraiN EngineeRed platform to eVAluate intestinal microflora impact on brain functionality)
Reporting period: 2021-11-01 to 2022-10-31
Brain neurodegeneration is a major challenging ISSUE for todays’ research: Alzheimer’s disease (AD) is expected to reach 80 million patients worldwide in 2050 but still its main triggers are mostly unknown. Recently, many biologists, gastroenterologists and clinicians have proposed an intriguing CONCEPT referred to as microbiota-gut-brain axis (MGBA) that states that there is a connection between the brain, the gut and our intestinal microflora, the group of microorganisms that lives in our gut and that are collectively named “microbiota”. A huge amount of data indicates that microbiota affects brain functionality: a reliable, fascinating HYPOTESYS states that microbiota can trigger neurodegeneration through the MGBA, suggesting new revolutionary therapeutic approaches aimed at slowing down neurodegenerative disease progression by modulating microbiota composition with a huge beneficial IMPACT on SOCIETY worldwide. Unfortunately, the lack of a comprehensive model of the MGBA able to bridge current in vivo model complexity with the in vitro tools’ simplicity represents still a GAP that keeps researchers far from fully clarify microbiota-neurodegeneration potential mechanisms. MINERVA project has targeted this gap using an INNOVATIVE bioengineered APPROACH, to help in exploring in a completely NEW way the relation between neurodegeneration and human intestinal microflora.
MINERVA final GOAL has been to develop the first engineered multi-organ platform recapitulating in vitro the connections among the main players of the MGBA: the gut microbiota; the gut epithelium; the immune system; the blood-brain barrier; the brain.
The PLATFORM relies on three compartments, one for each component of the axis, that host multiple units of an innovative, engineered organ-on-a-chip device, developed within MINERVA project, serially interconnected in order to represent the MGBA players’ interaction.
In the “Microbiota-compartment”, human microbiota strains can be cultured and produce a mix of molecules named “secretoma” that, once transported to the “Gut-compartment”, interacts with human gut epithelial cells and cells from the immune system, that modify it as occurs in vivo, giving the so-called “metabolised secretome”. It reaches the “Brain-compartment”, that hosts a complete Blood-Brain-Barrier model followed by two human brain cell models, where neurons, astrocytes and microglia, the three main cellular populations of the brain, can be: (a) co-cultured to explore microbiota effect on brain cells interconnected as in the real tissue; (b) cultured individually to investigate microbiota impact on each cell type.
MINERVA final GOAL has been to develop the first engineered multi-organ platform recapitulating in vitro the connections among the main players of the MGBA: the gut microbiota; the gut epithelium; the immune system; the blood-brain barrier; the brain.
The PLATFORM relies on three compartments, one for each component of the axis, that host multiple units of an innovative, engineered organ-on-a-chip device, developed within MINERVA project, serially interconnected in order to represent the MGBA players’ interaction.
In the “Microbiota-compartment”, human microbiota strains can be cultured and produce a mix of molecules named “secretoma” that, once transported to the “Gut-compartment”, interacts with human gut epithelial cells and cells from the immune system, that modify it as occurs in vivo, giving the so-called “metabolised secretome”. It reaches the “Brain-compartment”, that hosts a complete Blood-Brain-Barrier model followed by two human brain cell models, where neurons, astrocytes and microglia, the three main cellular populations of the brain, can be: (a) co-cultured to explore microbiota effect on brain cells interconnected as in the real tissue; (b) cultured individually to investigate microbiota impact on each cell type.
The main goal of the MINERVA project has been to develop in five Work Packages (WPs) the first microbiota-gut-brain engineered platform to investigate the interaction between the intestinal microbiota and brain functionality in healthy and neurodegenerative scenarios.
A summary of the activities and main results follows.
We have selected the hydrogel formulations for neural 3D cell model development and optimised the protocols for neural cell culturing and biochemical assessment (WP1). We have set up the protocols for gut epithelium and immune system device exploitation (WP2) and set up the culturing conditions for microbiota strains, developing a new hydrogel formulation to model the gut intestinal mucus in the microbiota device (WP3). We have developed a highly versatile organ-on-a-chip device as “basic unit” to be tailored according to the requirement of the Brain device, Blood-Brain Barrier device, Gut epithelium device, Immune system device, and Microbiota device (WP1-3), and then we have functionally and biologically validated them (WP1-3). We have serially connected the organ-on-a-chip devices implementing MINERVA multi-organ platform, assessing its functional and biological performance once loaded with the in vitro models delivered before and characterised according to selected parameters (WP4). Finally, we have cultured the human microbiota collected from AD patients and healthy donors and allowed the resulting secretoma flow through the multi-organ platform loaded with the in vitro models obtained, and addressing brain cell response to the resulting healthy and neurodegenerative scenarios (WP5).
During the project lifetime, we have shared MINERVA aims and efforts with both the scientific community and the general public, implementing many open access peer-reviewed publications, several press release actions, reported also in social media, several dedicated pages on the Host Institution website. Furthermore, the setting-up ex novo of a dedicated “MINERVA” website, together with the participation to many scientific congresses and events targeting general public have also helped in sharing our main achievements (see www.minerva.polimi.it).
A summary of the activities and main results follows.
We have selected the hydrogel formulations for neural 3D cell model development and optimised the protocols for neural cell culturing and biochemical assessment (WP1). We have set up the protocols for gut epithelium and immune system device exploitation (WP2) and set up the culturing conditions for microbiota strains, developing a new hydrogel formulation to model the gut intestinal mucus in the microbiota device (WP3). We have developed a highly versatile organ-on-a-chip device as “basic unit” to be tailored according to the requirement of the Brain device, Blood-Brain Barrier device, Gut epithelium device, Immune system device, and Microbiota device (WP1-3), and then we have functionally and biologically validated them (WP1-3). We have serially connected the organ-on-a-chip devices implementing MINERVA multi-organ platform, assessing its functional and biological performance once loaded with the in vitro models delivered before and characterised according to selected parameters (WP4). Finally, we have cultured the human microbiota collected from AD patients and healthy donors and allowed the resulting secretoma flow through the multi-organ platform loaded with the in vitro models obtained, and addressing brain cell response to the resulting healthy and neurodegenerative scenarios (WP5).
During the project lifetime, we have shared MINERVA aims and efforts with both the scientific community and the general public, implementing many open access peer-reviewed publications, several press release actions, reported also in social media, several dedicated pages on the Host Institution website. Furthermore, the setting-up ex novo of a dedicated “MINERVA” website, together with the participation to many scientific congresses and events targeting general public have also helped in sharing our main achievements (see www.minerva.polimi.it).
MINERVA RESULTS are INNOVATIVE, because rely on the delivery of a novel, patented organ-on-a-chip device and of the first engineered platform representing the MGBA main players. It embeds new-engineered in vitro models developed ad hoc to better simulate the main players of the axis and their interconnection, a key requirement to collect new insights to clarify microbiota-neurodegeneration biochemical mechanisms.
MINERVA is BREAKTROUGH: its new bioengineering APPROACH opens the way to a completely new FIELD at the boundary between neuroscience and engineering where the investigation of neurodegeneration causes can be shifted from the brain to the body periphery.
MINERVA has a potential HUGE IMPACT: its INNOVATIVE PLATFORM, thanks to its VERSATILE design, might address microbiota impact and therapeutic potential on brain but also not-brain centred diseases.
Finally, MINERVA represents by itself a major advancement in the field of multi-organ-on-chip devices but also in vitro brain disease models to explore new THERAPIES for brain disorders.
MINERVA is BREAKTROUGH: its new bioengineering APPROACH opens the way to a completely new FIELD at the boundary between neuroscience and engineering where the investigation of neurodegeneration causes can be shifted from the brain to the body periphery.
MINERVA has a potential HUGE IMPACT: its INNOVATIVE PLATFORM, thanks to its VERSATILE design, might address microbiota impact and therapeutic potential on brain but also not-brain centred diseases.
Finally, MINERVA represents by itself a major advancement in the field of multi-organ-on-chip devices but also in vitro brain disease models to explore new THERAPIES for brain disorders.