The lack of appropriate in vitro models of the human nervous system hinders the understanding of pathological mechanisms and the development of drugs. This project, part of the IMI2 PAEAN project, aims at generating, through innovative methods based on guided generation of organoids and 3D bioprinting, new 3D models of the human nervous system composed by brain cells derived from human induced pluripotent stem cells (iPSC). iPSCs can be derived from any individual by reprogramming the body's cells, and then differentiated to any cell type of interest. We will produce 3D models of neurodevelopmental and neurodegenerative disorders, as well as and brain tumors. We will analyze these in vitro disease models to study disease mechanisms, and to evaluate whether these models can be used for drug tests and for the development of diagnostic tools. The ultimate goal of the project is that these 3D models characterized by being patient-specific, become reliable platforms for drug screening for personalized medicine. These human, patient-specific preclinical tools might also represent a frontier in the development of new drugs and for the prediction of new and old drug neurotoxicity, and will be provided to pharmacological industry and research organization as an integrated risk assessment approach for better decision-points throughout the R&D process, and better protection of human volunteers and patients, shortening drug development timelines, reducing drug development costs and animal testing.
This project aims to develop and characterize new tools for the study of the biological complexity of the human brain, which can be used to understand the mechanisms of the physiological and pathological functioning of the brain in basic research, and which can become models for translational research.
In his vision, the project, articulated over 36 months, aims not only to generate and characterize 3D brain models, but also to optimize 3D bioprinting protocols that can also be extended to other human organs and other pathologies.
In a longer-term view, 3D models of human body organs can be integrated into microfluidic chips to obtain a flexible, realistic and reproducible platform aimed at replicating the function of the human body in a holistic approach. These platforms can be used by both researchers and pharmaceutical players to understand the fundamental mechanisms of action and the side effects of drugs on human organs, increasing our knowledge of the mechanisms of action and toxicity. In a long-term perspective, personalized therapies can be developed by reducing adverse events during clinical studies and improving development costs.
The vision behind the project is that the development of organoids from patient cells is a very powerful and innovative tool for the development of personalized treatments. The development of 3D printed models with bioprinting technology for R&D applications is expanding and is supported by a strong business drive. This is also thanks to the fact that 3D bioprinting allows the multiscale arrangement of cells, biomolecules and materials similar to the microenvironment of native tissues.
The organoids produced by the project will be derived from a particular type of human stem cell called iPSC, generated from non-stem cells (somatic) of the blood or skin taken from the patient, without health risks, and without manipulation of embryos. The project aims to provide added value by combining their use with another recent but consolidated technology, namely the 3D printing of biological material (3D Bioprinting). 3D bioprinting allows to produce 3D structures that recapitulate the organization of individual cells in the real organ. The procedure is based on the robotic manipulation of cellular material and organic encapsulation matrices, which are organized in space in an extremely controlled manner, similarly to the ink of a conventional 3D printer.
By combining the two technologies, the project will be able to provide extremely realistic 3D models, starting from patient cells. An added value is the possibility of providing tools for the study and pharmacological development for rare diseases for which pharmacological research is slowed down by the lack of animal models and by the high cost compared to the low profitability.
In addition, as regards cancer research and the development of patient-centered cancer therapies, to reduce resistance to the therapy itself, 3D-printed constructs starting from cells derived from brain tumor resected tissue, can reproduce, in a controlled way, the complexity of the tumor microenvironment (TME), fundamental for the efficacy of the therapy. In fact, although animal models have improved our knowledge of cancer biology and provided the scientific basis for numerous clinical studies, they are not able to fully recapitulate human TME. Recently, the development of standardized patient-derived xenograft models represents a powerful tool for predicting the effectiveness of cancer therapy. These models, however, devoid of immune cells, are not suitable for the study of the immune TME, unless they are grafted with the functional human immune system. The 3D bioprinting ensuring great repeatability, and the possibility of a precise design of the TME, also overcomes the barriers inherent in the tumor spheroids, which, based on the self-assembly of the cells, limit the control on the 3D culture environment, which is certainly necessary for the methodical investigation of specific TME functions.
Gender dimension
We will develop gender specific 3D brain models using iPSC-derived cells, enabling to test drugs in equal measure on "male" and "female" models, using somatic cells both from male and female patients. This responds to a major need of the society in the pharmaceutical research: to date, most preclinical tests are carried out on male laboratory animals and more men than women are generally enrolled in clinical trials. With respect to this male prevalence in preclinical and clinical trials, most adverse drug reactions affect women. We are sensitive to the problem of gender pharmacology: not always a drug developed for a man is also suitable for a woman. On the other hand, besides the hormonal factors that affect the whole person in its development, there are several characteristics related to the chromosomal difference (XY vs XX) that should not be underestimated and that we can emphasize by producing both "male" and "female" 3D models.