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Heart On chip based on induced pluripotent Stem cell Technology for personalized Medicine

Periodic Reporting for period 1 - CISTEM (Heart On chip based on induced pluripotent Stem cell Technology for personalized Medicine)

Reporting period: 2018-01-01 to 2019-12-31

Duchenne muscular dystrophy (DMD) is a serious genetic disorder, occurring in 1/3 500- 5000 children and develops due to mutation in dystrophin gene. Cardiomyopathy in DMD develops in the second decade of life in the significant proportion of patients (more than 60% at age of more than 18 years) together with the respiratory failure and both ultimately lead to death in second or third decade of life. Understanding the mechanisms of cardiomyopathy can provide better chance for patients, improving their conditions of life and prolonging its extent. However, the available animal models only partially reflect the severity of human diseases. In parallel, due to the variability in disease mechanisms, medications and dosages may vary among patients, making it difficult the provision of universally efficacious treatment, creating a tremendous burden on the healthcare system. Microfluidic technology-based “Organs-on-a-chip” like “Heart-on-a-chip” that mimic tailored micro-environment architecture inspired by organ-level functions in vivo represents a powerful tool for investigating DMD disease mechanisms and testing new drug and treatment. Its conjunction with hiPSCs (Human-induced pluripotent stem cells) might offer an excellent opportunity to engineer heart-on-a-chip in a patient-matched manner and overcome the variability in DMD disease mechanisms that can occur from one patient to another. CISTEM project aims to merge recent progress in stem cell biology and advanced microfluidic and MEMS technologies in order to develop a novel “heart-on-chips” platform based on iPSCs-derived cardiomyocytes and to propose a new model of DMD-cardiomyopathy. To achieve this objective, CISTEM has defined two main steps:

• Engineering Heart –on-chip from Cardiomyocytes-derived hiPSCs using microfluidic and microsystem technologies
• Use of the developed platform to model and investigate the cardiomyopathy coming from the Duchenne muscular dystrophy.

In addition, CISTEM project aims also to implement a high quality staff exchange programme in order to :

• Connect high-quality research infrastructures using them as training facilities for developing and completing the skills of the CISTEM’participants
• Contribute to bridging the gap between industry and high education institutions by implementing a structured secondment-based exchange programme
CISTEM has established a novel and strong multidisciplinary and intersectoral network of European and non-European researchers benefitting considerably from the exchange of expertise and resources. During the first 24 months of the project, scientific exchange has allowed the achievement of significant progress towards the scientific objectives and the network has produced a large number of results in the area of heart-on-chip which will be published in high impact journals in the fields. The main scientific results achieved so far are:

• The establishment of a protocol for 3D surface functionalization
• Optimisation of the co-culture condition with fibroblasts and endothelial cells inside a microfluidic device.
• Differentiation of iPSCs into cardiomyocytes
• Development of a perfusion station for investigating the shear stress effect on the maturation of cardiomyocytes
• Preliminary investigation of the shear stress effect on the maturation of CM-iPSC
• Development of electronic modules for TEER measurement and electrical stimulation
• Fabrication of heart membrane for mechanical stretch for mechanical actuation

The scientific results have been generated through the secondments/return phases of CISTEM secondees. The secondees have participated in many training activities for acquiring knowledge related to the project research activities such as cell culture protocols and procedures, surface functionalisation, cell culture characterization, microfluidic and microfabrication.
Career development of secondees is also an important objective of CISTEM which has been achieved through training, transfer of knowledge and networking activities, both inter-sectoral and intercontinental, mainly by means of secondment visits and workshop. CISTEM has offered a good opportunity for the seconded persons to reinforce their respective CV, and to consolidate their laboratory and general skills.
Genetic Rare Disorders such as Cardiomyopathy coming from Duchenne muscular dystrophy still represent an unmet medical need. Developing a heart model in a patient-match manner might contribute to a better understanding of mechanisms of cardiomyopathy that can provide better chance for patients, improving their conditions of life and prolonging its extent.
After these 2 years, CISTEM became a sustainable international network of academics and SMEs who are collectively overcoming obstacles in the development of Heart-on-chip for personalised medicine. As mentioned before the network has already provided clear results towards this objective: The CISTEM work has been focused on the optimisation of the culture condition of cardiomyocytes derived from iPSCs inside a microfluidic device. Different approaches have been evaluated and combined to improve cardiomyocyte viability and to reproduce the in-vivo environment inside the microfluidic device. Firstly, a robust protocol has been designed for 3D Surface functionalization of the device to reinforce the hydrogel attachment to the device and maintain the hydrogel architecture for a week. Secondly, iPSC-CM co-culture with human endothelial cells and fibroblasts was optimized to provide the CM-iPSc a more realistic cell culture environment which has shown to improve CM-iPSCs viability. Furthermore, to reproduce the dynamic condition of the in-vivo environment, a microfluidic set-up has been developed to generate controllable shear stress through the microfluidic device. Preliminary investigation has shown that the shear stress conditioning results in a change in cell morphology and an increase in vinculin production (protein needed for proper electrochemical signal transduction through cell-cell adhesion). We expect that the developed techniques will offer significant advantage for accurately modelling rare diseases.
Apart from the research activities, CISTEM has already promoted joint research initiatives and new collaborative projects are stemming out of the network. In addition, the researchers involved in the project have established a new network that reaches far beyond their home organization and gathered additional skills and knowledge that will significantly contribute to advance their careers by opening new professional opportunities.
Cell-culture training
Viability staining of iPSC-CM inside the microfluidic chip B-Gradiant
Business lesson