Periodic Reporting for period 1 - Tumor-LN-oC (Tumor and Lymph Node on Chip for cancer studies)
Okres sprawozdawczy: 2021-05-01 do 2022-10-31
Tumor-LN-oC aims to offer a comprehensive solution for a robust, automated tumor-lymph node-on-chip platform that will connect primary surgically removed human tumors and LN tissue from the same cancer patient. This will serve as a “biological twin” of the patient and allow to study the interaction of primary tumors with lymph nodes.
The overall objective is to generate a Tumor-LN-oC platform that allows monitoring of the LN metastasis process, the characterization of signalling ques facilitating LN metastasis and the identification of spectral and molecular signatures in metastasizing cells. This information could lead to novel diagnostic tools and therapeutic approaches. Moreover, the developed platform will serve as a preclinical setting for parallel testing of drugs for individual cancer lung cancer patients aiming at Precision Medicine approaches.
The project pursues the following specific objectives:
1) To introduce novel designs and develop robust, automated microfluidic chips optimized for tumor cell and LN culture enabling the study of their crosstalk,
2) To integrate Quantum Cascade Laser based mid- IR spectroscopy for specific chemical signatures,
3) To molecularly characterize both migrating tumor-derived cells attracted to the LN and the soluble signals driving migration,
4) To demonstrate an advanced image analysis and signal processing platform using deep learning algorithms facilitated by a micro-optics module to monitor in real time the cells migration,
5) To integrate all Tumor-LN-oC technologies in an automated platform prototype incorporating interfaces compatible with existing laboratory equipment.
6) To demonstrate the Tumor-LN-oC TRL5 platform and validate it with real patient samples
7) To establish regulatory pathways and assure regulatory standards and requirements compliance during the development of the Tumor-LN-oC in order to facilitate exploitation and early market entry
The multidisciplinary Tumor-LN-oC platform will rely on a novel multi-compartment microfluidic chip which will mimick the tumor microenvironment and its connection to the lymphatic system. Using sensitive proteomic and molecular approaches, the soluble signals that neutralize the immune response and allow tumor cells to metastasize to the LNs will be characterized. This will enable the use of existing drugs, or the development of new ones that could reverse this process and inhibit tumor growth and dissemination. Moreover, by employing novel imaging approaches (mid-IR Photothermal (MIP) spectroscopy) and imaging sensors (microlens array-based micro-optics), a spectral “fingerprint” of migrating/metastasizing cells will be generated, which could be used for diagnostic purposes in tumor and lymph node biopsies. Laser based bioprinting of LN and tumor cell lines and tumor organoids will be introduced to enable precise placement of cells within the microfluidic chip with unprecedented cell viability.
The Tumor-LN-oC platform will be developed at TRL5 and will be validated using real patient samples. Regulatory pathways, standards and requirements compliance will be considered in order to facilitate exploitation and early market entry.
The overall objective is to generate a Tumor-LN-oC platform that allows monitoring of the LN metastasis process, the characterization of signalling ques facilitating LN metastasis and the identification of spectral and molecular signatures in metastasizing cells. This information could lead to novel diagnostic tools and therapeutic approaches. Moreover, the developed platform will serve as a preclinical setting for parallel testing of drugs for individual cancer lung cancer patients aiming at Precision Medicine approaches.
The project pursues the following specific objectives:
1) To introduce novel designs and develop robust, automated microfluidic chips optimized for tumor cell and LN culture enabling the study of their crosstalk,
2) To integrate Quantum Cascade Laser based mid- IR spectroscopy for specific chemical signatures,
3) To molecularly characterize both migrating tumor-derived cells attracted to the LN and the soluble signals driving migration,
4) To demonstrate an advanced image analysis and signal processing platform using deep learning algorithms facilitated by a micro-optics module to monitor in real time the cells migration,
5) To integrate all Tumor-LN-oC technologies in an automated platform prototype incorporating interfaces compatible with existing laboratory equipment.
6) To demonstrate the Tumor-LN-oC TRL5 platform and validate it with real patient samples
7) To establish regulatory pathways and assure regulatory standards and requirements compliance during the development of the Tumor-LN-oC in order to facilitate exploitation and early market entry
The multidisciplinary Tumor-LN-oC platform will rely on a novel multi-compartment microfluidic chip which will mimick the tumor microenvironment and its connection to the lymphatic system. Using sensitive proteomic and molecular approaches, the soluble signals that neutralize the immune response and allow tumor cells to metastasize to the LNs will be characterized. This will enable the use of existing drugs, or the development of new ones that could reverse this process and inhibit tumor growth and dissemination. Moreover, by employing novel imaging approaches (mid-IR Photothermal (MIP) spectroscopy) and imaging sensors (microlens array-based micro-optics), a spectral “fingerprint” of migrating/metastasizing cells will be generated, which could be used for diagnostic purposes in tumor and lymph node biopsies. Laser based bioprinting of LN and tumor cell lines and tumor organoids will be introduced to enable precise placement of cells within the microfluidic chip with unprecedented cell viability.
The Tumor-LN-oC platform will be developed at TRL5 and will be validated using real patient samples. Regulatory pathways, standards and requirements compliance will be considered in order to facilitate exploitation and early market entry.
During the first 18 months of the project, Tumor-LN-oC research and development activities were focused on defining specifications and requirements for the platform and its individual components. Considerable advancements were made on all platform modules. Following results were achieved:
Objective 1 (WP5,6,7): The Microfluidic chip optimized for tumor cell and LN culture was designed and fabricated in two generations. Artificial cilia which mimic lymph node were successfully integrated to flow, tested by other partners who successfully observed cell migration between the two channels, reflecting the crosstalk. The chips are designed to be reusable and materials are suitable for industrial-scale production. Tests and optimizations are ongoing.
Objective 2 (WP9): The EC-QCL module was designed and the voice coil actuator selected. The first version of the mid-IR spectroscopy stand-alone system was assembled and its functionality in water confirmed (WP9)
Objective 3 (WP4): Optimal media for both co culture in a transwell and separate supply on the microfluidic chip have been identified and cell viability confirmed. Migrating tumor cells have been observed in the channels. Samples from 23 lung cancer patients have been obtained from a hospital in Greece, processes. Molecular characterization is ongoing.
Objective 4 (WP11,12): The first micro-optics module setup was completed and is currently being optimized. The first version of the software is ready, image analysis and signal processing platform using deep learning algorithms for cell detection and monitoring, machine learning models, preparations for real time monitoring of cell migration in the chip
Objective 5 (WP13): Discussions on how to best integrate all Tumor-LN-oC technologies in an automated platform prototype have started and first design were ideas circulated.
Objective 7 (WP16,17): A regulatory roadmap was developed which outlines various regulatory scenarios relevant to Tumor-LN-oC. Contact with regulatory authorities will be established shortly. An initial commercialisation plan provides a good foundation for future exploitation of project results and commercialisation of the platform.
Objective 1 (WP5,6,7): The Microfluidic chip optimized for tumor cell and LN culture was designed and fabricated in two generations. Artificial cilia which mimic lymph node were successfully integrated to flow, tested by other partners who successfully observed cell migration between the two channels, reflecting the crosstalk. The chips are designed to be reusable and materials are suitable for industrial-scale production. Tests and optimizations are ongoing.
Objective 2 (WP9): The EC-QCL module was designed and the voice coil actuator selected. The first version of the mid-IR spectroscopy stand-alone system was assembled and its functionality in water confirmed (WP9)
Objective 3 (WP4): Optimal media for both co culture in a transwell and separate supply on the microfluidic chip have been identified and cell viability confirmed. Migrating tumor cells have been observed in the channels. Samples from 23 lung cancer patients have been obtained from a hospital in Greece, processes. Molecular characterization is ongoing.
Objective 4 (WP11,12): The first micro-optics module setup was completed and is currently being optimized. The first version of the software is ready, image analysis and signal processing platform using deep learning algorithms for cell detection and monitoring, machine learning models, preparations for real time monitoring of cell migration in the chip
Objective 5 (WP13): Discussions on how to best integrate all Tumor-LN-oC technologies in an automated platform prototype have started and first design were ideas circulated.
Objective 7 (WP16,17): A regulatory roadmap was developed which outlines various regulatory scenarios relevant to Tumor-LN-oC. Contact with regulatory authorities will be established shortly. An initial commercialisation plan provides a good foundation for future exploitation of project results and commercialisation of the platform.
The Tumor-LN-oC platform introduces the first artificial model which can realistically mimic tumor and LN crosstalk. Therefore it is offering a novel personalised paradigm (biological twin) to which no alternative currently exists: an automated OoC platform for studies towards the understanding of tumor biology, but also for more personalized approaches in biomarker and drug target identification and preclinical testing of candidate drugs/therapeutic schemes.
Until the end of the project, an integrated Tumor-LN-oC platform will be delivered which addresses various unmet needs and achieved following expected impacts:
1. Verifiable progress in the application of Organ-on-Chip technologies for in-vitro research;
2. Reduction of the need for animal and clinical testing;
3. Lowering of barriers for application of Organ-on-Chip technology;
4. Improved competitiveness and attractiveness of the European biomedical and healthcare sector
5. Increased awareness and knowledge about medical regulatory policies and requirements, especially by academics and SMEs.
A user-friendly software and interfaces with standard lab equipment will ensure that the platform can be used by non-experts in the field of microfluidics, thus offering this technology to a broader user base. Moreover, by introducing key innovations in the fields of microfluidics, cell culture and tissue engineering, real-time detection and monitoring of biological systems, Tumor-LN-oC will significantly contribute to the progress of these interdisciplinary technological fields. The seamless integration of the chip with spectroscopic detection tools and image analysis technologies will further increase the added value of the proposed platform.
A low cost of the platform will ensure accessibility and enable fast commercialization. A validation of the prototype in lab environment in preclinical setting at TRL5 is aimed for by 2024. It is anticipated that TRL9 will be reached by 2026 with upscaling and industrial manufacturing possible by 2025.
Until the end of the project, an integrated Tumor-LN-oC platform will be delivered which addresses various unmet needs and achieved following expected impacts:
1. Verifiable progress in the application of Organ-on-Chip technologies for in-vitro research;
2. Reduction of the need for animal and clinical testing;
3. Lowering of barriers for application of Organ-on-Chip technology;
4. Improved competitiveness and attractiveness of the European biomedical and healthcare sector
5. Increased awareness and knowledge about medical regulatory policies and requirements, especially by academics and SMEs.
A user-friendly software and interfaces with standard lab equipment will ensure that the platform can be used by non-experts in the field of microfluidics, thus offering this technology to a broader user base. Moreover, by introducing key innovations in the fields of microfluidics, cell culture and tissue engineering, real-time detection and monitoring of biological systems, Tumor-LN-oC will significantly contribute to the progress of these interdisciplinary technological fields. The seamless integration of the chip with spectroscopic detection tools and image analysis technologies will further increase the added value of the proposed platform.
A low cost of the platform will ensure accessibility and enable fast commercialization. A validation of the prototype in lab environment in preclinical setting at TRL5 is aimed for by 2024. It is anticipated that TRL9 will be reached by 2026 with upscaling and industrial manufacturing possible by 2025.