Periodic Reporting for period 2 - Tumor-LN-oC (Tumor and Lymph Node on Chip for cancer studies)
Berichtszeitraum: 2022-11-01 bis 2024-04-30
Tumor-LN-oC aims to develop a robust, automated tumor-lymph node-on-chip platform connecting primary surgically removed human tumors and lymph node (LN) tissue from the same cancer patient. This "biological twin" will facilitate the study of primary tumor interactions with lymph nodes.
The main objective is to create a Tumor-LN-oC platform to monitor LN metastasis, characterize signaling cues facilitating LN metastasis, and identify spectral and molecular signatures in metastasizing cells. This could lead to new diagnostic tools and therapeutic approaches. Additionally, the platform will serve as a preclinical setting for drug testing for individual lung cancer patients.
Specific objectives include:
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 Tumor-LN-oC platform will use a novel multi-compartment microfluidic chip mimicking the tumor microenvironment and its lymphatic connection. Employing mid-IR Photothermal (MIP) spectroscopy and microlens array-based micro-optics, it will generate spectral "fingerprints" of metastasizing cells for diagnostic purposes. Laser-based bioprinting will precisely place LN and tumor cells within the chip with high viability. The platform will be developed at TRL5 and validated with real patient samples, considering regulatory pathways to facilitate market entry.
The main objective is to create a Tumor-LN-oC platform to monitor LN metastasis, characterize signaling cues facilitating LN metastasis, and identify spectral and molecular signatures in metastasizing cells. This could lead to new diagnostic tools and therapeutic approaches. Additionally, the platform will serve as a preclinical setting for drug testing for individual lung cancer patients.
Specific objectives include:
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 Tumor-LN-oC platform will use a novel multi-compartment microfluidic chip mimicking the tumor microenvironment and its lymphatic connection. Employing mid-IR Photothermal (MIP) spectroscopy and microlens array-based micro-optics, it will generate spectral "fingerprints" of metastasizing cells for diagnostic purposes. Laser-based bioprinting will precisely place LN and tumor cells within the chip with high viability. The platform will be developed at TRL5 and validated with real patient samples, considering regulatory pathways to facilitate market entry.
In the first 36 months, Tumor-LN-oC focused on developing and optimizing individual platform components and preparing for their integration. Significant advancements were made in all modules:
Objective 1 (WP5,6,7): Tumor-LN-oC microfluidic chips were continuously fabricated and tested. The chips, fitted with artificial cilia for continuous recirculatory flow, were validated for long-term cell culture with cell lines and surgical samples.
Objective 2 (WP9,10): The EC-QCL module, featuring rapid wavelength tunability (~30,000 cm-1/s), was designed, fabricated, and tested. Imaging will be performed at select wavelengths to minimize acquisition time. The EC-QCL prototype was integrated into the mid-IR spectroscopy system, and tests are ongoing.
Objective 3 (WP4,7,13): Optimal media for co-culture in transwells and separate supply on the microfluidic chip were identified, and cell viability was confirmed. Migrating tumor cells were observed in the channels. Samples from 60 lung cancer patients were obtained, processed, and stored in a biobank. Analyses showed successful isolation of medium from co-cultures and detection of cytokines with patient-specific expression patterns. Ongoing experiments aim to identify biomarkers for personalized cancer treatment. The first bioprinter prototype is ready.
Objective 4 (WP11,12): The micro-optics module, based on microlens arrays, creates images with an FOV of 8.4 x 6 mm and an optical resolution of 3.9 μm. The high-level control software integrates all system functionalities and is ready for extension. Machine learning was used for cell segmentation and motion quantification, enabling real-time monitoring of directional cell motion. Acquisition and processing time is around 2 minutes with potential for further acceleration.
Objective 5 (WP13): Integration of all modules into the Tumor-LN-oC platform is a key priority. The design concept includes two modules: one with an integrated micro-optics module for monitoring six chips, and one with an integrated MIP module for molecular characterization.
Objective 6 (WP13): The first prototype of the integrated Tumor-LN-oC platform is anticipated by M48, with validation using patient samples in the last six months of the project.
Objective 7 (WP14,15,16,17,18): The regulatory roadmap for metastasis diagnosis and drug testing applications was generated, identifying applicable guidelines and technical standards. A meeting with the EMA’s innovation task force was held in November 2023, with future meetings planned. An updated commercialization plan provides a foundation for future exploitation and continued cooperation of partners. A first patent application was submitted.
Objective 1 (WP5,6,7): Tumor-LN-oC microfluidic chips were continuously fabricated and tested. The chips, fitted with artificial cilia for continuous recirculatory flow, were validated for long-term cell culture with cell lines and surgical samples.
Objective 2 (WP9,10): The EC-QCL module, featuring rapid wavelength tunability (~30,000 cm-1/s), was designed, fabricated, and tested. Imaging will be performed at select wavelengths to minimize acquisition time. The EC-QCL prototype was integrated into the mid-IR spectroscopy system, and tests are ongoing.
Objective 3 (WP4,7,13): Optimal media for co-culture in transwells and separate supply on the microfluidic chip were identified, and cell viability was confirmed. Migrating tumor cells were observed in the channels. Samples from 60 lung cancer patients were obtained, processed, and stored in a biobank. Analyses showed successful isolation of medium from co-cultures and detection of cytokines with patient-specific expression patterns. Ongoing experiments aim to identify biomarkers for personalized cancer treatment. The first bioprinter prototype is ready.
Objective 4 (WP11,12): The micro-optics module, based on microlens arrays, creates images with an FOV of 8.4 x 6 mm and an optical resolution of 3.9 μm. The high-level control software integrates all system functionalities and is ready for extension. Machine learning was used for cell segmentation and motion quantification, enabling real-time monitoring of directional cell motion. Acquisition and processing time is around 2 minutes with potential for further acceleration.
Objective 5 (WP13): Integration of all modules into the Tumor-LN-oC platform is a key priority. The design concept includes two modules: one with an integrated micro-optics module for monitoring six chips, and one with an integrated MIP module for molecular characterization.
Objective 6 (WP13): The first prototype of the integrated Tumor-LN-oC platform is anticipated by M48, with validation using patient samples in the last six months of the project.
Objective 7 (WP14,15,16,17,18): The regulatory roadmap for metastasis diagnosis and drug testing applications was generated, identifying applicable guidelines and technical standards. A meeting with the EMA’s innovation task force was held in November 2023, with future meetings planned. An updated commercialization plan provides a foundation for future exploitation and continued cooperation of partners. A first patent application was submitted.
The Tumor-LN-oC platform introduces the first artificial model realistically mimicking tumor and LN crosstalk, offering a novel personalized paradigm (biological twin) with no existing alternatives. It is an automated platform for studying tumor biology and personalized approaches in biomarker and drug target identification and preclinical testing of therapeutic schemes.
By the end of the project, an integrated Tumor-LN-oC platform will address unmet needs and achieve the following 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.
User-friendly software and interfaces with standard lab equipment will ensure accessibility to non-experts in microfluidics, broadening the user base. Innovations in microfluidics, cell culture, tissue engineering, real-time detection, and monitoring will significantly advance these interdisciplinary fields. Integrating the chip with spectroscopic detection tools and image analysis technologies will further enhance the platform's value.
By the end of the project, an integrated Tumor-LN-oC platform will address unmet needs and achieve the following 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.
User-friendly software and interfaces with standard lab equipment will ensure accessibility to non-experts in microfluidics, broadening the user base. Innovations in microfluidics, cell culture, tissue engineering, real-time detection, and monitoring will significantly advance these interdisciplinary fields. Integrating the chip with spectroscopic detection tools and image analysis technologies will further enhance the platform's value.