Periodic Reporting for period 2 - BIOCDx (A miniature Bio-photonics Companion Diagnostics platform for reliable cancer diagnosis and treatment monitoring.)
Periodo di rendicontazione: 2018-07-01 al 2020-09-30
In the medical diagnostics industry, there is an ever-increasing need for the development of robust, reliable, accurate and fast devices for early diagnosis, screening and monitoring of diseases allowing for the presently emerging paradigm shift of Personalized Medicine and Companion Diagnostics. There is a special interest in cancer.
BIOCDx aims to develop a miniaturized, ultra-sensitive and reliable Point-of-Care (PoC) device with disposable microfluidic cartridge for the monitoring of cancer biomarkers in body fluids and specifically in whole blood samples. The proposed PoC device will aid primary tumour and metastases detection, as well as monitoring of drug efficacy as a companion diagnostic. It will provide clinicians, caretakers and patients with a more sensitive, robust and selective tool for improved clinical decisions through the early and fast diagnosis of the disease, as well as monitoring of therapeutic response, reducing the cost of the healthcare system. Besides the obvious health benefit for the patient, it will contribute to the sustainability of the European health care system by decreasing the expenditure associated with pharmaceutical treatments and with hospitalization.
One of the scientific breakthroughs of BIOCDx is the development of a “cancer stem cells” detection platform by virtue of expression of the cancer stem cell-specific transcription factor TWIST1, which controls the expression of the bloodstream circulating biomarkers like the periostin (POSTN) protein. Cancer stem cells represent the most aggressive/tumorigenic cell compartment within tumours and are known to support primary tumour growth and to migrate to distant tissues and establish secondary tumours (metastases). They are also believed to survive chemotherapy and upon completion of treatment, they grow back leading to tumour recurrence. These recurrences evolve to ever more resistant tumours with fatal outcome.
To do so, BIOCDx will combine advanced concepts from the photonic, nano-biochemical and fluidic parts and reader/packaging platforms aiming to overcome limitations related to detection reliability, sensitivity, specificity, compactness and cost issues. BIOCDx will use ultrasensitive, photonic elements based on an array of 8 asymmetric MZI waveguides fabricated by TriPlex technology on silicon nitride substrates and aiming to achieve a 100 fold improvement of the sensitivity (<10-8 RIU) with respect to current technologies. A sandwich assay, enhanced with nanoparticles, will be developed, based on the use of two antibodies per protein, to detect all three circulating proteins. This will enhance the limit of detection (LOD) close to femtomolar. BIOCDx photonic, nano-biochemical, fluidics and packaging platforms will be integrated into a POC device which will be validated in preclinical and clinical settings for three cancer types: breast cancer, hormone-independent prostate cancer and melanoma.
BIOCDx aims to develop a miniaturized, ultra-sensitive and reliable Point-of-Care (PoC) device with disposable microfluidic cartridge for the monitoring of cancer biomarkers in body fluids and specifically in whole blood samples. The proposed PoC device will aid primary tumour and metastases detection, as well as monitoring of drug efficacy as a companion diagnostic. It will provide clinicians, caretakers and patients with a more sensitive, robust and selective tool for improved clinical decisions through the early and fast diagnosis of the disease, as well as monitoring of therapeutic response, reducing the cost of the healthcare system. Besides the obvious health benefit for the patient, it will contribute to the sustainability of the European health care system by decreasing the expenditure associated with pharmaceutical treatments and with hospitalization.
One of the scientific breakthroughs of BIOCDx is the development of a “cancer stem cells” detection platform by virtue of expression of the cancer stem cell-specific transcription factor TWIST1, which controls the expression of the bloodstream circulating biomarkers like the periostin (POSTN) protein. Cancer stem cells represent the most aggressive/tumorigenic cell compartment within tumours and are known to support primary tumour growth and to migrate to distant tissues and establish secondary tumours (metastases). They are also believed to survive chemotherapy and upon completion of treatment, they grow back leading to tumour recurrence. These recurrences evolve to ever more resistant tumours with fatal outcome.
To do so, BIOCDx will combine advanced concepts from the photonic, nano-biochemical and fluidic parts and reader/packaging platforms aiming to overcome limitations related to detection reliability, sensitivity, specificity, compactness and cost issues. BIOCDx will use ultrasensitive, photonic elements based on an array of 8 asymmetric MZI waveguides fabricated by TriPlex technology on silicon nitride substrates and aiming to achieve a 100 fold improvement of the sensitivity (<10-8 RIU) with respect to current technologies. A sandwich assay, enhanced with nanoparticles, will be developed, based on the use of two antibodies per protein, to detect all three circulating proteins. This will enhance the limit of detection (LOD) close to femtomolar. BIOCDx photonic, nano-biochemical, fluidics and packaging platforms will be integrated into a POC device which will be validated in preclinical and clinical settings for three cancer types: breast cancer, hormone-independent prostate cancer and melanoma.
The highlights of the considered reporting period are summarized below:
• An “Expert Opinion Check” BIOCDx survey has been done. The main needs and requirements of end-users have been identified and consolidated. A provisional market analysis has been performed. BIOCDx PoC device is focused on cancer treatment monitoring for 2 cancer types. Designs of the individual components and platforms of the BIOCDx system. The system specifications of the BIOCDx system have been identified. A preliminary cost analysis of the BIOCDx device has been done.
• Samples of cancer patients obtained. A process to block the cartridge has been established. Plasma has been generated in the final filter cartridge using different hematocrit blood values. Successful biochemical multiplexing in the printed nanoparticle assay and the BIOCDx lab set-up. Amplification of the signal on the BIOCDx lab set-up by using nanoparticles. POSTN in 10% plasma on the BIOCDx lab set-up was successfully and specifically measured. Optimized protocol for the material-selective modification of the aMZIs. Development of hydrogels for 3D surface functionalization. Combination of laser printing technique with various chemical approaches. Successful transfer and photoactivation of 3D hydrogels by the use of laser-based technique.
• Lionix finalized hybrid integration of VCSELs and Photodiode array. Selective surface modification of G1 chips and hybrid chips. Validation experiments with real and spiked samples. LIFT of the BIOCDx relevant monoclonal antibodies on chemically modified sensors.
• Material selection and coatings for cartridge finalized. Fabrication process for hybrid chip integration finalized. Functional hybrid chips have been integrated into cartridges. Final validation of hybrid-chip cartridge and intermediate bread board performed.
• Measurement system for fluidic control finalized. Housing industrial design and fabrication finished and measurement electronic integrated. GUI design and implementation are finished. Heating of sensor chip fully implemented and tested. Tests with hybrid chip were done. Mechanical, electrical and software integration is finished.
• Biobank has been enriched and measures 122 cancer patient samples and 44 samples from healthy individuals. Metastatic breast and prostate cancers have been determined. A Western Blot protocol for the relevant quantification of the expression levels of the biomarkers in human sera has been employed. Validation of the significance of the biomarkers in humans performed. Statistical analysis was run with the use of WB to show a clear correlation between the levels of these two biomarkers in both breast and prostate cancer patients. Mice treatment with chemotherapeutic drugs has been concluded. Reductions in tumor growth correlate with decreases in the expression levels of the biomarkers. Data collected for mice treated with chemotherapeutic agents have been statistically analyzed and the relevance of POSTN and TGFBI in tracking tumor growth has been confirmed. Two samples from cancer patients and one sample from a healthy volunteer analyzed with the BIOCDx lab setup.
• The manufacturability and costs of the BIOCDx were determined and documented. An inventory of market size and opportunities was made for the BIOCDx device. A detailed roadmap to commercialization for the BIOCDx device was established. Four press releases, two journal articles and seven publications in conference proceedings, four participations in workshops, seven booth organizations at major international conferences, eighteen presentations in international and national conferences and one event other than workshop or conference have been published.
• An “Expert Opinion Check” BIOCDx survey has been done. The main needs and requirements of end-users have been identified and consolidated. A provisional market analysis has been performed. BIOCDx PoC device is focused on cancer treatment monitoring for 2 cancer types. Designs of the individual components and platforms of the BIOCDx system. The system specifications of the BIOCDx system have been identified. A preliminary cost analysis of the BIOCDx device has been done.
• Samples of cancer patients obtained. A process to block the cartridge has been established. Plasma has been generated in the final filter cartridge using different hematocrit blood values. Successful biochemical multiplexing in the printed nanoparticle assay and the BIOCDx lab set-up. Amplification of the signal on the BIOCDx lab set-up by using nanoparticles. POSTN in 10% plasma on the BIOCDx lab set-up was successfully and specifically measured. Optimized protocol for the material-selective modification of the aMZIs. Development of hydrogels for 3D surface functionalization. Combination of laser printing technique with various chemical approaches. Successful transfer and photoactivation of 3D hydrogels by the use of laser-based technique.
• Lionix finalized hybrid integration of VCSELs and Photodiode array. Selective surface modification of G1 chips and hybrid chips. Validation experiments with real and spiked samples. LIFT of the BIOCDx relevant monoclonal antibodies on chemically modified sensors.
• Material selection and coatings for cartridge finalized. Fabrication process for hybrid chip integration finalized. Functional hybrid chips have been integrated into cartridges. Final validation of hybrid-chip cartridge and intermediate bread board performed.
• Measurement system for fluidic control finalized. Housing industrial design and fabrication finished and measurement electronic integrated. GUI design and implementation are finished. Heating of sensor chip fully implemented and tested. Tests with hybrid chip were done. Mechanical, electrical and software integration is finished.
• Biobank has been enriched and measures 122 cancer patient samples and 44 samples from healthy individuals. Metastatic breast and prostate cancers have been determined. A Western Blot protocol for the relevant quantification of the expression levels of the biomarkers in human sera has been employed. Validation of the significance of the biomarkers in humans performed. Statistical analysis was run with the use of WB to show a clear correlation between the levels of these two biomarkers in both breast and prostate cancer patients. Mice treatment with chemotherapeutic drugs has been concluded. Reductions in tumor growth correlate with decreases in the expression levels of the biomarkers. Data collected for mice treated with chemotherapeutic agents have been statistically analyzed and the relevance of POSTN and TGFBI in tracking tumor growth has been confirmed. Two samples from cancer patients and one sample from a healthy volunteer analyzed with the BIOCDx lab setup.
• The manufacturability and costs of the BIOCDx were determined and documented. An inventory of market size and opportunities was made for the BIOCDx device. A detailed roadmap to commercialization for the BIOCDx device was established. Four press releases, two journal articles and seven publications in conference proceedings, four participations in workshops, seven booth organizations at major international conferences, eighteen presentations in international and national conferences and one event other than workshop or conference have been published.
The progress beyond the state of the art achieved so far for the 2nd period of the project includes the:
- Design and fabrication of hybrid photonic chips.
- Novel functionalization approaches for the immobilization of the biomaterials on the aMZIs.
- Design and testing of materials for the fabrication of a disposable, miniaturized, low-cost microfluidics cartridge with integrated sensor and blood filtering module.
- Validation of real samples with the photonic chips
- Design and fabrication of hybrid photonic chips.
- Novel functionalization approaches for the immobilization of the biomaterials on the aMZIs.
- Design and testing of materials for the fabrication of a disposable, miniaturized, low-cost microfluidics cartridge with integrated sensor and blood filtering module.
- Validation of real samples with the photonic chips