Leader: Dr A. Odysseos
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Personal Medicine Unleashed:
Using Zebrafish Avatars of BRAIN Cancer Patients to Predict Therapeutic Outcomes and Explore Novel Therapies Cyprus Research & Innovation Foundation
Excellence Hub, EXCELLENCE/0524/0397 |
Zebrafish Avatars: a functional, living, surrogate model, used for studying a patient's tumor in real-time, in order to predict a patient response to therapy and guide personalized treatment decisions.
What is the zBRAIN project ?
- The zBRAIN project is a clinical study leveraging zebrafish patient-derived xenografts of glioblastoma (Zebrafish Avatars) in order to predict the patient clinical outcomes and their response to standard and novel treatment through a combined biomarker analysis.
- This innovative approach integrates animal modelling into clinical setting for the prediction of tumor behavior, marking a significant advancement in translational personalized medicine in neuro-oncology.
- zBRAIN addresses the critical need for more effective treatment outcome prediction and new treatments by introducing zebrafish avatars as a novel translational research tool. This project stands out for its unique approach to integrating personalized medicine with high-throughput drug screening, providing actionable insights into tumor behavior and therapeutic responses. By pioneering the use of zebrafish avatars in a clinical setting, zBRAIN aims to significantly improve patient-specific treatment strategies, enhance the understanding of glioblastoma, and reduce the time and cost associated with drug development.
- Glioblastoma is one of the most aggressive and difficult-to-treat cancers. Traditional models often fail to capture the complexity and heterogeneity of human tumors, leading to limited success in translating preclinical findings to clinical practice. zBRAIN’s innovative approach promises to bridge this gap by providing a more accurate and scalable model for studying glioblastoma, ultimately leading to better patient outcomes and advancing the field of cancer pharmacology. The zBRAIN project aligns with the Programme and Call objectives.
General Objectives
1. To transform glioblastoma research by establishing a pioneering Zebrafish Avatar model that can simulate a patients’ glioblastoma response to therapy, providing a scalable and cost-effective alternative to traditional models and driving innovative research methodologies.
2. To develop and validate predictive models using patient-specific biomarkers to enhance treatment.
1. To transform glioblastoma research by establishing a pioneering Zebrafish Avatar model that can simulate a patients’ glioblastoma response to therapy, providing a scalable and cost-effective alternative to traditional models and driving innovative research methodologies.
2. To develop and validate predictive models using patient-specific biomarkers to enhance treatment.
Project outline
The zBRAIN summary image (left) illustrates how the project connects the patient’s normal clinical pathway with advanced laboratory modelling and predictive data analysis.
In the clinic, each participating brain cancer patient continues to receive their standard diagnostic work-up, treatment, imaging, and follow-up care. zBRAIN does not replace or alter this clinical sequence; instead, it collects and integrates the related clinical information shown in the image, including histology, sequencing, treatment data, radiological imaging, and clinical progression.
In parallel, in the lab, tumor and blood samples from the same patient are transferred to the laboratory to generate patient-specific Zebrafish Avatars. These are patient-derived cancer xenografts, where the patient’s own glioblastoma cells are transplanted into transparent zebrafish embryos, allowing real-time observation of tumor growth, invasion, spread, and treatment response in vivo. In addition to standard Zebrafish Avatars, zBRAIN applies a double-xenograft system: after isolation, priming, and stimulation, the patient’s own Natural Killer cells are introduced into the zebrafish model to study how the patient’s innate immune system behaves against their own cancer cells.
The platform also explores innovative therapeutic strategies. One approach involves injecting the Zebrafish Avatars with genetically engineered human cells that produce anticancer micro-RNA molecules, allowing the team to test whether this cell-based miRNA delivery strategy can affect the patient’s tumor in vivo. In parallel, zBRAIN performs personalized drug repurposing screening using clinical drugs with potential anticancer activity, selected and interpreted in the context of the patient’s own molecular and genetic tumor profile.
Finally, zBRAIN brings together all generated data (clinical data, in vitro cell-based data, and in vivo Zebrafish Avatar data) for comprehensive bioinformatics and machine-learning-based analyses. The aim is not only to test whether Zebrafish Avatars can predict patient-specific therapeutic responses, but also to determine how much each individual parameter contributes to the predictive power of the model. Through this integrated clinic-to-lab-to-data workflow, zBRAIN aims to advance personalized translational strategies for brain cancer research and, ultimately, support more precise therapeutic decision-making.
The zBRAIN summary image (left) illustrates how the project connects the patient’s normal clinical pathway with advanced laboratory modelling and predictive data analysis.
In the clinic, each participating brain cancer patient continues to receive their standard diagnostic work-up, treatment, imaging, and follow-up care. zBRAIN does not replace or alter this clinical sequence; instead, it collects and integrates the related clinical information shown in the image, including histology, sequencing, treatment data, radiological imaging, and clinical progression.
In parallel, in the lab, tumor and blood samples from the same patient are transferred to the laboratory to generate patient-specific Zebrafish Avatars. These are patient-derived cancer xenografts, where the patient’s own glioblastoma cells are transplanted into transparent zebrafish embryos, allowing real-time observation of tumor growth, invasion, spread, and treatment response in vivo. In addition to standard Zebrafish Avatars, zBRAIN applies a double-xenograft system: after isolation, priming, and stimulation, the patient’s own Natural Killer cells are introduced into the zebrafish model to study how the patient’s innate immune system behaves against their own cancer cells.
The platform also explores innovative therapeutic strategies. One approach involves injecting the Zebrafish Avatars with genetically engineered human cells that produce anticancer micro-RNA molecules, allowing the team to test whether this cell-based miRNA delivery strategy can affect the patient’s tumor in vivo. In parallel, zBRAIN performs personalized drug repurposing screening using clinical drugs with potential anticancer activity, selected and interpreted in the context of the patient’s own molecular and genetic tumor profile.
Finally, zBRAIN brings together all generated data (clinical data, in vitro cell-based data, and in vivo Zebrafish Avatar data) for comprehensive bioinformatics and machine-learning-based analyses. The aim is not only to test whether Zebrafish Avatars can predict patient-specific therapeutic responses, but also to determine how much each individual parameter contributes to the predictive power of the model. Through this integrated clinic-to-lab-to-data workflow, zBRAIN aims to advance personalized translational strategies for brain cancer research and, ultimately, support more precise therapeutic decision-making.
Specific Objectives
Objective 1: Develop and Validate Zebrafish Avatars for Glioblastoma Modeling. Aim: To create zebrafish avatars using patient-derived glioblastoma cells and NK cells. Expected Results: Successful establishment of orthotopic zebrafish xenografts that accurately represent patient-specific glioblastoma profiles. Quantitative Measure: Achieve at least 80% engraftment success rate in zebrafish larvae within the first 6 months.
Objective 2: High-Throughput Drug Screening Using Zebrafish Avatars. Aim: To utilize zebrafish avatars for high-throughput screening (HTS) of clinical and novel therapeutic compounds. Expected Results: Identification of at least 5 drug candidates showing significant anti-tumor activity. Quantitative Measure: Screen a minimum of 200 compounds within the first 12 months, achieving a 95% assay reproducibility rate.
Objective 3: Integration of Biomarkers for Predictive Modeling. Aim: To integrate multiple biomarkers (tumor size, invasion, metastasis, IDH status, MGMT status, NK cell activity) into a comprehensive predictive model.
Expected Results: Development of a robust predictive model to forecast clinical outcomes and optimize personalized treatment plans. Quantitative Measure: Achieve a predictive accuracy of 85% in correlating zebrafish avatar responses with patient clinical outcomes by M18.
Objective 4: In Vitro and In Vivo Synergistic Studies of NK Cells and Drug Therapeutics against Glioblastoma.
Aim: To study the synergistic effects of primed NK cells and pharmacological treatments in zebrafish avatars.
Expected Results: Demonstration of enhanced tumor reduction in zebrafish avatars treated with NK cells and selected drug combinations. Quantitative Measure: Achieve at least 30% greater tumor reduction in combination treatment groups compared to monotherapy by M18.
Objective 5: Dissemination and Knowledge Transfer. Aim: To disseminate the project’s findings through publications, conferences, and workshops, ensuring knowledge transfer and stakeholder engagement.
Expected Results: Publication of at least 3 peer-reviewed articles, presentation at 5 international conferences, and organization of 3 stakeholder workshops. Quantitative Measure: Reach an audience of at least 1000 stakeholders through various dissemination activities by the end of the project.
Objective 1: Develop and Validate Zebrafish Avatars for Glioblastoma Modeling. Aim: To create zebrafish avatars using patient-derived glioblastoma cells and NK cells. Expected Results: Successful establishment of orthotopic zebrafish xenografts that accurately represent patient-specific glioblastoma profiles. Quantitative Measure: Achieve at least 80% engraftment success rate in zebrafish larvae within the first 6 months.
Objective 2: High-Throughput Drug Screening Using Zebrafish Avatars. Aim: To utilize zebrafish avatars for high-throughput screening (HTS) of clinical and novel therapeutic compounds. Expected Results: Identification of at least 5 drug candidates showing significant anti-tumor activity. Quantitative Measure: Screen a minimum of 200 compounds within the first 12 months, achieving a 95% assay reproducibility rate.
Objective 3: Integration of Biomarkers for Predictive Modeling. Aim: To integrate multiple biomarkers (tumor size, invasion, metastasis, IDH status, MGMT status, NK cell activity) into a comprehensive predictive model.
Expected Results: Development of a robust predictive model to forecast clinical outcomes and optimize personalized treatment plans. Quantitative Measure: Achieve a predictive accuracy of 85% in correlating zebrafish avatar responses with patient clinical outcomes by M18.
Objective 4: In Vitro and In Vivo Synergistic Studies of NK Cells and Drug Therapeutics against Glioblastoma.
Aim: To study the synergistic effects of primed NK cells and pharmacological treatments in zebrafish avatars.
Expected Results: Demonstration of enhanced tumor reduction in zebrafish avatars treated with NK cells and selected drug combinations. Quantitative Measure: Achieve at least 30% greater tumor reduction in combination treatment groups compared to monotherapy by M18.
Objective 5: Dissemination and Knowledge Transfer. Aim: To disseminate the project’s findings through publications, conferences, and workshops, ensuring knowledge transfer and stakeholder engagement.
Expected Results: Publication of at least 3 peer-reviewed articles, presentation at 5 international conferences, and organization of 3 stakeholder workshops. Quantitative Measure: Reach an audience of at least 1000 stakeholders through various dissemination activities by the end of the project.
The zBRAIN's innovation!
The hybridization of Zebrafish Avatars (patient-derived orthotopic zebrafish xenografts of glioblastoma) with cancer patients, in a co-clinical study, with the aim to validate the predictive power of this model against the patients’ clinical outcome (cancer progression), is a ground-breaking approach that introduces not only one, but a number of innovations that aim to address the challenges we face in the fields of neuro-oncology, cancer pharmacology and animal modeling.
The funding of zBRAIN will bring a number of “firsts” :
✦ The first research project in Cyprus that will use the concept of Animal Avatars of Cancer Patients.
✦ The first clinical study in Cyprus that uses patient-derived xenografts to predict clinical outcomes.
✦ The first research project, globally, that will test patient-derived NK-cell priming as an innate immunotherapy option against glioblastoma.
✦ The first research project, globally, that will test the pharmacological efficacy of clinical drugs against Zebrafish Avatars of Glioblastoma Patients.
The zBRAIN project is a prime example of translational research integrated with personalized medical modelling, marking a significant leap in translational cancer research. By integrating multiple biomarkers to predict clinical outcomes and provide real-time feedback on effective drug combinations, zBRAIN will create new knowledge and technologies previously unknown in the field.
The use of Zebrafish Avatars for modeling human glioblastoma represents a novel application of existing technology, combining patient-derived glioblastoma cells and NK cells. This model is pioneering in its ability to mimic patient-specific tumor behavior in vivo. The zebrafish, as an animal model, facilitates high-throughput drug screenings and the assessment of tumor behavior, offering a scalable and cost-effective alternative to traditional rodent patient-derived models. The development of a predictive model integrating multiple biomarkers (known biomarkers such as tumor size, invasion, metastasis, IDH status, MGMT status, with novel ones such as NK-killing efficiency) is a significant innovation.
The hybridization of Zebrafish Avatars (patient-derived orthotopic zebrafish xenografts of glioblastoma) with cancer patients, in a co-clinical study, with the aim to validate the predictive power of this model against the patients’ clinical outcome (cancer progression), is a ground-breaking approach that introduces not only one, but a number of innovations that aim to address the challenges we face in the fields of neuro-oncology, cancer pharmacology and animal modeling.
The funding of zBRAIN will bring a number of “firsts” :
✦ The first research project in Cyprus that will use the concept of Animal Avatars of Cancer Patients.
✦ The first clinical study in Cyprus that uses patient-derived xenografts to predict clinical outcomes.
✦ The first research project, globally, that will test patient-derived NK-cell priming as an innate immunotherapy option against glioblastoma.
✦ The first research project, globally, that will test the pharmacological efficacy of clinical drugs against Zebrafish Avatars of Glioblastoma Patients.
The zBRAIN project is a prime example of translational research integrated with personalized medical modelling, marking a significant leap in translational cancer research. By integrating multiple biomarkers to predict clinical outcomes and provide real-time feedback on effective drug combinations, zBRAIN will create new knowledge and technologies previously unknown in the field.
The use of Zebrafish Avatars for modeling human glioblastoma represents a novel application of existing technology, combining patient-derived glioblastoma cells and NK cells. This model is pioneering in its ability to mimic patient-specific tumor behavior in vivo. The zebrafish, as an animal model, facilitates high-throughput drug screenings and the assessment of tumor behavior, offering a scalable and cost-effective alternative to traditional rodent patient-derived models. The development of a predictive model integrating multiple biomarkers (known biomarkers such as tumor size, invasion, metastasis, IDH status, MGMT status, with novel ones such as NK-killing efficiency) is a significant innovation.
Operational workflow
Our interdisciplinary, innovative, translational paradigm of personalized medicine, integrates clinical and laboratory research with the aim to tackle glioblastoma. Our workflow demonstrates how our diverse research teams, our advanced equipment, and innovative experimental design seamlessly cooperate to enhance the scientific, technical, and social impact of our research. The figure below illustrates this comprehensive workflow, encompassing tasks from all Work Packages which describe in detail the implementation plan of this project.
The project's operation has four major sections: a) Preparation & expansion of primary cells from patient samples and generation of two glioblastoma reporter cell lines, b) Generation of Zebrafish Avatars c) Screening of Avatars and cells against drugs and drug combinations, d) Generation of predictive models of patient outcomes based on multiple biomarkers.
Our interdisciplinary, innovative, translational paradigm of personalized medicine, integrates clinical and laboratory research with the aim to tackle glioblastoma. Our workflow demonstrates how our diverse research teams, our advanced equipment, and innovative experimental design seamlessly cooperate to enhance the scientific, technical, and social impact of our research. The figure below illustrates this comprehensive workflow, encompassing tasks from all Work Packages which describe in detail the implementation plan of this project.
The project's operation has four major sections: a) Preparation & expansion of primary cells from patient samples and generation of two glioblastoma reporter cell lines, b) Generation of Zebrafish Avatars c) Screening of Avatars and cells against drugs and drug combinations, d) Generation of predictive models of patient outcomes based on multiple biomarkers.
The workflow begins with the implementation of our sample processing and primary cell culturing (WP3), a key component of zBRAIN that describes patient recruitment and sampling at Nicosia General Hospital (OKYPY), where tumor and blood samples are collected and sent to EPOS labs. Here, primary glioblastoma cells (differentiated; DGCs and stem cell enriched; GSCs) and NK cells are extracted and expanded, forming the foundation for zebrafish xenografts. In addition, we create two transgenic glioblastoma double-reporter cell lines to use comparison controls for quality assurance.
At UCY, the zebrafish facility creates orthotopic xenografts using labelled glioblastoma and NK cells. The creation of Zebrafish Avatars (WP4) is the core of zBRAIN and its implementation is based on our previous work with orthotopic glioblastoma zebrafish xenografts (UCY) and our high-tech equipment which provides a solid basis for productivity. This part of the project establishes clinically relevant in vivo models that can be used to evaluate therapeutic agents against glioblastoma. The high-throughput drug screenings (WP5) conducted by UCY and EPOS, will focus on selected pre-ranked drugs and drug combinations against the generated Zebrafish Avatars and the patient- derived primary cell cultures. Part of this phase is the administration of pre-activated, stimulated, patient derived NK-cells, as an innate-immunotherapy option, is one of the most innovative aspects of zBRAIN. Its implementation is based on the UCY high-quality expertise in drug repurposing screening and the state-of-the- art equipment of UCY. Efficacy is evaluated using advanced imaging and real-time monitoring (described below). Collected data from these screenings will be used to evaluate tumor behavior and drug efficacy. The final phase of zBRAIN (WP6) will analyze the data from all biomarker readouts for their collective and combining predictive value against patient outcomes. Our partners (EUC-RC and CING) have extensive experience in complex analysis of pro-clinical and clinical data. The 10 studied biomarkers will be analyzed as a combined predictive model using ROC and decision tree analysis.The produced model will aim to enhance the predictability of clinical responses, guiding therapeutic decisions and optimizing patient care.
Our integrated workflow ensures:
►Strong collaboration among Research Team members.
►Higher utilization of advanced equipment.
►Significant contributions from research groups to project outcomes.
►A robust research output incorporating diverse expertise and equipment. This structured approach underscores zBRAIN's commitment to pioneering glioblastoma research, advancing the science of animal Avatars.
At UCY, the zebrafish facility creates orthotopic xenografts using labelled glioblastoma and NK cells. The creation of Zebrafish Avatars (WP4) is the core of zBRAIN and its implementation is based on our previous work with orthotopic glioblastoma zebrafish xenografts (UCY) and our high-tech equipment which provides a solid basis for productivity. This part of the project establishes clinically relevant in vivo models that can be used to evaluate therapeutic agents against glioblastoma. The high-throughput drug screenings (WP5) conducted by UCY and EPOS, will focus on selected pre-ranked drugs and drug combinations against the generated Zebrafish Avatars and the patient- derived primary cell cultures. Part of this phase is the administration of pre-activated, stimulated, patient derived NK-cells, as an innate-immunotherapy option, is one of the most innovative aspects of zBRAIN. Its implementation is based on the UCY high-quality expertise in drug repurposing screening and the state-of-the- art equipment of UCY. Efficacy is evaluated using advanced imaging and real-time monitoring (described below). Collected data from these screenings will be used to evaluate tumor behavior and drug efficacy. The final phase of zBRAIN (WP6) will analyze the data from all biomarker readouts for their collective and combining predictive value against patient outcomes. Our partners (EUC-RC and CING) have extensive experience in complex analysis of pro-clinical and clinical data. The 10 studied biomarkers will be analyzed as a combined predictive model using ROC and decision tree analysis.The produced model will aim to enhance the predictability of clinical responses, guiding therapeutic decisions and optimizing patient care.
Our integrated workflow ensures:
►Strong collaboration among Research Team members.
►Higher utilization of advanced equipment.
►Significant contributions from research groups to project outcomes.
►A robust research output incorporating diverse expertise and equipment. This structured approach underscores zBRAIN's commitment to pioneering glioblastoma research, advancing the science of animal Avatars.
Project Work Packages
WP1 - Project Management
WP2 - Dissemination/Exploitation
WP3 - Patient Sample Processing
WP4 - Zebrafish Avatars
WP5 - miRNA & Drug Screening
WP6 - Data Analysis
WP1 - Project Management
WP2 - Dissemination/Exploitation
WP3 - Patient Sample Processing
WP4 - Zebrafish Avatars
WP5 - miRNA & Drug Screening
WP6 - Data Analysis
The Vision of zBRAIN
To revolutionize glioblastoma treatment by integrating Zebrafish Avatars into clinical research, ultimately improving patient outcomes through personalized therapeutic strategies.
The Mission of zBRAIN
To establish a pioneering platform that combines patient-derived glioblastoma cells and NK cells in Zebrafish Avatars, enabling the real-time study of tumor behavior and therapeutic responses.
The Primary Aim of zBRAIN
To validate the predictive power of Zebrafish Avatars for clinical outcomes in glioblastoma patients, using an innovative predictive model of combined biomarkers.
To revolutionize glioblastoma treatment by integrating Zebrafish Avatars into clinical research, ultimately improving patient outcomes through personalized therapeutic strategies.
The Mission of zBRAIN
To establish a pioneering platform that combines patient-derived glioblastoma cells and NK cells in Zebrafish Avatars, enabling the real-time study of tumor behavior and therapeutic responses.
The Primary Aim of zBRAIN
To validate the predictive power of Zebrafish Avatars for clinical outcomes in glioblastoma patients, using an innovative predictive model of combined biomarkers.
RESEARCH TEAM - COLLABORATORS
