Periodic Reporting for period 1 - PROTECT (Protease Profiling and Triggered Drug Delivery for Personalized Cancer Therapy)
Okres sprawozdawczy: 2022-09-01 do 2025-02-28
Cancer progression is significantly influenced by the activity of proteases, enzymes that break down proteins and play a critical role in tumor growth, metastasis, and the formation of new blood vessels (angiogenesis). Despite extensive research, clinical efforts to target proteases for cancer therapy have largely failed. Existing methods for detecting and analyzing protease activity in biological samples struggle with low sensitivity and poor selectivity, making it difficult to translate protease-targeting approaches into clinical applications. As a result, new, innovative solutions are urgently needed to improve the detection and use of protease activity in cancer diagnostics and treatment.
PROTECT aims to revolutionize the way we monitor and target protease activity in cancer by developing a comprehensive platform for protease profiling. This platform will address the limitations of current strategies, dramatically increasing sensitivity and the ability to simultaneously detect multiple proteases. By doing so, PROTECT will create new tools for personalized cancer therapy, tailoring treatments based on the specific protease activity profile of each patient’s tumor.
In PROTECT project two groundbreaking concepts are developed:
Liposomal Activity-Based Sensors (LABS) – These sensors will localize protease-responsive peptides to the tumor microenvironment, amplifying the protease activity by releasing synthetic biomarkers.
Protease Profile Specific Drug Delivery Vehicles (PROVES) – These liposomal systems will deliver encapsulated drugs directly to the tumor site, with high efficiency and specificity, triggered by the unique protease profile of the tumor.
PROTECT is structured around three core research tasks:
Development of Liposome-Conjugated proMAPs: The first task focuses on developing protease-responsive membrane-active peptides (proMAPs) that will be linked to liposomes. These peptides will trigger the release of synthetic biomarkers—encoded molecular barcodes—as a direct response to protease activity. This innovative system will allow us to track the activity of proteases in the tumor environment in real-time, with greater accuracy than current methods.
Multiplexed Protease Profiling: The second task involves developing methods to detect and analyze the release patterns of these barcodes, allowing us to correlate them with the activity of multiple cancer-associated proteases. This will enable robust multiplexed protease profiling, meaning that several proteases can be monitored simultaneously, providing a detailed picture of the tumor’s protease activity.
Precision Medicine Through Protease-Triggered Drug Delivery: The third task will focus on using the tumor’s unique protease profile to guide the release of drugs from liposomal delivery vehicles. These systems will be designed to release therapeutic agents specifically in response to the protease activity within the tumor microenvironment, ensuring targeted and efficient drug delivery. The effectiveness of this system will be evaluated in advanced in vitro 3D breast cancer models, which simulate the complex biological environment of a tumor.
The PROTECT project is expected to go far beyond current state-of-the-art technologies in cancer diagnostics and therapy. By integrating advanced peptide design with innovative lipid-nanoparticle systems, this project will develop highly sensitive and specific tools for monitoring protease activity and delivering drugs with unprecedented precision. This breakthrough could transform cancer treatment by enabling personalized therapies tailored to the protease activity profiles of individual tumors. In the long term, PROTECT has the potential to improve patient outcomes, reduce side effects of cancer treatments, and open new pathways for research into protease-targeted therapies. The success of PROTECT depends on a truly interdisciplinary approach, combining expertise in peptide chemistry, nanotechnology, cancer biology, and bioengineering. By working at the intersection of these disciplines, PROTECT is positioned to make significant contributions to both cancer diagnostics and therapeutics.
PROTECT aims to revolutionize the way we monitor and target protease activity in cancer by developing a comprehensive platform for protease profiling. This platform will address the limitations of current strategies, dramatically increasing sensitivity and the ability to simultaneously detect multiple proteases. By doing so, PROTECT will create new tools for personalized cancer therapy, tailoring treatments based on the specific protease activity profile of each patient’s tumor.
In PROTECT project two groundbreaking concepts are developed:
Liposomal Activity-Based Sensors (LABS) – These sensors will localize protease-responsive peptides to the tumor microenvironment, amplifying the protease activity by releasing synthetic biomarkers.
Protease Profile Specific Drug Delivery Vehicles (PROVES) – These liposomal systems will deliver encapsulated drugs directly to the tumor site, with high efficiency and specificity, triggered by the unique protease profile of the tumor.
PROTECT is structured around three core research tasks:
Development of Liposome-Conjugated proMAPs: The first task focuses on developing protease-responsive membrane-active peptides (proMAPs) that will be linked to liposomes. These peptides will trigger the release of synthetic biomarkers—encoded molecular barcodes—as a direct response to protease activity. This innovative system will allow us to track the activity of proteases in the tumor environment in real-time, with greater accuracy than current methods.
Multiplexed Protease Profiling: The second task involves developing methods to detect and analyze the release patterns of these barcodes, allowing us to correlate them with the activity of multiple cancer-associated proteases. This will enable robust multiplexed protease profiling, meaning that several proteases can be monitored simultaneously, providing a detailed picture of the tumor’s protease activity.
Precision Medicine Through Protease-Triggered Drug Delivery: The third task will focus on using the tumor’s unique protease profile to guide the release of drugs from liposomal delivery vehicles. These systems will be designed to release therapeutic agents specifically in response to the protease activity within the tumor microenvironment, ensuring targeted and efficient drug delivery. The effectiveness of this system will be evaluated in advanced in vitro 3D breast cancer models, which simulate the complex biological environment of a tumor.
The PROTECT project is expected to go far beyond current state-of-the-art technologies in cancer diagnostics and therapy. By integrating advanced peptide design with innovative lipid-nanoparticle systems, this project will develop highly sensitive and specific tools for monitoring protease activity and delivering drugs with unprecedented precision. This breakthrough could transform cancer treatment by enabling personalized therapies tailored to the protease activity profiles of individual tumors. In the long term, PROTECT has the potential to improve patient outcomes, reduce side effects of cancer treatments, and open new pathways for research into protease-targeted therapies. The success of PROTECT depends on a truly interdisciplinary approach, combining expertise in peptide chemistry, nanotechnology, cancer biology, and bioengineering. By working at the intersection of these disciplines, PROTECT is positioned to make significant contributions to both cancer diagnostics and therapeutics.
The project is divided into three main research tasks (RTs).
In RT1, we worked on creating protease-responsive membrane-active peptides (proMAPs) and integrating them with liposomal systems to enhance drug delivery and biomarker release. We tested two strategies for inhibiting membrane activity and controlling the release of these proMAPs. Although the first approach did not achieve the desired inhibition, the second strategy, where we modified the peptide structure, has shown promising results. Additionally, we investigated how different liposomal compositions affect the activity of proMAPs, giving us more control over the release of synthetic biomarkers in the tumor microenvironment. A novel enzyme-assisted method was also developed to improve peptide-lipid conjugation, solving some of the technical challenges with oxidation during the process.
In RT2, we focused on identifying target proteases and developing in vitro breast cancer models that mimic real tissue conditions. We created 3D breast cancer models using engineered hydrogels that replicate the stiffness of human breast tissue. These models allowed us to observe how increased tissue stiffness impacts cancer progression, promoting tumor growth by enhancing protease activity and interactions between cancer cells and the surrounding stromal cells. In parallel, we used 3D bioprinting to study how cancer cells and fibroblasts communicate and remodel the tumor microenvironment, providing key insights into protease-driven cancer progression.
RT3, which will focus on protease-triggered drug delivery systems, has not yet begun but will build on the advances from RT1 and RT2.
The work performed so far sets a strong foundation for future developments in precision cancer treatments and diagnostics.
In RT1, we worked on creating protease-responsive membrane-active peptides (proMAPs) and integrating them with liposomal systems to enhance drug delivery and biomarker release. We tested two strategies for inhibiting membrane activity and controlling the release of these proMAPs. Although the first approach did not achieve the desired inhibition, the second strategy, where we modified the peptide structure, has shown promising results. Additionally, we investigated how different liposomal compositions affect the activity of proMAPs, giving us more control over the release of synthetic biomarkers in the tumor microenvironment. A novel enzyme-assisted method was also developed to improve peptide-lipid conjugation, solving some of the technical challenges with oxidation during the process.
In RT2, we focused on identifying target proteases and developing in vitro breast cancer models that mimic real tissue conditions. We created 3D breast cancer models using engineered hydrogels that replicate the stiffness of human breast tissue. These models allowed us to observe how increased tissue stiffness impacts cancer progression, promoting tumor growth by enhancing protease activity and interactions between cancer cells and the surrounding stromal cells. In parallel, we used 3D bioprinting to study how cancer cells and fibroblasts communicate and remodel the tumor microenvironment, providing key insights into protease-driven cancer progression.
RT3, which will focus on protease-triggered drug delivery systems, has not yet begun but will build on the advances from RT1 and RT2.
The work performed so far sets a strong foundation for future developments in precision cancer treatments and diagnostics.
The PROTECT project has made significant progress in developing new technologies for cancer diagnostics and treatment. One of the key breakthroughs is the creation of special peptides (proMAPs) that can be controlled by enzymes, allowing precise activation for drug delivery and biomarker detection. This new method overcomes issues with existing techniques and is a major step forward in creating systems that respond to specific cancer-related enzyme activity. These peptides are critical to the success of our two core concepts: LABS (Liposomal Activity-Based Sensors) and PROVES (Protease-Responsive Vesicles), which aim to improve cancer treatment and diagnostics by targeting the unique environment of tumors.
We have also developed advanced breast cancer models that mimic human tissue stiffness. These models help us understand how the physical properties of tissue influence cancer growth and can lead to new strategies for cancer therapy. The success of these models is already opening new opportunities for research into cancer progression.
To fully realize the potential of these innovations, further validation, collaboration with pre-clinical and clinical researchers, and securing intellectual property will be key steps in moving toward clinical application. These advancements could lead to more precise and effective cancer treatments, benefiting patients by reducing side effects and improving outcomes.
We have also developed advanced breast cancer models that mimic human tissue stiffness. These models help us understand how the physical properties of tissue influence cancer growth and can lead to new strategies for cancer therapy. The success of these models is already opening new opportunities for research into cancer progression.
To fully realize the potential of these innovations, further validation, collaboration with pre-clinical and clinical researchers, and securing intellectual property will be key steps in moving toward clinical application. These advancements could lead to more precise and effective cancer treatments, benefiting patients by reducing side effects and improving outcomes.