Periodic Reporting for period 1 - MAGNIFICO (MAGNIFICO-Pre-commercialization of multifunctional targeted MRI-contrast enhancing agents for brain research)
Okres sprawozdawczy: 2022-10-01 do 2024-03-31
The human brain stands as the pinnacle of complexity among human organs, serving as a pivotal domain for research within medicine and science. Within Europe, the prevalence of brain disorders imposes a significant burden, affecting approximately 165 million individuals. Notably, certain neurological conditions such as Alzheimer's disease, dementia, stroke, and Parkinson's exhibit heightened prevalence due to the demographic shift towards an aging population, given their increased incidence with advancing age. To alleviate the burden of brain disorders, an imperative initial step involves enhancing our comprehension of the brain and its intricate processes. This endeavor holds promise for the development of novel treatments and preventive measures.
Brain imaging emerges as a cornerstone in both research and clinical realms. In research, imaging offers crucial insights into the manifestation and progression of brain disorders such as Alzheimer's disease, aiding in the delineation of their spatial and temporal dynamics. Furthermore, imaging serves as a fundamental asset in drug discovery endeavors, facilitating the establishment of inclusion criteria, safety markers, and outcome measures for therapeutic trials. Enhanced understanding of the brain promises improved diagnostic capabilities and treatment modalities, thereby fostering more effective management of brain disorders. Given the substantial economic burden associated with the care costs of brain diseases, investments in advancing diagnostic and therapeutic avenues are poised to yield substantial returns through the reduction of direct non-medical costs and indirect burdens on healthcare systems.
Technological advancements in brain imaging hold immense potential for driving progress in brain research, paving the way for more efficacious treatments aimed at alleviating strain on healthcare systems already under duress.
Imaging modalities represent invaluable tools for elucidating the intricacies of the brain, with a particular emphasis on non-invasive, non-destructive methodologies capable of longitudinally monitoring the brain in its entirety. While light microscopy (LM) presents certain advantages, including high resolution, it is constrained by limited fields-of-view and depth penetration, necessitating invasive procedures. In contrast, Magnetic Resonance Imaging (MRI) stands out for its non-invasive and non-destructive nature, offering mesoscale images of the entire brain at various depths.
Within the framework of our ERC-PoC proposal, we aim to augment the existing arsenal of brain imaging tools by integrating light microscopy with enhanced MRI detection capabilities targeting specific entities within the brain. This involves leveraging genetically encoded targets combined with specific chemical labeling facilitated by multimodal imaging agents.
Our specific objectives encompass:
• Production and optimization of multimodal imaging molecules, alongside the development of exogenously-expressed genetic targets and their delivery to target cells.
• Demonstration of the suitability of labeling for multimodal imaging in vivo.
• Assessment of the commercial feasibility of the proposed approach.
Brain imaging emerges as a cornerstone in both research and clinical realms. In research, imaging offers crucial insights into the manifestation and progression of brain disorders such as Alzheimer's disease, aiding in the delineation of their spatial and temporal dynamics. Furthermore, imaging serves as a fundamental asset in drug discovery endeavors, facilitating the establishment of inclusion criteria, safety markers, and outcome measures for therapeutic trials. Enhanced understanding of the brain promises improved diagnostic capabilities and treatment modalities, thereby fostering more effective management of brain disorders. Given the substantial economic burden associated with the care costs of brain diseases, investments in advancing diagnostic and therapeutic avenues are poised to yield substantial returns through the reduction of direct non-medical costs and indirect burdens on healthcare systems.
Technological advancements in brain imaging hold immense potential for driving progress in brain research, paving the way for more efficacious treatments aimed at alleviating strain on healthcare systems already under duress.
Imaging modalities represent invaluable tools for elucidating the intricacies of the brain, with a particular emphasis on non-invasive, non-destructive methodologies capable of longitudinally monitoring the brain in its entirety. While light microscopy (LM) presents certain advantages, including high resolution, it is constrained by limited fields-of-view and depth penetration, necessitating invasive procedures. In contrast, Magnetic Resonance Imaging (MRI) stands out for its non-invasive and non-destructive nature, offering mesoscale images of the entire brain at various depths.
Within the framework of our ERC-PoC proposal, we aim to augment the existing arsenal of brain imaging tools by integrating light microscopy with enhanced MRI detection capabilities targeting specific entities within the brain. This involves leveraging genetically encoded targets combined with specific chemical labeling facilitated by multimodal imaging agents.
Our specific objectives encompass:
• Production and optimization of multimodal imaging molecules, alongside the development of exogenously-expressed genetic targets and their delivery to target cells.
• Demonstration of the suitability of labeling for multimodal imaging in vivo.
• Assessment of the commercial feasibility of the proposed approach.
Throughout the project duration, we conducted a systematic exploration of the synthetic chemistry involved in the creation of multimodal imaging molecules. As a result, we successfully developed a comprehensive suite of molecules tailored for utilization in light microscopy. Furthermore, we engineered a prototype agent capable of facilitating both MRI and light microscopy, exhibiting high specificity towards our target proteins, denoted as eTag.
In parallel, we established a diverse array of target proteins featuring adaptable genetic elements conducive to cellular surface trafficking, fluorescent molecule labeling, and precise manipulation of expression within specific cell types. The fusion of imaging molecules with these genetically engineered target proteins demonstrated exceptional specificity and exclusivity, yielding the anticipated multimodal labeling discernible through fluorescence and MRI signal detection.
A pivotal achievement of our endeavor involves the formulation of a novel protocol for ex vivo MRI imaging of intact tissues previously rendered suitable for ex vivo light microscopy imaging via tissue clearing techniques. This innovative ex vivo multimodal imaging protocol facilitates the comprehensive visualization of large, intact tissues utilizing both imaging modalities concurrently. By circumventing the necessity for tissue sectioning and subsequent image reconstruction, as well as mitigating potential artifacts such as tissue deformations, this approach offers users an unbiased methodology for image registration. Moreover, it facilitates rapid orientation within voluminous tissues and provides validation of MRI signal integrity through correlation with the fluorescent signal emitted by the multifunctional contrast agent.
In parallel, we established a diverse array of target proteins featuring adaptable genetic elements conducive to cellular surface trafficking, fluorescent molecule labeling, and precise manipulation of expression within specific cell types. The fusion of imaging molecules with these genetically engineered target proteins demonstrated exceptional specificity and exclusivity, yielding the anticipated multimodal labeling discernible through fluorescence and MRI signal detection.
A pivotal achievement of our endeavor involves the formulation of a novel protocol for ex vivo MRI imaging of intact tissues previously rendered suitable for ex vivo light microscopy imaging via tissue clearing techniques. This innovative ex vivo multimodal imaging protocol facilitates the comprehensive visualization of large, intact tissues utilizing both imaging modalities concurrently. By circumventing the necessity for tissue sectioning and subsequent image reconstruction, as well as mitigating potential artifacts such as tissue deformations, this approach offers users an unbiased methodology for image registration. Moreover, it facilitates rapid orientation within voluminous tissues and provides validation of MRI signal integrity through correlation with the fluorescent signal emitted by the multifunctional contrast agent.
In accordance with Objective 1, we have successfully achieved the production, optimization, and characterization of multiple multifunctional imaging agent (MFS-agents) variants alongside engineered target proteins (eTags). Extensive characterization studies have been conducted, encompassing labeling assessments in cell culture environments through fluorescence analysis, as well as in cell pellets through MRI signal evaluation.
Addressing Objective 2, our efforts have focused on demonstrating the delivery of eTags to brain tissues utilizing adeno-associated virus (AAV) vectors administered through various injection routes, including direct brain injection, cisterna magna infusion, and intravenous delivery via the tail vein. Additionally, we have showcased the manipulation of gene expression using a plethora of genetic tools, such as promoters, self-cleavable peptides, and viral capsids. A novel methodology for dual MRI and light microscopy (LM) imaging ex vivo has been developed, underscoring our commitment to advancing imaging capabilities.
However, a significant challenge emerged in the quest for enhancing MRI signal in vivo. Despite the successful synthesis of a prototype MFS molecule bearing a single MRI moiety, subsequent endeavors to synthesize a compound incorporating multiple MRI moieties encountered setbacks. Our internal chemistry team encountered difficulties in the conjugation process with the MRI agents, and subsequent collaboration with a subcontracting company, WuXi AppTec, also proved unsuccessful in overcoming this hurdle.
Regrettably, at present, we are unable to demonstrate dual MRI and LM imaging in vivo due to the unresolved challenges encountered during the synthesis and conjugation of multiple MRI moieties. Nonetheless, we remain committed to overcoming these obstacles and advancing the project towards its intended objectives.
Addressing Objective 2, our efforts have focused on demonstrating the delivery of eTags to brain tissues utilizing adeno-associated virus (AAV) vectors administered through various injection routes, including direct brain injection, cisterna magna infusion, and intravenous delivery via the tail vein. Additionally, we have showcased the manipulation of gene expression using a plethora of genetic tools, such as promoters, self-cleavable peptides, and viral capsids. A novel methodology for dual MRI and light microscopy (LM) imaging ex vivo has been developed, underscoring our commitment to advancing imaging capabilities.
However, a significant challenge emerged in the quest for enhancing MRI signal in vivo. Despite the successful synthesis of a prototype MFS molecule bearing a single MRI moiety, subsequent endeavors to synthesize a compound incorporating multiple MRI moieties encountered setbacks. Our internal chemistry team encountered difficulties in the conjugation process with the MRI agents, and subsequent collaboration with a subcontracting company, WuXi AppTec, also proved unsuccessful in overcoming this hurdle.
Regrettably, at present, we are unable to demonstrate dual MRI and LM imaging in vivo due to the unresolved challenges encountered during the synthesis and conjugation of multiple MRI moieties. Nonetheless, we remain committed to overcoming these obstacles and advancing the project towards its intended objectives.