Periodic Reporting for period 1 - GlioLighT (Next Generation Glioma Treatments using Direct Light Therapy)
Période du rapport: 2024-01-01 au 2024-12-31
Glioma is an extremely lethal cancer, due largely to the inaccessible nature of the brain and diffusion of cells from the tumour site.
These diffuse cells are usually too deeply embedded in the brain to safely remove by current means. Targeted Reactive Oxygen Species
(ROS) generation is a promising form of glioma treatment to selectively eliminate glioma, including diffuse cells. However, the only
current means of targeted ROS generation is photodynamic therapy (PDT) which generates ROS using expensive and potentially toxic
photosensitisers (PS) which are ineffective against distant diffused cells and introduce many treatment limitations. GlioLighT proposes
a novel alternative form of targeted ROS generation: Direct Light Therapy (DLT). DLT uses 1267nm light to generate 1O2 species in
glioma cells without dependency on a PS. The removal of PS will revolutionise glioma treatment, enabling novel treatment modalities
to vastly improve efficacy, earlier intervention options, all at reduced cost and complexity. However, whilst the principles of DLT have
been demonstrated, little is known about how DLT achieves its anti-cancer effects, or the extent of its therapeutic benefits. Leveraging
decades of accumulated PDT knowledge and technology development, GlioLighT will study DLT technology both independently and
compared to PDT. The effect of DLT on glioma and the brain, focusing on immunogenicity, will be studied to determine DLT’s efficacy,
safety, and mechanisms of action. Novel ultrashort pulse (USP) light sources will be developed to maximise optical penetration and
minimise safety risk, ensuring DLT is suited for clinical adoption. Lastly, the development of the preclinical GlioLighT delivery and
sensing system (pcGlio-DSS) ready for the next steps of clinical translation, will bring DLT a leap closer to vastly improving glioma
treatment in Europe and worldwide.
These diffuse cells are usually too deeply embedded in the brain to safely remove by current means. Targeted Reactive Oxygen Species
(ROS) generation is a promising form of glioma treatment to selectively eliminate glioma, including diffuse cells. However, the only
current means of targeted ROS generation is photodynamic therapy (PDT) which generates ROS using expensive and potentially toxic
photosensitisers (PS) which are ineffective against distant diffused cells and introduce many treatment limitations. GlioLighT proposes
a novel alternative form of targeted ROS generation: Direct Light Therapy (DLT). DLT uses 1267nm light to generate 1O2 species in
glioma cells without dependency on a PS. The removal of PS will revolutionise glioma treatment, enabling novel treatment modalities
to vastly improve efficacy, earlier intervention options, all at reduced cost and complexity. However, whilst the principles of DLT have
been demonstrated, little is known about how DLT achieves its anti-cancer effects, or the extent of its therapeutic benefits. Leveraging
decades of accumulated PDT knowledge and technology development, GlioLighT will study DLT technology both independently and
compared to PDT. The effect of DLT on glioma and the brain, focusing on immunogenicity, will be studied to determine DLT’s efficacy,
safety, and mechanisms of action. Novel ultrashort pulse (USP) light sources will be developed to maximise optical penetration and
minimise safety risk, ensuring DLT is suited for clinical adoption. Lastly, the development of the preclinical GlioLighT delivery and
sensing system (pcGlio-DSS) ready for the next steps of clinical translation, will bring DLT a leap closer to vastly improving glioma
treatment in Europe and worldwide.
WP1: Continuous Wave (CW) 1267nm laser systems were successfully produced, achieving all requested parameters. The CW laser system was distributed to partners and training was delivered to partners. Prototypes of the two Ultra Short Pulse (USP) laser systems (Distributed Feedback (DFB) and Fibre-Bragg Grating (FBG)) have been developed and characterised. So far, the Fibre-Bragg Grating structure system performs better in terms of pulse duration and does not suffer from the excess heating problem experienced with the DFB laser. Pulse duration has been reduced to 80-90ps with a FBG USP laser, which should be short enough that no further heating issues can arise; however, AU will proceed with semiconductor laser research in RP2 to reduce the pulse duration further, which would provide a larger margin of safety against unforeseen heating issues.
WP2: The right conditions of cell illumination and range of irradiations cytotoxic for human glioblastoma (GB) cell lines but not for normal human brain cells (astrocytes) or immune cells (macrophages) are identified. Neuronal astrocyte co-cultures and cell lines have been set up, and training provided to the scientific team on neuronal network calcium imaging recording and analysis. Preliminary information has been obtained on the cytotoxic effect of the laser on the human glioblastoma NCH82 cell line and its effect on the invasion capability of spheroids made of primary tumor material. Preliminary analysis has been performed on the effect of CW-DLT on inflammatory (gene expression) profiles of immune cells.
WP3: Optical phantoms for mimicking inhomogeneous brain tissue at 635nm were developed. Spectral Online Monitoring (SOM) measurements were performed and evaluated. Optical properties of human brain tissue at 1267nm were collected from literature, and due to discrepancy between results, plans were made to measure these properties at LMU. 3 different cylindrical fiber diffusors in 3 different media at 2 different wavelengths (1267 nm and 635 nm) were characterized regarding their emission characteristics by direct imaging. The ML8500 high-throughput illumination system was prepared for in vitro study to compare effectiveness of DLT and PDT in cytotoxicity & singlet oxygen generation. The ML6600 stand-alone laser for preclinical studies was manufactured as a DLT and ALA-PDT light source. Additionally, commercial light diffusers for both 635nm and 1267nm light were tested, and a cylindrical diffuser with a FBG-based temperature sensor was designed, for use in the pcGlio-DSS system.
WP4: Preparatory work has begun ahead of WP4 beginning in M13. VMDK embryos have been transferred from UM and a new mouse colony grown at UB for the in-vivo work planned. Preparation for the financial and bidding processes for future MRI recordings has been undertaken. Ethical licenses have been submitted for these in-vivo experiments, and licenses to work with mice for trainees have been collected. Training has been delivered in stereotaxic surgery, light stimulation, fibre photometry, and MRI scanning to the relevant staff.
WP6: A site for document sharing was established, and regular online and in-person consortium meetings were held. This allowed effective progress and risk monitoring. All deliverables due in the first period have been successfully completed.
WP2: The right conditions of cell illumination and range of irradiations cytotoxic for human glioblastoma (GB) cell lines but not for normal human brain cells (astrocytes) or immune cells (macrophages) are identified. Neuronal astrocyte co-cultures and cell lines have been set up, and training provided to the scientific team on neuronal network calcium imaging recording and analysis. Preliminary information has been obtained on the cytotoxic effect of the laser on the human glioblastoma NCH82 cell line and its effect on the invasion capability of spheroids made of primary tumor material. Preliminary analysis has been performed on the effect of CW-DLT on inflammatory (gene expression) profiles of immune cells.
WP3: Optical phantoms for mimicking inhomogeneous brain tissue at 635nm were developed. Spectral Online Monitoring (SOM) measurements were performed and evaluated. Optical properties of human brain tissue at 1267nm were collected from literature, and due to discrepancy between results, plans were made to measure these properties at LMU. 3 different cylindrical fiber diffusors in 3 different media at 2 different wavelengths (1267 nm and 635 nm) were characterized regarding their emission characteristics by direct imaging. The ML8500 high-throughput illumination system was prepared for in vitro study to compare effectiveness of DLT and PDT in cytotoxicity & singlet oxygen generation. The ML6600 stand-alone laser for preclinical studies was manufactured as a DLT and ALA-PDT light source. Additionally, commercial light diffusers for both 635nm and 1267nm light were tested, and a cylindrical diffuser with a FBG-based temperature sensor was designed, for use in the pcGlio-DSS system.
WP4: Preparatory work has begun ahead of WP4 beginning in M13. VMDK embryos have been transferred from UM and a new mouse colony grown at UB for the in-vivo work planned. Preparation for the financial and bidding processes for future MRI recordings has been undertaken. Ethical licenses have been submitted for these in-vivo experiments, and licenses to work with mice for trainees have been collected. Training has been delivered in stereotaxic surgery, light stimulation, fibre photometry, and MRI scanning to the relevant staff.
WP6: A site for document sharing was established, and regular online and in-person consortium meetings were held. This allowed effective progress and risk monitoring. All deliverables due in the first period have been successfully completed.
The GlioLight project is progressing well and has adhered effectively to the activities planned in the grant agreement. The first year of the project has largely been focused on establishing Direct Light Therapy (DLT) technology at the partner institutions, generating preliminary results that provide indications of the potential of DLT, and making the necessary preparations for work planned for the latter stages of the project.
Initial results on the effects of DLT in vitro provide some evidence that DLT can achieve specific cancer cell killing, in line with the project’s overarching hypothesis. However, the partners understand that these results generated during RP1 are extremely preliminary and suboptimal and do not yet prove that DLT achieves the intended effects. Extensive further work is required during RP2 to validate these very preliminary results. This is in accordance with the progress anticipated by this time point.
At the end of RP1, the consortium is now well-positioned to enter the latter phase of the project and generate the key results the project aims to produce.
Based on the results achieved so far, it is likely that the GlioLighT project will achieve the impacts it set out to achieve. None of the results produced to date give cause to believe that DLT is not capable of performing as anticipated.
Initial results on the effects of DLT in vitro provide some evidence that DLT can achieve specific cancer cell killing, in line with the project’s overarching hypothesis. However, the partners understand that these results generated during RP1 are extremely preliminary and suboptimal and do not yet prove that DLT achieves the intended effects. Extensive further work is required during RP2 to validate these very preliminary results. This is in accordance with the progress anticipated by this time point.
At the end of RP1, the consortium is now well-positioned to enter the latter phase of the project and generate the key results the project aims to produce.
Based on the results achieved so far, it is likely that the GlioLighT project will achieve the impacts it set out to achieve. None of the results produced to date give cause to believe that DLT is not capable of performing as anticipated.