Periodic Reporting for period 1 - SILORGS (Sensor islet organoids (SILORGS) for in vivo identification of anti-diabetic drugs)
Okres sprawozdawczy: 2024-07-01 do 2025-12-31
Diabetes affects millions worldwide due to loss of functional insulin-producing β-cells. Current drug development is slow and costly, as preclinical models cannot continuously monitor human β-cell function in living organisms. The SILORGS project overcomes this hurdle by transplanting genetically engineered pancreatic islet organoids into the anterior chamber of the eye (ACE), a natural body window allowing longitudinal islet/organoid monitoring in vivo, non-invasively, at high-resolution. The organoids are equipped with biosensors that report β-cell activity in real time through fluorescent Ca2+ signals. Human islets and sensor organoids were transplanted into immunocompromised mice, with diabetes induced by a high-fat, high-sucrose diet (HFHSD) and selective β-cell ablation of in situ islets. Validation of the platform using the GLP-1 analogue liraglutide, as a proof-of-concept medication, demonstrated the ability of the platform to detect functional changes in β-cells, assess graft morphology and vascularization, and monitor treatment effects over time. SILORGS establishes a novel, integrated platform for early-stage drug testing, combining human tissue, genetic biosensors, and longitudinal in vivo imaging, with strong potential for commercial preclinical screening of anti-diabetic therapies.
Current preclinical models of diabetes primarily rely on systemic metabolic measurements, endpoint histology, or in vitro assays, which fail to capture the dynamic behavior of human pancreatic β-cells in vivo. This limitation reduces predictive accuracy, contributing to high failure rates in clinical translation and slowing the development of innovative therapies. A key unmet need is the ability to monitor human β-cell function longitudinally and non-invasively using sensor-enabled islet organoids, while simultaneously assessing systemic metabolic responses.
The SILORGS Proof of Concept project aimed at establishing and validating an industry-oriented in vivo imaging platform using genetically engineered sensor islet organoids transplanted into the ACE of recipient mice. The central milestone, achieved within 18 months, was the in vivo validation of these organoids as functional reporters for human β-cell activity in humanized diabetic mouse models, enabling standardized protocols for drug testing.
The project successfully generated fluorescent biosensor-integrated islet organoids, established humanized mouse models of diet-induced diabetes, and validated the platform using the GLP-1 analogue liraglutide. Functional in vivo imaging was combined with metabolic testing and longitudinal intravital analyses to evaluate β-cell activity, islet morphology, and vascularization.
The expected impact of SILORGS is threefold:
1. Scientific impact: Establishment of a novel in vivo functional imaging platform using sensor islet organoids as high-resolution reporters of human β-cell function.
2. Translational impact: Enhanced predictive power in early-stage drug development, potentially reducing costs and failure rates.
3. Commercial impact: Development of a scalable screening service for pharmaceutical companies focused on validating anti-diabetic lead compounds.
By enabling direct visualization of functional human β-cell responses in living organisms, SILORGS addresses a critical bottleneck in diabetes drug development and strengthens Europe’s position in translational biomedical innovation.
Current preclinical models of diabetes primarily rely on systemic metabolic measurements, endpoint histology, or in vitro assays, which fail to capture the dynamic behavior of human pancreatic β-cells in vivo. This limitation reduces predictive accuracy, contributing to high failure rates in clinical translation and slowing the development of innovative therapies. A key unmet need is the ability to monitor human β-cell function longitudinally and non-invasively using sensor-enabled islet organoids, while simultaneously assessing systemic metabolic responses.
The SILORGS Proof of Concept project aimed at establishing and validating an industry-oriented in vivo imaging platform using genetically engineered sensor islet organoids transplanted into the ACE of recipient mice. The central milestone, achieved within 18 months, was the in vivo validation of these organoids as functional reporters for human β-cell activity in humanized diabetic mouse models, enabling standardized protocols for drug testing.
The project successfully generated fluorescent biosensor-integrated islet organoids, established humanized mouse models of diet-induced diabetes, and validated the platform using the GLP-1 analogue liraglutide. Functional in vivo imaging was combined with metabolic testing and longitudinal intravital analyses to evaluate β-cell activity, islet morphology, and vascularization.
The expected impact of SILORGS is threefold:
1. Scientific impact: Establishment of a novel in vivo functional imaging platform using sensor islet organoids as high-resolution reporters of human β-cell function.
2. Translational impact: Enhanced predictive power in early-stage drug development, potentially reducing costs and failure rates.
3. Commercial impact: Development of a scalable screening service for pharmaceutical companies focused on validating anti-diabetic lead compounds.
By enabling direct visualization of functional human β-cell responses in living organisms, SILORGS addresses a critical bottleneck in diabetes drug development and strengthens Europe’s position in translational biomedical innovation.
Experimental strategy: 400 human islets were transplanted into immunocompromised Rag1-/- mice in one eye to maintain metabolic control, while 10–15 human reporter islets were transplanted into the contralateral eye. After engraftment and streptozotocin-mediated ablation of endogenous murine β-cells, mice relied on the transplanted human islets for glucose homeostasis. Diabetes was established by feeding mice a high-fat-high-sucrose diet (HFHSD). Generation of human sensor islet organoids by viral GCaMP6s transduction showed insufficient efficiency. Therefore, murine GCaMP3-expressing sensor islet organoids, resembling the human-like intermingling of endocrine cells, were produced and co-transplanted into the ACE containing the human reporter islets.
Main technical achievements: A robust humanized diabetic mouse model was established by combining human islet transplantation, murine β-cell ablation, and HFHSD-induced metabolic stress. Sensor islet organoids were successfully generated from dissociated mouse GCaMP3 islets and re-aggregated into organoids suitable for ACE transplantation. A longitudinal in vivo imaging pipeline enabling repeated confocal measurements of β-cell intracellular Ca2+ dynamics in the same grafts over several months was developed. The biosensor system detected functional changes in Ca2+ oscillations in response to metabolic stress and treatment with the GLP-1 analogue liraglutide, with imaging results integrated with glucose tolerance tests, insulin tolerance tests, human C-peptide measurements, and vascular imaging. Liraglutide treatment improved metabolic and functional readouts, demonstrating feasibility of the ACE platform for preclinical drug efficacy testing. The system also enabled long-term monitoring of graft morphology, vascularization, and functional stability, supporting its suitability for extended efficacy and safety studies.
Outcomes: The primary milestone (validation of an in vivo sensor-organoid imaging platform for drug testing) was achieved, delivering a functional prototype suitable for further industrial development.
Main technical achievements: A robust humanized diabetic mouse model was established by combining human islet transplantation, murine β-cell ablation, and HFHSD-induced metabolic stress. Sensor islet organoids were successfully generated from dissociated mouse GCaMP3 islets and re-aggregated into organoids suitable for ACE transplantation. A longitudinal in vivo imaging pipeline enabling repeated confocal measurements of β-cell intracellular Ca2+ dynamics in the same grafts over several months was developed. The biosensor system detected functional changes in Ca2+ oscillations in response to metabolic stress and treatment with the GLP-1 analogue liraglutide, with imaging results integrated with glucose tolerance tests, insulin tolerance tests, human C-peptide measurements, and vascular imaging. Liraglutide treatment improved metabolic and functional readouts, demonstrating feasibility of the ACE platform for preclinical drug efficacy testing. The system also enabled long-term monitoring of graft morphology, vascularization, and functional stability, supporting its suitability for extended efficacy and safety studies.
Outcomes: The primary milestone (validation of an in vivo sensor-organoid imaging platform for drug testing) was achieved, delivering a functional prototype suitable for further industrial development.
Advancement beyond the state of the art: SILORGS establishes a first-of-its-kind translational sensor-organoid platform for real-time, longitudinal monitoring of human β-cell function in vivo. Unlike conventional preclinical approaches, SILORGS combines continuous in vivo Ca2+ imaging with assessment of graft morphology, vascularization, and whole-body metabolic measurements. This integrated approach increases precision, reduces animal numbers, and strengthens translational relevance, enabling more accurate preclinical validation of anti-diabetic compounds.
Potential impact: SILORGS accelerates drug discovery by providing mechanism-based functional validation of lead compounds at the β-cell level, supports evaluation of regenerative and replacement therapies, improves translational predictability with humanized models, reduces animal use via longitudinal monitoring, and offers extensibility to other cell types and disease areas, creating new scientific and commercial opportunities.
Key needs for uptake: Expanded validation in larger cohorts, optimization of human sensor-organoid production, strong intellectual property management, industrial partnerships for contract-based screening, access to follow-on funding, and alignment with regulatory and standardization frameworks will ensure scalable commercial deployment.
Overview of results: The project achieved in vivo validation of a humanized diabetic mouse platform with GCaMP-based sensor islet organoids. Longitudinal imaging captured β-cell Ca2+ dynamics, graft morphology, and vascularization. The system detected functional decline under diabetic conditions and improvement with GLP-1 analogue treatment, confirming feasibility for preclinical assessment of efficacy and safety of anti-diabetic drugs.
Potential impact: SILORGS accelerates drug discovery by providing mechanism-based functional validation of lead compounds at the β-cell level, supports evaluation of regenerative and replacement therapies, improves translational predictability with humanized models, reduces animal use via longitudinal monitoring, and offers extensibility to other cell types and disease areas, creating new scientific and commercial opportunities.
Key needs for uptake: Expanded validation in larger cohorts, optimization of human sensor-organoid production, strong intellectual property management, industrial partnerships for contract-based screening, access to follow-on funding, and alignment with regulatory and standardization frameworks will ensure scalable commercial deployment.
Overview of results: The project achieved in vivo validation of a humanized diabetic mouse platform with GCaMP-based sensor islet organoids. Longitudinal imaging captured β-cell Ca2+ dynamics, graft morphology, and vascularization. The system detected functional decline under diabetic conditions and improvement with GLP-1 analogue treatment, confirming feasibility for preclinical assessment of efficacy and safety of anti-diabetic drugs.