Periodic Reporting for period 1 - halt-RONIN (Discovering chronic inflammation biomarkers that define key stages in the Healthy-to-NASH (non-alcoholic steatohepatitis) transition to inform early prevention and treatment strategies)
Periodo di rendicontazione: 2022-12-01 al 2024-05-31
Metabolic-dysfunction associated liver disease (MASLD, before NAFLD), is a multifactorial chronic inflammatory disease that is prevalent in 1 of 4 individuals with a significant personal, socioeconomic and healthcare burden, especially at the later, more severe inflammatory stage of disease - non alcoholic steatohepatitis (NASH) or MASH. Despite the severe negative impact of the disease on society, MASLD remains difficult to diagnose and treat. The molecular mechanisms underlying the transition from health to fatty liver to MASH remain poorly understood due to the lack of models that faithfully reflect the complexity of human disease. Hence, Halt-RONIN aims to uncover the early triggers of disease initiation and complex mechanistic drivers of disease progression by implementing a systems biology approach with integrative disease modelling resulting in opportunities for the improvement of the existing detection methods, providing a blueprint to inform personalized intervention strategies and drug discovery for MASLD. To achieve this goal, Halt-RONIN will combine experimental data from advanced in vitro and in vivo models with multimodal data from extensive human MAFLD cohorts and biobanks and use in silico machine learning approaches, to discover new biomarkers and molecular targets specific to each stage of the health-to-disease transition. By validating preclinical experimental findings with real-world data, Halt-RONIN will allow for the discovery of novel biomarkers and molecular targets that are specific to the individual patient’s pathology. Consequently, healthcare professionals will gain the tools and knowledge required to diagnose and establish guidelines for the prevention and treatment of inflammation-driven health to disease. As such, in the long-term Halt-RONIN will decrease the number of MAFLD patients who progress into MASH and provide disease-modifying strategies to improve patient outcomes.
Halt-RONIN aim is Discovering chronic inflammation biomarkers that define key stages in the Healthy-to-MASH transition to inform early prevention and treatment strategies.
Halt-RONIN will develop novel models that can be used to advance the understanding of MAFLD-to-MASH progression, the discovery of novel biomarkers and the development of targeted and (cost-) effective drugs against the disease. A better understanding of the disease progression and risk factors will lead to the reduction of healthcare costs and shorter treatment times by enabling a faster drug development, facilitating the development of personalized treatment options and finally limiting the prevalence of MALFD/MASH.
The WP2-WP5 are focused on advancing the understanding of the transition from healthy liver to metabolic dysfunction-associated MASLD and MASH through in vitro and in vivo models.
WP2: Develope advanced human liver models, including 3D spheroids and patient-derived iPSCs, to simulate the MASLD-MASH transition. Activities include creating knockout cell lines, developing liver-on-a-chip systems, and characterizing models with phenotypic and functional markers. SOPs are established to study transitions, identify early biomarkers, and explore molecular mechanisms.
WP3: Focused on in vivo models using humanized FRGN mice and zebrafish to study STARD1's role in MASH progression and drug-induced liver injury (DILI). The team induce MASH through dietary interventions, study oxidative stress via Keap1-Nrf2 regulation, and performe omics analyses to validate biomarkers.
WP4: Integrate data from WP2 and WP3 with public datasets using systems biology to build in silico patient models. These models mapp disease pathways, identify key biomarkers, and use machine learning to uncover therapeutic targets.
WP5: Translate findings into clinical practice by analyzing cohort data to identify blood-based biomarkers for predicting liver disease progression. The team adapte to new MASLD definitions, improving patient stratification and relevance to current medical practice.
Halt-RONIN will develop novel models that can be used to advance the understanding of MAFLD-to-MASH progression, the discovery of novel biomarkers and the development of targeted and (cost-) effective drugs against the disease. A better understanding of the disease progression and risk factors will lead to the reduction of healthcare costs and shorter treatment times by enabling a faster drug development, facilitating the development of personalized treatment options and finally limiting the prevalence of MALFD/MASH.
The WP2-WP5 are focused on advancing the understanding of the transition from healthy liver to metabolic dysfunction-associated MASLD and MASH through in vitro and in vivo models.
WP2: Develope advanced human liver models, including 3D spheroids and patient-derived iPSCs, to simulate the MASLD-MASH transition. Activities include creating knockout cell lines, developing liver-on-a-chip systems, and characterizing models with phenotypic and functional markers. SOPs are established to study transitions, identify early biomarkers, and explore molecular mechanisms.
WP3: Focused on in vivo models using humanized FRGN mice and zebrafish to study STARD1's role in MASH progression and drug-induced liver injury (DILI). The team induce MASH through dietary interventions, study oxidative stress via Keap1-Nrf2 regulation, and performe omics analyses to validate biomarkers.
WP4: Integrate data from WP2 and WP3 with public datasets using systems biology to build in silico patient models. These models mapp disease pathways, identify key biomarkers, and use machine learning to uncover therapeutic targets.
WP5: Translate findings into clinical practice by analyzing cohort data to identify blood-based biomarkers for predicting liver disease progression. The team adapte to new MASLD definitions, improving patient stratification and relevance to current medical practice.
Currently, patients are stratified based on general metabolic risk factors: obesity, type 2 diabetes mellitus for steatohepatitis or fibrosis with confirmation of steatosis by liver function tests, fibrosis scores and elastographic techniques. This makes the differential diagnosis achieved in approximately 2 months.
In Halt-RONIN, patients are stratified based on transition stage-specific biomarkers through minimally invasive blood or urine testing, allowing for personalized interventions and optimal clinical management. Allowing for the definitive personal diagnosis to be obtained in 2 days.
This headline achievement will be underpinned by a number of key technical achievements:
-At present no biomarkers exist that are capable of tracking the transition of an individual’s liver from healthy to the severe inflammation-driven disease, MASH. Halt-RONIN will identify a panel of biomarkers that are capable of rapidly describing liver state with the specificity and sensitivity essential for their use as a clinical diagnostic tool.
- Current mechanistic understanding of inflammation-driven liver pathology is limited given the enormous complexity of the inflammatory response and lack of representative preclinical models. Halt-RONIN will generate multi-dimensional data using advanced data integration approaches with mechanistic systems analysis of healthy-disease transition, contributing to the scientific field of liver pathology and nabling drug target discovery.
- Combining separate liver models do not accurately translate the complexity of the human healthy-to MASLD-to-MASH transition due to model-specific limitations. A comprehensive comparative analysis of existing and new in vivo (i.e. first-of-its-kind humanized zebrafish) and in vitro liver (i.e. human liver-on-a-chip) models allows for identification of molecular events that are conserved across species and models to enable more efficient discovery and drug development using these models.
- Current molecular diagnostics do not consider the role of extracellular vesicles in the aetiology of liver disease, although detectable in liquid biopsies. Halt-RONIN will perform deep phenotyping of extracellular vesicles in preclinical models and clinical samples to identify novel low invasive biomarkers.
- The use of iPSC-derived liver cells for organotypic cultures and liver-on-a-chip in vitro systems is not fully developed and standardized. Halt-RONIN will deliver highly standardized and validated in vitro models with increasing complexity able to faithfully mimic health to disease transition.
In Halt-RONIN, patients are stratified based on transition stage-specific biomarkers through minimally invasive blood or urine testing, allowing for personalized interventions and optimal clinical management. Allowing for the definitive personal diagnosis to be obtained in 2 days.
This headline achievement will be underpinned by a number of key technical achievements:
-At present no biomarkers exist that are capable of tracking the transition of an individual’s liver from healthy to the severe inflammation-driven disease, MASH. Halt-RONIN will identify a panel of biomarkers that are capable of rapidly describing liver state with the specificity and sensitivity essential for their use as a clinical diagnostic tool.
- Current mechanistic understanding of inflammation-driven liver pathology is limited given the enormous complexity of the inflammatory response and lack of representative preclinical models. Halt-RONIN will generate multi-dimensional data using advanced data integration approaches with mechanistic systems analysis of healthy-disease transition, contributing to the scientific field of liver pathology and nabling drug target discovery.
- Combining separate liver models do not accurately translate the complexity of the human healthy-to MASLD-to-MASH transition due to model-specific limitations. A comprehensive comparative analysis of existing and new in vivo (i.e. first-of-its-kind humanized zebrafish) and in vitro liver (i.e. human liver-on-a-chip) models allows for identification of molecular events that are conserved across species and models to enable more efficient discovery and drug development using these models.
- Current molecular diagnostics do not consider the role of extracellular vesicles in the aetiology of liver disease, although detectable in liquid biopsies. Halt-RONIN will perform deep phenotyping of extracellular vesicles in preclinical models and clinical samples to identify novel low invasive biomarkers.
- The use of iPSC-derived liver cells for organotypic cultures and liver-on-a-chip in vitro systems is not fully developed and standardized. Halt-RONIN will deliver highly standardized and validated in vitro models with increasing complexity able to faithfully mimic health to disease transition.