It is our pleasure to participate to the ECM2022 congress the 22th, 23th and 24th of June in Copenhagen with two contributions (abstracts at the end):
- Our project leader in iPSc and Metabolic diseases Meryl Roudaut will present his works about the development of innovative hiPSC-based model including an innovative 3D modified hyaluronic acid hydroscaffold for phenotypic screening, on Friday 24 June, 13:15-14:45 in Blomstersalen during poster session (P074)
- Our project leader in pharmacology Veronique De Conto will present her works about the importance of microenvironment in cerebral in vitro models for phenotypic screening, on Saturday 25 June, 12:45-13:45 in Blomstersalen.
The aim of this congress is to focus on development of new drugs targeting the extracellular matrix and
improve our understanding of changes to the extracellular matrix across chronic diseases. Indeed, alterations to the extracellular matrix are a common denominator in most chronic diseases. It is not merely a consequence of the diseases but an active driver.
We look forward to meeting you, feel free to book a meeting now!
The importance of microenvironment in cerebral in vitro models for phenotypic screening
Veronique De Conto Saturday 25 June, 12:45-13:45 in Blomstersalen.
About 90% of drug candidates fail in clinical trials, for efficacy- and toxicity-related reasons, which often involve the Central Nervous System (CNS). This high failure rate highlights a lack of relevance in experimental models used upstream, including human in vitro models. Indeed, they do not take into account the complexity of the CNS, in which neurons are organized in 3 dimensions (3D) and interact with their microenvironment, composed of cells and extracellular matrix (ECM). The objectives of this work were i) to study the influence of the microenvironment on neuronal cells in cerebral in vitro models by automatized cellular imaging, and ii) to develop more relevant cerebral in vitro models for phenotypic screening, to assess neurotoxic or therapeutic effects, in the frame of Parkinson’s Disease (PD).
In this aim, we developed a hyaluronic acid-based hydroscaffold in 96-well plate, which allows to cultivated cells in 3D in a ECM-like microenvironment. First, the sensitivity of Luhmes cells, a dopaminergic neuronal cell line, to PD inducers has been studied in this model. Then, we performed a co-culture of Luhmes cells and primary human astrocytes in this matrix, to form a complex model including both the glial and the matricial microenvironments. All the results were acquired by confocal fluorescent microscopy and automatized quantitative image analysis.
In the hydroscaffold, the neuronal cells were organized in clusters and formed neurites. They displayed a lower sensitivity in 3D compared to cells cultured in 2 dimensions (2D). This difference was explained by two phenomena: a partial retention of toxic molecules in the matrix, and a difference in neuronal protein expression compared to cells cultured in 2D. In the co-culture, we observed spheroids containing both neurons and astrocytes. The analysis of matrix component expression in the co-culture, in healthy and pathological conditions, is ongoing.
This work highlighted that the microenvironment of neurons can modify the neuronal response in vitro, and should thus be considered carefully in academic research and as early as possible in the drug discovery industrial process.
The development of innovative hiPSC-based model including an innovative 3D modified hyaluronic acid hydroscaffold for phenotypic screening
Meryl Roudaut – Friday 24 June, 13:15-14:45 in Blomstersalen during poster session (P074)
We previously showed that human pluripotent stem cells (hiPSCs) provide a suitable model to study
metabolic diseases upon hepatocyte-like cell (HLC) differentiation. With a non-invasive approach, hiPSCs can be generated from urine samples of patients and HLCs have been used to model cholesterol metabolism regulation, by the study of LDLR- and PCSK9-mediated autosomal dominant hypercholesterolemia (ADH) as well as PCSK9-mediated familial hypobetalipoproteinemia (FHBL). This model provides promising advantages with a direct link to the patient and with an unlimited source of HLCs. But like all models, there are limitations, mainly by the neonatal characteristic of HLCs lead to difficulties for pharmacological investigations.
Therefore, to overcome these burdens, we chose to 1. Differentiate hiPSCs into HLCs in an innovative
3D hyaluronic acid-based hydroscaffold, BIOMIMESYS® produces by HCS Pharma to enhance their maturation. 2. Adapt our 3D differentiation process to a 96-well format to make it compatible for drug screening. 3. Characterization of the 3D HLCs model by metabolism tests and compare to primary human hepatocyte (PHH).
We gathered 3’ SRP data all along the differentiation process and RNAseq has been performed by comparing 2D and 3D differentiation conditions to characterize hiPSCs differentiation into liver organoids. We observed an enhanced expression of most hepatic genes and genes expressed by non-parenchymal cells such as stellate cells. Immunofluorescence data confirmed the co-localization of albumin-positive
hepatocytes, desmin-positive stellate cells and LYVE1-positive endothelial cells in liver organoids. Finally, at a functional level, several CYP activities including CYP3A4 were detected at the basal level and successfully induced. Liver organoids responded to pharmacological treatments as shown by their ability to accumulate lipids upon amiodarone treatment or uptake LDL-bodipy upon statin treatment.
Altogether, our development gave rise to functional liver organoids generated with a unique and common procedure, in a process of automating for future high throughput screening.