Cambridge Healthtech Institute’s Ann Nguyen recently interviewed Noushin Dianat, Ph.D., Postdoctoral Fellow, Laboratoire Colloïdes et Matériaux Divisés, ESPCI Paris.
In the leadup to her presentation on “Human Hepatocyte 3D Spheroids: A Breakthrough Technology for High-Throughput Drug-Drug Interaction Screening” at the 3D Cellular Models conference (part of World Preclinical Congress Europe 2017, 16-17 November in Lisbon, Portugal), Dr. Dianat shared some thoughts about her work on:
3D Liver Spheroids And Their Integration With High-Throughput Screening
Dr. Dianat has an interest in high-throughput technology for fabrication of miniaturized liver 3D spheroids. As she describes, in 2014, “a collaborative research project was launched between Sanofi and ESPCI to establish a new pipeline for high-throughput drug-drug interaction on a 3D liver model. Currently, unlike other critical ADME factors, there is no existing high-throughput screening (HTS) assay for Cytochrome (CYP) P450 family induction screening. Besides, the 2D cultured hepatocytes traditionally used for such tests fail to maintain their metabolic activities after only a few days of culture.”
Given those issues, “The goal of this project was to develop a whole workflow for high-throughput DDI screening at gene level on a miniaturized physiologically relevant 3D liver model.”
Challenges have persisted during the integration of such emerging 3D cellular models HTS. Dr. Dianat observes, “One of the limitations of current 3D spheroid models is the cell quantity/well (generally one spheroid/well) that narrows applicable downstream assays to imaging or highly sensitive biochemical kits capable of detecting low signals coming from around 1000 cells. With this few number of cells, measuring gene expression for example by classic methods such as qPCR is not conceivable.”
How did Dr. Dianat’s team address this hurdle?
“Our solution was to design a miniaturized, customizable and metabolically functional 3D liver model and to combine it with the highly sensitive next generation sequencing technology to measure the CYP induction gene expression.”
She elaborates on approaches intended to overcome such challenges: “Since 2010, an innovative technology for mammalian 3D cell culture named BioPearl has been developed in the Laboratory of Colloids and Divided Materials (LCMD) at ESPCI, where the HepatoPearl project was performed. During this two years of collaborative project, we have developed and characterized the HepatoPearl, a novel liver spheroid model with a lifespan of more than 45 days. This new 3D model displays key hepato-specific features and morphological structures found in the liver. Besides HepatoPearls are so easy to handle and offer the possibility of customizing the spheroid number/well according to the type of assay. They have shown a high and stable activity of CYP3A4, 1A2, 2B6 and 2C9 enzymes. These enzymes were shown to be inducible during six weeks in the HepatoPearls.”
Meanwhile, “In parallel we have developed an automated DDI screening pipeline combining molecular barcoding method with gene targeted RNA sequencing technology. In this way we could increase the number of screened drugs, decrease the cell quantity used per assay thanks to the miniaturization and customizability of HepatoPearls, and obtain more predictive results from this novel liver-mimicking model.”
Noushin DianatNoushin Dianat, Ph.D., Postdoctoral Fellow, Laboratoire Colloïdes et Matériaux Divisés, ESPCI Paris
Having graduated from a Biotherapy master program focalized on cell, gene and tissue therapy, at Université Paris Est, Noushin Dianat started her Ph.D. in Université Paris-Sud in 2010 and worked on in vitro liver metabolic disease modelling with patient-specific iPSCs and their differentiation into hepatocytes. During her thesis, she could also establish for the first time a guided protocol for differentiation of pluripotent stem cells into biliary epithelial cells, cholangiocytes. Afterwards, she was interested in tissue engineering and establishment of a high-throughput technology for fabrication of miniaturized liver 3D spheroids. In 2014, she joined LCMD lab at ESPCI Paris as postdoctoral fellow on a collaborative project with Sanofi. Since then she is working with her team on elaboration of this novel technology named “BioPearl”.
Learn more about Dr. Dianat’s presentation during the 3D Cellular Models conference