Single-cell morphological tracking of liver cell states to identify small-molecule modulators of liver differentiation

Alternative therapeutic strategies are urgently required to treat liver disease, which is responsible for 2 million deaths anually. By combining Cell Painting, a morphological profiling assay that captures diverse cellular states, with the bi-potent HepaRG liver progenitor cell line, we have developed a high-throughput, single-cell technique, to track liver cell fate and map small-molecule induced changes using a morphological atlas of bi-lineage liver cell differentiation. To our knowledge this is the first-time single-cell trajectory inference has been applied to image-based Cell Painting data and leveraged for drug screening. The overarching goal of this new method is to aid research into understanding liver cell regeneration mechanisms and facilitate the development of cell-based and small-molecule therapies. Using this approach, we have identified a class of small-molecule SRC family kinase inhibitors that promote differentiation of HepaRG single-cells towards the hepatocyte-like lineage and promotes differentiation of primary human hepatic progenitor cells towards a hepatocyte-like phenotype in vitro. ### Competing Interest Statement A.U-B and N.O.C disclose the following patents, EP3298015B1, JP6684831B2, US10294227B2, CN107849050B, and CA3021550A1 pertaining to the discovery of eCF506/NXP900 that have been licensed to Nuvectis Pharma Inc. A.U-B and N.O.C hold grants from Nuvectis Pharma to study eCF06/NXP900 outside of the submitted work. D.C.H is a co-founder, director and shareholder in Stimuliver ApS and Stemnovate Limited.