FCF Funded Projects

Development of a human-derived liver progenitor cell line with DNAJB1-PRKACA fusion gene

Status: Completed

Timeframe: 2019 – 2021

Goal: Development of a novel human-derived liver progenitor cell line model of fibrolamellar carcinoma

Principal Investigator: Khashayar Vakili, MD

Study overview: Prior to this effort, the study team’s lab had previously engineered a kidney cell line (HEK-DP) which contains the DNAJB1-PRKACA fusion gene found in FLC tumors. This cell line demonstrated interesting similarities to FLC tumors and served as a proof of concept for the development of additional cell lines. This effort applied apply the same strategy used to engineer the HEK-DP model to engineer normal human liver progenitor cells to express the DNAJB1-PRKACA fusion gene. These liver progenitor cells were to be grown as three-dimensional “organoid” cultures to better replicate in vivo conditions.

If successful, the study aimed to create a novel model to facilitate understanding how the fusion protein reprograms normal liver progenitor cells to become cancerous. Such understanding of the precise mechanisms underlying the formation of FLC could then provide insights to future therapeutic strategies.

Results: Following the successful expression of the fusion protein in HEK cells, the investigators introduced the CRISPR/CAS9 strategy in human-derived cholangiocyte cell cultures to create organoids carrying the fusion protein. However, the efficiency of the expression of the fusion protein was low. The team tried several strategies to increase the expression.

Once the most efficient method is identified, the team plans to conduct a single cell analysis to compare the fusion protein carrying cells with the normal cells in the same organoid.

Implications: This study demonstrated the successful adaptation of a strategy used in non-liver cells to generate specific mutation carrying organoids to liver progenitor cells. While much work remained to develop a usable model, the work was continued. Once these organoids are well characterized, they will be shared with the FLC community.