Generation of Patient-Specific Autologous Liver and Hepatocellular Carcinoma for Development of Cancer Therapies

Abstract

Introduction: The dire prognosis for patients with advanced Hepatocellular Carcinoma (HCC) is due to paucity of therapeutic options. For optimal patient care, it is critical to develop novel therapies tailored in a patient-specific fashion. This personalized approach requires preclinical models that faithfully recapitulate a patient's cancer and its microenvironment. For accurate evaluation of HCC therapies, we have developed conditions that allow us to grow HCC and non-cancerous hepatocytes in culture in a patient-specific fashion. Hypothesis: Patient-derived proliferating normal hepatocytes and HCC cell lines will retain the genomic landscapes of the patient's liver and tumor, respectively. Methods: Cancer and surrounding non-cancerous tissue from clinical HCC resections were collected. From these tissues, we isolated and expanded cancerous and non-cancerous cells. We developed culture methods to induce proliferation of primary human hepatocytes via generation of small hepatocyte proliferating cells (SHPCs). During cellular expansion, the SHPCs underwent gene expression profiling via RNA-sequencing at passages 0 and 3. Results: We isolated cancerous (Fig.1A) and non-cancerous hepatocytes from HCC resection specimens and cultured these cells to grow HCC and SHPCs. After in vitro expansion, SHPCs maintained expression of hepatocyte specific genes after passaging (Fig.1B). Conclusions: Expansion of human hepatocytes is an innovative technology to generate a proliferating hepatocyte population that maintains its gene expression profile over passages. Our goal is to humanize mouse livers with these non-cancerous hepatocytes followed by transplantation of tumor cells, from the same patient, to recreate HCC and tumor microenvironment to develop personalized therapies.