Crispr/cas9 gene therapy platform for inborn metabolic liver diseases using humanized models and 3D organoids

Overview

This platform addresses inborn metabolic liver diseases, conditions for which organ or cell transplantation remains the only curative treatment despite limitations such as immunosuppression requirements and scarce donor availability. The proposed solution leverages CRISPR/Cas9 gene editing to correct the underlying genetic defects in patient-derived hepatocytes, offering a less invasive therapeutic pathway.

The approach combines two complementary strategies. The first involves ex vivo correction of patient hepatocytes using 3D spheroid structures that overcome challenges associated with cell immortalization and re-differentiation, a known limitation when using induced pluripotent stem cells for liver applications. The second strategy targets direct in vivo correction through injection of CRISPR/Cas9 vectors into humanized mouse models bearing patient-derived liver cells. Together, these strategies aim to establish a reproducible preclinical pipeline for gene therapy in metabolic liver disease.

Technical specifications

Core technologies:

• CRISPR/Cas9 gene editing applied to correct monogenic mutations responsible for inherited metabolic liver disorders
• 3D human liver organoids and spheroids generated from patient-derived cells to closely reproduce pathophysiological conditions and overcome iPS maturation inefficiencies
• Humanized FRGN mice with repopulated human hepatocytes serving as patient-relevant xenograft models for in vivo validation

Validation capabilities:

• Functional assays including Phase 1 and Phase 2 metabolism, ammonia detoxification, and synthetic capacity measurements (albumin, alpha-1-antitrypsin) at both protein and transcriptional levels
• Proof-of-concept correction of metabolic disorders already demonstrated in vitro
• Established protocols for generating 3D liver structures and humanized mice, validated over three years

Technology readiness level

The platform is at a preclinical stage, with proof-of-concept already validated in vitro. The research team has established and optimized the foundational technologies, including 3D spheroid generation from patient cells and humanized mouse models with repopulated patient-derived hepatocytes. The next phase focuses on validating CRISPR/Cas9 correction in more relevant in vitro liver structures and evaluating direct in vivo correction strategies for the most common inherited metabolic liver diseases. The work is positioned to advance toward translational studies, supported by access to biobanked patient materials and established human-relevant disease models.