Nanoparticle Delivery of Cancer-Activated DNA Constructs for the Diagnosis of Liver Tumors

Early detection of liver cancer has the potential to substantially improve survival rates. Yet, diagnosis of hepatocellular carcinoma (HCC) is hampered by poor sensitivity of traditional imaging techniques - such as ultrasound, CT, and MRI - which often miss small or atypical tumors, particularly in cirrhotic livers with altered anatomy. As a result, more than a third of early-stage HCCs are missed. Earli is addressing these limitations by developing a platform technology that utilizes LNP-delivered recombinant DNA constructs containing cancer-activated synthetic promoters driving the selective expression of proteins such as well defined epitopes for the imaging of tumors or cytokines that have therapeutic potential to treat the tumor. Although non-viral systems like lipid nanoparticles (LNPs) have proven effective for RNA delivery, DNA delivery to tumor cells remains challenging. Earli-derived LNP formulation FRM237 was used to encapsulate a recombinant DNA comprised of a non-specific CAG promoter to drive the expression of a firefly luciferase reporter and was evaluated by ex vivo bioluminescence imaging (BLI) in a subcutaneous Hep3B (liver adenocarcinoma) xenograft model. The FRM237 formulation produced high levels of expression and achieved BLI levels that were 5-fold higher than the same DNA formulated in a multi-component LNP used in a commercial siRNA product. More importantly, engineered LNP enabled 88-fold greater tumor-to-liver BLI ratio, revealing liver tumor tropism. Pharmacokinetic studies show a large increase in circulation LNP-delivered DNA vector copies were consistently 10-fold higher in serum as compared to the competitor formulation. Effective extrahepatic delivery and expression to HCC was further verified in an orthotopic liver xenograft model using a recombinant DNA driven by a synthetic promoter known to be highly active in liver cancer cells. Administered via IV injection to mice with Hep3B tumors, this product produced 96-fold higher expression from those malignant tissues as compared to naïve liver tissues. Likewise, immunohistochemistry (IHC) analyses revealed selective expression of the protein throughout the orthotopic Hep3B tumors with little or no corresponding staining in normal hepatocytes. Finally, intravenously dosed lead formulations were tolerated as evident by modest but transient body weight loss and clinical observations. Overall, our data collectively points to successful delivery of DNA to hepatic tumors via engineered LNP formulations following iv administration. When combined with cancer-activated expression of PET reporter genes and further validation in preclinical mouse models and toxicology studies, this represents a promising diagnostic imaging platform for early detection of liver cancer. Nadege Morisot, Blaine McCarthy, Sushil Lathwal, Dariusz Wodziak, Suthara Ramachandran, Anderson Lee, Dang Dang, Blair Cain, Xiaobin Wu, Robby Chandra, Badriprasad Ananthanarayanan, David Suhy, Christine Peterson. Nanoparticle delivery of cancer-activated DNA constructs for the diagnosis of liver tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 60.