A novel auto-fluorescent porphyrin-lipid nanoparticle strategy for CTNNB1 gene silencing in hepatocellular carcinoma

Jeffrey C To; Alyssa Apilan; Elisa Pasini; Yulin Mo; Juan Chen; Anita Bakrania; Jiachuan Bu; Anni Pan; Oyedele Adeyi; Arndt Vogel; Gang Zheng; Mamatha Bhat

doi:10.3389/fonc.2026.1779803

A novel auto-fluorescent porphyrin-lipid nanoparticle strategy for CTNNB1 gene silencing in hepatocellular carcinoma

Front Oncol. 2026 May 8:16:1779803. doi: 10.3389/fonc.2026.1779803. eCollection 2026.

Authors

Jeffrey C To^{1

2}, Alyssa Apilan^{1

2}, Elisa Pasini^{1

2}, Yulin Mo^{2

3}, Juan Chen², Anita Bakrania^{1

2}, Jiachuan Bu², Anni Pan², Oyedele Adeyi^{4

5}, Arndt Vogel^{2

3

6}, Gang Zheng^{1

2

7}, Mamatha Bhat^{1

2

3

8

9}

Affiliations

¹ Ajmera Transplant Centre, University Health Network, Toronto, ON, Canada.
² Princess Margaret Cancer Centre, University Health Network, Toronto, ON, Canada.
³ Division of Gastroenterology and Hepatology, Toronto General Hospital, Health Network, Toronto, ON, Canada.
⁴ Department of Pathology and University of Minnesota Medical Center, University of Minnesota, Minneapolis, MN, United States.
⁵ Anatomic Pathology, The University of Alabama at Birmingham, Alabama, AL, United States.
⁶ Department of Gastroenterology, Hepatology, Infectious Diseases and Endocrinology, Hannover Medical School, Hannover, Germany.
⁷ Department of Medical Biophysics, University of Toronto, Toronto, ON, Canada.
⁸ Department of Medicine, University of Toronto., Toronto, ON, Canada.
⁹ Division of Gastroenterology & Hepatology, University of Toronto, Toronto, ON, Canada.

Abstract

Background: Hepatocellular carcinoma (HCC) remains a high-fatality cancer with limited effective therapies. CTNNB1 mutations, frequently observed in HCC, are associated with poor prognosis and immune evasion. CTNNB1 has long been considered an undruggable target due to its structural characteristics and extensive protein interactions. Porphyrin lipid nanoparticles (porphyrin-LNPs) are capable of targeting liver tumor cells, and their inherent autofluorescence allows evaluation of nanoparticle biodistribution in the liver. Our goal was to formulate a porphyrin-LNP encapsulating CTNNB1-targeting siRNA, as a novel strategy to target β-catenin-driven HCCs.

Methods: We developed porphyrin-LNPs for systemic delivery of CTNNB1-targeting siRNA. Porphyrin-LNPs were synthesized via microfluidic rapid mixing and characterized by cryo-TEM and dynamic light scattering. Their delivery efficacy was validated in HCC cell lines (Hep3B, HepG2), and their therapeutic potential was evaluated in a murine model of CTNNB1/KRAS-driven HCC.

Results: Porphyrin-LNPs showed high encapsulation efficiency (97%) and effective siRNA delivery in vitro. Treatment with porphyrin-LNP-si-CTNNB1 resulted in approximately 90% downregulation of CTNNB1 expression (p < 0.0001 in Hep3B at 50nM; p < 0.0001 in HepG2 at 10nM) and significantly reduced clonogenic survival in both Hep3B (50% reduction at 50nM, p < 0.0001) and HepG2 (75% reduction at 10nM, p < 0.001) cell lines. In vivo, porphyrin-LNP-si-CTNNB1 significantly reduced tumor burden by approximately 67% (p < 0.0001), liver-to-body weight ratio by 50% (p < 0.0001), histological tumor grade, and β-catenin expression in CTNNB1/KRAS-driven HCC mice by approximately 58% (p < 0.0001).

Conclusions: This study demonstrated that porphyrin-LNPs can effectively deliver siRNA to silence CTNNB1, an oncogene that has so far been undruggable in HCC. Future studies should explore biodistribution, immune modulation, and combination strategies to enhance clinical translatability of CTNNB1-targeted RNA interference in HCC.

Keywords: CTNNB1; RNA interference; hepatocellular carcinoma (HCC); lipid nanoparticles (LNPs); porphyrin-lipid nanoparticles; targeted gene therapy; β-catenin.