Ionizable Lipid Nanoparticle-Mediated TRAIL mRNA Delivery in the Tumor Microenvironment to Inhibit Colon Cancer Progression

Walison Nunes da Silva,1 Pedro Augusto Carvalho Costa,1 Sérgio Ricardo Aluotto Scalzo Júnior,1 Heloísa AS Ferreira,1 Pedro Henrique Dias Moura Prazeres,2 Caroline Leonel Vasconcelos Campos,3 Marco Túllio Rodrigues Alves,1 Natália Jordana Alves da Silva,1 Ana Luiza de Castro Santos,1 Lays Cordeiro Guimarães,1 Maria Eduarda Chen Ferris,1 Ajay Thatte,4 Alex Hamilton,4 Kelly Alves Bicalho,5 Anderson Oliveira Lobo,6 Helton da Costa Santiago,3 Lucíola da Silva Barcelos,1 Maria Marta Figueiredo,7 Mauro Martins Teixeira,3 Vivian Vasconcelos Costa,8 Michael J Mitchell,4 Frédéric Frézard,1 Pedro Pires Goulart Guimaraes1 1Department of Physiology and Biophysics, Federal University of Minas Gerais, Belo Horizonte, MG, Brazil; 2Department of Pathology, Federal University of Minas Gerais, Belo Horizonte, MG, 31270-901, Brazil; 3Department of Biochemistry and Immunology, Federal University of Minas Gerais, Belo Horizonte, MG, 31270-901, Brazil; 4Department of Bioengineering, University of Pennsylvania, Philadelphia, PA, USA; 5Instituto René Rachou, Fundação Oswaldo Cruz, Belo Horizonte, MG, Brazil; 6Department of Materials Engineering, Federal University of Piauí, Teresina, PI, Brazil; 7State University of Minas Gerais, Divinopolis, MG, Brazil; 8Department of Morphology, Federal University of Minas Gerais, Belo Horizonte, MG, BrazilCorrespondence: Pedro Pires Goulart Guimaraes, Department of Physiology and Biophysics, Federal University of Minas Gerais, Belo Horizonte, MG, Brazil, Email [email protected]: Immunotherapy has revolutionized cancer treatment by harnessing the immune system to enhance antitumor responses while minimizing off-target effects. Among the promising cancer-specific therapies, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) has attracted significant attention.Methods: Here, we developed an ionizable lipid nanoparticle (LNP) platform to deliver TRAIL mRNA (LNP-TRAIL) directly to the tumor microenvironment (TME) to induce tumor cell death. Our LNP-TRAIL was formulated via microfluidic mixing and the induction of tumor cell death was assessed in vitro. Next, we investigated the ability of LNP-TRAIL to inhibit colon cancer progression in vivo in combination with a TME normalization approach using Losartan (Los) or angiotensin 1– 7 (Ang(1– 7)) to reduce vascular compression and deposition of extracellular matrix in mice.Results: Our results demonstrated that LNP-TRAIL induced tumor cell death in vitro and effectively inhibited colon cancer progression in vivo, particularly when combined with TME normalization induced by treatment Los or Ang(1– 7). In addition, potent tumor cell death as well as enhanced apoptosis and necrosis was found in the tumor tissue of a group treated with LNP-TRAIL combined with TME normalization.Discussion: Together, our data demonstrate the potential of the LNP to deliver TRAIL mRNA to the TME and to induce tumor cell death, especially when combined with TME normalization. Therefore, these findings provide important insights for the development of novel therapeutic strategies for the immunotherapy of solid tumors.Keywords: immunotherapy, TRAIL, mRNA, lipid nanoparticle, losartan, angiotensin (1– 7