date: 2024-06-06T09:38:02Z pdf:unmappedUnicodeCharsPerPage: 0 pdf:PDFVersion: 1.7 pdf:docinfo:title: Optimizing mRNA-Loaded Lipid Nanoparticles as a Potential Tool for Protein-Replacement Therapy xmp:CreatorTool: LaTeX with hyperref Keywords: lipid nanoparticles; mRNA delivery; cytokines; PBMCs; in vivo biodistribution; metabolicdiseases access_permission:modify_annotations: true access_permission:can_print_degraded: true subject: Lipid nanoparticles (LNPs) tailored for mRNA delivery were optimized to serve as a platform for treating metabolic diseases. Four distinct lipid mixes (LMs) were formulated by modifying various components: LM1 (ALC-0315/DSPC/Cholesterol/ALC-0159), LM2 (ALC-0315/DOPE/Cholesterol/ALC-0159), LM3 (ALC-0315/DSPC/Cholesterol/DMG-PEG2k), and LM4 (DLin-MC3-DMA/DSPC/Cholesterol/ALC-0159). LNPs exhibited stability and homogeneity with a mean size of 75 to 90 nm, confirmed by cryo-TEM and SAXS studies. High mRNA encapsulation (95?100%) was achieved. LNPs effectively delivered EGFP-encoding mRNA to HepG2 and DC2.4 cell lines. LNPs induced cytokine secretion from human peripheral blood mononuclear cells (PBMCs), revealing that LM1, LM2, and LM4 induced 1.5- to 4-fold increases in IL-8, TNF-, and MCP-1 levels, while LM3 showed minimal changes. Reporter mRNA expression was observed in LNP-treated PBMCs. Hemotoxicity studies confirmed formulation biocompatibility with values below 2%. In vivo biodistribution in mice post intramuscular injection showed significant mRNA expression, mainly in the liver. The modification of LNP components influenced reactogenicity, inflammatory response, and mRNA expression, offering a promising platform for selecting less reactogenic carriers suitable for repetitive dosing in metabolic disease treatment. dc:creator: Rocío Gambaro, Ignacio Rivero Berti, María José Limeres, Cristián Huck-Iriart, Malin Svensson, Silvia Fraude, Leah Pretsch, Shutian Si, Ingo Lieberwirth, Stephan Gehring, Maximiliano Cacicedo and Germán Abel Islan dcterms:created: 2024-06-06T09:12:22Z Last-Modified: 2024-06-06T09:38:02Z dcterms:modified: 2024-06-06T09:38:02Z dc:format: application/pdf; version=1.7 title: Optimizing mRNA-Loaded Lipid Nanoparticles as a Potential Tool for Protein-Replacement Therapy Last-Save-Date: 2024-06-06T09:38:02Z pdf:docinfo:creator_tool: LaTeX with hyperref access_permission:fill_in_form: true pdf:docinfo:keywords: lipid nanoparticles; mRNA delivery; cytokines; PBMCs; in vivo biodistribution; metabolicdiseases pdf:docinfo:modified: 2024-06-06T09:38:02Z meta:save-date: 2024-06-06T09:38:02Z pdf:encrypted: false dc:title: Optimizing mRNA-Loaded Lipid Nanoparticles as a Potential Tool for Protein-Replacement Therapy modified: 2024-06-06T09:38:02Z cp:subject: Lipid nanoparticles (LNPs) tailored for mRNA delivery were optimized to serve as a platform for treating metabolic diseases. Four distinct lipid mixes (LMs) were formulated by modifying various components: LM1 (ALC-0315/DSPC/Cholesterol/ALC-0159), LM2 (ALC-0315/DOPE/Cholesterol/ALC-0159), LM3 (ALC-0315/DSPC/Cholesterol/DMG-PEG2k), and LM4 (DLin-MC3-DMA/DSPC/Cholesterol/ALC-0159). LNPs exhibited stability and homogeneity with a mean size of 75 to 90 nm, confirmed by cryo-TEM and SAXS studies. High mRNA encapsulation (95?100%) was achieved. LNPs effectively delivered EGFP-encoding mRNA to HepG2 and DC2.4 cell lines. LNPs induced cytokine secretion from human peripheral blood mononuclear cells (PBMCs), revealing that LM1, LM2, and LM4 induced 1.5- to 4-fold increases in IL-8, TNF-, and MCP-1 levels, while LM3 showed minimal changes. Reporter mRNA expression was observed in LNP-treated PBMCs. Hemotoxicity studies confirmed formulation biocompatibility with values below 2%. In vivo biodistribution in mice post intramuscular injection showed significant mRNA expression, mainly in the liver. The modification of LNP components influenced reactogenicity, inflammatory response, and mRNA expression, offering a promising platform for selecting less reactogenic carriers suitable for repetitive dosing in metabolic disease treatment. pdf:docinfo:subject: Lipid nanoparticles (LNPs) tailored for mRNA delivery were optimized to serve as a platform for treating metabolic diseases. Four distinct lipid mixes (LMs) were formulated by modifying various components: LM1 (ALC-0315/DSPC/Cholesterol/ALC-0159), LM2 (ALC-0315/DOPE/Cholesterol/ALC-0159), LM3 (ALC-0315/DSPC/Cholesterol/DMG-PEG2k), and LM4 (DLin-MC3-DMA/DSPC/Cholesterol/ALC-0159). LNPs exhibited stability and homogeneity with a mean size of 75 to 90 nm, confirmed by cryo-TEM and SAXS studies. High mRNA encapsulation (95?100%) was achieved. LNPs effectively delivered EGFP-encoding mRNA to HepG2 and DC2.4 cell lines. LNPs induced cytokine secretion from human peripheral blood mononuclear cells (PBMCs), revealing that LM1, LM2, and LM4 induced 1.5- to 4-fold increases in IL-8, TNF-, and MCP-1 levels, while LM3 showed minimal changes. Reporter mRNA expression was observed in LNP-treated PBMCs. Hemotoxicity studies confirmed formulation biocompatibility with values below 2%. In vivo biodistribution in mice post intramuscular injection showed significant mRNA expression, mainly in the liver. The modification of LNP components influenced reactogenicity, inflammatory response, and mRNA expression, offering a promising platform for selecting less reactogenic carriers suitable for repetitive dosing in metabolic disease treatment. Content-Type: application/pdf pdf:docinfo:creator: Rocío Gambaro, Ignacio Rivero Berti, María José Limeres, Cristián Huck-Iriart, Malin Svensson, Silvia Fraude, Leah Pretsch, Shutian Si, Ingo Lieberwirth, Stephan Gehring, Maximiliano Cacicedo and Germán Abel Islan X-Parsed-By: org.apache.tika.parser.DefaultParser creator: Rocío Gambaro, Ignacio Rivero Berti, María José Limeres, Cristián Huck-Iriart, Malin Svensson, Silvia Fraude, Leah Pretsch, Shutian Si, Ingo Lieberwirth, Stephan Gehring, Maximiliano Cacicedo and Germán Abel Islan meta:author: Rocío Gambaro, Ignacio Rivero Berti, María José Limeres, Cristián Huck-Iriart, Malin Svensson, Silvia Fraude, Leah Pretsch, Shutian Si, Ingo Lieberwirth, Stephan Gehring, Maximiliano Cacicedo and Germán Abel Islan dc:subject: lipid nanoparticles; mRNA delivery; cytokines; PBMCs; in vivo biodistribution; metabolicdiseases meta:creation-date: 2024-06-06T09:12:22Z created: 2024-06-06T09:12:22Z access_permission:extract_for_accessibility: true access_permission:assemble_document: true xmpTPg:NPages: 23 Creation-Date: 2024-06-06T09:12:22Z pdf:charsPerPage: 3971 access_permission:extract_content: true access_permission:can_print: true meta:keyword: lipid nanoparticles; mRNA delivery; cytokines; PBMCs; in vivo biodistribution; metabolicdiseases Author: Rocío Gambaro, Ignacio Rivero Berti, María José Limeres, Cristián Huck-Iriart, Malin Svensson, Silvia Fraude, Leah Pretsch, Shutian Si, Ingo Lieberwirth, Stephan Gehring, Maximiliano Cacicedo and Germán Abel Islan producer: pdfTeX-1.40.25 access_permission:can_modify: true pdf:docinfo:producer: pdfTeX-1.40.25 pdf:docinfo:created: 2024-06-06T09:12:22Z