Cationic Nanoliposomes Meet mRNA: Efficient Delivery of Modified mRNA Using Hemocompatible and Stable Vectors for Therapeutic Applications  Tatjana Michel,

Slides:



Advertisements
Similar presentations
Products > HUVEC Transfection Reagent (Human Umbilical Vein Endothelial Cells) Altogen Biosystems offers the HUVEC Transfection Reagent among a host of.
Advertisements

Products > bEnd-3 Transfection Reagent (Brain Endothelioma)
Altogen labs Leading Developer and Manufacturer of In Vivo and DNA Transfection Kits, Transfection Reagents and Electroporation Delivery Products Products.
Products > DI-TNC1 Transfection Reagent (Rat Brain Astrocytes)
Volume 50, Issue 3, Pages (March 2009)
Molecular Therapy - Nucleic Acids
Products > CHO Transfection Reagent (Chinese Hamster Ovary Cells)
Products > HCS-2 Transfection Reagent (Chondrosarcoma Cells)
Products > AsPC-1 Transfection Reagent (Pancreatic Beta Cells)
Products > 293 Transfection Reagent (Emb Kidney, CRL-1573)
Molecular Therapy - Methods & Clinical Development
Products > Keratinocyte Transfection Reagent (Keratinocytes)
Fluid Phase Endocytosis Contributes to Transfection of DNA by PEI-25
Products > HCAEC Transfection Reagent (Coronary Artery Endothelial)
In situ Delivery of Tumor Antigen– and Adjuvant-Loaded Liposomes Boosts Antigen- Specific T-Cell Responses by Human Dermal Dendritic Cells  Martine A.
Molecular Therapy - Nucleic Acids
Products > A375 Transfection Reagent (Melanoma Cells, CRL-1619)
Volume 21, Issue 10, Pages (October 2013)
Volume 44, Issue 3, Pages (March 2016)
Volume 26, Issue 6, Pages (June 2018)
Volume 21, Issue 2, Pages (February 2013)
Volume 14, Issue 4, Pages (October 2006)
Molecular Therapy - Nucleic Acids
The Efficacy of Cardiac Anti-miR-208a Therapy Is Stress Dependent
Insertion of the Type-I IFN Decoy Receptor B18R in a miRNA-Tagged Semliki Forest Virus Improves Oncolytic Capacity but Results in Neurotoxicity  Tina.
Blockade of Inflammation and Apoptosis Pathways by siRNA Prolongs Cold Preservation Time and Protects Donor Hearts in a Porcine Model  Jia Wei, Shiyou.
Human DMBT1-Derived Cell-Penetrating Peptides for Intracellular siRNA Delivery  Martina Tuttolomondo, Cinzia Casella, Pernille Lund Hansen, Ester Polo,
Molecular Therapy - Nucleic Acids
Development of Peptide-targeted Lipoplexes to CXCR4-expressing Rat Glioma Cells and Rat Proliferating Endothelial Cells  Wouter HP Driessen, Nobutaka.
Isoliquiritigenin Inhibits IL-1β-Induced Production of Matrix Metalloproteinase in Articular Chondrocytes  Lei Zhang, Shiyun Ma, Hang Su, Jiaxiang Cheng 
Volume 21, Issue 10, Pages (October 2013)
Induction of T-Cell Responses against Cutaneous T-Cell Lymphomas Ex Vivo by Autologous Dendritic Cells Transfected with Amplified Tumor mRNA  Xiao Ni,
Ribosomal Protein S3 Gene Silencing Protects Against Cigarette Smoke-Induced Acute Lung Injury  Jinrui Dong, Wupeng Liao, Hong Yong Peh, W.S. Daniel Tan,
Volume 13, Issue 12, Pages (December 2015)
Molecular Therapy - Nucleic Acids
Alexander Falkenhagen, Sadhna Joshi  Molecular Therapy - Nucleic Acids 
SUMO-2 Orchestrates Chromatin Modifiers in Response to DNA Damage
Codon-Optimized P1A-Encoding DNA Vaccine: Toward a Therapeutic Vaccination against P815 Mastocytoma  Alessandra Lopes, Kevin Vanvarenberg, Véronique Préat,
Thermoresponsive Bacteriophage Nanocarrier as a Gene Delivery Vector Targeted to the Gastrointestinal Tract  Katawut Namdee, Mattaka Khongkow, Suwimon.
Molecular Therapy - Nucleic Acids
Induced-Decay of Glycine Decarboxylase Transcripts as an Anticancer Therapeutic Strategy for Non-Small-Cell Lung Carcinoma  Jing Lin, Jia Hui Jane Lee,
VSV-G-Enveloped Vesicles for Traceless Delivery of CRISPR-Cas9
Molecular Therapy - Nucleic Acids
Volume 33, Issue 1, Pages (January 2009)
Volume 25, Issue 7, Pages (July 2017)
Induced-Decay of Glycine Decarboxylase Transcripts as an Anticancer Therapeutic Strategy for Non-Small-Cell Lung Carcinoma  Jing Lin, Jia Hui Jane Lee,
SUMO-2 Orchestrates Chromatin Modifiers in Response to DNA Damage
Products > Transfection Reagent for Chromaffin Cells
Molecular Therapy - Nucleic Acids
Modular Three-component Delivery System Facilitates HLA Class I Antigen Presentation and CD8+ T-cell Activation Against Tumors  Benjamin J Umlauf, Chin-Ying.
RNA Polymerase II Activity of Type 3 Pol III Promoters
Products > CLBPEC Transfection Reagent (Neuroblastoma Cells)
Volume 26, Issue 1, Pages (January 2018)
Gold Nanoparticles for BCR-ABL1 Gene Silencing: Improving Tyrosine Kinase Inhibitor Efficacy in Chronic Myeloid Leukemia  Raquel Vinhas, Alexandra R.
Molecular Therapy - Nucleic Acids
Molecular Therapy - Nucleic Acids
Molecular Therapy - Nucleic Acids
Volume 29, Issue 3, Pages e3 (March 2019)
Genetic Immunization With In Vivo Dendritic Cell-targeting Liposomal DNA Vaccine Carrier Induces Long-lasting Antitumor Immune Response  Arup Garu, Gopikrishna.
MRNA Vaccine with Antigen-Specific Checkpoint Blockade Induces an Enhanced Immune Response against Established Melanoma  Yuhua Wang, Lu Zhang, Zhenghong.
Zinc-Finger Nucleases Induced by HIV-1 Tat Excise HIV-1 from the Host Genome in Infected and Latently Infected Cells  Haiyan Ji, Panpan Lu, Baochi Liu,
Artificial Zinc-Finger Transcription Factor of A20 Suppresses Restenosis in Sprague Dawley Rats after Carotid Injury via the PPARα Pathway  Zhaoyou Meng,
Molecular Therapy - Nucleic Acids
Yoshinori Aragane, Akira Maeda, Chang-Yi Cui, Tadashi Tezuka 
Gemcitabine-Incorporated G-Quadruplex Aptamer for Targeted Drug Delivery into Pancreas Cancer  Jun Young Park, Ye Lim Cho, Ju Ri Chae, Sung Hwan Moon,
Systemic Administration of Platelets Incorporating Inactivated Sendai Virus Eradicates Melanoma in Mice  Tomoyuki Nishikawa, Li Yu Tung, Yasufumi Kaneda 
Volume 25, Issue 6, Pages (June 2017)
Molecular Therapy - Nucleic Acids
Volume 20, Issue 3, Pages (March 2012)
Aminoglycoside Enhances the Delivery of Antisense Morpholino Oligonucleotides In Vitro and in mdx Mice  Mingxing Wang, Bo Wu, Sapana N. Shah, Peijuan.
Presentation transcript:

Cationic Nanoliposomes Meet mRNA: Efficient Delivery of Modified mRNA Using Hemocompatible and Stable Vectors for Therapeutic Applications  Tatjana Michel, Daniel Luft, Meike-Kristin Abraham, Sabrina Reinhardt, Martha L. Salinas Medina, Julia Kurz, Martin Schaller, Meltem Avci-Adali, Christian Schlensak, Karlheinz Peter, Hans Peter Wendel, Xiaowei Wang, Stefanie Krajewski  Molecular Therapy - Nucleic Acids  Volume 8, Pages 459-468 (September 2017) DOI: 10.1016/j.omtn.2017.07.013 Copyright © 2017 The Author(s) Terms and Conditions

Figure 1 Preparation and Characterization of Liposomes (A) Schematic overview of the liposome production process including lipid preparation and mixing, drying under O2-free condition, and evaporation overnight in vacuum. The developed lipid cake was rehydrated with water followed by sonification, extrusion, and homogenization to get unilamellar lipid vesicles. (B) Visualization of the liposomes using TEM after negative staining. (C) Analysis of the mRNA encapsulation efficacy of liposomes compared to Lipofectamine 2000. Data are shown as mean ± SEM (n = 5). Molecular Therapy - Nucleic Acids 2017 8, 459-468DOI: (10.1016/j.omtn.2017.07.013) Copyright © 2017 The Author(s) Terms and Conditions

Figure 2 Transfection of Cells with Liposomes for Induced Expression of Different Proteins (A) The schematic overview shows the process of mRNA incorporation in liposomes and subsequent transfection and mRNA translation. During transfection, the lipoplexes are endocytosed by the cells of interest. Inside the cell, the lipid layer is degraded and mRNA is released. In the cytosol, the mRNA is translated by ribosomes and the protein is subsequently released intra- or extracellularly. (B) The presence of Cy3-labeled EGFP-encoding mRNA after liposome uptake and expression of EGFP protein in cells 24 hr posttransfection. Molecular Therapy - Nucleic Acids 2017 8, 459-468DOI: (10.1016/j.omtn.2017.07.013) Copyright © 2017 The Author(s) Terms and Conditions

Figure 3 Transfection Efficacy of Liposomes Containing EGFP mRNA at Different Time Points (A–D) Flow cytometric analysis showing % eGFP-expressing cells (A) and fluorescence intensity (B) of A549 cells transfected with different amount of liposomes containing 1 μg eGFP-encoding mRNA 48 h posttransfection. In addition, % eGFP-expressing cells (C) and fluorescence intensity (D) were measured 120 h after transfection. Data are shown as mean ± SEM (n = 5). *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. Molecular Therapy - Nucleic Acids 2017 8, 459-468DOI: (10.1016/j.omtn.2017.07.013) Copyright © 2017 The Author(s) Terms and Conditions

Figure 4 Investigations of Immunogenic and Toxic Effects of Liposomes on Cells after Different Time Points (A) Viability measurement of cells was performed after transfection for 72 or 120 hr with liposomes and lipoplexes. (B and C) Relative normalized gene expressions of IFN-α (B) or IFN-β (C) in cells after long-time transfection from 24 to 120 hr with liposomes were achieved using real-time qPCR. Data are shown as mean ± SEM (n = 5). Molecular Therapy - Nucleic Acids 2017 8, 459-468DOI: (10.1016/j.omtn.2017.07.013) Copyright © 2017 The Author(s) Terms and Conditions

Figure 5 Analysis of AAT Expression after Transfection with Liposomes or Lipofectamine 2000 Containing AAT-Encoding mRNA Concentration of AAT protein in cell supernatants after transfection with AAT mRNA encapsulated in liposomes or Lipofectamine 2000 for 24 or 72 hr. Data are shown as mean ± SEM (n = 5). **p < 0.01. Molecular Therapy - Nucleic Acids 2017 8, 459-468DOI: (10.1016/j.omtn.2017.07.013) Copyright © 2017 The Author(s) Terms and Conditions

Figure 6 Optimal Storage Conditions for Liposomes Containing mRNA EGFP expression was investigated in cells after transfection with fresh and stored lipoplexes. Fresh lipoplexes as well as lipoplexes, which were stored at RT for 80 days, have a significantly higher EGFP expression than the control samples. Lipoplexes, which were stored at 4°C, induce nearly the same level of EGFP expression as fresh-generated lipoplexes. Data are shown as mean ± SEM (n = 3). *p < 0.05; ****p < 0.001. Molecular Therapy - Nucleic Acids 2017 8, 459-468DOI: (10.1016/j.omtn.2017.07.013) Copyright © 2017 The Author(s) Terms and Conditions

Figure 7 Hemocompatibility of Liposomes and Lipoplexes Incubated in Human Whole Blood Markers for the activation of blood coagulation (A; TAT) and the complement system (B; SC5b-9), as well as for neutrophils (C; PMN elastase) and platelets (D; β-thromboglobulin), were quantified in untreated human whole blood or blood treated with liposomes and lipoplexes after incubation in a dynamic flow model using ELISA. Data are shown as mean ± SEM (n = 5). Molecular Therapy - Nucleic Acids 2017 8, 459-468DOI: (10.1016/j.omtn.2017.07.013) Copyright © 2017 The Author(s) Terms and Conditions

Figure 8 Blood Cell Counts in Human Whole Blood before and after Incubation with Liposomes Numbers of red blood cells (A), white blood cells (B), and platelets (C) per microliter of blood were measured. Additionally, hemoglobin (D) and hematocrit (E) values were quantified. Data are shown as mean ± SEM (n = 5). Molecular Therapy - Nucleic Acids 2017 8, 459-468DOI: (10.1016/j.omtn.2017.07.013) Copyright © 2017 The Author(s) Terms and Conditions