scholarly journals Enhanced target cell specificity and uptake of lipid nanoparticles using RNA aptamers and peptides

2021 ◽  
Vol 17 ◽  
pp. 891-907
Author(s):  
Roslyn M Ray ◽  
Anders Højgaard Hansen ◽  
Maria Taskova ◽  
Bernhard Jandl ◽  
Jonas Hansen ◽  
...  
Keyword(s):  
Target Cell ◽  
Cationic Lipid ◽  
Cost Effective ◽  
Cell Types ◽  
Lipid Nanoparticles ◽  
Cell Penetrating Peptides ◽  
Rna Aptamers ◽  
Target Tissue ◽  
Cell Specificity ◽  
Rnai Therapeutics

Lipid nanoparticles (LNPs) constitute a facile and scalable approach for delivery of payloads to human cells. LNPs are relatively immunologically inert and can be produced in a cost effective and scalable manner. However, targeting and delivery of LNPs across the blood–brain barrier (BBB) has proven challenging. In an effort to target LNPs composed of an ionizable cationic lipid (DLin-MC3-DMA), cholesterol, the phospholipid 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), and 1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol-2000 (DMG-PEG 2000) to particular cell types, as well as to generate LNPs that can cross the BBB, we developed and assessed two approaches. The first was centered on the BBB-penetrating trans-activator of transcription (Tat) peptide or the peptide T7, and the other on RNA aptamers targeted to glycoprotein gp160 from human immunodeficiency virus (HIV) or C-C chemokine receptor type 5 (CCR5), a HIV-1 coreceptor. We report herein a CCR5-selective RNA aptamer that acts to facilitate entry through a simplified BBB model and that drives the uptake of LNPs into CCR5-expressing cells, while the gp160 aptamer did not. We further observed that the addition of cell-penetrating peptides, Tat and T7, did not increase BBB penetration above the aptamer-loaded LNPs alone. Moreover, we found that these targeted LNPs exhibit low immunogenic and low toxic profiles and that targeted LNPs can traverse the BBB to potentially deliver drugs into the target tissue. This approach highlights the usefulness of aptamer-loaded LNPs to increase target cell specificity and potentially deliverability of central-nervous-system-active RNAi therapeutics across the BBB.

scholarly journals Enhanced cell target specificity and uptake of lipid nanoparticles using RNA aptamers and peptides

2021 ◽  
Author(s):  
Roslyn M Ray ◽  
Anders Højgaard Hansen ◽  
Maria Taskova ◽  
Bernhard Jandl ◽  
Jonas Hansen ◽  
...  
Keyword(s):  
Cost Effective ◽  
Cell Types ◽  
Lipid Nanoparticles ◽  
Cell Penetrating Peptides ◽  
Rna Aptamers ◽  
Target Tissue ◽  
Rna Aptamer ◽  
Cell Specificity ◽  
Cell Target ◽  
Hiv 1

Lipid nanoparticles (LNPs) constitute a facile and scalable approach for delivery of payloads to human cells. LNPs are relatively immunologically inert and can be produced in a cost effective and scalable manner. However, targeting and delivery of LNPs across the blood brain barrier (BBB) has proven challenging. In an effort to target LNPs to particular cell types as well as generating LNPs that can cross the BBB, we developed and assessed two approaches whereby BBB penetrating peptides Tat or T7, and RNA aptamers targeted to gp160 from HIV or CCR5, a HIV-1 co-receptor, were incorporated into LNPs. We report here that a CCR5 selective RNA aptamer acts to facilitate entry through a simplified BBB model and to drive the uptake of LNPs into CCR5 expressing cells, while the gp160 aptamer did not. We further observed that the addition of cell penetrating peptides, Tat, did not increase BBB penetration above the aptamer loaded LNPs alone. Moreover, we find that these targeted LNPs exhibit low immunogenic and low toxic profiles and that targeted LNPs can traverse the BBB to potentially deliver drugs into the target tissue. This approach highlights the usefulness of aptamer loaded LNPs to increase target cell specificity and potentially, deliverability across the BBB.

scholarly journals Advances in the Formulation and Assembly of Non-Cationic Lipid Nanoparticles for the Medical Application of Gene Therapeutics

Nanomaterials ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 825
Author(s):  
Richard K. Fisher ◽  
Phillip C. West ◽  
Samuel I. Mattern-Schain ◽  
Michael D. Best ◽  
Stacy S. Kirkpatrick ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Nucleic Acid ◽  
Cationic Lipid ◽  
Medical Application ◽  
Lipid Nanoparticles ◽  
Cell Penetrating Peptides ◽  
Cationic Lipids ◽  
Injection Technique ◽  
Ethanol Injection ◽  
Assembly Technique

Lipid nanoparticles have become increasingly popular delivery platforms in the field of gene therapy, but bench-to-bedside success has been limited. Many liposomal gene vectors are comprised of synthetic cationic lipids, which are associated with lipid-induced cytotoxicity and immunogenicity. Natural, non-cationic PEGylated liposomes (PLPs) demonstrate favorable biocompatibility profiles but are not considered viable gene delivery vehicles due to inefficient nucleic acid loading and reduced cellular uptake. PLPs can be modified with cell-penetrating peptides (CPPs) to enhance the intracellular delivery of liposomal cargo but encapsulate leakage upon CPP-PLP assembly is problematic. Here, we aimed to identify parameters that overcome these performance barriers by incorporating nucleic acid condensers during CPP-PLP assembly and screening variable ethanol injection parameters for optimization. CPP-PLPs were formed with R8-amphiphiles via pre-insertion, post-insertion and post-conjugation techniques and liposomes were characterized for size, surface charge, homogeneity, siRNA encapsulation efficiency and retention and cell associative properties. Herein we demonstrate that pre-insertion of stearylated R8 into PLPs is an efficient method to produce non-cationic CPP-PLPs and we provide additional assembly parameter specifications for a modified ethanol injection technique that is optimized for siRNA encapsulation/retention and enhanced cell association. This assembly technique could provide improved clinical translation of liposomal based gene therapy applications.

Nanotheranostic agents for neurodegenerative diseases

2020 ◽  
Vol 4 (6) ◽  
pp. 645-675
Author(s):  
Parasuraman Padmanabhan ◽  
Mathangi Palanivel ◽  
Ajay Kumar ◽  
Domokos Máthé ◽  
George K. Radda ◽  
...  
Keyword(s):  
Drug Delivery ◽  
Neurodegenerative Diseases ◽  
Cell Types ◽  
Specific Cell ◽  
Ageing Population ◽  
Target Tissue ◽  
Therapeutic Approaches ◽  
Surface Receptors ◽  
Theranostic Agents ◽  
Preclinical Animal Models

Neurodegenerative diseases (NDDs), including Alzheimer's disease (AD) and Parkinson's disease (PD), affect the ageing population worldwide and while severely impairing the quality of life of millions, they also cause a massive economic burden to countries with progressively ageing populations. Parallel with the search for biomarkers for early detection and prediction, the pursuit for therapeutic approaches has become growingly intensive in recent years. Various prospective therapeutic approaches have been explored with an emphasis on early prevention and protection, including, but not limited to, gene therapy, stem cell therapy, immunotherapy and radiotherapy. Many pharmacological interventions have proved to be promising novel avenues, but successful applications are often hampered by the poor delivery of the therapeutics across the blood-brain-barrier (BBB). To overcome this challenge, nanoparticle (NP)-mediated drug delivery has been considered as a promising option, as NP-based drug delivery systems can be functionalized to target specific cell surface receptors and to achieve controlled and long-term release of therapeutics to the target tissue. The usefulness of NPs for loading and delivering of drugs has been extensively studied in the context of NDDs, and their biological efficacy has been demonstrated in numerous preclinical animal models. Efforts have also been made towards the development of NPs which can be used for targeting the BBB and various cell types in the brain. The main focus of this review is to briefly discuss the advantages of functionalized NPs as promising theranostic agents for the diagnosis and therapy of NDDs. We also summarize the results of diverse studies that specifically investigated the usage of different NPs for the treatment of NDDs, with a specific emphasis on AD and PD, and the associated pathophysiological changes. Finally, we offer perspectives on the existing challenges of using NPs as theranostic agents and possible futuristic approaches to improve them.

scholarly journals Cationic and Biocompatible Polymer/Lipid Nanoparticles as Immunoadjuvants

Pharmaceutics ◽  
2021 ◽  
Vol 13 (11) ◽  
pp. 1859
Author(s):  
Yunys Pérez-Betancourt ◽  
Péricles Marques Araujo ◽  
Bianca de Carvalho Lins Fernandes Távora ◽  
Daniele Rodrigues Pereira ◽  
Eliana Lima Faquim-Mauro ◽  
...  
Keyword(s):  
Immune Responses ◽  
Methyl Methacrylate ◽  
Cationic Lipid ◽  
Lipid Nanoparticles ◽  
Hybrid Nanoparticles ◽  
Ammonium Bromide ◽  
The Novel ◽  
Biocompatible Polymer ◽  
Dimethyl Ammonium ◽  
Oppositely Charged

Nanostructures have been of paramount importance for developing immunoadjuvants. They must be cationic and non-cytotoxic, easily assembling with usually oppositely charged antigens such as proteins, haptens or nucleic acids for use in vaccines. We obtained optimal hybrid nanoparticles (NPs) from the biocompatible polymer poly(methyl methacrylate) (PMMA) and the cationic lipid dioctadecyl dimethyl ammonium bromide (DODAB) by emulsion polymerization of methyl methacrylate (MMA) in the presence of DODAB. NPs adsorbed ovalbumin (OVA) as a model antigen and we determined their adjuvant properties. Interestingly, they elicited high double immune responses of the cellular and humoral types overcoming the poor biocompatibility of DODAB-based adjuvants of the bilayer type. The results suggested that the novel adjuvant would be possibly of use in a variety of vaccines.

scholarly journals Two Modes of Toxicity of Lipid Nanoparticles Containing a pH-Sensitive Cationic Lipid on Human A375 and A375-SM Melanoma Cell Lines

BPB Reports ◽  
2019 ◽  
Vol 2 (4) ◽  
pp. 48-55
Author(s):  
Ahmed Y. AlBaloul ◽  
Yusuke Sato ◽  
Nako Maishi ◽  
Kyoko Hida ◽  
Hideyoshi Harashima
Keyword(s):  
Cell Lines ◽  
Melanoma Cell ◽  
Cationic Lipid ◽  
Lipid Nanoparticles ◽  
Ph Sensitive ◽  
Melanoma Cell Lines

scholarly journals Transparent PDMS Bioreactors for the Fabrication and Analysis of Multi-Layer Pre-vascularized Hydrogels Under Continuous Perfusion

2020 ◽  
Vol 8 ◽  
Author(s):  
Juan Liu ◽  
Huaiyuan Zheng ◽  
Xinyi Dai ◽  
Patrina S. P. Poh ◽  
Hans-Günther Machens ◽  
...  
Keyword(s):  
Functional Organization ◽  
Umbilical Vein ◽  
Cost Effective ◽  
Practical Method ◽  
Cell Types ◽  
Collagen Gels ◽  
Vascular Walls ◽  
Novel Strategy ◽  
Umbilical Vein Endothelial Cells ◽  
Continuous Perfusion

Tissue engineering in combination with stem cell technology has the potential to revolutionize human healthcare. It aims at the generation of artificial tissues that can mimic the original with complex functions for medical applications. However, even the best current designs are limited in size, if the transport of nutrients and oxygen to the cells and the removal of cellular metabolites waste is mainly dependent on passive diffusion. Incorporation of functional biomimetic vasculature within tissue engineered constructs can overcome this shortcoming. Here, we developed a novel strategy using 3D printing and injection molding technology to customize multilayer hydrogel constructs with pre-vascularized structures in transparent Polydimethysiloxane (PDMS) bioreactors. These bioreactors can be directly connected to continuous perfusion systems without complicated construct assembling. Mimicking natural layer-structures of vascular walls, multilayer vessel constructs were fabricated with cell-laden fibrin and collagen gels, respectively. The multilayer design allows functional organization of multiple cell types, i.e., mesenchymal stem cells (MSCs) in outer layer, human umbilical vein endothelial cells (HUVECs) the inner layer and smooth muscle cells in between MSCs and HUVECs layers. Multiplex layers with different cell types showed clear boundaries and growth along the hydrogel layers. This work demonstrates a rapid, cost-effective, and practical method to fabricate customized 3D-multilayer vascular models. It allows precise design of parameters like length, thickness, diameter of lumens and the whole vessel constructs resembling the natural tissue in detail without the need of sophisticated skills or equipment. The ready-to-use bioreactor with hydrogel constructs could be used for biomedical applications including pre-vascularization for transplantable engineered tissue or studies of vascular biology.

scholarly journals Delivery of self-amplifying mRNA vaccines by cationic lipid nanoparticles: The impact of cationic lipid selection

2020 ◽  
Vol 325 ◽  
pp. 370-379 ◽  
Author(s):  
Gustavo Lou ◽  
Giulia Anderluzzi ◽  
Signe Tandrup Schmidt ◽  
Stuart Woods ◽  
Simona Gallorini ◽  
...  
Keyword(s):  
Cationic Lipid ◽  
Lipid Nanoparticles ◽  
The Impact

scholarly journals Internalization mechanisms of cell-penetrating peptides

2020 ◽  
Vol 11 ◽  
pp. 101-123 ◽  
Author(s):  
Ivana Ruseska ◽  
Andreas Zimmer
Keyword(s):  
Cellular Uptake ◽  
Cell Types ◽  
Cell Penetrating Peptides ◽  
Uptake Mechanism ◽  
Black And White ◽  
Cell Penetrating ◽  
Basic Peptides ◽  
Different Cell Types ◽  
Immense Potential

In today’s modern era of medicine, macromolecular compounds such as proteins, peptides and nucleic acids are dethroning small molecules as leading therapeutics. Given their immense potential, they are highly sought after. However, their application is limited mostly due to their poor in vivo stability, limited cellular uptake and insufficient target specificity. Cell-penetrating peptides (CPPs) represent a major breakthrough for the transport of macromolecules. They have been shown to successfully deliver proteins, peptides, siRNAs and pDNA in different cell types. In general, CPPs are basic peptides with a positive charge at physiological pH. They are able to translocate membranes and gain entry to the cell interior. Nevertheless, the mechanism they use to enter cells still remains an unsolved piece of the puzzle. Endocytosis and direct penetration have been suggested as the two major mechanisms used for internalization, however, it is not all black and white in the nanoworld. Studies have shown that several CPPs are able to induce and shift between different uptake mechanisms depending on their concentration, cargo or the cell line used. This review will focus on the major internalization pathways CPPs exploit, their characteristics and regulation, as well as some of the factors that influence the cellular uptake mechanism.

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