Rational Design of a siRNA Delivery System: ALOX5 and Cancer Stem Cells as Therapeutic Targets

The search for an ideal gene delivery system is a long and laborious process in which several factors from the first idea to final formulation, including main challenges, peaks and troughs, should be deeply taken into consideration to ensure adequate biological safety and in vivo efficacy endpoints. Arachidonate 5-lipoxygenase (ALOX5), a crucial player related with cancer development and in particular with cancer stem cells malignancy. In this work we describe the process behind the development of a small interfering RNA (siRNA) delivery system to inhibit ALOX5 in cancer stem cells (CSC), as a model target gene. We started by screening chitosan polyplexes, among different types of chitosan in different complexation conditions. Due to the low silencing efficacy obtained, chitosan polyplexes were combined with Pluronic®-based polymeric micelles with recognized advantages regarding gene transfection. We tested different types of polymeric particles to improve the biological efficacy of chitosan polyplexes. Nevertheless, limited transfection efficiency was still detected. The well-established polyethyleneimine (PEI) cationic polymer was used in substitution of chitosan, in combination with polymeric micelles, originating PEI-siRNA-Pluronic® systems. The presence of Pluronic® F127 in the formulation showed to be of utmost importance because not only the silencing activity of the polyplexes was improved, but also PEI-associated toxicity was clearly reduced. This, allowed to increase the amount of PEI inside the system and its overall efficacy. Indeed, different types of PEI, N/P ratios and preparation methods were tested until an optimal formulation composed by PEI 10k branched-based polyplexes at an N/P ratio of 50 combined with micelles of Pluronic® F127 was selected. This combined micelle presented adequate technological properties, safety profile, and biological efficacy, resulting in high ALOX5 gene silencing and strong reduction of invasion and transformation capabilities of a stem cell subpopulation isolated from MDA-MB-231 triple negative breast cancer cells.

Download Full-text

AKT2 siRNA delivery with amphiphilic-based polymeric micelles show efficacy against cancer stem cells

Drug Delivery ◽

10.1080/10717544.2018.1461276 ◽

2018 ◽

Vol 25 (1) ◽

pp. 961-972 ◽

Cited By ~ 15

Author(s):

Diana Rafael ◽

Petra Gener ◽

Fernanda Andrade ◽

Joaquin Seras-Franzoso ◽

Sara Montero ◽

...

Keyword(s):

Stem Cells ◽

Cancer Stem Cells ◽

Polymeric Micelles ◽

Sirna Delivery

Download Full-text

Higher Order Architecture of Designer Peptides Forms Bioinspired 10 nm siRNA Delivery System

Scientific Reports ◽

10.1038/s41598-019-53462-1 ◽

2019 ◽

Vol 9 (1) ◽

Author(s):

Alicia Gamboa ◽

Selina F. Urfano ◽

Katrina Hernandez ◽

Deborah A. Fraser ◽

Luladey Ayalew ◽

...

Keyword(s):

Nucleic Acid ◽

Delivery System ◽

Rational Design ◽

Sirna Delivery ◽

Higher Order ◽

Collagen Peptide ◽

Complex Forms ◽

Quaternary Structures ◽

Quaternary Complex ◽

Sirna Delivery System

AbstractThe higher-order architecture observed in biological systems, like viruses, is very effective in nucleic acid transport. The replications of this system has been attempted with both synthetic and naturally occurring polymers with mixed results. Here we describe a peptide/siRNA quaternary complex that functions as an siRNA delivery system. The rational design of a peptide assembly is inspired by the viral capsids, but not derived from them. We selected the collagen peptide (COL) to provide the structural stability and the folding framework, and hybridize it with the cell penetrating peptide (CPP) that allows for effective penetration of biological barriers. The peptide/siRNA quaternary complex forms stoichiometric, 10 nm nanoparticles, that show fast cellular uptake (<30 min), effective siRNA release, and gene silencing. The complex provides capsid-like protection for siRNA against nucleases without being immunostimulatory, or cytotoxic. Our data suggests that delivery vehicles based on synthetic quaternary structures that exhibit higher-order architecture may be effective in improving delivery and release of nucleic acid cargo.

Download Full-text

Targeted Delivery of siRNA Therapeutics using Ligand Mediated Biodegradable Polymeric Nanocarriers

Current Pharmaceutical Design ◽

10.2174/1381612824666180702113345 ◽

2018 ◽

Vol 24 (16) ◽

pp. 1788-1800 ◽

Cited By ~ 1

Author(s):

Kye-Soo Cho ◽

Seo-Jin Hong ◽

Min-Hye Ahn ◽

Sukdeb Pal ◽

Pill-Hoon Choung ◽

...

Keyword(s):

Cancer Treatment ◽

Delivery System ◽

Targeted Delivery ◽

Sirna Delivery ◽

Public Health Issue ◽

Magic Bullet ◽

Bodily Fluids ◽

Low Cytotoxicity ◽

Genetic And Environmental Factors ◽

Sirna Delivery System

Background: Cancer poses a major public health issue, is linked with high mortality rates across the world, and shows a strong interplay between genetic and environmental factors. To date, common therapeutics, including chemotherapy, immunotherapy, and radiotherapy, have made significant contributions to cancer treatment, although diverse obstacles for achieving the permanent “magic bullet” cure have remained. Recently, various anticancer therapeutic agents designed to overcome the limitations of these conventional cancer treatments have received considerable attention. One of these promising and novel agents is the siRNA delivery system; however, poor cellular uptake and altered siRNA stability in physiological environments have limited its use in clinical trials. Therefore, developing the ideal siRNA delivery system with low cytotoxicity, improved siRNA stability in the body’s circulation, and prevention of its rapid clearance from bodily fluids, is rapidly emerging as an innovative therapeutic strategy to combat cancer. Moreover, active targeting using ligand moieties which bind to over-expressed receptors on the surface of cancer cells would enhance the therapeutic efficiency of siRNA. Conclusion: In this review, we provide 1) an overview of the non-viral carrier associated with siRNA delivery for cancer treatment, and 2) a description of the five major cancer-targeting ligands.

Download Full-text

Novel Cell type-specific aptamer-siRNA delivery system for HIV-1 therapy

Nature Precedings ◽

10.1038/npre.2007.1299.1 ◽

2007 ◽

Cited By ~ 1

Author(s):

Jiehua Zhou ◽

Haitang Li ◽

Shirley Li ◽

John Zaia ◽

John Rossi

Keyword(s):

Delivery System ◽

Sirna Delivery ◽

Cell Type ◽

Cell Type Specific ◽

Hiv 1 ◽

Sirna Delivery System

Download Full-text

Biocompatible, chimeric peptide-condensed supramolecular nanoparticles for tumor cell-specific siRNA delivery and gene silencing

Chemical Communications ◽

10.1039/c4cc01061b ◽

2014 ◽

Vol 50 (58) ◽

pp. 7806-7809 ◽

Cited By ~ 27

Author(s):

Hangxiang Wang ◽

Wei Chen ◽

Haiyang Xie ◽

Xuyong Wei ◽

Shengyong Yin ◽

...

Keyword(s):

Tumor Cell ◽

Gene Silencing ◽

Delivery System ◽

Self Assembly ◽

Sirna Delivery ◽

Single Step ◽

Chimeric Peptide ◽

Supramolecular Nanoparticles ◽

Sirna Delivery System

A practical and tumor cell-specific siRNA delivery system was developedviasingle-step self-assembly of an arginine-rich chimeric peptide with siRNA.

Download Full-text

Mesenchymal Stem Cells and Cancer Stem Cells: An Overview of Tumor-Mesenchymal Stem Cell Interaction for therapeutic interventions

Current Drug Targets ◽

10.2174/1389450122666210824142247 ◽

2021 ◽

Vol 22 ◽

Author(s):

Soheila Montazersaheb ◽

Ezzatollah Fathi ◽

Ayoub Mamandi ◽

Raheleh Farahzadi ◽

Hamid Reza Heidari

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Tumor Microenvironment ◽

Cancer Stem Cells ◽

Tumor Cells ◽

Cell Types ◽

Cell Interaction ◽

Therapeutic Interventions ◽

Tumorigenic Potential ◽

Different Types

: Tumors are made up of different types of cancer cells that contribute to tumor heterogeneity. Among these cells, cancer stem cells (CSCs) have a significant role in the onset of cancer and development. Like other stem cells, CSCs are characterized by the capacity for differentiation and self-renewal. A specific population of CSCs is constituted by mesenchymal stem cells (MSCs) that differentiate into mesoderm-specific cells. The pro-or anti-tumorigenic potential of MSCs on the proliferation and development of tumor cells has been reported as contradictory results. Also, tumor progression is specified by the corresponding tumor cells like the tumor microenvironment. The tumor microenvironment consists of a network of reciprocal cell types such as endothelial cells, immune cells, MSCs, and fibroblasts as well as growth factors, chemokines, and cytokines. In this review, recent findings related to the tumor microenvironment and associated cell populations, homing of MSCs to tumor sites, and interaction of MSCs with tumor cells will be discussed.

Download Full-text

Nasal Administration of Cationic Nanoemulsions as CD73-siRNA Delivery System for Glioblastoma Treatment: a New Therapeutical Approach

Molecular Neurobiology ◽

10.1007/s12035-019-01730-6 ◽

2019 ◽

Vol 57 (2) ◽

pp. 635-649 ◽

Cited By ~ 11

Author(s):

J. H. Azambuja ◽

R. S. Schuh ◽

L. R. Michels ◽

N. E. Gelsleichter ◽

L. R. Beckenkamp ◽

...

Keyword(s):

Delivery System ◽

Sirna Delivery ◽

Nasal Administration ◽

Sirna Delivery System

Download Full-text

The Role of Breast Cancer Stem Cells as a Prognostic Marker and a Target to Improve the Efficacy of Breast Cancer Therapy

Cancers ◽

10.3390/cancers11071021 ◽

2019 ◽

Vol 11 (7) ◽

pp. 1021 ◽

Cited By ~ 12

Author(s):

Maria Giovanna Scioli ◽

Gabriele Storti ◽

Federico D’Amico ◽

Pietro Gentile ◽

Giulia Fabbri ◽

...

Keyword(s):

Breast Cancer ◽

Stem Cells ◽

Cancer Stem Cells ◽

Tumor Cells ◽

The Body ◽

Cell Subpopulation ◽

Phenotypic Characterization ◽

Quiescent State ◽

Breast Cancer Stem Cells ◽

Mesenchymal Transition

Breast cancer is the most common form of tumor in women and the leading cause of cancer-related mortality. Even though the major cellular burden in breast cancer is constituted by the so-called bulk tumor cells, another cell subpopulation named cancer stem cells (CSCs) has been identified. The latter have stem features, a self-renewal capacity, and the ability to regenerate the bulk tumor cells. CSCs have been described in several cancer types but breast cancer stem cells (BCSCs) were among the first to be identified and characterized. Therefore, many efforts have been put into the phenotypic characterization of BCSCs and the study of their potential as prognostic indicators and therapeutic targets. Many dysregulated pathways in BCSCs are involved in the epithelial–mesenchymal transition (EMT) and are found up-regulated in circulating tumor cells (CTCs), another important cancer cell subpopulation, that shed into the vasculature and disseminate along the body to give metastases. Conventional therapies fail at eliminating BCSCs because of their quiescent state that gives them therapy resistance. Based on this evidence, preclinical studies and clinical trials have tried to establish novel therapeutic regimens aiming to eradicate BCSCs. Markers useful for BCSC identification could also be possible therapeutic methods against BCSCs. New approaches in drug delivery combined with gene targeting, immunomodulatory, and cell-based therapies could be promising tools for developing effective CSC-targeted drugs against breast cancer.

Download Full-text