scholarly journals Evaluation of Cytochalasin B-Induced Membrane Vesicles Fusion Specificity with Target Cells

2018 ◽  
Vol 2018 ◽  
pp. 1-6 ◽  
Author(s):  
Marina Gomzikova ◽  
Sevindzh Kletukhina ◽  
Sirina Kurbangaleeva ◽  
Albert Rizvanov
Keyword(s):  
Targeted Delivery ◽  
Cytochalasin B ◽  
Membrane Vesicles ◽  
Membrane Receptors ◽  
Surface Proteins ◽  
Vector System ◽  
Target Cells ◽  
Significant Preference ◽  
Induced Membrane ◽  
Pc3 Cells

Extracellular vesicles (EV) represent a promising vector system for biomolecules and drug delivery due to their natural origin and participation in intercellular communication. As the quantity of EVs is limited, it was proposed to induce the release of membrane vesicles from the surface of human cells by treatment with cytochalasin B. Cytochalasin B-induced membrane vesicles (CIMVs) were successfully tested as a vector for delivery of dye, nanoparticles, and a chemotherapeutic. However, it remained unclear whether CIMVs possess fusion specificity with target cells and thus might be used for more targeted delivery of therapeutics. To answer this question, CIMVs were obtained from human prostate cancer PC3 cells. The diameter of obtained CIMVs was 962,13 ± 140,6 nm. We found that there is no statistically significant preference in PC3 CIMVs fusion with target cells of the same type. According to our observations, the greatest impact on CIMVs entry into target cells is by the heterophilic interaction of CIMV membrane receptors with the surface proteins of target cells.

scholarly journals Angiogenic Activity of Cytochalasin B-Induced Membrane Vesicles of Human Mesenchymal Stem Cells

Cells ◽  
2019 ◽  
Vol 9 (1) ◽  
pp. 95 ◽  
Author(s):  
Marina O. Gomzikova ◽  
Margarita N. Zhuravleva ◽  
Vyacheslav V. Vorobev ◽  
Ilnur I. Salafutdinov ◽  
Alexander V. Laikov ◽  
...  
Keyword(s):  
Cytochalasin B ◽  
Therapeutic Potential ◽  
Membrane Vesicles ◽  
Membrane Receptors ◽  
Molecular Composition ◽  
Target Cells ◽  
Human Mscs ◽  
Angiogenic Activity ◽  
Induced Membrane

The cytochalasin B-induced membrane vesicles (CIMVs) are suggested to be used as a vehicle for the delivery of therapeutics. However, the angiogenic activity and therapeutic potential of human mesenchymal stem/stromal cells (MSCs) derived CIMVs (CIMVs-MSCs) remains unknown. Objectives: The objectives of this study were to analyze the morphology, size distribution, molecular composition, and angiogenic properties of CIMVs-MSCs. Methods: The morphology of CIMVs-MSC was analyzed by scanning electron microscopy. The proteomic analysis, multiplex analysis, and immunostaining were used to characterize the molecular composition of the CIMVs-MSCs. The transfer of surface proteins from a donor to a recipient cell mediated by CIMVs-MSCs was demonstrated using immunostaining and confocal microscopy. The angiogenic potential of CIMVs-MSCs was evaluated using an in vivo approach of subcutaneous implantation of CIMVs-MSCs in mixture with Matrigel matrix. Results: Human CIMVs-MSCs retain parental MSCs content, such as growth factors, cytokines, and chemokines: EGF, FGF-2, Eotaxin, TGF-α, G-CSF, Flt-3L, GM-CSF, Fractalkine, IFNα2, IFN-γ, GRO, IL-10, MCP-3, IL-12p40, MDC, IL-12p70, IL-15, sCD40L, IL-17A, IL-1RA, IL-1a, IL-9, IL-1b, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IP-10, MCP-1, MIP_1a, MIP-1b, TNF-α, TNF-β, VEGF. CIMVs-MSCs also have the expression of surface receptors similar to those in parental human MSCs (CD90+, CD29+, CD44+, CD73+). Additionally, CIMVs-MSCs could transfer membrane receptors to the surfaces of target cells in vitro. Finally, CIMVs-MSCs can induce angiogenesis in vivo after subcutaneous injection into adult rats. Conclusions: Human CIMVs-MSCs have similar content, immunophenotype, and angiogenic activity to those of the parental MSCs. Therefore, we believe that human CIMVs-MSCs could be used for cell free therapy of degenerative diseases.

scholarly journals Cytochalasin B-induced membrane vesicles convey angiogenic activity of parental cells

Oncotarget ◽  
2017 ◽  
Vol 8 (41) ◽  
pp. 70496-70507 ◽  
Author(s):  
Marina O. Gomzikova ◽  
Margarita N. Zhuravleva ◽  
Regina R. Miftakhova ◽  
Svetlana S. Arkhipova ◽  
Vladimir G. Evtugin ◽  
...  
Keyword(s):  
Cytochalasin B ◽  
Membrane Vesicles ◽  
Angiogenic Activity ◽  
Induced Membrane

scholarly journals Biomimetic Membrane Vesicles: Evaluation of Inflammatory Response In Vivo

Blood ◽  
2020 ◽  
Vol 136 (Supplement 1) ◽  
pp. 1-1
Author(s):  
Olga Vasileva ◽  
Ekaterina E Garanina ◽  
Albert A Rizvanov ◽  
Marina Gomzikova
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Russian Federation ◽  
Cytochalasin B ◽  
Membrane Vesicles ◽  
Host Cells ◽  
Vector System ◽  
Gm Csf ◽  
Biomimetic Membrane ◽  
The Russian Federation

Introduction: Biomimetic membrane vesicles are produced from live cells using cytochalasin B which disrupts the structure of the cytoskeleton and facilitate generation of membrane vesicles under subsequent vortexing. Membrane vesicles are pinched off from the cell surface, surrounded by a cytoplasmic membrane and contain the cytoplasm of parental cell. It is known that mesenchymal stem cells (MSCs) are immunoprivileged. The aim of our study was to determine whether Cytochalasin B-induced membrane vesicles (CIMVs) derived from mesenchymal stem cells retain the immunoprivileged properties of parental cells. Method: All experiments were carried out in compliance with the procedure protocols approved by Kazan Federal University and local ethics committee (protocol #5, date 27.05.2014). To analyze the immunogenicity, murine MSCs (7.5x104 cells) either CIMVs-MSCs (15µg) were injected i.v. in 8 week old mice (Mus musculus, C57Bl/6). CIMVs were used at a concentration equivalent to 7.5×104 MSCs based on total protein concentration. Serum isolation was performed after 2 hours post-administration. Secretion of inflammatory cytokines was evaluated using multiplex analysis BioPlex Pro Mouse 23 Plex kit. (BioRad, USA). Results: We detected all investigated cytokines in serum of control and experimental mice: Eotaxin, G-CSF, GM-CSF, IFN-g, IL-10, IL-13, IL-17A, IL-1a, IL-1b, IL-2, IL-3, IL-4, IL-5, IL-6, IL-9, IL12p40, KC, MCP-1, MIP-1a, MIP-1b, RANTES, TNFa. Allogenic MSCs but not CIMVs increased concentration of Eotaxin, G-CSF, IL-17A and IL-9. The level of GM-CSF, IFN-g, IL-10, IL-1a, IL-1b, IL-2, IL-3, IL-4, IL-5, IL-6, IL12p40, KC, MCP-1, MIP-1a, MIP-1b, RANTES, TNFa in mice serum were not affected by murine MSCs injection. Injection of CIMVs did not induce any statistically significant changes in cytokines level. Conclusion: Elevated levels of Eotaxin, G-CSF, IL-17A and IL-9 cytokines after the i.v.injection of murine MSCs suggest that moderate allergy inflammation was developed after the MSCs allotransplantation. CIMVs injection did not induced increase of cytokines level in mice serum indicating absence of immunogenicity. Taken together our results demonstrate that CIMVs show less/no immunogenicity compared to parental MSCs. We believe that small diameter, better biodistribution and fusion with host cells lead to the non-immunogenicity of CIMVs. Thus, CIMVs are confirmed to be a perspective, new biomimetic vector system. This study was funded by the grant of the President of the Russian Federation for state support of the leading scientific schools of the Russian Federation НШ-2603.2020.4. Kazan Federal University was supported by the Russian Government Program of Competitive Growth. Disclosures No relevant conflicts of interest to declare.

scholarly journals Storage stability and delivery potential of cytochalasin B induced membrane vesicles

2021 ◽  
Vol 30 ◽  
pp. e00616
Author(s):  
Sevindzh K. Kletukhina ◽  
Olga A. Neustroeva ◽  
Sirina V. Kurbangaleeva ◽  
Ilnur I. Salafutdinov ◽  
Alexey M. Rogov ◽  
...  
Keyword(s):  
Cytochalasin B ◽  
Storage Stability ◽  
Membrane Vesicles ◽  
Induced Membrane

scholarly journals Angiogenic activity of cytochalasin B-induced membrane vesicles of human mesenchymal stem cells

2019 ◽  
Author(s):  
M.O. Gomzikova ◽  
M.N. Zhuravleva ◽  
V.V. Vorobev ◽  
I.I. Salafutdinov ◽  
A.V. Laikov ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Cytochalasin B ◽  
Human Mesenchymal Stem Cells ◽  
Membrane Vesicles ◽  
Molecular Composition ◽  
Human Mscs ◽  
Angiogenic Activity ◽  
Induced Membrane

AbstractBackgroundThe cytochalasin B-induced membrane vesicles (CIMVs) are suggested to be used as a vehicle for the delivery of therapeutics. However, the angiogenic activity and therapeutic potential of human mesenchymal stem cells (MSCs) derived CIMVs (CIMVs-MSCs) remains unknown.ObjectivesThe objectives of this study were to analyzed the morphology, size distribution, molecular composition and angiogenic properties of CIMVs-MSCs.MethodsThe morphology of CIMVs-MSC was analyzed by scanning electron microscopy. The proteomic analysis, multiplex analysis and immunostaining were used to characterize the molecular composition of the CIMVs-MSCs. The transfer of surface proteins from a donor to a recipient cell mediated by CIMVs-MSCs was demonstrated using immunostaining and confocal microscopy. The angiogenic potential of CIMVs-MSCs was evaluated using in vivo approach of subcutaneous implantation of CIMVs-MSCs in mixture with Matrigel matrix.ResultsHuman CIMVs-MSCs retain parental MSCs content such as growth factors, cytokines, chemokines: EGF, FGF-2, Eotaxin, TGF-α, G-CSF, Flt-3L, GM-CSF, Fractalkine, IFNα2, IFN-γ, GRO, IL-10, MCP-3, IL-12p40, MDC, IL-12p70, IL-15, sCD40L, IL-17A, IL-1RA, IL-1a, IL-9, IL-1b, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IP-10, MCP-1, MIP_1a, MIP-1b, TNF-α, TNF-β, VEGF. CIMVs-MSCs also have the expression of surface receptors similar to those in parental human MSCs (CD90+, CD29+, CD44+, CD73+). Additionally, CIMVs-MSCs could transfer membrane receptors to the surfaces of target cellsin vitro. Finally, CIMVs-MSCs can induce angiogenesisin vivoafter subcutaneous injection into adult rats.ConclusionsHuman CIMVs-MSCs have similar content, immunophenotype and angiogenic activity to those of the parental MSCs. Therefore, we believe that human CIMVs-MSCs could be used for cell free therapy of degenerative diseases.

Cytochalasin B-induced Membrane Vesicles from Human Mesenchymal Stem Cells Overexpressing TRAIL, PTEN and IFN-β1 Can Kill Carcinoma Cancer Cells

2021 ◽  
pp. 101664
Author(s):  
Daria S. Chulpanova ◽  
Zarema E. Gilazieva ◽  
Elvira R. Akhmetzyanova ◽  
Sevindzh K. Kletukhina ◽  
Albert A. Rizvanov ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Cancer Cells ◽  
Cytochalasin B ◽  
Human Mesenchymal Stem Cells ◽  
Membrane Vesicles ◽  
Induced Membrane

Stepwise Development of Biomimetic Chimeric Peptides for Gene Delivery

2020 ◽  
Vol 27 (8) ◽  
pp. 698-710
Author(s):  
Roya Cheraghi ◽  
Mahboobeh Nazari ◽  
Mohsen Alipour ◽  
Saman Hosseinkhani
Keyword(s):  
Gene Delivery ◽  
Targeted Delivery ◽  
Viral Vectors ◽  
Large Scale ◽  
Transfection Efficiency ◽  
Cationic Lipids ◽  
Target Cells ◽  
Scale Production ◽  
Stepwise Development ◽  
Chimeric Peptides

Gene-based therapy largely relies on the vector type that allows a selective and efficient transfection into the target cells with maximum efficacy and minimal toxicity. Although, genes delivered utilizing modified viruses transfect efficiently and precisely, these vectors can cause severe immunological responses and are potentially carcinogenic. A promising method of overcoming this limitation is the use of non-viral vectors, including cationic lipids, polymers, dendrimers, and peptides, which offer potential routes for compacting DNA for targeted delivery. Although non-viral vectors exhibit reduced transfection efficiency compared to their viral counterpart, their superior biocompatibility, non-immunogenicity and potential for large-scale production make them increasingly attractive for modern therapy. There has been a great deal of interest in the development of biomimetic chimeric peptides. Biomimetic chimeric peptides contain different motifs for gene translocation into the nucleus of the desired cells. They have motifs for gene targeting into the desired cell, condense DNA into nanosize particles, translocate the gene into the nucleus and enhance the release of the particle into the cytoplasm. These carriers were developed in recent years. This review highlights the stepwise development of the biomimetic chimeric peptides currently being used in gene delivery.

scholarly journals Avian sarcoma leukosis virus receptor-envelope system for simultaneous dissection of multiple neural circuits in mammalian brain

2015 ◽  
Vol 112 (22) ◽  
pp. E2947-E2956 ◽  
Author(s):  
Makoto Matsuyama ◽  
Yohei Ohashi ◽  
Tadashi Tsubota ◽  
Masae Yaguchi ◽  
Shigeki Kato ◽  
...  
Keyword(s):  
Gene Transfer ◽  
Gene Delivery ◽  
Cross Reactivity ◽  
Vector System ◽  
Target Cells ◽  
Specific Gene ◽  
Multiple Target ◽  
Neuronal Systems ◽  
Avian Sarcoma

Pathway-specific gene delivery is requisite for understanding complex neuronal systems in which neurons that project to different target regions are locally intermingled. However, conventional genetic tools cannot achieve simultaneous, independent gene delivery into multiple target cells with high efficiency and low cross-reactivity. In this study, we systematically screened all receptor–envelope pairs resulting from the combination of four avian sarcoma leukosis virus (ASLV) envelopes (EnvA, EnvB, EnvC, and EnvE) and five engineered avian-derived receptors (TVA950, TVBS3, TVC, TVBT, and DR-46TVB) in vitro. Four of the 20 pairs exhibited both high infection rates (TVA–EnvA, 99.6%; TVBS3–EnvB, 97.7%; TVC–EnvC, 98.2%; and DR-46TVB–EnvE, 98.8%) and low cross-reactivity (<2.5%). Next, we tested these four receptor–envelope pairs in vivo in a pathway-specific gene-transfer method. Neurons projecting into a limited somatosensory area were labeled with each receptor by retrograde gene transfer. Three of the four pairs exhibited selective transduction into thalamocortical neurons expressing the paired receptor (>98%), with no observed cross-reaction. Finally, by expressing three receptor types in a single animal, we achieved pathway-specific, differential fluorescent labeling of three thalamic neuronal populations, each projecting into different somatosensory areas. Thus, we identified three orthogonal pairs from the list of ASLV subgroups and established a new vector system that provides a simultaneous, independent, and highly specific genetic tool for transferring genes into multiple target cells in vivo. Our approach is broadly applicable to pathway-specific labeling and functional analysis of diverse neuronal systems.

scholarly journals Subunit assembly of hemoglobin: an important determinant of hematologic phenotype

Blood ◽  
1987 ◽  
Vol 69 (1) ◽  
pp. 1-6 ◽  
Author(s):  
HF Bunn
Keyword(s):  
Electrostatic Interactions ◽  
Hemoglobin Concentration ◽  
Cytoskeletal Proteins ◽  
Membrane Receptors ◽  
Target Cells ◽  
Red Cell Membrane ◽  
Alpha Beta ◽  
The Difference ◽  
Polypeptide Chains ◽  
Positively Charged

Hemoglobin's physiologic properties depend on the orderly assembly of its subunits in erythropoietic cells. The biosynthesis of alpha- and beta-globin polypeptide chains is normally balanced. Heme rapidly binds to the globin subunit, either during translation or shortly thereafter. The formation of the alpha beta-dimer is facilitated by electrostatic attraction of a positively charged alpha-subunit to a negatively charged beta-subunit. The alpha beta-dimer dissociates extremely slowly. The difference between the rate of dissociation of alpha beta- and alpha gamma-dimers with increasing pH explains the well-known alkaline resistance of Hb F. Two dimers combine to form the functioning alpha 2 beta 2-tetramer. This model of hemoglobin assembly explains the different levels of positively charged and negatively charged mutant hemoglobins that are encountered in heterozygotes and the effect of alpha-thalassemia and heme deficiency states in modifying the level of the variant hemoglobin as well as Hb A2. Electrostatic interactions also affect the binding of hemoglobin to the cytoplasmic surface of the red cell membrane and may underlie the formation of target cells. Enhanced binding of positively charged variants such as S and C trigger a normally dormant pathway for potassium and water loss. Thus, the positive charge on beta c is responsible for the two major contributors to the pathogenesis of Hb SC disease: increased proportion of Hb S and increased intracellular hemoglobin concentration. It is likely that electrostatic interactions play an important role in the assembly of a number of other multisubunit macromolecules, including membrane receptors, cytoskeletal proteins, and DNA binding proteins.

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