scholarly journals Intercellular mRNA trafficking via membrane nanotubes in mammalian cells

2017 ◽  
Author(s):  
Gal Haimovich ◽  
Christopher M. Ecker ◽  
Margaret C. Dunagin ◽  
Elliot Eggan ◽  
Arjun Raj ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Genetic Information ◽  
Quantitative Data ◽  
Mammalian Cells ◽  
Full Length ◽  
Cell Contact ◽  
Cell Physiology ◽  
Membrane Nanotubes ◽  
Donor And Acceptor ◽  
Direct Cell

AbstractRNAs have been shown to undergo transfer between mammalian cells, though the mechanism behind this phenomenon and its overall importance to cell physiology is not well understood. Numerous publications have suggested that RNAs (microRNAs and incomplete mRNAs) undergo transfer via extracellular vesicles (e.g. exosomes). However, in contrast to a diffusion-based transfer mechanism, we find that full-length mRNAs undergo direct cell-cell transfer via cytoplasmic extensions, called membrane nanotubes (mNTs), which connect donor and acceptor cells. By employing a simple co-culture experimental model and using single-molecule imaging, we provide quantitative data showing that mRNAs are transferred between cells in contact. Examples of mRNAs that undergo transfer include those encoding GFP, mouse β-actin, and human Cyclin D1, BRCA1, MT2A, and HER2. We show that intercellular mRNA transfer occurs in all co-culture models tested (e.g. between primary cells, immortalized cells, and in co-cultures of immortalized human and murine cells). Rapid mRNA transfer is dependent upon actin, but independent of de novo protein synthesis, and is modulated by stress conditions and gene expression levels. Hence, this work supports the hypothesis that full-length mRNAs undergo transfer between cells through a refined structural connection. Importantly, unlike the transfer of miRNA or RNA fragments, this process of communication transfers genetic information that could potentially alter the acceptor cell proteome. This phenomenon may prove important for the proper development and functioning of tissues, as well as host-parasite or symbiotic interactions.SignificanceMessenger RNA (mRNA) molecules convey genetic information within cells, beginning from genes in the nucleus to ribosomes in the cell body, where they are translated into proteins. Here, we show a novel mode of transferring genetic information from one cell to another. Contrary to previous publications suggesting that mRNAs transfer via extracellular vesicles, we provide visual and quantitative data showing that mRNAs transfer via membrane nanotubes and direct cell-to-cell contact. We predict that this process has a major role in regulating local cellular environments with respect to tissue development and maintenance, cellular responses to stress, interactions with parasites, tissue transplants, and the tumor microenvironment.Author contributionsG.H., A.R. and R.H.S. conceived the research and designed the experiments; C.M.E. performed and analyzed the experiments with WM983b+/-GFP, including transwell and exosomes; M.C.D. and E.E. performed and analyzed the WM983b/NIH393 co-culture experiments; G.H. performed and analyzed all other experiments; and G.H., J.E.G, A.R. and R.H.S. wrote the paper.

scholarly journals Intercellular mRNA trafficking via membrane nanotube-like extensions in mammalian cells

2017 ◽  
Vol 114 (46) ◽  
pp. E9873-E9882 ◽  
Author(s):  
Gal Haimovich ◽  
Christopher M. Ecker ◽  
Margaret C. Dunagin ◽  
Elliott Eggan ◽  
Arjun Raj ◽  
...  
Keyword(s):  
Single Molecule ◽  
Mammalian Cells ◽  
De Novo ◽  
Full Length ◽  
Cell Physiology ◽  
Donor And Acceptor ◽  
Immortalized Cells ◽  
Rna Fragments ◽  
Direct Cell ◽  
Structural Connection

RNAs have been shown to undergo transfer between mammalian cells, although the mechanism behind this phenomenon and its overall importance to cell physiology is not well understood. Numerous publications have suggested that RNAs (microRNAs and incomplete mRNAs) undergo transfer via extracellular vesicles (e.g., exosomes). However, in contrast to a diffusion-based transfer mechanism, we find that full-length mRNAs undergo direct cell–cell transfer via cytoplasmic extensions characteristic of membrane nanotubes (mNTs), which connect donor and acceptor cells. By employing a simple coculture experimental model and using single-molecule imaging, we provide quantitative data showing that mRNAs are transferred between cells in contact. Examples of mRNAs that undergo transfer include those encoding GFP, mouse β-actin, and human Cyclin D1, BRCA1, MT2A, and HER2. We show that intercellular mRNA transfer occurs in all coculture models tested (e.g., between primary cells, immortalized cells, and in cocultures of immortalized human and murine cells). Rapid mRNA transfer is dependent upon actin but is independent of de novo protein synthesis and is modulated by stress conditions and gene-expression levels. Hence, this work supports the hypothesis that full-length mRNAs undergo transfer between cells through a refined structural connection. Importantly, unlike the transfer of miRNA or RNA fragments, this process of communication transfers genetic information that could potentially alter the acceptor cell proteome. This phenomenon may prove important for the proper development and functioning of tissues as well as for host–parasite or symbiotic interactions.

scholarly journals Gap junctional protein Cx43 is involved in the communication between extracellular vesicles and mammalian cells

2015 ◽  
Vol 5 (1) ◽  
Author(s):  
Ana Rosa Soares ◽  
Tania Martins-Marques ◽  
Teresa Ribeiro-Rodrigues ◽  
Joao Vasco Ferreira ◽  
Steve Catarino ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Intercellular Communication ◽  
Mammalian Cells ◽  
Connexin 43 ◽  
Biological Effects ◽  
Biological Information ◽  
Target Cells ◽  
New Paradigm ◽  
Donor And Acceptor ◽  
Messenger Molecules

Abstract Intercellular communication is vital to ensure tissue and organism homeostasis and can occur directly, between neighbour cells via gap junctions (GJ), or indirectly, at longer distances, through extracellular vesicles, including exosomes. Exosomes, as intercellular carriers of messenger molecules, mediate the transfer of biological information between donor and acceptor cells. Although the biological effects of exosomes in target cells have been intensively studied, the mechanisms that govern exosomal uptake are not fully understood. Here, we show that Connexin 43 (Cx43), the most widely expressed GJ protein, is present in exosomes in the form of hexameric channels and, more importantly, that exosomal Cx43 is able to modulate the interaction and transfer of information between exosomes and acceptor cells. This study envisions a new paradigm where Cx43-containing channels mediate the release of exosomal content into cells, which constitutes a novel and unanticipated mechanism to modulate intercellular communication.

Mouse Jagged2 is differentially expressed in hematopoietic progenitors and endothelial cells and promotes the survival and proliferation of hematopoietic progenitors by direct cell-to-cell contact

Blood ◽  
2000 ◽  
Vol 96 (3) ◽  
pp. 950-957 ◽  
Author(s):  
Schickwann Tsai ◽  
Jutta Fero ◽  
Steve Bartelmez
Keyword(s):  
Stem Cells ◽  
Endothelial Cells ◽  
Hematopoietic Stem Cells ◽  
Full Length ◽  
Differentially Expressed ◽  
Cell Contact ◽  
Proliferative Potential ◽  
Hematopoietic Progenitors ◽  
Hematopoietic Stem ◽  
Direct Cell

To study the regulation of the early stages of hematopoiesis, cDNA representational difference analysis was used to isolate genes that were differentially expressed in primitive hematopoietic progenitors. The reasoning was that such genes were more likely to provide functions important to hematopoietic stem cells and progenitors. One of the genes identified through this approach encodes mouse Jagged2(mJagged2). Using quantitative reverse transcription–polymerase chain reaction, it was shown that mJagged2 was differentially expressed in c-kit+ hematopoietic progenitors, including those with the phenotypes of Lin− c-kit+Rhlo Holo and Lin−c-kit+ Rhhi Holo, and that they have been shown to be highly enriched for long-term and short-term repopulating hematopoietic stem cells, respectively. Western blot analyses showed that endothelial cells also expressed high levels of Jagged2, but stromal fibroblasts did not. Using a coculture system we found that exogenous, full-length mJagged2 promoted the survival and proliferation of hematopoietic progenitors, including the high-proliferative potential colony-forming cells. Direct cell-to-cell contact was required for this effect. Taken together, these findings indicate that both c-kit+ hematopoietic progenitors and endothelial cells express Jagged2 and that exogenous, full-length Jagged2 promotes the survival and proliferation of hematopoietic progenitors.

scholarly journals Extracellular Vesicles: From Biomarkers to Therapeutic Tools

Biology ◽  
2020 ◽  
Vol 9 (9) ◽  
pp. 258
Author(s):  
Simona Bernardi ◽  
Carolina Balbi
Keyword(s):  
Extracellular Vesicles ◽  
Intercellular Communication ◽  
Cell Contact ◽  
Apoptotic Bodies ◽  
Therapeutic Tool ◽  
The Status ◽  
Direct Cell ◽  
Therapeutic Tools ◽  
Multicellular Organisms ◽  
Target Cancer

Intercellular communication is an essential hallmark of multicellular organisms and can be mediated through direct cell–cell contact or transfer of secreted molecules. In the last two decades, a third mechanism for intercellular communication has emerged that involves intercellular transfer of extracellular vesicles (EVs). EVs are membranous vesicles of 30–5000 nm in size. Based on their dimension and biogenesis, EVs can be divided into different categories, such as microvesicles, apoptotic bodies, ectosomes, and exosomes. It has already been demonstrated that protein changes, expressed on the surfaces or in the content of these vesicles, may reflect the status of producing cells. For this reason, EVs, and exosomes in particular, are considered ideal biomarkers in several types of disease—from cancer diagnosis to heart rejection. This aspect opens different opportunities in EVs clinical application, considering the importance given to liquid biopsy in the recent years. Furthermore, extracellular vesicles can be natural or engineered carriers of cytoprotective or cytotoxic factors and applied, as a therapeutic tool, from regenerative medicine to target cancer therapy. This is of pivotal importance in the so called “era of the 4P medicine”. This Editorial focuses on recent findings pertaining to EVs in different medical areas, from biomarkers to therapeutic applications.

scholarly journals Reshaping the tumor microenvironment: extracellular vesicles as messengers of cancer cells

2020 ◽  
Vol 41 (11) ◽  
pp. 1461-1470
Author(s):  
Bibek Bhatta ◽  
Tomer Cooks
Keyword(s):  
Tumor Microenvironment ◽  
Cancer Cells ◽  
Extracellular Vesicles ◽  
Cancer Progression ◽  
Cell Contact ◽  
Long Distance ◽  
Donor Cells ◽  
Distance Communication ◽  
Direct Cell ◽  
Cancer Types

Abstract The tumor microenvironment (TME) comprises an assortment of immune and non-immune cells. The interactions between the cancer cells and their surrounding TME are known to be a cardinal factor in all stages of cancer progression, from initiation to metastasis. Tumor-associated macrophages (TAMs) and cancer-associated fibroblasts (CAFs) are considered two of the most abundant TME members associated with poor prognosis in various cancer types. Intercellular communication between the cancer cells and TME cells might occur via direct cell–cell contact or achieved through secreted factors such as cytokines, growth factors and extracellular vesicles (EVs). EVs are released by almost every cell type and by cancer cells in particular. EVs are loaded with unique molecular cargos that might include DNA, proteins, RNA and lipids, commonly reflecting the physiological traits of their donor cells. Once released, EVs are capable of initiating short- and long-distance communication in an autocrine, paracrine and endocrine fashion. The molecular cargos within the EVs are able to impart phenotypic changes at the receiving end thus allowing EV-releasing cancer cells to deliver messages to TME cells and tighten their grasp over the cancerous tissue. In this concise review, we aim to document the bidirectional EV-based communication between cancer cell, TAMs and CAFs, tilting the balance in favor of cancer progression and metastasis.

scholarly journals Highly efficient intercellular spreading of protein misfolding mediated by viral ligand - receptor interactions

2020 ◽  
Author(s):  
Shu Liu ◽  
Andre Hossinger ◽  
Annika Hornberger ◽  
Oleksandra Buravlova ◽  
Stephan Müller ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Protein Misfolding ◽  
Protein Aggregates ◽  
Cell Contact ◽  
List Type ◽  
Viral Glycoprotein ◽  
Receptor Ligand ◽  
Cellular Models ◽  
Ligand Interactions ◽  
Direct Cell

SUMMARYPathological protein aggregates associated with neurodegenerative diseases have the ability to transmit to unaffected cells, thereby templating their own aberrant conformation onto soluble proteins of the same kind. Proteopathic seeds can be released into the extracellular space, secreted in association with extracellular vesicles (EV) or exchanged by direct cell-to-cell contact. The extent to which each of these pathways contributes to the prion-like spreading of protein misfolding is unclear. Exchange of cellular cargo by both direct cell-to-cell contact as well as via EV depends on receptor-ligand interactions and subsequent release of cargo into the cytosol. We hypothesized that enabling these interactions through viral ligands enhances the aggregate-inducing capacity of EV-associated proteopathic seeds. Using different cellular models propagating model prion-like protein aggregates, mouse-adapted prions or pathogenic Tau aggregates, we demonstrate that vesicular stomatitis virus glycoprotein and SARS-CoV-2 spike S increase protein aggregate induction by direct cell-to-cell contact or via viral glycoprotein-decorated EV. Thus, receptor-ligand interactions are major determinants of intercellular aggregate dissemination. Further, our data raise the intriguing possibility that acute or latent viral infections contribute to proteopathic seed spreading by facilitating intercellular cargo transfer.HIGHLIGHTSDifferent types of proteopathic seeds are secreted in association with extracellular vesiclesReceptor-ligand interactions are important drivers of direct cell-to-cell and extracellular vesicle-mediated spreading of protein misfoldingViral glycoproteins mediating attachment and membrane fusion strongly enhance aggregate inducing capacity in recipient cellsGRAPHICAL ABSTRACT

Mouse Jagged2 is differentially expressed in hematopoietic progenitors and endothelial cells and promotes the survival and proliferation of hematopoietic progenitors by direct cell-to-cell contact

Blood ◽  
2000 ◽  
Vol 96 (3) ◽  
pp. 950-957 ◽  
Author(s):  
Schickwann Tsai ◽  
Jutta Fero ◽  
Steve Bartelmez
Keyword(s):  
Stem Cells ◽  
Endothelial Cells ◽  
Hematopoietic Stem Cells ◽  
Full Length ◽  
Differentially Expressed ◽  
Cell Contact ◽  
Proliferative Potential ◽  
Hematopoietic Progenitors ◽  
Hematopoietic Stem ◽  
Direct Cell

Abstract To study the regulation of the early stages of hematopoiesis, cDNA representational difference analysis was used to isolate genes that were differentially expressed in primitive hematopoietic progenitors. The reasoning was that such genes were more likely to provide functions important to hematopoietic stem cells and progenitors. One of the genes identified through this approach encodes mouse Jagged2(mJagged2). Using quantitative reverse transcription–polymerase chain reaction, it was shown that mJagged2 was differentially expressed in c-kit+ hematopoietic progenitors, including those with the phenotypes of Lin− c-kit+Rhlo Holo and Lin−c-kit+ Rhhi Holo, and that they have been shown to be highly enriched for long-term and short-term repopulating hematopoietic stem cells, respectively. Western blot analyses showed that endothelial cells also expressed high levels of Jagged2, but stromal fibroblasts did not. Using a coculture system we found that exogenous, full-length mJagged2 promoted the survival and proliferation of hematopoietic progenitors, including the high-proliferative potential colony-forming cells. Direct cell-to-cell contact was required for this effect. Taken together, these findings indicate that both c-kit+ hematopoietic progenitors and endothelial cells express Jagged2 and that exogenous, full-length Jagged2 promotes the survival and proliferation of hematopoietic progenitors.

scholarly journals Extracellular vesicles mediate the intercellular exchange of nanoparticles

2021 ◽  
Author(s):  
Xian Wu ◽  
Tang Tang ◽  
Yushuang Wei ◽  
Katherine A. Cummins ◽  
David K. Wood ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Cell Contact ◽  
Therapeutic Effects ◽  
Cellular Receptors ◽  
Transcellular Transport ◽  
Direct Cell ◽  
Intercellular Exchange

AbstractIn order to exert their therapeutic effects, nanoparticles (NPs) often need to travel into the tissues composed of multilayered cells. Accumulative evidence has revealed the central role of transcellular transport route (entry into one cell, exocytosis, and re-entry into another) in this process. While NP endocytosis and subcellular transport have been intensively characterized, the exocytosis and re-entry steps are poorly understood, which becomes a barrier to improve NP delivery into complex tissues. Here, we termed the exocytosis and re-entry steps together as intercellular exchange. We developed a novel collagen-based 3D cellular assay to specifically monitor and quantify the intercellular exchange events of NPs and distinguish the contributions of several potential mechanisms. Our results showed that NPs can be exocytosed freely or enclosed inside extracellular vesicles (EVs) for re-entry, while direct cell-cell contact is hardly involved. EVs account for a significant fraction of NP intercellular exchange, and its importance in NP delivery was demonstrated in vitro and in vivo. Intriguingly, while freely released NPs engage with the same cellular receptors for re-entry, EV-enclosed ones bypass this dependence. These studies provide an easy and precise system to investigate the intercellular exchange stage of NP delivery, and shed the first light in the importance of EVs in NP transport between cells and across complex tissues.

Sign in / Sign up

Export Citation Format

Share Document