Cultured human amniocytes express hTERT, which is distributed between nucleus and cytoplasm and is secreted in extracellular vesicles

Radeghieri, Annalisa; Savio, Giulia; Zendrini, Andrea; Noto, G.; Salvi, Alessandro; Bergese, Paolo; Piovani, Giovanna

doi:10.1016/j.bbrc.2016.12.077

Cited by 25 publications

(20 citation statements)

References 32 publications

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“…The purity, size distribution and number of separated EVs were revealed by Atomic Force Microscopy (AFM) and Colloidal nanoplasmonic (CONAN) assay ( Fig 2F, 2G and 2H). Western blot analysis, using antibodies against surface and internal makers, excluded contamination by other membrane fragments (GM-130), confirmed EV-specific marker-expression (Annexin XI, Annexin V and TSG-101), TERT-expression (in accordance with a recently-discovered biomarker on amniocyte-derived EVs [24]) and characteristic CD29 protein-expression of the parental MSCs (Fig 2I).…”

Section: Resultssupporting

confidence: 66%

Extracellular vesicles from rat-bone-marrow mesenchymal stromal/stem cells improve tendon repair in rat Achilles tendon injury model in dose-dependent manner: A pilot study

et al. 2020

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Mesenchymal stromal/stem cells (MSCs) are increasingly employed for tissue regeneration, largely mediated through paracrine actions. Currently, extracellular vesicles (EVs) released by MSCs are major mediators of these paracrine effects. We evaluated whether rat-bonemarrow-MSC-derived EVs (rBMSCs-EVs) can ameliorate tendon injury in an in vivo rat model. Pro-collagen1A2 and MMP14 protein are expressed in rBMSC-EVs, and are important factors for extracellular-matrix tendon-remodeling. In addition, we found pro-col-lagen1A2 in rBMSC-EV surface-membranes by dot blot. In vitro on cells isolated from Achilles tendons, utilized as rBMSC-EVs recipient cells, EVs at both low and high doses induce migration of tenocytes; at higher concentration, they induce proliferation and increase expression of Collagen type I in tenocytes. Pretreatment with trypsin abrogate the effect of EVs on cell proliferation and migration, and the expression of collagen I. When either low-or high-dose rBMSCs-EVs were injected into a rat-Achilles tendon injury-model (immediately after damage), at 30 days, rBMSC-EVs were found to have accelerated the remodeling stage of tendon repair in a dose-dependent manner. At histology and histomorphology evaluation, high doses of rBMSCs-EVs produced better restoration of tendon architecture, with optimal tendon-fiber alignment and lower vascularity. Higher EV-concentrations demonstrated greater expression of collagen type I and lower expression of

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Section: Resultssupporting

confidence: 66%

Extracellular vesicles from rat-bone-marrow mesenchymal stromal/stem cells improve tendon repair in rat Achilles tendon injury model in dose-dependent manner: A pilot study

et al. 2020

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“…This increase could have significant functions in the viral life cycle of HPV, therefore playing an essential role in cell immortalization and in the process of malignant transformation (2,22). Regarding the cellular localization of the hTERT protein, the reported findings in different tissues are few (14,15,(22)(23)(24)(25)(26)(27)(28)(29). In some reports, it has been shown that although hTERT is expressed in the nucleus of benign tissues, it is gradually transferred to the cytoplasm in the process of malignant transformation (2,22,23).…”

Section: Discussionmentioning

confidence: 99%

hTERT Protein Expression in Cytoplasm and Nucleus and its Association With HPV Infection in Patients With Cervical Cancer

Moreno‐Acosta

Molano

Morales

et al. 2020

Cancer Genomics Proteomics

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Background: Few studies have analyzed the association between human telomerase reverse transcriptase (hTERT) protein expression (nuclear and cytoplasmic localization), hTERT methylation status, and human papillomavirus (HPV) genotype infection in cervical cancer. Patients and Methods: One hundred seventy-three patients with cervical cancer were analyzed. hTERT protein expression was detected by immunohistochemistry. hTERT DNA methylation analysis was performed using a PCR-RLB-hTERT assay, targeting two regions of the hTERT promoter. Type specific HPV infection was detected by using GP5+/GP6+PCR-RLB. Results: hTERT protein expression was found in both cytoplasm and nucleus (78.0% of the samples showed a cytoplasmic localization and 79.8% had a nuclear localization). A statistically significant association was found between alpha 9 and 7 HPV species with a nonmethylation pattern of the hTERT promoter and between these species and high expression of hTERT protein with nuclear localization. Conclusion: hTERT protein is found in both the nucleus and cytoplasm of patients with cervical cancer and confirm the relationship between the nonmethylated status of hTERT promoter and some HPV species as well as the relationship between these species and hTERT protein expression. Epidemiological studies have demonstrated a strong causal relationship between persistent infection with human papillomavirus (HPV), especially high-risk HPV (HRHPV) types and the development of cervical cancer (1). In vitro studies have shown that neoplastic transformation of cells infected with HRHPV is mainly due to telomerase activation by the action of HPV E6 oncoprotein (2). Telomerase is a ribonucleoprotein complex with terminal transferase activity and is made up by an RNA component (hTR), a catalytic protein subunit (hTERT) and the telomerase associated protein TEP1 (3). Some studies have shown a significant telomerase expression and telomerase activity levels in different types of cancer (e.g. lung, pancreas, hepatocellular, prostate, skin and certain gastrointestinal tumors) and malignant cell lines, while their levels are very low in 615 This article is freely accessible online.

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“…To obtain conditioned media for exosome isolation, HaCaT cells were grown in complete DMEM until the reach of 70% confluence, and then the media were removed and renewed with DMEM without FBS for 48 hours. Afterward, conditioned media were collected and processed with a serial centrifugation protocol (37,38). Briefly, conditioned media were centrifuged firstly at 800 g for 30 minutes to pellet and discard possibly contained apoptotic bodies and cellular debris.…”

Section: Methodsmentioning

confidence: 99%

Exosome-delivered microRNAs promote IFN-α secretion by human plasmacytoid DCs via TLR7

Salvi¹,

Gianello²,

Busatto³

et al. 2018

JCI Insight

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The excessive production of type I IFNs is a hallmark and a main pathogenic mechanism of many autoimmune diseases, including systemic lupus erythematosus (SLE). In these pathologies, the sustained secretion of type I IFNs is dependent on the improper activation of plasmacytoid DCs (pDCs) by self-nucleic acids. However, the nature and origin of pDC-activating self-nucleic acids is still incompletely characterized. Here, we report that exosomes isolated from the plasma of SLE patients can activate the secretion of IFN-α by human blood pDCs in vitro. This activation requires endosomal acidification and is recapitulated by microRNAs isolated from exosomes, suggesting that exosome-delivered microRNAs act as self-ligands of innate single-stranded endosomal RNA sensors. By using synthetic microRNAs, we identified an IFN induction motif that is responsible for the TLR7-dependent activation, maturation, and survival of human pDCs. These findings identify exosome-delivered microRNAs as potentially novel TLR7 endogenous ligands able to induce pDC activation in SLE patients. Therefore, microRNAs may represent novel pathogenic mediators in the onset of autoimmune reactions and potential therapeutic targets in the treatment of type I IFN-mediated diseases.

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Cultured human amniocytes express hTERT, which is distributed between nucleus and cytoplasm and is secreted in extracellular vesicles

Cited by 25 publications

References 32 publications

Extracellular vesicles from rat-bone-marrow mesenchymal stromal/stem cells improve tendon repair in rat Achilles tendon injury model in dose-dependent manner: A pilot study

Extracellular vesicles from rat-bone-marrow mesenchymal stromal/stem cells improve tendon repair in rat Achilles tendon injury model in dose-dependent manner: A pilot study

hTERT Protein Expression in Cytoplasm and Nucleus and its Association With HPV Infection in Patients With Cervical Cancer

Exosome-delivered microRNAs promote IFN-α secretion by human plasmacytoid DCs via TLR7

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