Therapy free of cells vs human mesenchymal stem cells from umbilical cord stroma to treat the inflammation in OA

[Abstract] Osteoarthritis (OA) is closely linked to the increase in the number of senescent cells in joint tissues, and the senescence-associated secretory phenotype (SASP) is implicated in cartilage degradation. In the last decade, extracellular vesicles (EV) in combination with the use of miRNAs to modify post-transcriptional expressions of multiple genes have shown their utility in new therapies to treat inflammatory diseases. This work delves into the anti-inflammatory effect of extracellular vesicles derived from mesenchymal stem cells (MSC) previously modified to inhibit the expression of miR-21. We compare the efficacy of two treatments, MSC with their miR-21 inhibited through lentiviral transfection and their EV, against inflammation in a new OA animal model. The modified MSC and their EV were intraperitoneally injected in an OA animal model twice. One month after treatment, we checked which therapy was the most effective to reduce inflammation compared with animals untreated. Treated OA model sera were analyzed for cytokines and chemokines. Subsequently, different organs were analyzed to validate the results obtained. EV were the most effective treatment to reduce chemokines and cytokines in serum of OA animals as well as SASP, in their organs checked by proteomic and genomic techniques, compared with MSC alone in a statistically significant way. In conclusion, MSC-miR-21--derived EV showed a higher therapeutic potential in comparison with MSCs-miR-21-. They ameliorate the systemic inflammation through inactivation of ERK1/2 pathway in OA in vivo model. Workflow of the realization of the animal model of OA by injecting cells into the joint cavity of the left knee of the animals, which produces an increase in serum cytokines and chemokines in the animals in addition to the increase in SASP and markers of inflammation. Inhibition of miR-21 in MSCs, from the stroma of the human umbilical cord, by lentivirus and extraction of their EVs by ultracentrifugation. Finally, application of MSC therapy with its miR-21 inhibited or its EVs produces a decrease in serum cytokines and chemokines in the treated animals, in addition to an increase in SASP and markers of inflammation. The cell-free therapy being the one that produces a greater decrease in the parameters studied.Xunta de Galicia; ED481D-2021-020Instituto de Salud Carlos III; PI20/0049

Describing Polyps Behavior of a Deep-Sea Gorgonian, Placogorgia sp., Using a Deep-Learning Approach

Gorgonians play a fundamental role in the deep sea (below 200 m depth), composing three-dimensional habitats that are characterized by a high associated biodiversity and playing an important part in biogeochemical cycles. Here we describe the use of a benthic lander to monitoring polyps activity, used as a proxy of gorgonian feeding activity of three colonies of Placogorgia sp. Images cover a period of 22 days with a temporal resolution of 30 min. In addition, this seafloor observatory is instrumented with oceanographic sensors that allows continuous monitoring of the hydrographic conditions in the site. Deep-learning is used for automatic detection of the state of the polyps registered in the images. More than 1000 images of 3 large specimens of gorgonians are analyzed, annotating polyps as extended or retracted, using the semantic segmentation algorithm ConvNeXt. The segmentation results are used to describe the feeding patterns of this species. Placogorgia sp. shows a daily pattern of feeding conduct, depending on the hours of day and night. Using a Singular Spectrum Analysis approach, feeding activity is related to currents dynamics and Acoustic Doppler Current Profile (ADCP) return signal intensity, as proxy of suspended matter, achieving a linear correlation of 0.35 and 0.11 respectively. This is the first time that the behavior of the Placogorgia polyps, directly related to their feeding process, is described

Action Mechanisms of Small Extracellular Vesicles in Inflammaging [Review]

This article belongs to the Special Issue Extracellular Vesicles Research in Inflamm-Aging[Abstract] The accumulation process of proinflammatory components in the body due to aging influences intercellular communication and is known as inflammaging. This biological mechanism relates the development of inflammation to the aging process. Recently, it has been reported that small extracellular vesicles (sEVs) are mediators in the transmission of paracrine senescence involved in inflammatory aging. For this reason, their components, as well as mechanisms of action of sEVs, are relevant to develop a new therapy called senodrugs (senolytics and senomorphic) that regulates the intercellular communication of inflammaging. In this review, we include the most recent and relevant studies on the role of sEVs in the inflammatory aging process and in age-related diseases such as cancer and type 2 diabetes.J.F.L. was funded by Xunta de Galicia, Grant Number ED481D-2021-020. M.C.A. received a grant from the Spanish National Health Institute Carlos III (PI20/00497)Xunta de Galicia; ED481D-2021-02

Deep learning-assisted high resolution mapping of vulnerable habitats within the Capbreton Canyon System, Bay of Biscay

The Capbreton Canyon System is an area currently under study for its proposal as a Site of Community Importance under the EU Habitats Directive in the context of the LIFE IP INTEMARES project. Identifying and mapping benthic Vulnerable Marine Ecosystems (VMEs) plays a key role in this process. Although obtaining information on species distribution in deep sea rocky habitats has traditionally been a complicated task, the development of underwater remote sensing techniques resulted in a massive increase in the collection of digital imagery; however, processing all this information has led to another bottleneck due to the time-consuming nature of biota manual annotation. At this point, the use of computer vision and deep learning to automate image processing has substantial benefits but has rarely been adopted within the field of marine ecology. This study presents the integration of deep learning techniques for benthic fauna identification, high resolution multibeam echosounder (MBES) data and Species Distribution Models (SDMs), to map the potential habitat of the yellow coral Dendrophyllia cornigera, a representative species of the VME 1170 Reef habitat, on the circalitoral area of the Capbreton Canyon System. The localization and identification of the coral colonies was based on more than 7500 photographs taken during the INTEMARES-CapBreton 0619 and 0620 surveys using the photogrammetric ROTV Politolana. For the automatic annotation of the image set a deep learning based framework was developed by testing two different deep neural networks architectures; a FasterRCNN+Resnet101 model, accomplishing a precision of 100% over human expert annotation for presence/absence discrimination, was selected. Environmental data included different quantitative terrain attributes derived from high resolution MBES bathymetry data. A presence-only species distribution model, Maximum Entropy (MaxEnt), was used to infer the spatial distribution of D. cornigera over the study area. Predicted occurrences corresponded mainly to relevant topographic structures with significant slope, mainly associated to the edge of the continental shelf. These results are consistent with the ecological knowledge on the species and validate the use of deep learning tools to assist in the identification and mapping of VME for management and conservation purposes. This study provides a baseline for the protection of vulnerable habitats of the Capbreton Canyon System in the context of the Natura 2000 Network.The authors would like to thank the crew and scientific team aboard the R/V Ramón Margalef from the Spanish Institute of Oceanography and also appreciate the helpful assistance of the technicians of the ROTV Politolana for their skill in executing the dangerous visual transects very close to the rough bottoms of the study area. This research has been performed in the scope of the INTEMARES project. INTEMARES was partially funded by the European Commission LIFE + “Nature and Biodiversity” call (LIFE15 IPE ES 012). The Biodiversity Foundation of the Spanish Ministry for Ecological Transition, was the institution responsible for coordination this project. Deep-learning advances presented here are part of Deep-RAMP (Deep learning to improve the management of marine protected area network in the North Atlantic region) project funded in the frame of the Pleamar Program of the Biodiversity Foundation and is co-financed by the European Maritime and Fisheries Fund (EMFF)

Effect of miR-21 in mesenchymal stem cells-derived extracellular vesicles behavior

Abstract Background A challenging new branch of research related to aging-associated diseases is the identification of miRNAs capable of modulating the senescence-associated secretory phenotype (SASP) which characterizes senescent cells and contributes to driving inflammation. Methods Mesenchymal stem cells (MSC) from human umbilical cord stroma were stable modified using lentivirus transduction to inhibit miR-21-5p and shotgun proteomic analysis was performed in the MSC-derived extracellular vesicles (EV) to check the effect of miR-21 inhibition in their protein cargo. Besides, we studied the paracrine effect of those modified extracellular vesicles and also their effect on SASP. Results Syndecan-1 (SDC1) was the most decreased protein in MSC-miR21−-derived EV, and it was involved in inflammation and EV production. MSC-miR21−-derived EV were found to produce a statistically significant inhibitory effect on SASP and inflammaging markers expression in receptor cells, and in the opposite way, these receptor cells increased their SASP and inflammaging expression statistically significantly when treated with MSC-miR-21+-derived EV. Conclusion This work demonstrates the importance of miR-21 in inflammaging and its role in SASP through SDC1. Graphical abstrac

Shotgun proteomics reveals senomorphic targets based on SASP-mediated by small extracellular vesicles

Poster.-- Human Proteome Organization World Congress, HUPO 2023, 17-21 SeptemberCells have the capacity to modulate the microenvironment through the secretion of molecules (cytokines, chemokines, matrix proteins…) and vesicles, which vary according to the pathology involved, such as cancer or natural process such as senescence. Cellular senescence is a process that enhance with ageing and its association with the onset of age-related diseases. The process is characterized by less proliferation, increased β-galactosidase activity and specific secretory phenotype known as SASP. SASP lead the microenvironment to a more pro-inflammatory one and has the capacity to induce paracrine senescence in neighbouring cells. The regulation of small extracellular vesicles (sEV), which are part of SASP, have a high potential to develop drugs that modulate the senescence transmission. Our aim is to find a proteomic signature of the SASP mediated by sEV to reveal pathways associated with the senescence transmissionN

Study of Ferroptosis Transmission by Small Extracellular Vesicles in Epithelial Ovarian Cancer Cells

Epithelial ovarian cancer (EOC) is the most lethal gynecological cancer. The current treatment for EOC involves surgical debulking of the tumors followed by a combination of chemotherapy. While most patients achieve complete remission, many EOCs will recur and develop chemo-resistance. The cancer cells can adapt to several stress stimuli, becoming resistant. Because of this, new ways to fight resistant cells during the disease are being studied. However, the clinical outcomes remain unsatisfactory. Recently, ferroptosis, a novel form of regulated cell death trigged by the accumulation of iron and toxic species of lipid metabolism in cells, has emerged as a promising anti-tumor strategy for EOC treatment. This process has a high potential to become a complementary treatment to the current anti-tumor strategies to eliminate resistant cells and to avoid relapse. Cancer cells, like other cells in the body, release small extracellular vesicles (sEV) that allow the transport of substances from the cells themselves to communicate with their environment. To achieve this, we analyzed the capacity of epithelial ovarian cancer cells (OVCA), treated with ferroptosis inducers, to generate sEV, assessing their size and number, and study the transmission of ferroptosis by sEV. Our results reveal that OVCA cells treated with ferroptotic inducers can modify intercellular communication by sEV, inducing cell death in recipient cells. Furthermore, these receptor cells are able to generate a greater amount of sEV, contributing to a much higher ferroptosis paracrine transmission. Thus, we discovered the importance of the sEV in the communication between cells in OVCA, focusing on the ferroptosis process. These findings could be the beginning form to study the molecular mechanism ferroptosis transmission through sEV

Speech Communication

Contains table of contents for Part V, table of contents for Section 1, reports on six research projects and a list of publications.C.J. Lebel FellowshipDennis Klatt Memorial FundNational Institutes of Health Grant R01-DC00075National Institutes of Health Grant R03-DC01721National Institutes of Health Grant R01-DC01291National Institutes of Health Grant R01-DC00261National Institutes of Health Grant P01-DC00361-06A1National Institutes of Health Grant R01-DC00776National Science Foundation Grant IRI 89-05439National Science Foundation Grant IRI 89-10561National Science Foundation Grant INT 90-2471