Journal of Extracellular Vesicles最新文献

MISEV2023 has been written and revised to stand on its own as a guide to EV studies . Starting with an introduction with a historical perspective and ‘user guide’, MISEV2023 next takes the reader through all of the main aspects of an EV study. Community-sourced, it has something to offer beginners and veterans alike. The nomenclature section introduces the diversity of EVs and the need to avoid misusing the term ‘exosome’. It also highlights the ‘non-vesicular extracellular particles’ (NVEPs) that abound in most EV preparations. Collection and pre-processing underscores the importance of pre-analytical variables, with input on specific EV sources from ISEV task forces. EV separation and concentration and EV characterization have new details on key approaches. A new section on technique-specific reporting for EV characterization focuses on leading commercially available methods. EV release and uptake examines opportunities and pitfalls, while functional studies reminds us of important controls that are needed to help attribute an outcome to EVS. Finally, there is a new section on studying EVs in vivo.

Despite the stand-alone nature of MISEV2023, it is nevertheless instructive to read and interpret this new document in the context of the previous MISEV publications, MISEV2014 and MISEV2018 (Lötvall et al., 2014; Théry et al., 2018). MISEV2014 was an editorial of the ISEV board, while MISEV2018 first gathered community input, with nearly 400 contributors. Interspersed with the previous MISEVs, numerous ISEV position papers, along with collaborative reviews and editorials, have guided the field in important ways. The ISEV position papers are summarized in Table 1 of MISEV2023.

MISEV2023 began, as did MISEV2018, with a pre-drafting survey (Witwer et al., 2021). The input from this 2020 survey identified existing aspects to emphasize and new areas to explore. After reviewing the results of the survey, the ISEV board met at the end of 2020 and assigned a five-author committee (J.A. Welsh, D.C. Goberdhan, L. O'Driscoll, C. Théry and K.W. Witwer) to prepare a draft and guide the community input for the next MISEV. Numerous drafts and refinements were then prepared and shared with the ISEV board for feedback. Approximately 70 authors contributed to section drafts during this phase. Then, in 2022, an intermediate draft and a survey were shared with the ISEV community, resulting in >1000 responses. The three co-first authors worked hard and long to update MISEV based on this input. Multiple rounds of internal review and additional invited contributions/revisions ensued, along with style ‘unification’. In mid-2023, the article received pre-submission comments from JEV, followed by more revisions, formal submission, and three post-submission rounds of scrutiny (the first, main round including comments from more than 30 individuals). A near-final dra

Extracellular vesicle (EV) research has expanded exponentially over the last 10–15 years, with less than 100 entries in PubMed in 2007 but almost 8000 in 2022, depending on the specifics of the search terms applied. As this meteoric rise began, the field moved at a high pace, with methodology and technology evolving swiftly. This evolution has had notable impact on the stringency of the research produced. When ISEV was established in 2011, it was evident that researchers in the field needed some guidance to improve the quality of published science, as well as advice regarding the technologies being developed to support their work.

The aim of the ISEV leadership has always been to support the positive development of EV research in general. To assist with this, ISEV has published the so-called MISEV guidelines (Lotvall et al., 2014 and Thery et al., 2018). In the first version, MISEV2014, the word “requirement” was used in the title of the document, which resulted in some criticism, and therefore in 2018 the term “information” was used to better represent the overall goal of MISEV. MISEV guidelines aim to represent the consensus of the field regarding the current state of the art, and they are NOT a set of requirements for EV researchers. MISEV2014 was a short and succinct advisory text on how EV research can provide relatively conclusive results, as well as identifying some caveats that can be avoided in this field of research. Four years later, in 2018, a vast technology development had occurred, with new methods and analytical tools having been introduced to the field. The MISEV guidelines grew significantly, and many authors were recruited to contribute to the document. This effort evolved to the massive MISEV 2018 publication, which is extensively referenced, sometimes as a handbook of EV research. Indeed, if the advice in this document is applied by researchers, the likelihood that the results observed in any study depend on the EVs, are relatively likely.

Since MISEV2018, there have been even further developments in the field, including new methods of EV isolation and technologies to characterize EVs at the single-EV level. ISEV initiated a process to develop a new and updated MISEV several years ago, and an enormous effort has been employed to develop texts and advice for the version being published in the current issue of Journal of Extracellular Vesicles. The document has 1051 authors, and the whole process of compiling the document has been immense and very challenging but ultimately rewarding.

The editorial approach taken towards the MISEV document, much like the process of compiling the manuscript itself, has been extensive. In the spring of 2023, a full draft of the MISEV 2023 guidelines had been compiled, and discussions between the ISEV board and the Editorial leadership of JEV on how, and when, to submit the document for consideration by the journal ensued. The journal clarified that,

Small-for-gestational age (SGA) neonates exhibit increased perinatal morbidity and mortality, and a greater risk of developing chronic diseases in adulthood. Currently, no effective maternal blood-based screening methods for determining SGA risk are available. We used a high-resolution MS/MS^ALL shotgun lipidomic approach to explore the lipid profiles of small extracellular vesicles (sEV) released from the placenta into the circulation of pregnant individuals. Samples were acquired from 195 normal and 41 SGA pregnancies. Lipid profiles were determined serially across pregnancy. We identified specific lipid signatures of placental sEVs that define the trajectory of a normal pregnancy and their changes occurring in relation to maternal characteristics (parity and ethnicity) and birthweight centile. We constructed a multivariate model demonstrating that specific lipid features of circulating placental sEVs, particularly during early gestation, are highly predictive of SGA infants. Lipidomic-based biomarker development promises to improve the early detection of pregnancies at risk of developing SGA, an unmet clinical need in obstetrics.

The COVID-19 pandemic highlighted the clear risk that zoonotic viruses pose to global health and economies. The scientific community responded by developing several efficacious vaccines which were expedited by the global need for vaccines. The emergence of SARS-CoV-2 breakthrough infections highlights the need for additional vaccine modalities to provide stronger, long-lived protective immunity. Here we report the design and preclinical testing of small extracellular vesicles (sEVs) as a multi-subunit vaccine. Cell lines were engineered to produce sEVs containing either the SARS-CoV-2 Spike receptor-binding domain, or an antigenic region from SARS-CoV-2 Nucleocapsid, or both in combination, and we tested their ability to evoke immune responses in vitro and in vivo. B cells incubated with bioengineered sEVs were potent activators of antigen-specific T cell clones. Mice immunised with sEVs containing both sRBD and Nucleocapsid antigens generated sRBD-specific IgGs, nucleocapsid-specific IgGs, which neutralised SARS-CoV-2 infection. sEV-based vaccines allow multiple antigens to be delivered simultaneously resulting in potent, broad immunity, and provide a quick, cheap, and reliable method to test vaccine candidates.

Extracellular vesicles (EVs), through their complex cargo, can reflect the state of their cell of origin and change the functions and phenotypes of other cells. These features indicate strong biomarker and therapeutic potential and have generated broad interest, as evidenced by the steady year-on-year increase in the numbers of scientific publications about EVs. Important advances have been made in EV metrology and in understanding and applying EV biology. However, hurdles remain to realising the potential of EVs in domains ranging from basic biology to clinical applications due to challenges in EV nomenclature, separation from non-vesicular extracellular particles, characterisation and functional studies. To address the challenges and opportunities in this rapidly evolving field, the International Society for Extracellular Vesicles (ISEV) updates its ‘Minimal Information for Studies of Extracellular Vesicles’, which was first published in 2014 and then in 2018 as MISEV2014 and MISEV2018, respectively. The goal of the current document, MISEV2023, is to provide researchers with an updated snapshot of available approaches and their advantages and limitations for production, separation and characterisation of EVs from multiple sources, including cell culture, body fluids and solid tissues. In addition to presenting the latest state of the art in basic principles of EV research, this document also covers advanced techniques and approaches that are currently expanding the boundaries of the field. MISEV2023 also includes new sections on EV release and uptake and a brief discussion of in vivo approaches to study EVs. Compiling feedback from ISEV expert task forces and more than 1000 researchers, this document conveys the current state of EV research to facilitate robust scientific discoveries and move the field forward even more rapidly.

Pluripotent stem cell-derived small extracellular vesicles (PSC-sEVs) have demonstrated great clinical translational potential in multiple aging-related degenerative diseases. Characterizing the PSC-sEVs is crucial for their clinical applications. However, the specific marker pattern of PSC-sEVs remains unknown. Here, the sEVs derived from two typical types of PSCs including induced pluripotent stem cells (iPSC-sEVs) and embryonic stem cells (ESC-sEVs) were analysed using proteomic analysis by liquid chromatography with tandem mass spectrometry (LC-MS/MS), and surface marker phenotyping analysis by nanoparticle flow cytometry (NanoFCM). A group of pluripotency-related proteins were found to be enriched in PSC-sEVs by LC-MS/MS and then validated by Western Blot analysis. To investigate whether these proteins were specifically expressed in PSC-sEVs, sEVs derived from seven types of non-PSCs (non-PSC-sEVs) were adopted for analysis. The results showed that PODXL, OCT4, Dnmt3a, and LIN28A were specifically enriched in PSC-sEVs but not in non-PSC-sEVs. Then, commonly used surface antigens for PSC identification (SSEA4, Tra-1-60 and Tra-1-81) and PODXL were gauged at single-particle resolution by NanoFCM for surface marker identification. The results showed that the positive rates of PODXL (>50%) and SSEA4 (>70%) in PSC-sEVs were much higher than those in non-PSC-sEVs (<10%). These results were further verified with samples purified by density gradient ultracentrifugation. Taken together, this study for the first time identified a cohort of specific markers for PSC-sEVs, among which PODXL, OCT4, Dnmt3a and LIN28A can be detected with Western Blot analysis, and PODXL and SSEA4 can be detected with NanoFCM analysis. The application of these specific markers for PSC-sEVs identification may advance the clinical translation of PSCs-sEVs.

Extracellular vesicles (EVs) exert a significant influence not only on the pathogenesis of diseases but also on their therapeutic interventions, contingent upon the variances observed in their originating cells. Mitochondria can be transported between cells via EVs to promote pathological changes. In this study, we found that EVs derived from M1 macrophages (M1-EVs), which encapsulate inflammatory mitochondria, can penetrate pancreatic beta cells. Inflammatory mitochondria fuse with the mitochondria of pancreatic beta cells, resulting in lipid peroxidation and mitochondrial disruption. Furthermore, fragments of mitochondrial DNA (mtDNA) are released into the cytosol, activating the STING pathway and ultimately inducing apoptosis. The potential of adipose-derived stem cell (ADSC)-released EVs in suppressing M1 macrophage reactions shows promise. Subsequently, ADSC-EVs were utilized and modified with an F4/80 antibody to specifically target macrophages, aiming to treat ferroptosis of pancreatic beta cells in vivo. In summary, our data further demonstrate that EVs secreted from M1 phenotype macrophages play major roles in beta cell ferroptosis, and the modified ADSC-EVs exhibit considerable potential for development as a vehicle for targeted delivery to macrophages.

mRNA-based molecular subtypes have implications for bladder cancer prognosis and clinical benefit from certain therapies. Whether small extracellular vesicles (sEVs) can reflect bladder cancer molecular subtypes is unknown. We performed whole transcriptome RNA sequencing for formalin fixed paraffin embedded (FFPE) tumour tissues and sEVs separated from matched tissue explants, urine and plasma in patients with bladder cancer. sEVs were separated using size-exclusion chromatography, and characterized by transmission electron microscopy, nano flow cytometry and western blots, respectively. High yield of sEVs were obtained using approximately 1 g of tissue, incubated with media for 30 min. FFPE tumour tissue and tumour tissue-derived sEVs demonstrated good concordance in molecular subtype classification. All urinary sEVs were classified as luminal subtype, while all plasma sEVs were classified as Ba/Sq subtype, regardless of the molecular subtypes indicated by their matched FFPE tumour tissue. The comparison within urine sEVs, which may exclude the sample type specific background, could pick up the different biology between NMIBC and MIBC, as well as the signature genes related to molecular subtypes. Four candidate sEV-related bladder cancer-specific mRNA biomarkers, FAM71E2, OR4K5, FAM138F and KRTAP26-1, were identified by analysing matched urine sEVs, tumour tissue derived sEVs, and adjacent normal tissue derived sEVs. Compared to sEVs separated from biofluids, tissue-derived sEVs may reflect more tissue- or disease-specific biological features. Urine sEVs are promising biomarkers to be used for liquid biopsy-based molecular subtype classification, but the current algorithm needs to be modified/adjusted. Future work is needed to validate the four new bladder cancer-specific biomarkers in large cohorts.

Urzì, O., Bergqvist, M., Lässer, C., Moschetti, M., Johansson, J., D´Arrigo, D., Olofsson Bagge, R., & Crescitelli, R. (2024). Heat inactivation of foetal bovine serum performed after EV-depletion influences the proteome of cell-derived extracellular vesicles. Journal of Extracellular Vesicles, 13, e12408. https://doi.org/10.1002/jev2.12408

In the originally published article, a portion of Figure 3 was mislabeled. The section labeled “MML1” should be labeled “Medium + FBS”. The corrected figure is shown below. This has been corrected in the online version of the article.

We apologize for this error.

Sandau, U. S., Magaña, S. M., Costa, J., Nolan, J. P., Ikezu, T., Vella, L. J., Jackson, H. K., Moreira, L. R., Palacio, P. L., Hill, A. F., Quinn, J. F., Van Keuren-Jensen, K. R., McFarland, T. J., Palade, J., Sribnick, E. A., Su, H., Vekrellis, K., Coyle, B., Yang, Y., … Saugstad, J. A., International Society for Extracellular Vesicles Cerebrospinal Fluid Task Force. (2024). Recommendations for reproducibility of cerebrospinal fluid extracellular vesicle studies. Journal of Extracellular Vesicles, 13, e12397. https://doi.org/10.1002/jev2.12397

In the originally published article, author You Yang's affiliation is incorrect. The correct affiliation is given below:

You Yang⁵

⁵Department of Neuroscience, Mayo Clinic Florida, Jacksonville, Florida, USA

We apologize for this error.