scholarly journals Intraoperative visualization of the tumor microenvironment and quantification of extracellular vesicles by label-free nonlinear imaging

2018 ◽  
Vol 4 (12) ◽  
pp. eaau5603 ◽  
Author(s):  
Yi Sun ◽  
Sixian You ◽  
Haohua Tu ◽  
Darold R. Spillman ◽  
Eric J. Chaney ◽  
...  
Keyword(s):  
Tumor Microenvironment ◽  
Extracellular Vesicles ◽  
Cancer Progression ◽  
Spatial Information ◽  
Imaging System ◽  
Label Free ◽  
Breast Reduction Surgery ◽  
Diagnosis And Prognosis ◽  
Nonlinear Imaging

Characterization of the tumor microenvironment, including extracellular vesicles (EVs), is important for understanding cancer progression. EV studies have traditionally been performed on dissociated cells, lacking spatial information. Since the distribution of EVs in the tumor microenvironment is associated with cellular function, there is a strong need for visualizing EVs in freshly resected tissues. We intraoperatively imaged untreated human breast tissues using a custom nonlinear imaging system. Label-free optical contrasts of the tissue, correlated with histological findings, enabled point-of-procedure characterization of the tumor microenvironment. EV densities from 29 patients with breast cancer were found to increase with higher histologic grade and shorter tumor-to-margin distance and were significantly higher than those from 7 cancer-free patients undergoing breast reduction surgery. Acquisition and interpretation of these intraoperative images not only provide real-time visualization of the tumor microenvironment but also offer the potential to use EVs as a label-free biomarker for cancer diagnosis and prognosis.

scholarly journals Label-Free Prostate Cancer Detection by Characterization of Extracellular Vesicles Using Raman Spectroscopy

2018 ◽  
Vol 90 (19) ◽  
pp. 11290-11296 ◽  
Author(s):  
Wooje Lee ◽  
Afroditi Nanou ◽  
Linda Rikkert ◽  
Frank A. W. Coumans ◽  
Cees Otto ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Raman Spectroscopy ◽  
Extracellular Vesicles ◽  
Cancer Detection ◽  
Label Free ◽  
Prostate Cancer Detection

The cholesterol metabolite 27-hydroxycholesterol increases the secretion of extracellular vesicles which promote breast cancer progression

Endocrinology ◽  
2021 ◽  
Author(s):  
Amy E Baek ◽  
Natalia Krawczynska ◽  
Anasuya Das Gupta ◽  
Svyatoslav Victorovich Dvoretskiy ◽  
Sixian You ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Tumor Growth ◽  
Cancer Cells ◽  
Extracellular Vesicles ◽  
Cancer Progression ◽  
Myeloid Cells ◽  
Breast Cancer Progression ◽  
Label Free ◽  
Tumor Growth And Metastasis ◽  
Promote Breast Cancer

Abstract Cholesterol has been implicated in the clinical progression of breast cancer, a disease that continues to be the most commonly diagnosed cancer in women. Previous work has identified the cholesterol metabolite, 27-hydroxycholesterol (27HC), as a major mediator of the effects of cholesterol on breast tumor growth and progression. 27HC can act as an estrogen receptor (ER) modulator to promote the growth of ERα+ tumors, and a liver x receptor (LXR) ligand in myeloid immune cells to establish an immune-suppressive program. In fact, the metastatic properties of 27HC require the presence of myeloid cells, with neutrophils (PMNs) being essential for the increase in lung metastasis in murine models. In an effort to further elucidate the mechanisms by which 27HC alters breast cancer progression, we made the striking finding that 27HC promoted the secretion of extracellular vesicles (EVs), a diverse assortment of membrane bound particles that include exosomes. The resulting EVs had a size distribution that was skewed slightly larger, compared to EVs generated by treating cells with vehicle. The increase in EV secretion and size was consistent across three different subtypes: primary murine PMNs, RAW264.7 monocytic cells and 4T1 murine mammary cancer cells. Label-free analysis of 27HC-EVs indicated that they had a different metabolite composition to those from vehicle-treated cells. Importantly, 27HC-EVs from primary PMNs promoted tumor growth and metastasis in two different syngeneic models, demonstrating the potential role of 27HC induced EVs in the progression of breast cancer. EVs from PMNs were taken up by cancer cells, macrophages and PMNs, but not T cells. Since EVs did not alter proliferation of cancer cells, it is likely that their pro-tumor effects are mediated through interactions with myeloid cells. Interestingly, RNA-seq analysis of tumors from 27HC-EV treated mice do not display significantly altered transcriptomes, suggesting that the effects of 27HC-EVs occur early on in tumor establishment and growth. Future work will be required to elucidate the mechanisms by which 27HC increases EV secretion, and how these EVs promote breast cancer progression. Collectively however, our data indicate that EV secretion and content can be regulated by a cholesterol metabolite, which may have detrimental effects in terms of disease progression, important findings given the prevalence of both breast cancer and hypercholesterolemia.

scholarly journals Comprehensive label-free characterization of extracellular vesicles and their surface proteins

2020 ◽  
Author(s):  
E. Priglinger ◽  
J. Strasser ◽  
B. Buchroithner ◽  
F. Weber ◽  
S. Wolbank ◽  
...  
Keyword(s):  
Quality Control ◽  
Extracellular Vesicles ◽  
Impedance Measurement ◽  
Surface Proteins ◽  
Label Free ◽  
Neuroprotective Effects ◽  
Characterization Methods ◽  
Minimal Sample ◽  
Flow Filtration

AbstractInterest in mesenchymal stem cell derived extracellular vesicles (MSC-EVs) as therapeutic agents has dramatically increased over the last decade. Preclinical studies show that MSC-EVs have anti-apoptotic and neuroprotective effects, boost wound healing, and improve the integration of allogeneic grafts through immunomodulation. Current approaches to the characterization and quality control of EV-based therapeutics include particle tracking techniques, Western blotting, and advanced cytometry, but standardized methods are lacking. In this study, we established and verified quartz crystal microbalance (QCM) as highly sensitive label-free immunosensing technique for characterizing clinically approved umbilical cord MSC-EVs enriched by tangential flow filtration and ultracentrifugation. Using QCM in conjunction with common characterization methods, we were able to specifically detect EVs via EV (CD9, CD63, CD81) and MSC (CD44, CD49e, CD73) markers and gauge their prevalence. Additionally, we characterized the topography and elasticity of these EVs by atomic force microscopy (AFM), enabling us to distinguish between EVs and non-vesicular particles (NVPs) in a therapeutic formulation. This measurement modality makes it possible to identify EV sub-fractions, discriminate between EVs and NVPs, and to characterize EV surface proteins, all with minimal sample preparation and using label-free measurement devices with low barriers of entry for labs looking to widen their spectrum of characterization techniques. Our combination of QCM with impedance measurement (QCM-I) and AFM measurements provides a robust multi-marker approach to the characterization of clinically approved EV formulations and opens the door to improved quality control.

scholarly journals Immuno-Acoustic Sorting of Disease-Specific Extracellular Vesicles by Acoustophoretic Force

Micromachines ◽  
2021 ◽  
Vol 12 (12) ◽  
pp. 1534
Author(s):  
Junyuan Liu ◽  
Yuxin Qu ◽  
Han Wang
Keyword(s):  
Extracellular Vesicles ◽  
Processing Time ◽  
Life Science ◽  
Biological Fluids ◽  
Label Free ◽  
Her2 Positive ◽  
Diagnosis And Prognosis ◽  
Efficient Recovery ◽  
Downstream Analysis ◽  
Disease Specific

Methods for the isolation and analysis of extracellular vesicles (EVs) have been extensively explored in the field of life science and in clinical diagnosis in recent years. The separation and efficient recovery of high-purity target EVs from biological samples are important prerequisites in the study of EVs. So far, commonly used methods of EV separation include ultracentrifugation, filtration, solvent precipitation and immunoaffinity capturing. However, these methods suffer from long processing time, EV damage and low enrichment efficiency. The use of acoustophoretic force facilitates the non-contact label-free manipulation of cells based on their size and compressibility but lacks specificity. Additionally, the acoustophoretic force exerted on sub-micron substances is normally weak and insufficient for separation. Here we present a novel immuno-acoustic sorting technology, where biological substances such as EVs, viruses, and biomolecules, can be specifically captured by antibody/receptor coated microparticles through immunoaffinity, and manipulated by an acoustophoretic force exerted on the microparticles. Using immuno-acoustic sorting technology, we successfully separated and purified HER2-positive EVs for further downstream analysis. This method holds great potential in isolating and purifying specific targets such as disease-related EVs from biological fluids and opens new possibilities for the EV-based early diagnosis and prognosis of diseases.

scholarly journals Early Detection and Investigation of Extracellular Vesicles Biomarkers in Breast Cancer

2021 ◽  
Vol 8 ◽  
Author(s):  
Erika Bandini ◽  
Tania Rossi ◽  
Emanuela Scarpi ◽  
Giulia Gallerani ◽  
Ivan Vannini ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Tumor Microenvironment ◽  
Early Detection ◽  
Extracellular Vesicles ◽  
Cancer Progression ◽  
Poor Prognosis ◽  
Cancer Death ◽  
Female Population ◽  
Flow Cytometric ◽  
Metastatic Development

Breast cancer (BC) is the most commonly diagnosed malignant tumor in women worldwide, and the leading cause of cancer death in the female population. The percentage of patients experiencing poor prognosis along with the risk of developing metastasis remains high, also affecting the resistance to current main therapies. Cancer progression and metastatic development are no longer due entirely to their intrinsic characteristics, but also regulated by signals derived from cells of the tumor microenvironment. Extracellular vesicles (EVs) packed with DNA, RNA, and proteins, are the most attractive targets for both diagnostic and therapeutic applications, and represent a decisive challenge as liquid biopsy-based markers. Here we performed a study based on a multiplexed phenotyping flow cytometric approach to characterize BC-derived EVs from BC patients and cell lines, through the detection of multiple antigens. Our data reveal the expression of EVs-related biomarkers derived from BC patient plasma and cell line supernatants, suggesting that EVs could be exploited for characterizing and monitoring disease progression.

scholarly journals Multi-modal nonlinear optical and thermal imaging platform for label-free characterization of biological tissue

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Wilson R. Adams ◽  
Brian Mehl ◽  
Eric Lieser ◽  
Manqing Wang ◽  
Shane Patton ◽  
...  
Keyword(s):  
Thermal Imaging ◽  
Multimodal Imaging ◽  
Live Cells ◽  
Label Free ◽  
Rodent Brain ◽  
Potential Applications ◽  
Research Questions ◽  
Nonlinear Imaging ◽  
Excised Tissues

AbstractThe ability to characterize the combined structural, functional, and thermal properties of biophysically dynamic samples is needed to address critical questions related to tissue structure, physiological dynamics, and disease progression. Towards this, we have developed an imaging platform that enables multiple nonlinear imaging modalities to be combined with thermal imaging on a common sample. Here we demonstrate label-free multimodal imaging of live cells, excised tissues, and live rodent brain models. While potential applications of this technology are wide-ranging, we expect it to be especially useful in addressing biomedical research questions aimed at the biomolecular and biophysical properties of tissue and their physiology.

scholarly journals Platelets Extracellular Vesicles as Regulators of Cancer Progression—An Updated Perspective

2020 ◽  
Vol 21 (15) ◽  
pp. 5195 ◽  
Author(s):  
Magdalena Żmigrodzka ◽  
Olga Witkowska-Piłaszewicz ◽  
Anna Winnicka
Keyword(s):  
Tumor Microenvironment ◽  
Extracellular Vesicles ◽  
Cancer Progression ◽  
Circulatory System ◽  
Eukaryotic Cells ◽  
Cancer Growth ◽  
Metastasis Formation ◽  
Pathological Conditions ◽  
Membrane Bound ◽  
The Impact

Extracellular vesicles (EVs) are a diverse group of membrane-bound structures secreted in physiological and pathological conditions by prokaryotic and eukaryotic cells. Their role in cell-to-cell communications has been discussed for more than two decades. More attention is paid to assess the impact of EVs in cancer. Numerous papers showed EVs as tumorigenesis regulators, by transferring their cargo molecules (miRNA, DNA, protein, cytokines, receptors, etc.) among cancer cells and cells in the tumor microenvironment. During platelet activation or apoptosis, platelet extracellular vesicles (PEVs) are formed. PEVs present a highly heterogeneous EVs population and are the most abundant EVs group in the circulatory system. The reason for the PEVs heterogeneity are their maternal activators, which is reflected on PEVs size and cargo. As PLTs role in cancer development is well-known, and PEVs are the most numerous EVs in blood, their feasible impact on cancer growth is strongly discussed. PEVs crosstalk could promote proliferation, change tumor microenvironment, favor metastasis formation. In many cases these functions were linked to the transfer into recipient cells specific cargo molecules from PEVs. The article reviews the PEVs biogenesis, cargo molecules, and their impact on the cancer progression.

scholarly journals An Electrokinetically-Driven Microchip for Rapid Entrapment and Detection of Nanovesicles

Micromachines ◽  
2020 ◽  
Vol 12 (1) ◽  
pp. 11
Author(s):  
Leilei Shi ◽  
Leyla Esfandiari
Keyword(s):  
Electrical Impedance ◽  
Disease Diagnosis ◽  
Electrical Impedance Spectroscopy ◽  
Label Free ◽  
Early Disease ◽  
Non Invasive ◽  
Diagnosis And Prognosis ◽  
Rapid Characterization ◽  
Impedance Sensor

Electrical Impedance Spectroscopy (EIS) has been widely used as a label-free and rapid characterization method for the analysis of cells in clinical research. However, the related work on exosomes (40–150 nm) and the particles of similar size has not yet been reported. In this study, we developed a new Lab-on-a-Chip (LOC) device to rapidly entrap a cluster of sub-micron particles, including polystyrene beads, liposomes, and small extracellular vesicles (exosomes), utilizing an insulator-based dielectrophoresis (iDEP) scheme followed by measuring their impedance utilizing an integrated electrical impedance sensor. This technique provides a label-free, fast, and non-invasive tool for the detection of bionanoparticles based on their unique dielectric properties. In the future, this device could potentially be applied to the characterization of pathogenic exosomes and viruses of similar size, and thus, be evolved as a powerful tool for early disease diagnosis and prognosis.

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