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At least 649 records · Page 36

Extracellular vesicles promote epithelial-to-mesenchymal transition of lens epithelial cells under oxidative stress

Posterior capsule opacification (PCO), resulting from residual lens epithelial cell (LEC) epithelial–mesenchymal transition (EMT), abnormal proliferation, and migration, is the most common complication of cataract surgery. A recent study determined that extracellular vesicles (EVs) and reactive oxygen species (ROS) regulate the EMT process during cutaneous wound healing and tumour metastasis. However, their underlying mechanism in PCO is unclear. In this study, we examined the secreted EVs from a scratch model in vitro. We found that the production of ROS was increased after mechanical injury, especially at the wound edge, and there was an increased viability of LECs, which can be blocked by diphenyleneiodonium, an NADPH oxidase inhibitor. Cell viability and migration were increased upon treatment with 1 μM H{sub 2}O{sub 2}, but significantly reduced when the concentration of H{sub 2}O{sub 2} increased to 100 μM. Transwell assay showed that both post-surgery LECs and LECs treated with 1 μM H{sub 2}O{sub 2} significantly induced the migration of normal LECs by EV secretion. Extraction and quantification of EVs derived from injured and H{sub 2}O{sub 2}-treated LECs showed a similar increase in production. Co-incubation of EVs from both injured and H{sub 2}O{sub 2}-treated LECs with normal LECs and organ-cultured mouse lenses activated EMT, which was attenuated by a ROS inhibitor. These results suggest that EVs participate in ROS-induced lens EMT, making EVs a potential target for treating PCO.

60 APPLIED LIFE SCIENCES↗

Cultured cardiac fibroblasts and myofibroblasts express Sushi Containing Domain 2 and assemble a unique fibronectin rich matrix

Highlights: • Cultured cardiac fibroblasts and myofibroblasts derived from normal human hearts express the novel marker SUSD2. • Under high density culture conditions, human cardiac myofibroblasts deposit a matrix scaffold rich in fibronectin. • Cultured dermal fibroblasts do not express SUSD2 and did not for form scaffolds in vitro. Cardiac fibroblasts and myofibroblasts assemble and maintain extracellular matrix during normal development and following injury. Culture expansion of these cells yield a bioengineered matrix that could lead to intriguing therapeutic opportunities. For example, we reported that cultured rat cardiac fibroblasts form a matrix that can be used to delivery therapeutic stem cells. Furthermore, we reported that matrix derived from cultured human cardiac fibroblasts/myofibroblasts converted monocytes into macrophages that express interesting anti-inflammatory and pro-angiogenic properties. Expanding these matrix investigations require characterization of the source cells for quality control. In these efforts, we observed and herein report that Sushi Containing Domain 2 (SUSD2) is a novel and consistent marker for cultured human cardiac fibroblast and myofibroblasts.

60 APPLIED LIFE SCIENCES↗

Single-cell RNA-seq revealed diverse cell types in the mouse placenta at mid-gestation

The mammalian placenta consists of a set of cells to ensure normal placental functions throughout gestation. Dysfunctional placentae are considered as the origin of a series of pregnancy complications. Therefore, it is urgent for detailed information about the molecular recipes of the cell types within the normal placenta. In the past years, gene expression analysis via single-cell RNA-seq (scRNA-seq) offers opportunities to identify new cell types in a variety of organs and tissues. In this study, scRNA-seq was used to explore the cell heterogeneity within the E10.5 mouse placenta and unravel their discrepancies in cell composition and communications. We identified sixteen cell clusters, including some cell clusters that originated from the maternal tissue. Moreover, we traced the developmental trajectories of trophoblasts and Hofbauer-like cells. Further analysis revealed cell connections between the endothelial cells and pericytes, syncytiotrophoblasts, as well as decidual cells. Besides, we highlighted several signaling pathways, such as the EGF, FGF, canonical, and non-canonical WNT signaling pathways, which mediated the potential crosstalk between different cell types within placenta. Our research provides an in-depth understanding of placental development, cellular composition, and communications at the maternal-fetal interface.

60 APPLIED LIFE SCIENCES↗

ACBD3 is up-regulated in gastric cancer and promotes cell cycle G1-to-S transition in an AKT-dependent manner

Highlights: • ACBD3 is up-regulated in gastric cancer. • ACBD3 regulates cell-cycle of gastric cancer in an AKT-dependent manner. It has been reported that ACBD3 is closely related to the malignant process of cells, but its role in gastric cancer has not been elucidated. This study aims to investigate the expression and function of ACBD3 in human gastric cancer. The Cancer Genome Atlas (TCGA) database were selected to analyze mRNA levels of ACBD3 in gastric cancer tissues and normal gastric epithelial tissues. qPCR and Western blot were conducted to detect the expression of ACBD3 in two normal gastric epithelial cell lines and five gastric cancer cell lines which were cultured in our laboratory. To exclude differences in individual background between different patients, we further detected the expression of ACBD3 in 8 pairs of malignant/non-malignant clinical gastric tissues. Through the establishment of stable cells, in vitro cell experiments and in vivo xenotransplantation models in mice, the role of ACBD3 in the proliferation of gastric cancer cells has been further explored. AKT inhibitors were used to deeply explore the molecular regulation mechanism of ACBD3. The results showed that the elevated ACBD3 in gastric cancer tissue were positively correlated with the clinical grade and prognosis of gastric cancer. In terms of molecular function, we found that ACBD3 can enhance the production and growth of gastric cancer cells. At the same time, the activation of AKT kinase played an important role in ACBD3's promotion of G1-to-S transition. The experiments generally indicate that ACBD3 is expected to become a potential diagnostic molecule or therapeutic target for gastric cancer.

60 APPLIED LIFE SCIENCES↗

Heterogeneous response of cancer-associated fibroblasts to the glucose deprivation through mitochondrial calcium uniporter

Highlights: • CAFs from a tumor mass are heterogeneous under the glucose deficient condition. • CAFs survive under glucose deficient condition by activation of TGF-β and calcium. • Calcium influx to mitochondria is critical for survival of glucose-deficient CAFs. • MCU in CAFs could be a feasible target for the development of anticancer drugs. Cancer-associated fibroblasts (CAFs) are an abundant component of the tumor microenvironment and have distinct features from normal fibroblasts (NFs). However, the discriminative nature of heterogeneous CAFs under glucose starvation remains unknown. In this study, we investigated the changes in the mitochondrial calcium concentration and relevant intracellular machinery in CAFs under glucose-deficient conditions. Xenografted tumor masses were dissected into multiple pieces and subjected to the CAF isolation using magnetically activated cell sorting (MACS). NFs were separated from the normal lung and skin. Under glucose starvation, CAFs from the tumor mass exhibited heterogeneity in cell proliferation, ATP production and calcium concentration. Compared to NFs, mitochondrial calcium concentration was significantly higher in glucose-starved CAFs with upregulation of mitochondrial calcium uniporter (MCU) that led to enhancement of ATP production and cell growth. Intriguingly, treatment of glucose-starved CAFs with oligomycin increased apoptosis by disrupted calcium homeostasis following overactivation of the mPTP. Moreover, oligomycin-induced apoptosis was mitigated by calcium chelation. This study demonstrated that the discriminative calcium influx to mitochondria through MCU coordinated cell growth and apoptosis in glucose-starved CAFs but not in NFs.

60 APPLIED LIFE SCIENCES↗

The oncogenic role of meiosis-specific Aurora kinase C in mitotic cells

Highlights: • AURKC is expressed in meiotic cells and some cancers. • AURKC expression increases cell migration and transformation of human mitotic cells. • Deletion of AURKC in U2OS cells decreases cell migration and soft agar growth. • Transcriptome analysis reveals that AURKC alters the extracellular matrix. Aberrant expression of meiosis-specific genes in cancer has recently emerged as a driver of some cancer formation. Aurora kinase C (AURKC) is a member of the Aurora kinase family of proteins known to regulate chromosome segregation during cell divisions. AURKC is normally expressed in meiotic cells; however, elevated levels of AURKC mRNA and protein are frequently measured in cancer cells. To understand the function of AURKC in cancer cells, expression was induced in noncancerous, human retina pigmented epithelial cells. While AURKC expression did not alter cell proliferation over 72 h, it did increase cell migration and anchorage independent growth in soft agar suggesting an oncogenic role in mitotically dividing cells. To evaluate AURKC as a potential therapeutic target, a frameshift mutation in the gene was introduced in U2OS osteosarcoma cells using CRISPR-Cas9 technology resulting in a premature stop codon. Cancer cells lacking AURKC displayed no change in cell proliferation over 72 h but did migrate less and formed fewer colonies in soft agar. Whole transcriptome sequencing analysis uncovered over 400 differentially expressed genes in U2OS cells with and without AURKC. GO analysis revealed alterations in proteinaceous extracellular matrix genes including COL1A1. These data indicate that therapeutics targeting AURKC could decrease cancer cell metastasis and disease progression. Because AURKC is transcriptionally silenced in normal mitotic cells, its disruption could specifically target cancer cells limiting the toxic side effects associated with current therapeutics.

60 APPLIED LIFE SCIENCES↗

Collective effect of thermal plumes on temperature fluctuations in a closed Rayleigh–Benard convection cell

Here, we report a systematic study of the collective effect of thermal plumes on the probability density function (PDF) P(δT) of temperature fluctuations δT(t) in turbulent Rayleigh-Benard convection. By decomposing δT(t) into four basic fluctuation modes associated with single and multiple warm and cold plumes and a turbulent background, we derive an analytic form of P(δT) based on the convolutions of the five independent modes. To test the derived form of P(δT) in the multiple-plume regions, where the thermal plumes are heavily populated, we conduct time series measurements of temperature fluctuations in two convection cells; one is a vertical thin disk and the other is an upright cylinder of aspect ratio unity. For a given normalized position in most regions of the convection cell, all of the measured PDFs P(δT) for different Rayleigh numbers fall onto a single master curve, once δT is normalized by its rms value σ T . It is found that the measured P(δT / σT ) at different locations along the symmetric horizontal and vertical axes of the convection cells can all be well described by the derived form of P(δT / σT ). The fitted values of the parameters associated with the number of plumes in multiple plume clusters and their relative strengths and degrees of intermittency are closely linked to the spatial distribution of thermal plumes and local dynamics of the large-scale circulation in a closed convection cell. Our work thus provides a unified theoretical approach for understanding scalar PDFs in a turbulent field, which is very useful not only for the present study but also for the study of many turbulent mixing problems of practical interest.

42 ENGINEERING↗

Odd-viscosity-induced passivation of Moffatt vortices

Odd viscosity is associated with a number of striking physical phenomena: a disk rotating in a viscous liquid experiences a normal compressive or tensile stress depending on its sense of rotation; chiral active matter interfacial deformations level off in a manner akin to surface tension; and unstable thermocapillary films become stabilized. In this article we show that odd viscosity decreases the size and intensity of Moffatt vortices in a corner formed by a solid substrate and a free interface. The effect is present because odd viscosity creates a normal-stress-induced shear stress at the liquid–gas interface. The new shear stress is out of phase and oppositely directed with respect to the interfacial liquid velocity. It squeezes the vortices and damps their rotation. As a consequence, Moffatt vortices can now be observed with greater facility in an experiment. Odd viscosity also increases the range of corner angles over which Moffatt vortices can be observed, and, in general, it shows a tendency to moderate singular behaviour.

Mechanics↗

Transition to turbulence in viscoelastic channel flow of dilute polymer solutions

The transition to turbulence in a plane Poiseuille flow of dilute polymer solutions is studied by direct numerical simulations of a finitely extensible nonlinear elastic fluid with the Peterlin closure. The range of Reynolds number ($Re$)$2000 \le Re \le 5000$is studied but with the same level of elasticity in viscoelastic flows. The evolution of a finite-amplitude perturbation and its effects on the transition dynamics are investigated. A viscoelastic flow begins transition at an earlier time than its Newtonian counterparts, but the transition time appears to be insensitive to polymer concentration in the dilute and semi-dilute regimes studied. Increasing polymer concentration, however, decreases the maximum attainable energy growth during the transition process. The critical or minimum perturbation amplitude required to trigger transition is computed. Interestingly, both Newtonian and viscoelastic flows follow almost the same power-law scaling of$Re^\gamma$with the critical exponent$\gamma \approx -1.25$, which is in close agreement with previous studies. However, a shift downward is observed for viscoelastic flow, suggesting that smaller perturbation amplitudes are required for the transition. A mechanism of the early transition is investigated by the evolution of wall-normal and spanwise velocity fluctuations and flow structure. The early growth of these fluctuations and the formation of quasi-streamwise vortices around low-speed streaks are promoted by polymers, hence causing an early transition. These vortical structures are found to support the critical exponent$\gamma \approx -1.25$. Once the transition process is completed, polymers play a role in dampening the wall-normal and spanwise velocity fluctuations and vortices to attain a drag-reduced state in viscoelastic turbulent flows.

Mechanics↗

Kinetic-ballooning-mode turbulence in low-average-magnetic-shear equilibria

In this work, kinetic-ballooning-mode (KBM) turbulence is studied via gyrokinetic flux-tube simulations in three magnetic equilibria that exhibit small average magnetic shear: the Helically Symmetric eXperiment (HSX), the helical-axis Heliotron-J and a circular tokamak geometry. For HSX, the onset of KBM being the dominant instability at low wavenumber occurs at a critical value of normalized plasma pressure $\beta ^{\rm KBM}_{\rm crit}$ that is an order of magnitude smaller than the magnetohydrodynamic (MHD) ballooning limit $\beta ^{\rm MHD}_{\rm crit}$ when a strong ion temperature gradient (ITG) is present. However, $\beta ^{\rm KBM}_{\rm crit}$ increases and approaches the MHD ballooning limit as the ITG tends to zero. For these configurations, $\beta ^{\rm KBM}_{\rm crit}$ also increases as the magnitude of the average magnetic shear increases, regardless of the sign of the normalized magnetic shear. Simulations of Heliotron-J and a circular axisymmetric geometry display behaviour similar to HSX with respect to $\beta ^{\rm KBM}_{\rm crit}$ . Despite large KBM growth rates at long wavelengths in HSX, saturation of KBM turbulence with $\beta > \beta _{\rm crit}^{\rm KBM}$ is achievable in HSX and results in lower heat transport relative to the electrostatic limit by a factor of roughly five. Nonlinear simulations also show that KBM transport dominates the dynamics when KBMs are destabilized linearly, even if KBM growth rates are subdominant to ITG growth rates.

70 PLASMA PHYSICS AND FUSION TECHNOLOGY↗

Direct, simple and efficient computation of all components of the virtual-casing magnetic field in axisymmetric geometries with Kapur–Rokhlin quadrature

In a recent publication (Toler et al., J. Plasma Phys., vol. 89, issue 2, 2023, p. 905890210), we demonstrated that for axisymmetric geometries, the Kapur–Rokhlin quadrature rule provided an efficient and high-order accurate method for computing the normal component, on the plasma surface, of the magnetic field due to the toroidal current flowing in the plasma, via the virtual-casing principle. The calculation was indirect, as it required the prior computation of the magnetic vector potential from the virtual-casing principle, followed by the computation of its tangential derivative by Fourier differentiation, to obtain the normal component of the magnetic field. Our approach did not provide the other components of the virtual-casing magnetic field. In this letter, we show that a more direct and more general approach is available for the computation of the virtual-casing magnetic field. The Kapur–Rokhlin quadrature rule accurately calculates the principal value integrals in the expression for all the components of the magnetic field on the plasma boundary, and the numerical error converges at a rate nearly as high as the indirect method we presented previously.

70 PLASMA PHYSICS AND FUSION TECHNOLOGY↗

Isotopic Fractionation and Kinetic Isotope Effects of a Purified Bacterial Nitric Oxide Reductase (NOR)

Nitrous oxide (N 2 O) is a serious concern due to its role in global warming and ozone destruction. Agricultural practices account for ~80% of all anthropogenic N 2 O produced in the US, due in large part to the stimulation of microbial denitrification. Stable isotopes are uniquely suited to examine both microbial N 2 O sources and the mechanism of N 2 O biosynthesis through the use of 2 Site Preference (δ 15 N SP ; the difference in δ 15 N between the central and outer N atoms in N 2 O) and kinetic isotope effects (KIEs), respectively. Using trace gas isotope ratio mass spectrometry (TG-IRMS), we determined the δ 15 N, δ 15 N α , δ 15 N β , and δ 18 O of N 2 O produced by a purified cytochrome c nitric oxide reductase (cNOR) from Paracoccus denitrificans. We also calculated δ 15 N SP , the KIEs, and associated isotopic enrichment factors (ε) for N bulk , N α , and N β . A normal isotope effect was observed for bulk 15 N, with a KIE value of 1.0086 ± 0.0009 (ε = -8.6 ± 0.9‰). The isotope effects for both 15 N α and 15 N β were also normal, with position-specific KIEs of 1.0072 ± 0.0010 (ε = -7.2 ± 1.0‰) and 1.0100 ± 0.0010 (ε = -9.9 ± 1.0‰), respectively, and δ 15 N SP values ranged from 0.5 to 8.7‰ with no significant trend as the reaction proceeded. Values of δ 18 O increased with N 2 O production (slope of δ 18 O against [-flnf/(1-f)] = -19.9 ± 1.9‰). We present implications for the mechanism of N 2 O production from cNOR based on our data.

Rivett, Elise D. [Michigan State Univ., East Lansi↗

Benchmarking Concentration and Extraction Methods for Wastewater-Based Surveillance of Eight Human Respiratory Viruses: Implications for Rapid Application to Novel Pathogens

To provide early warning and support a rapid response to a novel virus through wastewater surveillance, it would be ideal to understand in advance which concentration and extraction methods are likely to be effective for dPCR-based methods, depending on virus characteristics. In this study, we spiked raw wastewater samples with eight human respiratory viruses and processed them with four methods that concentrate and/or extract nucleic acids from both liquid and solid fractions (Promega, Nanotrap, and InnovaPrep) or only the solid fraction of wastewater (Solids). Our findings provide encouraging evidence that all four methods combined with dPCR could detect an emerging virus in wastewater, although they differed in sensitivity. The pattern of recovery efficiency for adenoviruses, coronaviruses, and influenza A viruses was consistent across methods, with Promega producing higher median recovery efficiencies, while distinct patterns were observed for coxsackieviruses. We also normalized the concentration data with two endogenous fecal indicators, PMMoV and Carjivirus (formerly crAssphage). We found that normalization could reduce method-associated differences if the indicator exhibited a recovery pattern similar to that of the target virus. These findings can guide the selection of concentration and extraction methods for wastewater monitoring based on the properties of target viruses, thus enhancing pandemic preparedness.

Biological and medical sciences↗

Identification of KLHL12 Ligands Using Fragment-Based Methods

Targeted protein degradation can be induced by recruiting a protein of interest to an E3 ligase, resulting in its ubiquitination and subsequent proteasome-mediated degradation. However, only a small number of E3 ligases have been utilized for degradation. Expansion of the repertoire of useful E3 ligases via the identification of ligands to those ligases could broaden the scope and applicability of the degradation paradigm. We have identified KLHL12 as an E3 ligase with higher expression in cancer over normal tissues. We report here the use of NMR-based screening to identify fragments that bind to KLHL12, and X-ray structures of a fragment hit bound to KLHL12. Using this structural information, we optimized the hits, leading to the first reported small molecules that bind to KLHL12 with submicromolar affinity. Derivatives of these compounds may be useful for the construction of PROTACs to selectively degrade protein targets in tumors while sparing normal cells.

Amines↗

High Yield of B-Side Electron Transfer at 77 K in the Photosynthetic Reaction Center Protein from Rhodobacter sphaeroides

In this work, the primary electron transfer (ET) processes at 295 K and 77 K are compared for the Rhodobacter sphaeroides reaction center (RC) pigment-protein complex from thirteen mutants including a wild-type control. The engineered RCs bear mutations in the L and M polypeptides that largely inhibit ET from the excited state P* of the primary electron donor (P, a bacteriochlorophyll dimer) to the normally photoactive A-side cofactors and enhance ET to the C 2 -symmetry related, and normally photoinactive, B-side cofactors. P* decay is multi-exponential at both temperatures and modeled as arising from subpopulations that differ in contributions of two-step ET (e.g., P* → P + B B – → P + H B – ), one-step superexchange ET (e.g., P* → P + H B – ) and P* → ground state. [H B and B B are monomeric bacteriopheophytin and bacteriochlorophyll, respectively.] The relative abundances of the subpopulations and the inherent rate constants of the P* decay routes vary with temperature. Regardless, ET to produce P + H B – is generally faster at 77 K than at 295 K by about a factor of two. A key finding is that the yield of P + H B – , which ranges from ~5% to ~90% among the mutant RCs, is essentially the same at 77 K as at 295 K in each case. Overall, the results show that ET from P* to the B-side cofactors in these mutants does not require thermal activation and involves combinations of ET mechanisms analogous to those operative on the A side in the native RC.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Infrared Spectroscopy of Li + Solvation in EmimBF 4 and in Propylene Carbonate: Ab Initio Molecular Dynamics and Experiment

Infrared (IR) spectra of solutions of the lithium salt LiBF 4 in the ionic liquid 1-ethyl-3-methylimidazolium tetrafluoroborate (EmimBF 4 ) and in the organic solvent propylene carbonate (PC) are studied via infrared spectroscopy and ab initio molecular dynamics (AIMD) simulations. The measurements show that the major effects of LiBF 4 in both solutions, compared to their neat counterparts, are the appearance of a new broad band in the 300-450 cm -1 frequency region and a broadening of the IR structure in the 900-1200 cm -1 region with the development of a new peak at 980 cm -1 . Computational analysis indicates that hindered translational motions of Li + in its solvation cage are mainly responsible for the former, while the latter is due to Li + -induced structural changes and accompanying vibrational frequency shifts of constituent ions and molecules of the solutions. In addition, molecular motions in these and lower frequency regions are generally correlated, disclosing the collective nature of the vibrational dynamics, which involve multiple ions/molecules. Herein, detailed analysis of these features via AIMD simulations of the spectrum and its components arising from auto- and cross-correlations of motions of constituent molecular species, combined with generalized normal modes of the solutions and normal modes of small Li + -containing clusters, is presented. Furthermore, other minor spectral changes caused by the lithium salt as well as the interaction-induced effect on IR spectra are also discussed.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Observation of [VCu1–Ini2+VCu1–] Defect Triplets in Cu-Deficient CuInS2

Copper indium disulfide (CuInS2) is a semiconductor with a direct energy band gap of 1.53 eV, an optimal value for highly efficient thin-film solar cells. But it has reached only ~ 11 % power conversion efficiency, far less than the theoretically achievable value of ~ 30 %. The cause of this low performance is not understood. A single crystal grown from 1 mol% Cu-deficient melt was studied, using atomic resolution high-angle annular dark-field (HAADF) scanning transmission electron microscopy (STEM) and electron dispersive spectroscopy (EDS). While the bulk crystal is exactly stoichiometric CuInS2, it contains nanometer-thick, structurally coherent, Cu-deficient interphases that form along rotational twin boundaries in the {112} plane. Transition zones from bulk crystal to interphase are observed, where In is seen to move from its normal site InIn in the chalcopyrite structure to a tetrahedral interstitial site Ini, while Cu remains in its normal CuCu position. Two InIn rows of the bulk crystal merge into one row of Ini, causing excess Ini in the interphase. The concentrations of CuCu and Ini reflect a ratio of Cu vacancies, VCu, to excess Ini of ~ 2. Their relative lattice positions, and the high electrical resistivity of the crystal, suggests that VCu and excess Ini ‘precipitate’ as self-compensating, electrically neutral, [VCu1- Ini2+ VCu1-] defect triplets. This is the first atomic-level observation of the ordered defect that has been invoked as the basic structural modifier in chalcopyrite compound homologs. The interphases introduce an optical gap of 1.47 eV. Electron trapping in band tail states, evident from a photoconductivity exponent of 0.54, is the likely cause of an unusually low electron mobility of 0.1 cm2V-1s-1. The overall result is that making CuInS2 slightly copper-poor inserts nanometer-thick layers of the interphase into the bulk crystal. This study shows that apparently conflicting results of the effect of Cu deficiency on CuInS2 thin-film solar cells may be resolved by analyzing structure and composition at nanometer spatial resolution.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

In-Depth Proteome Profiling of Small Extracellular Vesicles Isolated from Cancer Cell Lines and Patient Serum

Extracellular Vesicle (EV) secretion has been observed from most types of normal and tumor cells. These EVs contain a variety of distinctive cargo where tumor-derived serum proteins in EVs secreted into the bloodstream can be used to provide a minimally invasive method for clinical monitoring. We have therefore undertaken a comprehensive study of the protein content of the EVs from several cancer cell lines using direct Data Independent Analysis (DIA). Further, several thousand proteins were detected including many of the classic EV markers such as CD9, CD81, CD63, TSG101 and Syndecan-1 among others. Many distinctive cancer related proteins were detected and some of them are known to be potential markers of cancer detection and monitoring of cancer progression. We have further studied the protein content of EVs from patient serum for both normal controls and pancreatic (PC) and Hepatocellular Carcinoma (HCC) cancers. The EVs for these studies have been isolated by various methods for comparison including ultracentrifugation (UC) and CD9 immunoaffinity column (CD9-IC). Typically, 500-1,000 proteins were identified where most of them overlapped with the EV proteins identified from the cell lines studied. Many of the EV protein markers were identified in these serum EVs and large numbers of proteins related to pancreatic and HCC cancers were identified.

60 APPLIED LIFE SCIENCES↗