Serum and Lipoprotein Particle miRNA Profile in Uremia Patients

Axmann, Markus; Meier, Sabine M.; Karner, Andreas; Strobl, Wolfgang; Stangl, Herbert; Plochberger, Birgit

doi:10.3390/genes9110533

Cited by 18 publications

(15 citation statements)

References 43 publications

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“…At an epigenetic level, the presence of co-morbidities has also been shown to induce changes to the HDL-miRNA signature [60]. As such, an association between HDL-miRNAs profile and CAD stability has been established [97], and the content of HDL-miR-223 is found to be altered in the presence of insulin resistance and familial hypercholesterolemia subjects [26,[98][99][100][101][102]. Extending these previous findings, we have recently evidenced that hypercholesterolemic HDL particles transport higher levels of miR-126 as compared to native-HDL, which in turn is delivered to endothelial cells through an SRB1-dependent mechanism likely modulating HIF-1α expression [103].…”

Section: Impact Of Cardiovascular Risk Factor and Co-morbidities On Hmentioning

confidence: 99%

Advances in HDL: Much More than Lipid Transporters

Ben‐Aicha

Badimon

Vilahur

2020

IJMS

102

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High Density Lipoprotein (HDL) particles, beyond serving as lipid transporters and playing a key role in reverse cholesterol transport, carry a highly variable number of proteins, micro-RNAs, vitamins, and hormones, which endow them with the ability to mediate a plethora of cellular and molecular mechanisms that promote cardiovascular health. It is becoming increasingly evident, however, that the presence of cardiovascular risk factors and co-morbidities alters HDLs cargo and protective functions. This concept has led to the notion that metrics other than HDL-cholesterol levels, such as HDL functionality and composition, may better capture HDL cardiovascular protection. On the other hand, the potential of HDL as natural delivery carriers has also fostered the design of engineered HDL-mimetics aiming to improve HDL efficacy or as drug-delivery agents with therapeutic potential. In this paper, we first provide an overview of the molecules known to be transported by HDL particles and mainly discuss their functions in the cardiovascular system. Second, we describe the impact of cardiovascular risk factors and co-morbidities on HDL remodeling. Finally, we review the currently developed HDL-based approaches.

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Section: Impact Of Cardiovascular Risk Factor and Co-morbidities On Hmentioning

confidence: 99%

Advances in HDL: Much More than Lipid Transporters

Ben‐Aicha

Badimon

Vilahur

2020

IJMS

102

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“…MiR-16-5p is among the most abundant miRNA in EVs [ 22 ], while miR-21-5p is reported to be associated withs lipoproteins and could be involved in lipid metabolism [ 23 ]. We quantitatively detected miRNAs levels within NT, HT and S/D groups by ddPCR analysis; results are summarized in Figure 6 .…”

Section: Resultsmentioning

confidence: 99%

Extracellular Vesicles Analysis in the COVID-19 Era: Insights on Serum Inactivation Protocols towards Downstream Isolation and Analysis

Frigerio

Musicò

Brucale

et al. 2021

Cells

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Since the outbreak of the COVID-19 crisis, the handling of biological samples from confirmed or suspected SARS-CoV-2-positive individuals demanded the use of inactivation protocols to ensure laboratory operators’ safety. While not standardized, these practices can be roughly divided into two categories, namely heat inactivation and solvent-detergent treatments. These routine procedures should also apply to samples intended for Extracellular Vesicles (EVs) analysis. Assessing the impact of virus-inactivating pre-treatments is therefore of pivotal importance, given the well-known variability introduced by different pre-analytical steps on downstream EVs isolation and analysis. Arguably, shared guidelines on inactivation protocols tailored to best address EVs-specific requirements will be needed among the analytical community, yet deep investigations in this direction have not yet been reported. We here provide insights into SARS-CoV-2 inactivation practices to be adopted prior to serum EVs analysis by comparing solvent/detergent treatment vs. heat inactivation. Our analysis entails the evaluation of EVs recovery and purity along with biochemical, biophysical and biomolecular profiling by means of a set of complementary analytical techniques: Nanoparticle Tracking Analysis, Western Blotting, Atomic Force Microscopy, miRNA content (digital droplet PCR) and tetraspanin assessment by microarrays. Our data suggest an increase in ultracentrifugation (UC) recovery following heat treatment; however, it is accompanied by a marked enrichment in EVs-associated contaminants. On the other hand, solvent/detergent treatment is promising for small EVs (<150 nm range), yet a depletion of larger vesicular entities was detected. This work represents a first step towards the identification of optimal serum inactivation protocols targeted to EVs analysis.

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“…. Artificial enrichment increases the ratio by a factor of 10,000, which facilitates the estimation of the cellular lipoprotein uptake rate (no significant difference is detected using native lipoprotein particles 19 ). The high sensitivity of qPCR makes it possible to determine this uptake rate by measuring the number of miRNA strands after the incubation time and the miRNA/particle ratio.…”

Section: Discussionmentioning

confidence: 99%

Enrichment of Native Lipoprotein Particles with microRNA and Subsequent Determination of Their Absolute/Relative microRNA Content and Their Cellular Transfer Rate

Axmann¹,

Karner

Meier³

et al. 2019

JoVE

Self Cite

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Lipoprotein particles are predominately transporters of lipids and cholesterol in the bloodstream. Furthermore, they contain small amounts of strands of noncoding microRNA (miRNA). In general, miRNA alters the protein expression profile due to interactions with messenger-RNA (mRNA). Thus, knowledge of the relative and absolute miRNA content of lipoprotein particles is essential to estimate the biological effect of cellular particle uptake. Here, a quantitative real-time polymerase chain reaction (qPCR)-based protocol is presented to determine the absolute miRNA content of lipoprotein particles-exemplified shown for native and miRNA-enriched lipoprotein particles. The relative miRNA content is quantified using multiwell microfluidic array cards. Furthermore, this protocol allows scientists to estimate the cellular miRNA and, thus, the lipoprotein particle uptake rate. A significant increase of the cellular miRNA level is observable when using high-density lipoprotein (HDL) particles artificially loaded with miRNA, whereas incubation with native HDL particles yields no significant effect due to their rather low miRNA content. In contrast, the cellular uptake of low-density lipoprotein (LDL) particles-neither with native miRNA nor artificially loaded with it-did not alter the cellular miRNA level.

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Serum and Lipoprotein Particle miRNA Profile in Uremia Patients

Cited by 18 publications

References 43 publications

Advances in HDL: Much More than Lipid Transporters

Advances in HDL: Much More than Lipid Transporters

Extracellular Vesicles Analysis in the COVID-19 Era: Insights on Serum Inactivation Protocols towards Downstream Isolation and Analysis

Enrichment of Native Lipoprotein Particles with microRNA and Subsequent Determination of Their Absolute/Relative microRNA Content and Their Cellular Transfer Rate

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