Mass spectrometric detection of 27-hydroxycholesterol in breast cancer exosomes

Røberg-Larsen, Hanne; Lund, Kaja; Seterdal, Kristina Erikstad; Solheim, Stian; Vehus, Tore; Solberg, Nina; Krauß, Stefan; Lundanes, Elsa; Wilson, Steven Ray

doi:10.1016/j.jsbmb.2016.02.006

Cited by 69 publications

(48 citation statements)

References 32 publications

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“…Notably, EVs may also contain metabolites of cholesterol: 27-hydroxycholesterol was found in exosomes secreted by an estrogen-receptor-positive breast cancer cell line (21). Thus, hydroxysterol-containing EVs may serve as diagnostic markers for specific diseases.…”

Section: Cholesterol Content Of Evsmentioning

confidence: 99%

Thematic Review Series: Exosomes and Microvesicles: Lipids as Key Components of their Biogenesis and Functions, Cholesterol and the journey of extracellular vesicles

Pfrieger

Vitale

2018

Journal of Lipid Research

106

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Section: Cholesterol Content Of Evsmentioning

confidence: 99%

Thematic Review Series: Exosomes and Microvesicles: Lipids as Key Components of their Biogenesis and Functions, Cholesterol and the journey of extracellular vesicles

Pfrieger

Vitale

2018

Journal of Lipid Research

106

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“…Similar detection capabilities have been achieved in OT format regarding peptides10. Compared to a standard LC-system18, 100 times less sample could be introduced, and still achieving the same signal (Fig. 3).…”

Section: Resultsmentioning

confidence: 54%

“…Using a linear gradient on the attoLC-MS system, hydroxylated cholesterol (OHC) isomers could be baseline/satisfactory separated from each other (Supplementary Figure 5). Although the attoLC columns have meter-scale length, OHC retention times are similar to standard LC format18 (t R about 15 minutes in both cases), as the linear velocity of the attoLC mobile phase is significantly higher. Re-equilibration times (pressure-dependent when using Thermo nanoLC pumps) are also similar, as long, open columns generate approximately the same back-pressure as shorter, particle-packed columns.…”

Section: Resultsmentioning

confidence: 93%

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Versatile, sensitive liquid chromatography mass spectrometry – Implementation of 10 μm OT columns suitable for small molecules, peptides and proteins

Vehus

Røberg-Larsen

Waaler

et al. 2016

Sci Rep

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We have designed a versatile and sensitive liquid chromatographic (LC) system, featuring a monolithic trap column and a very narrow (10 μm ID) fused silica open tubular liquid chromatography (OTLC) separation column functionalized with C18-groups, for separating a wide range of molecules (from small metabolites to intact proteins). Compared to today’s capillary/nanoLC approaches, our system provides significantly enhanced sensitivity (up to several orders) with matching or improved separation efficiency, and highly repeatable chromatographic performance. The chemical properties of the trap column and the analytical column were fine-tuned to obtain practical sample loading capacities (above 2 μg), an earlier bottleneck of OTLC. Using the OTLC system (combined with Orbitrap mass spectrometry), we could perform targeted metabolomics of sub-μg amounts of exosomes with 25 attogram detection limit of a breast cancer-related hydroxylated cholesterol. With the same set-up, sensitive bottom-up proteomics (targeted and untargeted) was possible, and high-resolving intact protein analysis. In contrast to state-of-the-art packed columns, our platform performs chromatography with very little dilution and is “fit-for-all”, well suited for comprehensive analysis of limited samples, and has potential as a tool for challenges in diagnostics.

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“…Impressively, using EADSA and GT derivatisation Roberg-Larsen et al. were able to show that 26-HC is elevated in exosomes from an ER+ breast cancer cell line derived from only 200,000 cells [72]. This group use low flow-rate chromatography with ESI to enhance sensitivity and incorporate on-line solid-phase extraction for sample clean-up.…”

Section: Analytical Improvementsmentioning

confidence: 99%

Cholesterolomics: An update

Griffiths

Abdel-Khalik

Yutuc

et al. 2017

Analytical Biochemistry

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Cholesterolomics can be regarded as the identification and quantification of cholesterol, its precursors post squalene, and metabolites of cholesterol and of its precursors, in a biological sample. These molecules include 1,25-dihydroxyvitamin D3, steroid hormones and bile acids and intermediates in their respective biosynthetic pathways. In this short article we will concentrate our attention on intermediates in bile acid biosynthesis pathways, in particular oxysterols and cholestenoic acids. These molecular classes are implicated in the aetiology of a diverse array of diseases including autoimmune disease, Parkinson's disease, motor neuron disease, breast cancer, the lysosomal storage disease Niemann-Pick type C and the autosomal recessive disorder Smith-Lemli-Opitz syndrome. Mass spectrometry (MS) is the dominant technology for sterol analysis including both gas-chromatography (GC)-MS and liquid chromatography (LC)-MS and more recently matrix-assisted laser desorption/ionisation (MALDI)-MS for tissue imaging studies. Here we will discuss exciting biological findings and recent analytical improvements.

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Mass spectrometric detection of 27-hydroxycholesterol in breast cancer exosomes

Cited by 69 publications

References 32 publications

Thematic Review Series: Exosomes and Microvesicles: Lipids as Key Components of their Biogenesis and Functions, Cholesterol and the journey of extracellular vesicles

Thematic Review Series: Exosomes and Microvesicles: Lipids as Key Components of their Biogenesis and Functions, Cholesterol and the journey of extracellular vesicles

Versatile, sensitive liquid chromatography mass spectrometry – Implementation of 10 μm OT columns suitable for small molecules, peptides and proteins

Cholesterolomics: An update

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