Niemann Pick C2 protein enables cholesterol transfer from endo-lysosomes to the plasma membrane for efflux by shedding of extracellular vesicles

Juhl, Alice Dupont; Lund, Frederik Wendelboe; Jensen, Maria; Szomek, Maria; Heegaard, Christian W.; Werner, Stephan; McNally, James G.; Schneider, Gerd; Kapishnikov, Sergey; Wüstner, Daniel

doi:10.1016/j.chemphyslip.2020.105047

Cited by 25 publications

(50 citation statements)

References 82 publications

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“…In line with this conclusion is our recent observation that treating NPC2-deficient human fibroblasts with purified NPC2 mobilized cholesterol from endolysosomes towards the PM, which was paralleled by reallocation of endo-lysosomes to the periphery and direct sterol transfer to the PM (Juhl et al, 2021). The latter could be shown by bleaching TopFluor-cholesterol in a portion of the PM and measuring a decrease in fluorescence of this sterol probe in nearby LE/LYSs underneath the PM (Juhl et al, 2021). Using kinetic modeling of such a fluorescence loss in photobleaching (FLIP) experiment, we inferred a dynamic pool of sterol in endolysosomes in exchange with the PM, which has a residence time in LE/LYSs of about 40 s (Juhl et al, 2021).…”

Section: Lipids Npc Proteins and Atp-binding Cassette Transporters Co...supporting

confidence: 55%

“…These important observations suggest that NPC2 is needed to solubilize cholesterol inside of the lysosome and donate it to different transporters including NPC1 and ABCA1 for export from this compartment. In line with this conclusion is our recent observation that treating NPC2-deficient human fibroblasts with purified NPC2 mobilized cholesterol from endolysosomes towards the PM, which was paralleled by reallocation of endo-lysosomes to the periphery and direct sterol transfer to the PM (Juhl et al, 2021). The latter could be shown by bleaching TopFluor-cholesterol in a portion of the PM and measuring a decrease in fluorescence of this sterol probe in nearby LE/LYSs underneath the PM (Juhl et al, 2021).…”

Section: Lipids Npc Proteins and Atp-binding Cassette Transporters Co...mentioning

confidence: 58%

“… Visualization of extracellular vesicles and particles during cholesterol efflux. (A) Spinning disk microscopy image of extracellular vesicles from a NPC2-deficient fibroblasts treated with NPC2 protein, tagged with a red Alexa546 fluorophore (red), and labeled additionally with TopFluor-cholesterol (green) ( Juhl et al, 2021 ). (B) Confocal microscopy image of migrasomes from L929 cell transfected with TSPAN4-GFP ( Chen et al, 2018 ).…”

Section: Origin Structure and Function Of Cholesterol-containing Extr...mentioning

confidence: 99%

“…(D,D′,D′′) examples of extracellular vesicles from NPC2 deficient and healthy fibroblasts imaged with cryo-SXT. Scalebar 0.5 µm for (D) and 0.15 µm in (D′,D′′) ( Juhl et al, 2021 ). Red arrows point to microvesicles.…”

Section: Origin Structure and Function Of Cholesterol-containing Extr...mentioning

confidence: 99%

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Pathways and Mechanisms of Cellular Cholesterol Efflux—Insight From Imaging

Juhl

Wüstner

2022

Front. Cell Dev. Biol.

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Cholesterol is an essential molecule in cellular membranes, but too much cholesterol can be toxic. Therefore, mammalian cells have developed complex mechanisms to remove excess cholesterol. In this review article, we discuss what is known about such efflux pathways including a discussion of reverse cholesterol transport and formation of high-density lipoprotein, the function of ABC transporters and other sterol efflux proteins, and we highlight their role in human diseases. Attention is paid to the biophysical principles governing efflux of sterols from cells. We also discuss recent evidence for cholesterol efflux by the release of exosomes, microvesicles, and migrasomes. The role of the endo-lysosomal network, lipophagy, and selected lysosomal transporters, such as Niemann Pick type C proteins in cholesterol export from cells is elucidated. Since oxysterols are important regulators of cellular cholesterol efflux, their formation, trafficking, and secretion are described briefly. In addition to discussing results obtained with traditional biochemical methods, focus is on studies that use established and novel bioimaging approaches to obtain insight into cholesterol efflux pathways, including fluorescence and electron microscopy, atomic force microscopy, X-ray tomography as well as mass spectrometry imaging.

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Section: Lipids Npc Proteins and Atp-binding Cassette Transporters Co...supporting

confidence: 55%

Section: Lipids Npc Proteins and Atp-binding Cassette Transporters Co...mentioning

confidence: 58%

Section: Origin Structure and Function Of Cholesterol-containing Extr...mentioning

confidence: 99%

Section: Origin Structure and Function Of Cholesterol-containing Extr...mentioning

confidence: 99%

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Pathways and Mechanisms of Cellular Cholesterol Efflux—Insight From Imaging

Juhl

Wüstner

2022

Front. Cell Dev. Biol.

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“…The authors used these results to determine the surface fraction of endo-lysosomes in contact with mitochondria, providing clarity on how cholesterol can be delivered to the mitochondria. In a related study, the authors used SXT to observe the transfer of excess cholesterol from endo-lysosomes to the plasma membrane by shedding extracellular vesicles [ 47 ]. Taken together, these studies demonstrate the usefulness of correlative cryo-fluorescence microscopy and SXT in mapping cellular rearrangements and quantifying inter-organelle contacts.…”

Section: Use Of Sxt To Quantify Mitochondrial Network Structurementioning

confidence: 99%

The use of soft X-ray tomography to explore mitochondrial structure and function

Loconte

White

2022

Molecular Metabolism

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Background Mitochondria are cellular organelles responsible for energy production, and dysregulation of the mitochondrial network is associated with many disease states. To fully characterize the mitochondrial network's structure and function, a three-dimensional whole cell mapping technique is required. Scope of review This review highlights the use of soft X-ray tomography (SXT) as a relatively high-throughput approach to quantify mitochondrial structure and function under multiple cellular conditions. Major conclusions The use of SXT opens the door for mapping cellular rearrangements during critical processes such as insulin secretion, stem cell differentiation, or disease progression. SXT provides unique information such as biochemical compositions or molecular densities of organelles and allows for unbiased, label-free imaging of intact whole cells. Mapping mitochondria in the context of the near-native cellular environment will reveal more information regarding mitochondrial network functions within the cell.

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