Journal article
Two polymorphic cholesterol monohydrate crystal structures form in macrophage culture models of atherosclerosis
Proceedings Of The National Academy Of Sciences Of The United States Of America-Biological Sciences, Vol.115(30), pp.7662-7669
24/Jul/2018
Abstract
The formation of atherosclerotic plaques in the blood vessel walls is the result of LDL particle uptake, and consequently of cholesterol accumulation in macrophage cells. Excess cholesterol accumulation eventually results in cholesterol crystal deposition, the hallmark of mature atheromas. We followed the formation of cholesterol crystals in J774A.1 macrophage cells with time, during accumulation of LDL particles, using a previously developed correlative cryosoft X-ray tomography (cryo-SXT) and stochastic optical reconstruction microscopy (STORM) technique. We show, in the initial accumulation stages, formation of small quadrilateral crystal plates associated with the cell plasma membrane, which may subsequently assemble into large aggregates. These plates match crystals of the commonly observed cholesterol monohydrate triclinic structure. Large rod-like cholesterol crystals form at a later stage in intracellular locations. Using cryotransmission electron microscopy (cryo-TEM) and cryoelectron diffraction (cryo-ED), we show that the structure of the large elongated rods corresponds to that of monoclinic cholesterol monohydrate, a recently determined polymorph of the triclinic crystal structure. These monoclinic crystals form with an unusual hollow cylinder or helical architecture, which is preserved in the mature rodlike crystals. The rod-like morphology is akin to that observed in crystals isolated from atheromas. We suggest that the crystals in the atherosclerotic plaques preserve in their morphology the memory of the structure in which they were formed. The identification of the polymorph structure, besides explaining the different crystal morphologies, may serve to elucidate mechanisms of cholesterol segregation and precipitation in atherosclerotic plaques.
Details
- Title
- Two polymorphic cholesterol monohydrate crystal structures form in macrophage culture models of atherosclerosis
- Creators
- Neta Varsano (null) - 972WIS_INST___9Fabio Beghi (null) - University of Milan (Italy, Milan) - UNIMINadav Elad (null) - 972WIS_INST___100Eva Pereiro (null) - ALBA SynchrotronTali Dadosh (null) - 972WIS_INST___100Iddo Pinkas (null) - 972WIS_INST___100Ana J. Perez-Berna (null) - ALBA SynchrotronXueting Jin (null) - National Heart Lung and Blood Institute (United States, Bethesda) - NHLBIHoward S. Kruth (null) - National Heart Lung and Blood Institute (United States, Bethesda) - NHLBILeslie Leiserowitz (null) - 972WIS_INST___98Lia Addadi (Corresponding Author) - 972WIS_INST___9
- Resource Type
- Journal article
- Publication Details
- Proceedings Of The National Academy Of Sciences Of The United States Of America-Biological Sciences, Vol.115(30), pp.7662-7669; 24/Jul/2018
- Number of pages
- 8
- Language
- English
- DOI
- https://doi.org/10.1073/pnas.1803119115
- Grant note
- We thank Prof. Benjamin Geiger for his support in the cell culture growth and maintenance. Superresolution microscopy and EM studies were conducted at the Irving and Cherna Moskowitz Center for Nano and Bio-Nano Imaging at the Weizmann Institute of Science. Cryo-SXT experiments were performed at the MISTRAL beamline at ALBA Synchrotron with the collaboration of ALBA staff and CALIPSOplus Grant 730872. This research was supported by Binational Science Foundation Grant 2013045 (to L.A. and H.S.K.). L.A. is the incumbent of the Dorothy and Patrick Gorman Professorial Chair of Biological Ultrastructure at the Weizmann Institute of Science. Author contributions: N.V., H.S.K., L.L., and L.A. designed research; N.V., F.B., N.E., E.P., T.D., I.P., and A.J.P.-B. performed research; E.P. and X.J. contributed new reagents/analytic tools; N.V., F.B., N.E., E.P., T.D., I.P., L.L., and L.A. analyzed data; and N.V., F.B., H.S.K., L.L., and L.A. wrote the paper.
- Record Identifier
- 993264770603596
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