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Monocytes of patients with familial hypercholesterolemia show alterations in cholesterol metabolism

Purpose

Elevated plasma cholesterol promotes the formation of atherosclerotic lesions in which monocyte-derived lipid-laden macrophages are frequently found. To analyze, if circulating monocytes already show increased lipid content and differences in lipoprotein metabolism, we compared monocytes from patients with Familial Hypercholesterolemia (FH) with those from healthy individuals.

Hypothesis

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Experimental Design

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Experimental Variables

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Controls

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Methods

Cholesterol and oxidized cholesterol metabolite serum levels of FH and of healthy, gender/age matched control subjects were measured by combined gas chromatography ? mass spectroscopy. Monocytes from patients with FH and from healthy subjects were isolated by antibody-assisted density centrifugation. Gene expression profiles of isolated monocytes were measured using Affymetrix HG-U 133 Plus 2.0 microarrays. We compared monocyte gene expression profiles from FH patients with healthy controls using a Welch T-test with correction for multiple testing (p < 0.05; Benjamini Hochberg correction, False Discovery Rate = 0.05). The differential expression of FH associated genes was validated at the mRNA level by qRT-PCR and/or at the protein level by Western Blot or flow cytometry. Functional validation of monocyte scavenger receptor activities were done by binding assays and dose/time dependent uptake analysis using native and oxidized LDL.

Additional Information
  • Mosig S, Rennert K, Büttner P, Krause S et al. Monocytes of patients with familial hypercholesterolemia show alterations in cholesterol metabolism. BMC Med Genomics 2008 Nov 28;1:60. PMID: 19040724
  • Microarray
    Affymetrix HG-U133_Plus_2
    23 Samples Loaded: 23
    Human (Homo sapiens)
    Monocytes
    Hypercholesterolemia
    Sample Set Spreadsheet
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    Samples Preview
    Sample ID !Sample_title cell type disease state
    GSM140232 heterozygot FH P249 monocyte heterozygote FH
    GSM140233 homozygot FH P251 monocyte homozygote FH
    GSM140234 homozygot FH P253 monocyte homozygote FH
    GSM140235 homozygot FH P262 monocyte homozygote FH
    GSM140236 heterozygot FH P266 monocyte heterozygote FH
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    Sample ID !Sample Title Cell type Disease state
    GSM140232
    heterozygot FH P249
    monocyte
    heterozygote FH
    GSM140233
    homozygot FH P251
    monocyte
    homozygote FH
    GSM140234
    homozygot FH P253
    monocyte
    homozygote FH
    GSM140235
    homozygot FH P262
    monocyte
    homozygote FH
    GSM140236
    heterozygot FH P266
    monocyte
    heterozygote FH
    GSM140237
    homozygot FH P270wh
    monocyte
    homozygote FH
    GSM140239
    heterozygot FH P279
    monocyte
    heterozygote FH
    GSM140240
    heterozygot FH P282
    monocyte
    heterozygote FH
    GSM140241
    heterozygot FH P560
    monocyte
    heterozygote FH
    GSM140244
    control participant P303
    monocyte
    control participant
    GSM140245
    control participant P307
    monocyte
    control participant
    GSM140246
    control participant P309
    monocyte
    control participant
    GSM140247
    control participant P311
    monocyte
    control participant
    GSM140248
    control participant P313
    monocyte
    control participant
    GSM140249
    control participant P317
    monocyte
    control participant
    GSM140250
    control participant P319
    monocyte
    control participant
    GSM140251
    control participant P327
    monocyte
    control participant
    GSM140252
    control participant P329
    monocyte
    control participant
    GSM140253
    control participant P331
    monocyte
    control participant
    GSM140254
    control participant P333
    monocyte
    control participant
    GSM140255
    control participant P335
    monocyte
    control participant
    GSM140256
    control participant P351
    monocyte
    control participant
    GSM140257
    heterozygot FH P387neu
    monocyte
    heterozygote FH

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