Comparison of the genetic defect with LDL-receptor activity in cultured cells from patients with a clinical diagnosis of heterozygous familial hypercholesterolemia
XM Sun, DD Patel, BL Knight… - … , thrombosis, and vascular …, 1997 - Am Heart Assoc
XM Sun, DD Patel, BL Knight, AK Soutar
Arteriosclerosis, thrombosis, and vascular biology, 1997•Am Heart AssocIn this study we have analyzed the genetic defect in 42 patients with a diagnosis of
heterozygous familial hypercholesterolemia (FH) by Southern blotting, SSCP, and
sequencing of PCR-amplified fragments of genomic DNA or sequencing of RT-PCR
products from mRNA in cultured cells. The apoB Arg3500Gln mutation was identified in five
patients. A molecular defect in the LDL-receptor gene was confirmed in 23 patients; 16 of
these mutations have not been described before. No defect in the coding region, intron …
heterozygous familial hypercholesterolemia (FH) by Southern blotting, SSCP, and
sequencing of PCR-amplified fragments of genomic DNA or sequencing of RT-PCR
products from mRNA in cultured cells. The apoB Arg3500Gln mutation was identified in five
patients. A molecular defect in the LDL-receptor gene was confirmed in 23 patients; 16 of
these mutations have not been described before. No defect in the coding region, intron …
Abstract
In this study we have analyzed the genetic defect in 42 patients with a diagnosis of heterozygous familial hypercholesterolemia (FH) by Southern blotting, SSCP, and sequencing of PCR-amplified fragments of genomic DNA or sequencing of RT-PCR products from mRNA in cultured cells. The apoB Arg3500Gln mutation was identified in five patients. A molecular defect in the LDL-receptor gene was confirmed in 23 patients; 16 of these mutations have not been described before. No defect in the coding region, intron:exon junctions or proximal promoter of the LDL-receptor gene or in the region of the apoB gene coding for the LDL-receptor binding domain was found in the remaining 14 patients. LDL-receptor activity and protein content of cultured lymphoblasts from the patients was significantly lower in cells from patients with severe rather than mild LDL-receptor mutations. Cells from four patients with no detectable defect showed reduced LDL receptor activity compared with eight normal cell lines, whereas six others had reduced LDL-receptor activity but LDL-receptor protein content within the normal range. Cells from four patients appeared to have normal LDL-receptor function. Cells from two patients with a defined defect also had LDL-receptor activity within the normal range. The findings demonstrate the problems involved in the genetic diagnosis of FH in patients. .
Am Heart Assoc