GENE DIAGNOSIS AND GENE THERAPY OF CHOLESTEROL TRANSPORT AND CHOLESTEROL REVERSE TRANSPORT DISORDERS

家族性高胆固醇血症的LDL受体异常：全世界已报道LDL受体基因有230多种不同的突变。我们收集了20例FH的纯合子和1,500多例FH的杂合子，在我们的实验室中已经鉴定出9种LDL受体基因变异。Southern杂交检测到4个突变体存在大片段缺失，SSCP分析和PCR产物直接测序检测到5个突变体存在点突变。这9种突变株仅占FH的17.5%，其中FH敦贺株在第6外显子（280个Asp-Tyr）发生点突变。FH金泽-2显示395 Arg-Trp的点突变。FH Morioka在第9外显子有一个由C到T的点突变（395 Arg-Trp），FH南尾在第2外显子有23个终止点突变，FH Yokote在第15外显子有718个终止点突变。家族性高密度脂蛋白胆固醇血症的胆固醇酯转移蛋白（CETP）缺乏症 关于我们 在内含子14的5 ′剪接供体（+1位）处，G → A的突变使严格保守的G-T内含子剪接供体变为A-T。一种剪接供体位点突变体是在内含子14的+3位置插入胸苷，这将再次导致剪接缺陷。另一个新的突变体是外显子15的错义突变，导致天冬氨酸变为甘氨酸。这种突变也很常见，几乎十分之一。在718名普通受试者中，这两种常见突变体至少产生68个CETP杂合子，可能提高日本人的HDL-胆固醇水平，降低冠心病发病率。先证者为29例男性，血清胆固醇（CHOL）33 mg/dl，甘油三酯（TG）0 mg/dl，高密度脂蛋白胆固醇（HDL-C）28 mg/dl。基因分析显示第10外显子和第9内含子连接处的点突变（G至A），这将产生剪接异常，并且没有MTP蛋白。少

LDL-RECEPTOR ABNORMALITIES IN FAMILIAL HYPERCHOLESTEROLEMIA.More than 230 different mutations in the LDL receptor gene have been reported in the world. We have collected 20 homozygotes and more than 1,500 heterozygotes of FH.Nine variants of LDL receptor gene have been identified in our laboratory. Four mutants showed large deletions detected by Southern blot analysis, and 5 mutants were point mutations detected by SSCP analysis and direct sequencing of PCR products. These 9 mutants accounted for only 17.5% of FH.FH Tsuruga showed a point mutation in exon 6 (280 Asp-Tyr). FH Kanazawa-2 showed a point mutaiopn of 395 Arg-Trp. FH Morioka shwoed a point mutation from C to T (395 Arg-Trp) in exon 9.FH Nanao showed 23 stop mutant in exon 2.FH Yokote showed a mutation of 718 stop in exon 15.CHOLESTERYL-ESTER TRANSFER PROTEIN (CETP) DEFICIENCY IN FAMILIAL HYPER-HDL-CHOLESTEROLEMIA.The genomic DNA of patients with CETP deficiency was used as a substrate for amplification of the CETP gene by PC … More R.At the 5'splice donor of intron 14 (position+1) there was a G to A change altering the strictly conserved G-T intron splice donor to A-T.We found two novel mutants of CETP gene. One splice donor site mutant is a thymidine insertion in +3 position in intron 14, which will, again, result in splicing defect. Another new mutant is a missense mutation in exon 15, producing change of aspartic acid into glycine. This mutant is also highly frequent, almost 1 in 10. Thus, these two common mutants produce at least 68 CETP heterozygotes in 718 general subjects, and might raise the HDL-cholesterol levels and reduce coronary heart disease in the Japanese.LCATDEFICIENCY.The proband is 37 male patient, and his CHOL level was 228mg/dl, HDL-C was 34 mg/dl, his LCAT activity was 0%, and the gene analysis showed a point mutant of 30 Ser-Gly.MICROSOMAL TRANSFER PROTEIN (MTP) GENE MUTATION IN ABETALIPOPROTEINEMIA.The proband was 29 male patient, and his CHOL level was 33mg/dl, TG was 0mg/dl, and HDL-C was 28mg/dl. The gene analysis showed a point mutation in the junction of exon 10 and intron 9 (G to A), which would produce splicing abnormalities and no MTP protein. Less