• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Molecular Basis of the Etiology and Pathogenesis of Hyperlipoproteinemia

Research Project

Project/Area Number 61440053
Research Category

Grant-in-Aid for General Scientific Research (A)

Allocation TypeSingle-year Grants
Research Field 内分泌・代謝学
Research InstitutionNational Cardiovascular Center Research Institute (NCVC)

Principal Investigator

YAMAMOTO Akira  NCVC, Vice Director, 副所長 (00028408)

Co-Investigator(Kenkyū-buntansha) 高木 敦子  国立循環器病センター研究所, 病因部, 室長 (90179416)
IKEDA Yasuyuki  NCVC, Dept. Etiology/Pathogenesis, Staff, 病因部, 室長 (90176107)
TAJIMA Shoji  NCVC, Dept. Etiology/Pathogenesis, Chief, 病因部, 室長 (50132931)
MIYAKE Yasuko  NCVC, Dept. Etiology/Pathogenesis, Chief, 病因部, 室長 (00132936)
YAMAMURA Taku  NCVC, Dept. Etiology/Pathogenesis, Chief, 病因部, 室長 (20132938)
YAKOYAMA Shinji  NCVC, Dept. Etiology/Pathogenesis, Chief (10142192)
高市 成子  国立循環器病センター研究所, 病因部, 室員 (00093930)
Project Period (FY) 1986 – 1989
Project Status Completed (Fiscal Year 1989)
Budget Amount *help
¥14,000,000 (Direct Cost: ¥14,000,000)
Fiscal Year 1989: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1988: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1987: ¥4,000,000 (Direct Cost: ¥4,000,000)
Fiscal Year 1986: ¥7,000,000 (Direct Cost: ¥7,000,000)
KeywordsPlasma lipoprotein / Hyperlipidemia / LDL receptor / Apolipoprotein E / Lipoprotein lipase / Familial hypercholesterolemia / Cholesterol / Atherosclerosis / リポタンパク質リパ-ゼ / 粥状動脈硬化 / 血漿リポ蛋白 / 家族性高コンステロール血症 / 低密度リポ蛋白レセプター(LDLレセプター) / アポリポ蛋白E5とE7 / 動脈硬化 / マクロファージ / 家族性高コレステロール血症 / リポ蛋白リパーゼ / 虚血性心疾患
Research Abstract

Hyperlipoproteinemia is one of the most important risk factors for atherosclerotic vascular diseases. Hyperchylomicronemia often results in fatal pancreatitis. Although the plasma lipid level is strongly influenced by dietary conditions,most cases of hyperlipoproteinemia are based upon the genetic of abnormalities in plasms lipoprotein metabolism. Depending upon such hereditary predispositions,the response to dietary management is different. The present study is aimed at elucidating the etiology and pathogenesis of hyperlipoproteinemia on the molecular basis and establishing the criteria for discriminating of trait of hyperlipoproteinemia.
1) Analysis of the LDL-receptor defect in familial hypercholesterolemia (FH): Fibrobasts were cultured from skin biopsy samples from 23 cases pf FH homozygotes belonging to 17 different families. Functional activities of LDL-receptor proteins were estimated by measuring ^<125>I-LDL binding and uptake. LDL- receptor proteins were analyzed by ^<35>S-met … More hionine pulse labeling followed by the immunoprocipitation and SDS electophoresis. Ten families were diagnosed as a receptor-negative type. Among them, receptor proteins were completely lacking in 5 families and markedly reduced in 2 families. Processing of the precursor to mature protein was disturbed in 2 families and a receptor protein 5kd smaller in size was formed in one family. Among 5 families with receptor-defective type, two showed a reduction in number of the receptors and three showed a delay in processing from precursor to the mature form. In one family, a peculiar mutation was found. The EGF precursor homology domain was completely lacking due to the Alu-Alu recombination. Although the receptor protein appeared on the cell surface, it could not bind LDL. The receptor could bind beta-VLDL but recycling was impaired. Our results show that a variety of mutants exist in LDL receptors of PH among Japanese population. [Ref.1-3]
2) Purification and Characterization of lipoprotein lipase(LpL) and the analysis of its deficiency in type I hyperlipoproteinemia: LpL and hepatic triglyceride lipase(H-TGL) were purified from human postheparin plasma(PHP) and specific antibodies (polyclonal and monoclonal) were prepared. Assay kits for LPL and H-TGL with selective immunoinactivation method (for enzyme activities) and sandwich EIA method (for mass) were developed. Using THP-1 cells, a human monocytic leukemia cell line, the synthesis and secretion of LpL were studied. It was synthesized as a precursor with 55kd molecular weight and processed by the addition of sugar moiety into 61kd mature protein. In one of the patient with type I hyperlipoproteinemia, no LpL nor MRNA was detected in monocute-macrophages. In another patient a small amount of LPL with normal molecular weight was detected in monocute-macrophages, while there was no mass nor activity of the enzyme was found in his PHP. [Ref.4, 5]
3) Apolipoprotein E (apo E) polymorphism and its relation to hyperlipoproteinemia: Two new mutants of apo E were found and the site of mutation was determined; apo E5 with the change of the 3rd amino acid residue from Glu to Lys and apo E7 with the change at 244 and 245 both from Glu to Lys. The substitution occurred by the same base exchange G->A. The frequency distribution of the major apo E isoforms and these new mutants was investigated among 199 patients with ischemic heart diseases (mainly myocardial infarction) administered to CCU of our hospital. The frequency of E4,5, and 7 was significantly higher in relatively young male patients aged below 60 years and in female patients compared to the healthy population. [Ref.6-8]
4) Purification and characterization of lipid transfer protein (cholesterol ester transfer protein CETP): A rapid purification procedure using 3 steps gel chromatography was established and CETP purified 43,000 fold with a yield of 30% from human plasma. CETP-catalyzed cholesterol ester transfer (trig ly-ceride transfer into the reverse direction) was estimated by using LDL, HDL and artificial triolein particle covered by phosphatidylcholine monolayer and stabilized by apolipoprotein A-I,A-II, C-II, C-III, or E. The kinetics was consistent with the model that the reaction is bidirectional in equilibrium, taking both non-polar lipids as substrate in a single pool. In one patient with strong hyperalphalipoproteinemia with HDL-cholesterol 239 mg/dl, a reduction in CETP activity was observed by using patient's HDL as a substrate suggesting the existence of an inhibitor. [Ref.9, 10] Less

Report

(5 results)
  • 1989 Annual Research Report   Final Research Report Summary
  • 1988 Annual Research Report
  • 1987 Annual Research Report
  • 1986 Annual Research Report
  • Research Products

    (47 results)

All Other

All Publications (47 results)

  • [Publications] Funahashi,T.,Miyake,Y.,Yamamoto,A.,Matsuzawa,Y.Kishino,B.: "Mutations of the low density lipoprotein receptor in Japanese kindreds with familial hypercholesterolemia" Human Genetics. 79. 103-108 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Miyake,Y.,Tajima,S.,Yamamoto,A.: "Analysis of a recycling-impaired mutant of low density lipoprotein receptor in familial hypercholesterolemia" J.Biol.Chem.264. 16584-16590 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Ikeda,Y.,Takagi,A.Yamamoto,A.: "Purification and characterization of lipoprotein lipase and hepatic triglyceride lipase from human postheparin plasma:Production of monospecific antibody to the individual lipase." Biochim.Biophys.Acta. 1003. 643-655 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Tajima,S.,Yamamura,T.,Yamamoto,A.: "Analysis of apolipoprotein E5 gene from a patient with hyperlipoproteinemia" J.Biochem.104. 730-734 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Tajima,S.,Yamamura,T.,Menju,M.,Yamamoto,A.: "Analysis of apolipoprotein E7(apolipoprotein E-Suita)gene from a patient with hyperlipoproteinemia" J.Biochem.105. 249-253 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Yokoyama,S.,Kurasawa,T.Nishikawa,O.,Yamamoto,A.: "High density lipoproteins with poor reactivity to cholesteryl ester transfer reaction observed in a homozygote of familial hyperalphalipoproteinemia" Artery. 14. 43-51 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] 山本章: "血清脂質ー標準値の設定" 山本章、中村治雄、八杉忠男編 協和企画通信社, 147 pp17-28 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] 山本章: "高脂血症の最近の治療「新内科学大系年刊版'87-B」" 鎭目和夫、石川誠他4名編 中山書店, 279 pp177-194 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Funahashi, T., Miyake, Y., Yamamoto, A., Matsuzawa, Y. and Kishino, B.: "Mutations of the low density lipoprotein receptor in Japanese kindreds with familial hypercholesterolemia." Human Genetics. 79. 103-108 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Miyake, Y., Tajima, S. and Yamamoto, A.: "Analysis of a recycling-impaired mutant of low density lipoprotein receptor in familial hypercholesterolemia." J. Biol. Chem. 264:16584-16590, 1989.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Yamamoto, A., Kamuya, T., Yamamura, T., Yokoyama, S., Horiguchi, Y., Funahashi, T., Kawaguchi, A., Miyake, Y., Beppu, S., Ishikawa, K., Matsuzawa, Y. and Takaichi, S.: "Clinical features of familial hypercholesterolemia." Arteriosclerosis 9 (Suppl 1):66-74, 1989.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Hayashi, R., Tajima, S. and Yamamoto, A.: "Purification and characterization of lipoprotein lipase from human postheparin plasma and its comparison with purified bovine milk lipoprotein lipase." J. Biochem. 100:319-331, 1986.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Ikeda, Y., Takagi, A. and Yamamoto, A.: "Purification and characterization of lipoprotein lipase and hepatic triglyceride lipase from human postheparin plasma: Production of monospecific antibody to the individual lipase." Biochim. Biophys. Acta 1003:254-269, 1989.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Tajima, S., Yamamura, T. and Yamamoto, A.: "Analysis of apolipoprotein E5 gene from a patient with hyperlipoproteinemia." J. Biochem. 104:730-734, 1988.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Tajima, S., Yamamura, T., Menju, M. and Yamamoto, A.: "Analysis of apolipoprotein E7 (apolipoprotein E-Suita) gene from a patient with hyperlipoproteinemia." J. Biochem. 105:249-253, 1989.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Dong, Li-M., Yamamura, T. and Yamamoto, A.: "Enhanced binding activity of an apolipoptoerin E mutant." Biochem. Biophys. Res. Commun.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Nishikawa, O., Yokoyama, S., Okabe, H. and Yamamoto, A.: "Enhancement of non-polar lipid transfer reaction through stabilization of substrate lipid particles with apolipoproteins." J. Biochem. 103:188-194, 1988.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Yokoyama, S., Kurasawa, T., Nishikawa, O. and Yamamoto, A.: "High density lipoproteins with poor reactivity to cholesterol ester transfer reaction observed in a homozygote of familial hyperalphalipoproteinemia." Artery 14:43-51, 1986.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1989 Final Research Report Summary
  • [Publications] Ikeda,Y.,Takagi,A.,Yamamoto,A.: "Purification and characterization of lipoprotein lipase and hepatic triglycericle lipase from human postheparin plasma" Biochim.Biophys.Acta. 1003. 254-269 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] 池田康行,高木敦子,山本章: "ヒトリポ蛋白リパ-ゼ異常症の分子免疫学的および遺伝子解析" Medical Immunology. 18. 643-655 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Miyake,Y.,Tajima,S.,Yamamoto,A.: "Analysis of a recycling-impaired mutant of low density lipoprotein receptor in familial hypercholesterolemia" J.Biol.Chem.264. 16584-16590 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] 山村卓,田嶋正二,山本章: "アポリポ蛋白Eの遺伝素因と動脈硬化" 動脈硬化. 16. 1047-1051 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Yamamoto,A.,Kamiya,T.,Yomamura,T.,Yokoyama,S.,Horiguchi,Y.et al: "Clinical features of familial hypercholesterolemia" Arteriosclerosis. 9(supl I). 66-74 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Menju,M.,Tajima,S.,Yamamoto,A.: "Expression of the apolipoprotein E gene in a human macrophage-like cell line,THP-1." J.Biochem.106. 505-510 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] 山本章: "總合脂質科学(のうち「リポプロテイン」の項)" 鹿山光編,恒星社厚生閣, 934 pp361-374 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Funahashi,T.;Miyake,Y.;Yamanoto,A.;Matsuzawa,Y.;Kishino,B.: Human Genetics. 79. 103-108 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] Tajima,S.;Yamamura,T.;Yamamoto,A.: J.Biochem.(Tokyo). 104. 48-52 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] Tajima,S.;Yamamura,T.;Menju,M.;Yamamoto,A.: J.Biochem.(Tokyo). 105. 249-253 (1989)

    • Related Report
      1988 Annual Research Report
  • [Publications] Nishikawa,O.;Yokoyama,S.;Okabe,H.;Yamamoto,A.: J.Biochem.(Tokyo). 103. 188-194 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] Yamamoto,A.;Horibe,H.;Sawada,S.;Uyama,M.;Matsuzawa,Y.;Yamamura,T.;Yokoyama,S.;Kasagi,F.;Kameda,K.: Preventive Medicine. 17. 93-108 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] Yamamoto,A.;Hara,H.;Takaichi,S.;Wakasugi,J.;Tomikawa,M.: Am.J.Cardiol.62. 31B-36B (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] Miyake Y., Funahashi, T, Yamamoto, A.: J. Biochem. (Tokyo). 101. 1355-1360 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] Yokoyama, S., Yamamoto, A. Hayashi, R. Satani, M.: Jap. Circul. J.51. 1116-1122 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] Yutani, C., Go., S., Imakita, M., Ishibashi-Ueda, H., Hatanaka, K., Yamamoto, A.: Acta Pathol. Japn.37. 1489-1504 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] Funahashi, T., Miyake, Y., Yamamoto, A., Matsuzawa, Y., Kishino, B.: Human Genetics. (1988)

    • Related Report
      1987 Annual Research Report
  • [Publications] Tajima, S., Yamamura T., Yamamoto, A.: J. Biochem. (Tokyo). (1988)

    • Related Report
      1987 Annual Research Report
  • [Publications] 池田康行,高木敦子,山本章: 脂質生化学研究. 29. 149-152 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] Yamamoto,A., Sawada,S., Uyama,M., Matsuzawa,Y., Yamamura,S., Kameda,K., Kasagi,F.,Horibe,H: "Recent Aspects of Diagnosis and Treatment of Lipoprotein Disorders: Impact on Prevention of Atherosclerotic Diseases,: Serum lipid levels in elementary and junior high school children and their relationship to obesity pp107-116" Alan R. Liss, Inc., New York Editors: Kurt Widhalm and Herbert K. Naito, 383 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] Yamamoto, A., Yokoyama, S., Yamamura, T.: "Drugs Affecting Lipid Metabolism: Intensive drug treatment for familial hypercholesterolemia pp 269-273" Springer Verlag , Berlin-Heidelberg Editors: Rodolfo Paoletti and David Kritchevsky, 451 (1987)

    • Related Report
      1987 Annual Research Report
  • [Publications] Yamamoto,A.;Takaichi,S.;Hara,H.;Nishikawa,O.;Yokoyama,S.;Yamamura,T.;Yamaguchi,T.: Atherosclerosis. 62. 209-217 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] Yamamoto,A.;Matsuzawa,Y.;Yokoyama,S.;Funahashi,T.;Yamamura,T.: Am.J.Cardiol.57. 29H-35H (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] Miyake,Y.;Funahashi,T.;Yamamoto,A.: J.Biochem.(Tokyo). (1987)

    • Related Report
      1986 Annual Research Report
  • [Publications] Funahashi,T.;Miyake,Y.;Yamamoto,A.;Matsuzawa,Y.;Kishino,B.: J.Clin.Invest.(1987)

    • Related Report
      1986 Annual Research Report
  • [Publications] 船橋徹,三宅康子,山本章: 代謝. 23. 617-626 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] 山村卓,山本章: 臨床病理. 34. 408-414 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] Yamamoto,A.;Yokoyama,S.;Yamamura,T. Editor:R.Fears: "Pharmacological Control of Hyperlipidemia" J.R.Prous Science Publishers, 2)510(333-34 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] 山本章,三宅康子,船橋徹 著,永井克孝,野沢義則,林恭三 編: "レセプターの化学" 化学同人, 270(109-120) (1986)

    • Related Report
      1986 Annual Research Report

URL: 

Published: 1987-03-31   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi