公开(公告)号：US20050223420A1

公开(公告)日：2005-10-06

申请号：US11099343

申请日：2005-04-05

申请人： Monty Krieger ,  Songwen Zhang ,  Sharon Karackattu

发明人： Monty Krieger ,  Songwen Zhang ,  Sharon Karackattu

IPC分类号： A01K67/027 ,  A61K49/00 ,  C07K14/705 ,  C07K14/775 ,  C12N15/85

CPC分类号： A01K67/0275 ,  A01K67/0276 ,  A01K2217/05 ,  A01K2217/075 ,  A01K2227/105 ,  A01K2267/03 ,  A01K2267/0362 ,  A01K2267/0375 ,  A61K49/0008 ,  C07K14/705 ,  C07K14/775 ,  C12N15/8509

摘要： An animal model of coronary heart disease has been developed where myocardial infarct can be induced by altering the animal's diet. In all embodiments, this animal model is a result of reduced activity of scavenger receptor class BI (SR-BI) and apolipoprotein E (ApoE). In a preferred embodiment, the model is a result of crossbreeding two transgenic mouse lines: a knockout of SR-BI (SR-BI−/−) and an impaired ApoE expressor (hypoE). The impaired ApoE gene results in only 2-5% expression of ApoE and a reduction in cholesterol homeostasis. Resulting animals are predisposed to hypercholesterolemia but can live longer than a year on a normal low fat diet. Serum plasma levels can be significantly elevated by changing the animal's diet to one containing high levels of fat and cholesterol. Within a month on a high fat, high cholesterol diet, animals develop atherosclerosis and myocardial infarction occurs. Survival depends on the nature of the diet and the conditions of animal husbandry and can typically be around 20-30 days after administration of the modified diet depending on the specific conditions. Housing the animals alone or in groups significantly affects survival of these animals on a high fat diet. Analysis of B- and T-cell deficient SR-BI/ApoE/RAG2 triple knockout mice established that B- and T-lymphocytes do not play a key role in the pathophysiology of the SR-BI ApoE dKO model of human disease. These animal models can be used to study mechanisms and progression of CHD as a function of diet, treatment with drugs to be screened for efficacy or undesirable side effects, and social environmental effects.

摘要翻译： 已经开发了冠状动脉心脏病的动物模型，其中可以通过改变动物的饮食来诱导心肌梗死。 在所有实施方案中，该动物模型是清道夫受体类BI（SR-BI）和载脂蛋白E（ApoE）的活性降低的结果。 在优选的实施方案中，该模型是杂交两种转基因小鼠品系的结果：SR-BI（SR-BI - / - ）的敲除和受损的ApoE表达（hypoE）。 受损的ApoE基因导致ApoE仅2-5％的表达和胆固醇体内平衡的降低。 导致的动物易发生高胆固醇血症，但在正常的低脂肪饮食中可以长于一年。 通过将动物的饮食改为含有高脂肪和胆固醇水平的饮食，可以显着提高血浆血浆水平。 在高脂肪，高胆固醇饮食的一个月内，动物会发生动脉粥样硬化并发生心肌梗塞。 生存取决于饮食的性质和畜牧业的条件，并且根据具体情况通常可以在施用改进的饮食之后约20-30天。 单独或分组饲养动物会显着影响这些动物在高脂肪饮食中的生存。 B细胞和T细胞缺陷型SR-BI / ApoE / RAG2三重敲除小鼠的分析表明，B细胞和T淋巴细胞在人类疾病的SR-BI ApoE dKO模型的病理生理学中不起关键作用。 这些动物模型可用于研究作为饮食功能的CHD的机制和进展，用待筛选的药物治疗有效性或不良副作用以及社会环境影响。

公开(公告)号：US20050136005A1

公开(公告)日：2005-06-23

申请号：US10933037

申请日：2004-09-02

申请人： Karen Kozarsky ,  Attilio Rigotti ,  Monty Krieger

发明人： Karen Kozarsky ,  Attilio Rigotti ,  Monty Krieger

IPC分类号： A61K38/00 ,  C07K14/705 ,  A61K49/00 ,  A61K48/00

CPC分类号： C07K14/705 ,  A01K2217/05 ,  A61K38/00 ,  C12N2799/022

摘要： Methods for regulation of lipid and cholesterol uptake are described which are based on regulation of the expression or function of the SR-BI HDL receptor. The examples demonstrate that estrogen dramatically downregulates SR-BI under conditions of tremendous upregulation of the LDL-receptor. The examples also demonstrate the upregulation of SR-BI in rat adrenal membranes and other non-placental steroidogenic tissues from animals treated with estrogen, but not in other non-placental non-steroidogenic tissues, including lung, liver, and skin. Examples further demonstrate the uptake of fluorescently labeled HDL into the liver cells of animal, which does not occur when the animals are treated with estrogen. Examples also demonstrate the in vivo effects of SR-BI expression on HDL metabolism, in mice transiently overexpressing hepatic SR-BI following recombinant adenovirus infection. Overexpression of the SR-BI in the hepatic tissue caused a dramatic decrease in cholesterol blood levels. These results demonstrate that modulation of SR-BI levels, either-directly or indirectly, can be used to modulate levels of cholesterol in the blood.

摘要翻译： 描述了调节脂质和胆固醇摄取的方法，其基于调节SR-BI HDL受体的表达或功能。 这些实施例证明雌激素在LDL-受体大量上调的条件下显着下调SR-BI。 这些实施例还证明了在用雌激素治疗的动物的大鼠肾上腺膜和其它非胎盘类固醇生成组织中SR-BI的上调，但在其他非胎盘非类固醇生成组织（包括肺，肝脏和皮肤）中的上调。 实施例进一步证明了将荧光标记的HDL摄取到动物的肝细胞中，当用雌激素治疗时不会发生。 实例还证明了在重组腺病毒感染后瞬时过表达肝SR-BI的小鼠中SR-BI表达对HDL代谢的体内作用。 肝组织中SR-BI的过度表达导致胆固醇血液水平急剧下降。 这些结果表明，直接或间接调节SR-BI水平可用于调节血液中的胆固醇水平。

公开(公告)号：US20050112723A1

公开(公告)日：2005-05-26

申请号：US10781012

申请日：2004-02-18

申请人： Judith Fridovich-Keil ,  Monty Krieger ,  Hazel Holden ,  James Thoden ,  Jenny Schulz

发明人： Judith Fridovich-Keil ,  Monty Krieger ,  Hazel Holden ,  James Thoden ,  Jenny Schulz

IPC分类号： C07H21/04 ,  C12N20060101 ,  C12N1/15 ,  C12N1/18 ,  C12N1/21 ,  C12N5/10 ,  C12N9/54 ,  C12N9/90 ,  C12N15/74

CPC分类号： C12N9/80 ,  C12N2800/102 ,  C12Y305/01003

摘要： Genetically modified cell lines that express a UDP-galactose 4-epimerase (GALE) capable of interconverting UDP-galactose (UDP-gal) and UDP-glucose (UDP-glc), but essentially incapable of interconverting UDP-N-acetylgalactosamine (UDP-galNAc) and UDP-N-acetylglucosamine (UDP-glcNAc).

摘要翻译： 表达能够相互转化UDP-半乳糖（UDP-gal）和UDP-葡萄糖（UDP-glc）的UDP-半乳糖4-差向异构酶（GALE）的基因修饰的细胞系，但基本上不能互相转化UDP-N-乙酰半乳糖胺（UDP- galNAc）和UDP-N-乙酰葡糖胺（UDP-glcNAc）。

公开(公告)号：US06350859B1

公开(公告)日：2002-02-26

申请号：US09241581

申请日：1999-02-02

申请人： Monty Krieger ,  Susan L. Acton ,  Attilio Rigotti

发明人： Monty Krieger ,  Susan L. Acton ,  Attilio Rigotti

IPC分类号： C07K1628

CPC分类号： C07K14/705 ,  A61K38/00

摘要： Two distinct scavenger receptor type proteins having high affinity for modified lipoproteins and other ligands have been isolated, characterized and cloned. HaSR-BI, an AcLDL and LDL binding scavenger receptor, which is distinct from the type I and type II macrophage scavenger receptors, has been isolated and characterized and DNA encoding the receptor cloned from a variant of Chinese Hamster Ovary Cells, designated Var-261. dSR-CI, a non-mammalian AcLDL binding scavenger receptor having high ligand affinity and broad specificity, was isolated from Drosophila melanogaster. The isolated receptors are useful in screening for drugs that inhibit uptake of cholesterol in endothelial or adipose cells or macrophages, respectively. They are also useful as probes for the isolation of other lipoprotein receptors and in research the roles of these receptors.

公开(公告)号：US06437215B1

公开(公告)日：2002-08-20

申请号：US09606324

申请日：2000-06-28

申请人： Monty Krieger ,  Jay M. Edelberg ,  Bernardo Trigatti

发明人： Monty Krieger ,  Jay M. Edelberg ,  Bernardo Trigatti

IPC分类号： A01K67027

CPC分类号： A61K31/10 ,  A01K67/0276 ,  A01K2217/075 ,  A01K2227/105 ,  A01K2267/03 ,  A01K2267/0375 ,  A61K31/34 ,  A61K49/0008 ,  C07K14/705 ,  C07K14/775 ,  C12N15/8509

摘要： Transgenic animals that do not express functional SR-BI and ApoE develop severe atherosclerosis, by age four weeks in transgenic mice. Moreover, these animals exhibit progressive heart block by age four weeks, and die by age nine weeks. Pathology shows extensive fibrosis of the heart and occlusion of coronary arteries. The occlusion appears to be due to clotting, since fat deposition is in the walls. These animals are good models for the following diseases, and for screening of drugs useful in the treatment and/or prevention of these disorders: cardiac fibrosis, myocardial infarction, defects in electrical conductance, atherosclerosis, unstable plaque, and stroke. In contrast to other known models for atherosclerosis, these animals do not have to be fed extreme diets for long periods before developing atherosclerosis. No other known model for heart attacks and stroke is known.

摘要翻译： 不表达功能性SR-BI和ApoE的转基因动物在转基因小鼠中4周龄时出现严重的动脉粥样硬化。 此外，这些动物在四周时间内呈现进行性心脏阻塞，并且死亡9周。 病理显示广泛的心脏纤维化和冠状动脉闭塞。 闭塞似乎是由于凝血，因为脂肪沉积在墙壁中。 这些动物是以下疾病的良好模型，并且用于筛选用于治疗和/或预防这些疾病的药物：心脏纤维化，心肌梗塞，电导缺陷，动脉粥样硬化，不稳定斑块和中风。 与其他已知的动脉粥样硬化模型相反，这些动物在发展动脉粥样硬化之前不必长时间喂食极限饮食。 没有其他已知的心脏病发作和中风模型已知。

公开(公告)号：US5962322A

公开(公告)日：1999-10-05

申请号：US749907

申请日：1996-11-15

申请人： Karen Kozarsky ,  Attilio Rigotti ,  Monty Krieger

发明人： Karen Kozarsky ,  Attilio Rigotti ,  Monty Krieger

IPC分类号： C07K14/705 ,  C12N5/06 ,  C12N5/10

CPC分类号： C07K14/705 ,  A01K2217/05 ,  C12N2799/022

摘要： Methods for regulation of lipid and cholesterol uptake are described which are based on regulation of the expression or function of the SR-BI HDL receptor. The examples demonstrate that estrogen dramatically downregulates SR-BI under conditions of tremendous upregulation of the LDL-receptor. The examples also demonstrate the upregulation of SR-BI in rat adrenal membranes and other non-placental steroidogenic tissues from animals treated with estrogen, but not in other non-placental non-steroidogenic tissues, including lung, liver, and skin. Examples further demonstrate the uptake of fluorescently labeled HDL into the liver cells of animal, which does not occur when the animals are treated with estrogen. Examples also demonstrate the in vivo effects of SR-BI expression on HDL metabolism, in mice transiently overexpressing hepatic SR-BI following recombinant adenovirus infection. Overexpression of the SR-BI in the hepatic tissue caused a dramatic decrease in cholesterol blood levels. These results demonstrate that modulation of SR-BI levels, either directly or indirectly, can be used to modulate levels of cholesterol in the blood.

摘要翻译： 描述了调节脂质和胆固醇摄取的方法，其基于调节SR-BI HDL受体的表达或功能。 这些实施例证明雌激素在LDL-受体大量上调的条件下显着下调SR-BI。 这些实施例还证明了在用雌激素治疗的动物的大鼠肾上腺膜和其它非胎盘类固醇生成组织中SR-BI的上调，但在其它非胎盘非类固醇生成组织（包括肺，肝脏和皮肤）中的上调。 实施例进一步证明了将荧光标记的HDL摄取到动物的肝细胞中，当用雌激素治疗时不会发生。 实例还证明了在重组腺病毒感染后瞬时过表达肝SR-BI的小鼠中SR-BI表达对HDL代谢的体内作用。 肝组织中SR-BI的过度表达导致胆固醇血液水平急剧下降。 这些结果表明，直接或间接调节SR-BI水平可用于调节血液中的胆固醇水平。

公开(公告)号：US07078511B1

公开(公告)日：2006-07-18

申请号：US08765108

申请日：1995-06-19

申请人： Monty Krieger ,  Susan L. Acton

发明人： Monty Krieger ,  Susan L. Acton

IPC分类号： C07K1/00 ,  C07H21/04 ,  C12N1/20 ,  C12P21/06 ,  G01N33/567

CPC分类号： C07K14/705 ,  A61K38/00

摘要： Two distinct scavenger receptor type proteins having high affinity for modified lipoproteins and other ligands have been isolated, characterized and cloned. HaSR-BI, an AcLDL and LDL binding scavenger receptor, which is distinct from the type I and type II macrophage scavenger receptors, has been isolated and characterized and DNA encoding the receptor cloned from a variant of Chinese Hamster Ovary Cells, designated Var-261. dSR-CI, a non-mammalian AcLDL binding scavenger receptor having high ligand affinity and broad specificity, was isolated from Drosophila melanogaster. The isolated receptors are useful in screening for drugs that inhibit uptake of cholesterol in endothelial or adipose cells or macrophages, respectively. They are also useful as probes for the isolation of other lipoprotein receptors and in research the roles of these receptors.

公开(公告)号：US07514592B2

公开(公告)日：2009-04-07

申请号：US11099343

申请日：2005-04-05

申请人： Monty Krieger ,  Songwen Zhang ,  Sharon L. Karackattu

发明人： Monty Krieger ,  Songwen Zhang ,  Sharon L. Karackattu

IPC分类号： G01N33/00 ,  A01K67/00 ,  C12N15/05

CPC分类号： A01K67/0275 ,  A01K67/0276 ,  A01K2217/05 ,  A01K2217/075 ,  A01K2227/105 ,  A01K2267/03 ,  A01K2267/0362 ,  A01K2267/0375 ,  A61K49/0008 ,  C07K14/705 ,  C07K14/775 ,  C12N15/8509

摘要： An animal model of coronary heart disease has been developed where myocardial infarct can be induced by altering the animal's diet. In all embodiments, this animal model is a result of reduced activity of scavenger receptor class BI (SR-BI) and apolipoprotein E (ApoE). In a preferred embodiment, the model is a result of crossbreeding two transgenic mouse lines: a knockout of SR-BI (SR-BI−/−) and an impaired ApoE expressor (hypoE). The impaired ApoE gene results in only 2-5% expression of ApoE and a reduction in cholesterol homeostasis. Resulting animals are predisposed to hypercholesterolemia but can live longer than a year on a normal low fat diet. Serum plasma levels can be significantly elevated by changing the animal's diet to one containing high levels of fat and cholesterol. Within a month on a high fat, high cholesterol diet, animals develop atherosclerosis and myocardial infarction occurs. Survival depends on the nature of the diet and the conditions of animal husbandry and can typically be around 20-30 days after administration of the modified diet depending on the specific conditions. Housing the animals alone or in groups significantly affects survival of these animals on a high fat diet. Analysis of B- and T-cell deficient SR-BI/ApoE/RAG2 triple knockout mice established that B- and T-lymphocytes do not play a key role in the pathophysiology of the SR-BI ApoE dKO model of human disease. These animal models can be used to study mechanisms and progression of CHD as a function of diet, treatment with drugs to be screened for efficacy or undesirable side effects, and social environmental effects.

摘要翻译： 已经开发了冠状动脉心脏病的动物模型，其中可以通过改变动物的饮食来诱导心肌梗死。 在所有实施方案中，该动物模型是清道夫受体类BI（SR-BI）和载脂蛋白E（ApoE）的活性降低的结果。 在优选的实施方案中，该模型是杂交两种转基因小鼠品系的结果：SR-BI（SR-BI - / - ）的敲除和受损的ApoE表达（hypoE）。 受损的ApoE基因导致ApoE仅2-5％的表达和胆固醇体内平衡的降低。 导致的动物易发生高胆固醇血症，但在正常的低脂肪饮食中可以长于一年。 通过将动物的饮食改为含有高脂肪和胆固醇水平的饮食，可以显着提高血浆血浆水平。 在高脂肪，高胆固醇饮食的一个月内，动物会发生动脉粥样硬化并发生心肌梗塞。 生存取决于饮食的性质和畜牧业的条件，并且根据具体情况通常可以在施用改进的饮食之后约20-30天。 单独或分组饲养动物会显着影响这些动物在高脂肪饮食中的生存。 B细胞和T细胞缺陷型SR-BI / ApoE / RAG2三重敲除小鼠的分析表明，B细胞和T淋巴细胞在人类疾病的SR-BI ApoE dKO模型的病理生理学中不起关键作用。 这些动物模型可用于研究作为饮食功能的CHD的机制和进展，用待筛选的药物治疗有效性或不良副作用以及社会环境影响。

公开(公告)号：US07208467B2

公开(公告)日：2007-04-24

申请号：US10164863

申请日：2002-06-07

申请人： Monty Krieger ,  Helena E. Miettinen

发明人： Monty Krieger ,  Helena E. Miettinen

IPC分类号： A01N37/18 ,  A01N43/16 ,  A01N43/08 ,  A01N31/00 ,  A61K38/00

CPC分类号： A61K31/375 ,  A61K31/00 ,  A61K31/095 ,  A61K31/355 ,  A61K31/365 ,  A61K2300/00

摘要： SR-BI is present at relatively high levels on the membranes of hepatocytes and steroidogenic tissues, including the adrenal gland, testes, and ovaries, where it mediates the uptake and transport of cholesteryl ester from high density lipoproteins. It has been demonstrated that transgenic animals which do not produce SR-BI are healthy, with the exception that the females are infertile. SR-BI KO females have abnormal HDLs, ovulate dysfunctional oocytes and are infertile. Surgical, genetic and pharmacologic methods were used to show that the fertility of SR-BI KO females (or their transplanted oocytes) can be restored in the absence of ovarian and/or extraovarian SR-BI expression by manipulations that modify the structure, composition and/or abundance of their abnormal plasma lipoproteins. These manipulations included inactivation of the apolipoprotein A-I gene and administration of the cholesterol-lowering drug PROBUCOL™. In the absence of treatment, female animals which do not express SR-BI have dramatically reduced levels of offspring, even though they are otherwise healthy and the males normal. Studies demonstrate that they do not produce viable eggs and have a defect involving implantation of normal eggs.

摘要翻译： SR-BI以相当高的水平存在于肝细胞和类固醇生成组织的膜上，包括肾上腺，睾丸和卵巢，其中介导胆固醇酯从高密度脂蛋白的摄取和转运。 已经证明，不产生SR-BI的转基因动物是健康的，除了女性是不育的。 SR-BI KO女性HDL异常，排卵功能异常卵母细胞不育。 使用外科学，遗传学和药理学方法表明，SR-BI KO女性（或其移植的卵母细胞）的生育力可以通过改变结构，组成和功能的操作在没有卵巢和/或外源SR-BI表达的情况下恢复 /或其异常血浆脂蛋白的丰度。 这些操作包括载脂蛋白A-1基因的失活和胆固醇降低药物PROBUCOL TM的给药。 在没有治疗的情况下，不表达SR-BI的雌性动物大大降低了后代的水平，即使它们健康，而雄性也正常。 研究表明，它们不产生活的卵，并且涉及植入正常卵的缺陷。

公开(公告)号：US5925333A

公开(公告)日：1999-07-20

申请号：US559505

申请日：1995-11-15

申请人： Monty Krieger ,  Susan L. Acton ,  Attilio Rigotti ,  Helen Haskell Hobbs ,  Katherine Tallman Landschulz

发明人： Monty Krieger ,  Susan L. Acton ,  Attilio Rigotti ,  Helen Haskell Hobbs ,  Katherine Tallman Landschulz

IPC分类号： C12N15/09 ,  A61K31/00 ,  A61K31/56 ,  A61K31/565 ,  A61K35/76 ,  A61K38/00 ,  A61K38/22 ,  A61K45/00 ,  A61K48/00 ,  A61P3/00 ,  A61P3/06 ,  C07K14/705 ,  C12N15/00 ,  C12N15/12 ,  C12N15/85 ,  C12N15/79

CPC分类号： C12N15/8509 ,  C07K14/705 ,  A01K2217/05 ,  A01K2227/105 ,  A01K2267/025 ,  A01K2267/03 ,  A01K2267/0362 ,  A61K38/00 ,  A61K48/00 ,  C12N2799/022

摘要： Methods for regulation of lipid and cholesterol uptake are described which are based on regulation of the expression or function of the SR-BI HDL receptor. The examples demonstrate that estrogen dramatically downregulates SR-BI under conditions of tremendous upregulation of the LDL-receptor. The examples also demonstrate the upregulation of SR-BI in rat adrenal membranes and other non-placental steroidogenic tissues from animals treated with estrogen, but not in other non-placental non-steroidogenic tissues, including lung, liver, and skin. Examples further demonstrate the uptake of fluorescently labeled HDL into the liver cells of animal, which does not occur when the animals are treated with estrogen.

摘要翻译： 描述了调节脂质和胆固醇摄取的方法，其基于调节SR-BI HDL受体的表达或功能。 这些实施例证明雌激素在LDL-受体大量上调的条件下显着下调SR-BI。 这些实施例还证明了在用雌激素治疗的动物的大鼠肾上腺膜和其它非胎盘类固醇生成组织中SR-BI的上调，但在其他非胎盘非类固醇生成组织（包括肺，肝脏和皮肤）中的上调。 实施例进一步证明了将荧光标记的HDL摄取到动物的肝细胞中，当用雌激素治疗时不会发生。