HERITABLE DISORDERS OF METABOLISM OF PHOSPHOLIPIDS AND LIPID-MODIFIED PROTEINS

磷脂和脂质修饰蛋白代谢的遗传性疾病

• 批准号: 6290198
• 负责人: ANIL B MUKHERJEE
• 金额: --
• 依托单位: EUNICE KENNEDY SHRIVER NATIONAL INSTITUTE OF CHILD HEALTH & HUMAN DEVELOPMENT
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6290198
• 关键词: 3T3 cells; antisense nucleic acid; asthma; chemokine; disease /disorder model; extracellular matrix; gene expression; gene targeting; genetic disorder; globulins; glomerulonephritis; human tissue; laboratory mouse; macular degeneration; mammary gland; neuronal ceroid lipofuscinosis; nuclear magnetic resonance spectroscopy; osteopontin; osteoporosis; phospholipase A2; protein structure function; protooncogene; restenosis; tissue /cell culture; vascular smooth muscle

During the past year we have accomplished the following : (i) by generating two different animal models (i.e. antisense- transgenic and gene-knockout), expressing a low level or lacking uteroglobin, a potent phospholipase A2 (PLA2) inhibitor, we uncovered that both models develop IgA nephropathy, the most common primary renal glomerular disease in the world. We determined the molecular mechanism by which lack of UG causes IgA deposition in the glomeruli of these animals and demonstrated that supplementation of recombinant UG prevents glomerular IgA deposition. Studies in progress will delineate whether the human disease is caused by UG deficiency; (ii) characterized a high-affinity receptor of uteroglobin and its inducible expression in various normal and tumor cell lines. We also found that via this receptor uteroglobin regulates the motility and ECM-invasiveness of many cell types. We also uncovered that via this receptor- mediated pathway uteroglobin may reverse the transformed phenotype of cancer cells expressing the receptor, and in this respect, it manifests a tumor-suppressor-like function; (iii) isolated and characterized mouse palmitoyl-protein thioesterase (mPPT) cDNA and the gene, respectively, and deciphered a temporal pattern suggesting that the expression of this gene in the retina precedes that in the developing brain, explaining in part, why the loss of vision occurs earlier than the deterioration of brain function in infantile neuronal ceroid lipofuscinosis (INCL); (iv) cloned and characterized the cDNA and the gene encoding murine and human palmitoyl-protein thioesterases (PPT, respectively, mutations in which causes infantile ceroid lipofuscinosis (INCL). INCL belongs to a relatively common (1/12,500) autosomal recessive heritable disorder without effective treatment; (v) the gene encoding human acid ceramidase, the mutation of which causes an autosomal recessive heritable disorder of lipid metabolism, Farber lipogranulomatosis, has been cloned and characterized. A corresponding murine cDNA and the gene have also been cloned and attempts being made to generate a mouse model for this disease in order to understand the pathology and to develop an effective treatment; (vi) characterized a high-affinity cell surface receptor for group I phospholipase A2 (PLA2) and found that via this receptor sPLA2-I induces cellular invasion of the extracellular matrix and this receptor is expressed at a high level in several tumor cell lines; (vii) demonstrated a critical role of nuclear factor IL6 (NF IL-6) in the induction of sPLA2-I receptor-mediated activation of the cyclooxygenase- 2 (COX-2) gene expression in MC-3T3 and human colorectal adenocarcinoma cell lines; (viii) cloned and characterized a cDNA and the gene for murine group I pancreatic PLA2 and its differential tissue-specific expression in the mouse; (ix) discovered that sPLA2-I gene expression is drastically down- regulated in colorectal cancer tissues and in the colon of Min (multiple intestinal neoplasia) mouse, an animal model for familial adenomatous polyposis (FAP) and heritable colorectal cancer in humans suggesting a modifier role of sPLA2-I in this disease; (x)) in collaboration with Claragen, Inc., animal studies are performed to determine the tissue distribution and toxicity of recombinant UG towards phase-I clinical trial of this protein in the treatment of inflammatory and fibrotic lung disorders. Patent applications for the use of recombinant UG as a potential therapeutic agent for IgA-nephropathy has been filed. - Infantile Neuronal Ceroid Lipofuscinosis, Batten Disease,IgA-Nephropathy, Farber disease

在过去的一年里，我们完成了以下工作：(i) 通过生成两种不同的动物模型（即反义转基因和基因敲除），表达低水平或缺乏子宫珠蛋白（一种有效的磷脂酶 A2 (PLA2) 抑制剂），我们发现两种模型都会出现 IgA 肾病，这是世界上最常见的原发性肾小球疾病。我们确定了缺乏 UG 导致这些动物肾小球 IgA 沉积的分子机制，并证明补充重组 UG 可防止肾小球 IgA 沉积。正在进行的研究将确定人类疾病是否是由 UG 缺乏引起的； (ii) 表征了子宫珠蛋白的高亲和力受体及其在各种正常和肿瘤细胞系中的诱导表达。我们还发现，子宫珠蛋白通过该受体调节许多细胞类型的运动性和 ECM 侵袭性。我们还发现，通过这种受体介导的途径，子宫珠蛋白可以逆转表达该受体的癌细胞的转化表型，在这方面，它表现出类似肿瘤抑制的功能； (iii) 分别分离和表征了小鼠棕榈酰蛋白硫酯酶 (mPPT) cDNA 和该基因，并破译了一种时间模式，表明该基因在视网膜中的表达先于发育中的大脑中的表达，部分解释了为什么缺失婴儿神经元蜡样质脂褐质沉积症 (INCL) 中视力的下降早于脑功能的恶化； (iv) 克隆并表征了编码鼠和人棕榈酰蛋白硫酯酶的 cDNA 和基因（分别为 PPT，突变导致婴儿蜡质脂褐质沉着症 (INCL)。 INCL 属于相对常见的 (1/12,500) 常染色体隐性遗传性疾病未经有效治疗；（v）编码人酸性神经酰胺酶的基因，其突变导致常染色体隐性遗传性脂质代谢疾病，Farber已克隆并鉴定了相应的鼠 cDNA 和基因，并尝试建立该疾病的小鼠模型，以了解病理学并开发有效的治疗方法； -I族磷脂酶A2（PLA2）的亲和细胞表面受体，并发现通过该受体sPLA2-I诱导细胞外基质的细胞侵袭，并且该受体在多种肿瘤细胞中高水平表达(vii) 证明核因子 IL6 (NF IL-6) 在诱导 MC-3T3 和人结直肠腺癌中 sPLA2-I 受体介导的环氧合酶-2 (COX-2) 基因表达激活中发挥关键作用细胞系； (viii) 克隆并表征了小鼠 I 组胰腺 PLA2 的 cDNA 和基因及其在小鼠中的差异组织特异性表达； (ix) 发现 sPLA2-I 基因表达在结直肠癌组织和 Min（多发性肠瘤）小鼠的结肠中急剧下调，Min（多发性肠瘤）小鼠是家族性腺瘤性息肉病 (FAP) 和人类遗传性结直肠癌的动物模型，这表明sPLA2-I 在该疾病中的修饰作用； (x)) 与 Claragen, Inc. 合作，进行动物研究以确定重组 UG 的组织分布和毒性，以进行该蛋白治疗炎症和纤维化肺部疾病的 I 期临床试验。使用重组 UG 作为 IgA 肾病潜在治疗剂的专利申请已提交。 - 婴儿神经元蜡质脂褐质沉积症、Batten病、IgA肾病、Farber病