Live-Cells Assays for Lysosomal Enzyme Activity

溶酶体酶活性的活细胞测定

基本信息

批准号：
7424029
负责人：
JOHN J NALEWAY
金额：
$ 19.98万
依托单位：
MARKER GENE TECHNOLOGIES
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-01-01 至 2009-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7424029
关键词：
Acid Phosphatase Acids Affect Ally Anabolism Animal Model Antibiotics Antiviral Agents Apoptosis Area Aspartylglucosaminuria Autophagocytosis Biochemistry Biological Assay Biology Carbohydrates Caspase Cathepsins Cell Culture System Cell Cycle Stage Cell Separation Cell Survival Cell membrane Cells Cellular Morphology Cellular biology Charge Class Communities Condition Cultured Cells Degenerative Disorder Detection Development Disease Drug Delivery Systems Environment Enzyme Induction Enzymes Evaluation Event Fabry Disease Fluorescence Fluorogenic Substrate Fucosidosis Gangliosidoses Gangliosidoses GM2 Gaucher Disease Gene Expression Gene Expression Regulation Gene Targeting Genes Glycoside Hydrolases Glycosphingolipids Goals Hereditary Disease In Vitro Individual Intervention Intervention Studies Investigation Knowledge Lead Life Lipase Lipoidosis Localized Lysosomal Storage Diseases Lysosomes Mammalian Cell Mannosidase Deficiency Diseases Measurement Measures Mediating Medical Medicine Methodology Methods Modality Modeling Molecular Monitor Morphology Mucolipidoses Mucopolysaccharidoses Mucopolysaccharidosis I Mucopolysaccharidosis I H Mucopolysaccharidosis III Mucopolysaccharidosis IV Mucopolysaccharidosis VI Mucopolysaccharidosis VII Mutation Myoclonus Cherry Red Spot Syndrome Normal Cell Organelles Peptide Hydrolases Phagocytosis Pharmaceutical Preparations Pharmacologic Substance Pharmacology Phase Phosphoric Monoester Hydrolases Physiological Preclinical Drug Evaluation Preparation Process Production Property Proteins Protocols documentation Reagent Reporter Research Route Sandhoff Disease Screening procedure Side Small Business Innovation Research Grant Solubility Staining method Stains Standards of Weights and Measures Syndrome Synthesis Chemistry System Tay-Sachs Disease Techniques Technology Testing Therapeutic Time Tissue Sample Tissues Translating Validation Viral Work analytical tool base cell type drug discovery enzyme activity enzyme substrate high throughput screening improved in vitro Assay in vivo inhibitor/antagonist innovation interest novel novel therapeutics prevent protein aminoacid sequence protein metabolism prototype response success tissue culture tissue/cell culture tool

项目摘要

DESCRIPTION (provided by applicant): This Small Business Innovation Research Phase I project aims to develop new targeted fluorogenic substrates capable of measuring lysosomal enzyme activity in living cells and tissues. If successful, the proposed research will provide breakthroughs needed to advance the discovery of promising new therapies and modulating drugs for lysosomal storage diseases and allied medical applications. In Phase I of this project, Marker Gene Technologies, Inc. will establish the feasibility of the technology by preparing new fluorogenic glycosidase substrates for lysosomal enzymes and demonstrating differential staining in living cells that are from normal or are of disease origin. In Phase II, these new substrates will be assayed in vitro and in vivo for their ability to measure specific and localized inhibition or induction of lysosomal enzymes in living cells as well as differentiate individual enzyme activities in a cell- or tissue-specific manner. The new substrates and the resulting detection systems will provide innovative methods to quantitate lysosomal enzyme function and to screen for the influence of secondary drug or protein administration, making them useful analytical tools for a variety of significant medical applications. The success of this project opens up enormous commercial possibilities in the fields of medical intervention in lysosomal storage diseases such as Sandhoff's disease, Tay-Sachs syndrome, Krabb¿'s disease and Gaucher's disease, as well as in the screening of new proteins and drugs in cell-culture systems for efficacy in modulating lysosomal enzyme activity in these diseases, and development of new, general and specific high-throughput lysosomal enzyme detection strategies. In addition, it will lead to commercial and licensable products in these areas.

描述（由应用提供）：这个小型企业创新研究阶段项目旨在开发能够测量活细胞和组织中溶酶体酶活性的新的靶向荧光底物。如果成功的话，拟议的研究将提供突破性的突破，以提高有望的新疗法并调节用于溶酶体储存疾病和相关医疗应用的药物。在该项目的第一阶段，Marker Gene Technologies，Inc。将通过为溶酶体酶准备新的荧光糖苷酶底物来确定该技术的可行性，并在活细胞中表现出来自正常或疾病起源的活细胞中的差异染色。在第二阶段，这些新底物将在体外和体内分配，以测量活细胞中溶酶体酶的特定和局部抑制或诱导，并以细胞或组织特异性方式区分单个酶活性。新的底物和结果检测系统将提供创新的方法来定量溶酶体酶功能并筛选二次药物或蛋白质给药的影响，从而使它们适用于各种重要的医疗应用。 The success of this project opens up enormous commercial possibilities in the fields of medical intervention in lysosomal storage diseases such as Sandhoff's disease, Tay-Sachs syndrome, Krabb¿ 's disease and Gaucher's disease, as well as in the screening of new proteins and drugs in cell-culture systems for efficiency in modulating lysosomal enzyme activity in these diseases, and development of new, general and specific高通量溶酶体酶检测策略。此外，这将导致这些领域的商业和有许可产品。

项目成果

期刊论文数量（2）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Fluorogenic Substrates for Visualizing Acidic Organelle Enzyme Activities.

DOI：
10.1371/journal.pone.0156312
发表时间：
2016
期刊：
PloS one
影响因子：
3.7
作者：
Harlan FK;Lusk JS;Mohr BM;Guzikowski AP;Batchelor RH;Jiang Y;Naleway JJ
通讯作者：
Naleway JJ

A long-wavelength fluorescent substrate for continuous fluorometric determination of alpha-mannosidase activity: resorufin alpha-D-mannopyranoside.

用于连续荧光测定 α-甘露糖苷酶活性的长波长荧光底物：试卤灵 α-D-吡喃甘露糖苷。

DOI：
10.1016/j.ab.2009.11.039
发表时间：
2010
期刊：
Analytical biochemistry
影响因子：
2.9
作者：
Coleman,DanielJ;Kuntz,DouglasA;Venkatesan,Meenakshi;Cook,GabrieleM;Williamson,StaciP;Rose,DavidR;Naleway,JohnJ
通讯作者：
Naleway,JohnJ