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Novel Intestinal Nutrient Membrane Transporters

新型肠道营养膜转运蛋白

基本信息

批准号：
RGPIN-2018-06027
负责人：
Eck, Peter
金额：
$ 5.83万
依托单位：
University of Manitoba
依托单位国家：
加拿大
项目类别：
Discovery Grants Program - Individual
财政年份：
2022
资助国家：
加拿大
起止时间：
2022-01-01 至 2023-12-31
项目状态：
已结题

来源：
https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=757185
关键词：
Novel Intestinal Nutrient Membrane Transporters

项目摘要

Background: This basic research program is set up to discover novel nutrient membrane transporters in order to understand the biology of transport pathways. Our research group has a track record of identifying novel transporters and we have developed a “Research Pipeline” for membrane transporters discovery. We have four current research leads on “missing links” in ascorbic acid and organic cation pathways, which we propose to explore using a novel methodology developed by us. Due to a key role of intestinal absorption as a determinant of systemic homoeostasis the long term focus of this research program lays on transport pathways in the absorbing epithelial cell, the enterocyte. Since we do not have a complete understanding of all the transporters expressed in the enterocyte, we propose two experimental approaches to identify future targets. Objectives: Four existing candidate genes in pathways of vitamin C and organic cation transport will be explored. Two subprojects will inform on future research targets. The individual objectives are: 1. Ascorbic Acid Uptake: Identification of additional candidates through differential gene expression in CaCo-2 cells exposed to varying concentration of ascorbic acid. 2. Ascorbic Acid Release: Characterization of slc23a3 and slc22a23 as “releaser”. 3. Dehydroascorbic Acid Uptake in the Small Intestine: Characterization of GLUT14's role in the pathway. 4. Organic Cations Uptake: Characterization of slc22a23 as multispecific “uptaker”.5. Organic Cations Release: Characterization of mfsd14b as “releaser” of primary amines. 6. The Transcriptome of the Enterocyte to generate future research targets. Design: Our molecular “Research Pipeline” utilizes common methodology to characterize individual membrane transporters in objectives 2-5. This will result in genes (a) annotation, (b) Subcloning of major protein isoforms, (c) determination of subcellular localization, (d) substrate studies. Two transcriptomics studies aid the identification of another elusive ascorbic acid uptake transporter and the transcriptome of the enterocyte and will inform on future targets. Significance: The program will discover novel membrane transporters. Its methodology will be a template for further studies. The basic discoveries will prime translation to enhance physiologic performance or environmental challenges.

背景资料：这个基础研究计划的目的是发现新的营养膜转运蛋白，以了解运输途径的生物学。我们的研究小组在识别新型转运蛋白方面有着良好的记录，我们已经为膜转运蛋白的发现开发了一个“研究管道”。我们有四个目前的研究线索“缺失环节”在抗坏血酸和有机阳离子途径，我们建议探索使用一种新的方法，我们开发的。由于肠道吸收作为全身体内平衡的决定因素的关键作用，本研究计划的长期重点在于吸收上皮细胞（肠上皮细胞）中的转运途径。由于我们没有一个完整的理解，所有的转运蛋白在肠上皮细胞中表达，我们提出了两个实验方法来确定未来的目标。目的：探索维生素C和有机阳离子转运途径中存在的4个候选基因。两个子项目将为今后的研究目标提供信息。个人目标是：1。抗坏血酸摄取：通过暴露于不同浓度的抗坏血酸的CaCo-2细胞中的差异基因表达鉴定其他候选物。2.抗坏血酸释放：slc 23 a3和slc 22 a23作为“抗坏血酸释放剂”的表征。 3.小肠中脱氢抗坏血酸的摄取：GLUT 14在通路中作用的表征。4.有机阳离子吸收：slc 22 a23作为多特异性“吸收剂”的表征。有机阳离子释放：mfsd 14 b作为伯胺的“释放剂”的表征。6.肠上皮细胞的转录组产生未来的研究目标。设计：我们的分子“研究管道”利用常见的方法来表征目标2-5中的单个膜转运蛋白。这将导致基因（a）注释，（B）主要蛋白质同种型的亚克隆，（c）亚细胞定位的确定，（d）底物研究。两个转录组学研究有助于确定另一个难以捉摸的抗坏血酸摄取转运蛋白和肠上皮细胞的转录组，并将告知未来的目标。意义：该计划将发现新的膜转运蛋白。其方法将成为进一步研究的模板。这些基本发现将引发翻译，以提高生理性能或环境挑战。