Mouse Models of Vitamin Transport Defects

描述(由申请人提供)： SLC19A3是维生素转运蛋白家族的成员，该家族包括还原型叶酸载体SLC19A1和高亲和力硫胺转运蛋白SLC19A2。这个溶质载体亚家族的成员是12膜跨膜蛋白，专门运输微量营养素，如叶酸(SLC19A1)或硫胺(SLC19A2和SLC19A3)。我们假设SLC19A3作为硫胺素进入体内的主要门户，作为胎盘硫胺素的主要(如果不是唯一的)转运蛋白。它也可能是肠道硫胺素的主要转运体。硫胺素和其他维生素一样，是通过饮食摄入获得的。全身性(饮食)硫胺素缺乏可导致一系列问题，包括神经退化、消瘦和死亡。本研究拟对SLCI9A3RNA和蛋白质在发育中的胚胎和成体组织中的表达模式进行研究。通过SLC19A3将硫胺素传递到组织中的功能作用将通过靶向缺失SLC19A3的小鼠模型进行研究。此外，将建立仅在某些组织中缺失SLC19A3的转基因小鼠系，以评估硫胺素对特定发育阶段和单个器官系统的重要性。这些研究将提供有关硫胺素在发育以及在正常和疾病系统中的作用的重要信息。 建议的工作将在Ellis Neufeld博士的实验室和儿童医院血液科提供的出色环境中进行。Neufeld博士在遗传学、血液学和生物化学方面的专业知识，以及与小鼠开发和转基因模型专家(例如Stuart Orkin博士和Nancy Andrews博士)的密切联系，将对申请者进一步发展成为独立的学术研究生涯产生重大影响。拟议的项目将在一流的培训环境中进行，这将提高申请者的技能和知识水平。建议的实验类型，使用转基因/基因敲除方法，可能会为申请者未来作为独立研究人员提供几个新的研究途径，并肯定会提供必要的初步数据，以竞争完全独立的资金。

DESCRIPTION (provided by applicant): SLC19A3 is a member of the vitamin transporter family that includes the reduced folate carrier, SLC 19A1 and the high-affinity thiamine transporter, SLC19A2. Members of this solute carrier sub-family are 12-membrane spanning proteins that specifically transport micronutrients such as folate (SLC19A1) or thiamine (SLC19A2 and SLC19A3). We hypothesize that SLC19A3 functions as a major portal for thiamine into the body, functioning as a major (if not the only) placental thiamine transporter. It is likely to also be a major intestinal thiamine transporter as well. Thiamine, like other vitamins, is obtained through dietary intake. Systemic (dietary) thiamine deficiency can lead to a multitude of problems including neurodegeneration, wasting, and death. This study proposes to characterize the expression pattern of SLCI9A3 RNA and protein in developing embryos and adult tissues. The functional role of thiamine delivery into tissues via SLC19A3 will be addressed using a mouse model with a targeted deletion of SLC19A3. Additionally, transgenic mouse lines in which SLC19A3 is deleted in only certain tissues will be created in order to assess the importance of thiamine to particular stages of development and to individual organ systems. These studies will provide important information concerning the contribution of thiamine in development and in normal and disease systems. The work proposed will be performed in the outstanding environment provided by Dr. Ellis Neufeld's laboratory and the Division of Hematology at Children's Hospital. Dr. Neufeld's expertise in genetics, hematology and biochemistry, as well as the close proximity of experts in mouse development and transgenic models (e.g. Drs. Stuart Orkin and Nancy Andrews) will have a great impact on the further development of the applicant towards an independent academic research career. The proposed project will be carried out in a superb training environment that will enhance the applicant's skill and knowledge repertoire. The types of experiments proposed, using a transgenic/knockout approach, will likely provide the applicant with several new research avenues to focus on in the future as an independent researcher, and will certainly provide the necessary preliminary data to compete for completely independent funding.