Identifying New Therapeutics and Molecular Mechanisms in Congenital Disorders of Glycosylation.

确定先天性糖基化疾病的新疗法和分子机制。

基本信息

批准号：
10644811
负责人：
Hans Martin Dalton
金额：
$ 11.61万
依托单位：
UNIVERSITY OF UTAH
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-06-01 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10644811
关键词：
Advisory Committees Affect Apoptosis Biological Biological Assay Candidate Disease Gene Cell Culture Techniques Cell Line Cell Proliferation Cell model Cells Cellular Metabolic Process Cessation of life Clinical Trials Congenital disorders of glycosylation Defect Development Developmental Delay Disorders Disease Disease model Down-Regulation Drosophila genus Drug Screening Ensure Enzymes Epilepsy Eye Faculty Future Gene Expression Genes Genetic Glycobiology Goals Health Human Human Cell Line In Vitro Inborn Errors of Metabolism Individual Institution Intervention Learning Link Mass Spectrum Analysis Mentors Mentorship Metabolism Microscopy Modeling Molecular Molecular Biology Mutation Organism Pathway interactions Patients Pharmaceutical Preparations Pre-Clinical Model Process Proliferating Proteins RNA Interference Rare Diseases Research Seizures Stress Techniques Testing Therapeutic Therapeutic Uses Training Translating Universities Utah Work Writing Yeasts career common symptom developmental disease disease phenotype drug repurposing drug use screening endoplasmic reticulum stress fly genetic manipulation glycosylation improved in vivo in vivo Model loss of function loss of function mutation member metabolomics new therapeutic target novel therapeutics patient population protein expression reduce symptoms small molecule small molecule libraries sugar therapeutic evaluation tool

项目摘要

Glycosylation is an essential biological pathway that involves the post- or co-translational addition of sugar moieties to proteins. Congenital Disorders of Glycosylation (CDGs) are rare developmental disorders caused by inborn errors of metabolism in glycosylation pathways. CDGs lack good treatment options, and this is typically due to poorly understood mechanisms and difficulty in establishing clinical trials in small patient populations. My long-term objectives are to determine mechanisms of CDGs and identify new therapeutics for CDG patients. One example is DPAGT1-CDG - a CDG caused by mutations in the gene DPAGT1 which encodes for the first enzyme used in N-linked glycosylation. Recently, I identified many modifier genes which can be perturbed to rescue a model of DPAGT1-CDG, but their mechanisms are not yet known. In Aim 1, I propose to determine the mechanisms of these modifier genes using human cell culture in order to characterize new therapeutic targets for this disorder. I will use a DPAGT1-CDG cell model to determine how these rescuing modifier genes affect patient-related health metrics of proliferation, stress, and their glycoproteome. In Aim 2, to identify new drugs that can rescue this disorder, I will use an in vivo Drosophila DPAGT1-CDG model to perform a repurposed drug screen using 1,500+ small molecules (98% FDA/EMA-approved). Using an in vivo model will ensure these drugs are safe during development, and this repurposed drug screen will help expedite the clinical trial process for new CDG therapies. In Aim 3, I will characterize a new finding that suggests that genes underlying CDGs ("CDG genes") themselves represent an enriched set of modifier genes for treating CDGs. Perturbation of CDGs can rescue models of DPAGT1-CDG, as well as a model of the most common CDG, PMM2-CDG. I will use RNA interference to perturb all 150+ CDG genes to identify any that are capable of rescuing both DPAGT1- and PMM2-CDG human cell models (with the same health metrics as in Aim 1). The discovery of new CDG gene modifier genes capable of rescuing these models could have the potential to translate into future therapies for many other CDGs. Finally, in Aim 4, I will synthesize the above Aims to test therapeutic drugs from Aim 2 in human cell culture models and CDG modifier genes from Aim 3 in Drosophila models. I will use high-throughput tools in cell culture and in vivo stress markers in Drosophila to determine the mechanisms of these new therapies. This multi-species approach will ensure a better transition from preclinical models into therapies for patients. In addition to the above, completion of this proposal will provide me with training to complete my career goals. I will learn new techniques in cell culture and small molecule screens while also taking formal courses in mentorship and writing. I have an outstanding mentor, co-mentor, and advisory committee consisting of faculty with expertise in CDGs, cell culture, drug screening, genetics, and molecular biology. I also have state-of-the-art facilities and staff at the University of Utah available to me. With my plan, committed faculty members, and excellent institution, completing this proposal will help me successfully transition to an independent research career.

糖基化是一种重要的生物途径，涉及糖的翻译后或共翻译添加蛋白质的部分。先天性糖基化障碍 (CDG) 是由以下原因引起的罕见发育障碍糖基化途径代谢的先天性错误。 CDG 缺乏良好的治疗选择，这通常是由于对机制知之甚少以及在小患者群体中开展临床试验存在困难。我的长期目标是确定 CDG 的机制并为 CDG 患者找到新的治疗方法。一个例子是 DPAGT1-CDG - 一种由编码第一个基因 DPAGT1 的突变引起的 CDG 用于 N-连接糖基化的酶。最近，我发现了许多可以扰乱的修饰基因拯救了 DPAGT1-CDG 模型，但其机制尚不清楚。在目标 1 中，我建议确定使用人类细胞培养物来表征这些修饰基因的机制，以表征新的治疗靶点对于这种疾病。我将使用 DPAGT1-CDG 细胞模型来确定这些拯救修饰基因如何影响与患者相关的增殖、应激及其糖蛋白组健康指标。目标 2：识别新药为了挽救这种疾病，我将使用体内果蝇 DPAGT1-CDG 模型来进行药物的重新利用使用 1,500 多种小分子进行筛选（98% 获得 FDA/EMA 批准）。使用体内模型将确保这些药物在开发过程中是安全的，这种重新利用的药物筛选将有助于加快新药的临床试验过程 CDG疗法。在目标 3 中，我将描述一项新发现，该发现表明 CDG 背后的基因（“CDG 基因”）本身代表了一组丰富的用于治疗 CDG 的修饰基因。CDG 的扰动可以 DPAGT1-CDG 的救援模型，以及最常见的 CDG 的模型 PMM2-CDG。我将使用RNA 干扰扰乱所有 150 多个 CDG 基因，以确定任何能够拯救 DPAGT1- 和 PMM2-CDG 人类细胞模型（具有与目标 1 相同的健康指标）。新CDG基因的发现能够拯救这些模型的修饰基因可能有潜力转化为未来的治疗方法许多其他 CDG。最后，在目标 4 中，我将综合上述目标来测试目标 2 中的治疗药物人类细胞培养模型和果蝇模型中 Aim 3 的 CDG 修饰基因。我将使用高吞吐量细胞培养工具和果蝇体内应激标记物，以确定这些新疗法的机制。这种多物种方法将确保更好地从临床前模型过渡到患者治疗。在除此之外，完成本提案将为我提供完成职业目标的培训。我会学习细胞培养和小分子筛选的新技术，同时参加正规的指导课程和写作。我有一位杰出的导师、共同导师和由具有专业知识的教师组成的咨询委员会 CDG、细胞培养、药物筛选、遗传学和分子生物学。我还拥有最先进的设施犹他大学的工作人员可以为我服务。凭借我的计划、忠诚的教职人员和优秀的机构，完成这个提案将帮助我成功过渡到独立研究生涯。