Targeting the Notch3 Mutation to Cure Lehman Syndrome

针对 Notch3 突变来治愈雷曼综合症

基本信息

批准号：
10684552
负责人：
Ernesto Canalis
金额：
$ 41.1万
依托单位：
UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-15 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10684552
关键词：
Affect Alleles Antisense Oligonucleotides Antisense Technology Award Bone remodeling Cell Line Cells Clinical DNA Sequence Alteration Development Disease Down-Regulation Exhibits Exons Gene Mutation Genes Genetic Diseases Goals Human In Vitro Intervention Knowledge Lateral Lead Liver Mendelian disorder Meningocele Molecular Abnormality Mus Mutant Strains Mice Mutation NOTCH3 gene Osteoblasts Osteoclasts Osteopenia Other Genetics Peripheral Nervous System Phase Phenotype Play Positioning Attribute Proteins Regulation Research Retina Role Secondary to Skeleton Syndrome TRANCE protein Testing Ubiquitination Work bone bone loss cell behavior cortical bone craniofacial effective intervention efficacy study gain of function human disease human model in vivo induced pluripotent stem cell mouse model mutant notch protein novel strategies novel therapeutic intervention osteoclastogenesis premature prevent skeletal skeletal disorder substantia spongiosa

项目摘要

PROJECT SUMMARY/ABSTRACT Notch receptors play a critical role in cell fate decisions and in the regulation of osteoblast and osteoclast differentiation and function. As a consequence, Notch plays an important role in bone remodeling. Lehman Syndrome or Lateral Meningocele Syndrome is a devastating disease characterized by craniofacial developmental abnormalities, bone loss and meningoceles. The syndrome is associated with mutations in exon 33 of NOTCH3 upstream of the PEST domain leading to NOTCH3 stabilization and presumably gain-of- function. We created a mouse model of Lehman Syndrome (Notch3tm1.1Ecan) that presents with osteopenia due to enhanced osteoclastogenesis secondary to an increase in receptor activator of nuclear factor Kappa B ligand (RANKL) expression by cells of the osteoblast lineage. The aim of the proposed research is to develop ways to correct the skeletal manifestations of the disease by targeting the mutation with Notch3 antisense oligonucleotides (ASO), a strategy that would be applicable to other genetic disorders of the skeleton. Our specific aims are: Phase R61 Aim 1) To establish the efficacy of Notch ASOs in vitro and in vivo. In this initial aim, we will test whether Notch3 can be downregulated in the skeleton and reverse the skeletal phenotype of Notch3tm1.1Ecan mice; Aim 2) To establish that the Notch3tm1.1Ecan mutation can be targeted. We will determine whether the Notch3tm1.1Ecan mutation can be downregulated specifically and the Notch3tm1.1Ecan skeletal phenotype reversed by the administration of Notch36691-TAATGA antisense oligonucleotides; and Phase R33 Aim 3) To validate the ASO approach in NOTCH3 mutant-induced pluripotent (iPS) cells. In the R33 phase, we intend to prove the utility of Notch3 ASOs in human cells by creating mutant iPS cell lines to study the efficacy of Notch3 ASOs in downregulating NOTCH3 mutant alleles. The goals of the proposed work are to develop specific antisense technology to treat skeletal manifestations of a devastating NOTCH3-associated disease, as an initial step in the treatment of genetic disorders of the skeleton.

项目摘要/摘要 Notch受体在细胞命运决策以及成骨细胞和破骨细胞的调节中起关键作用分化和功能。结果，Notch在骨骼重塑中起着重要作用。雷曼综合征或外侧脑膜酸综合征是一种毁灭性疾病，其特征是颅面发育异常，骨质流失和脑膜炎。该综合征与突变有关 Notch3的外显子33害虫域的上游导致Notch3稳定，并可能获得功能。我们创建了一个带有骨质减少的鼠标综合征（Notch3tm1.1can）的鼠标模型为了增强核因子kappa b受体激活剂的继发于骨质质发生成骨细胞谱系细胞的配体（RANKL）表达。拟议研究的目的是发展通过Notch3反义靶向突变，纠正疾病的骨骼表现的方法寡核苷酸（ASO），该策略适用于骨骼的其他遗传疾病。我们的具体目的是：R61 AIM 1）确定Notch ASOS在体外和体内的功效。在此最初目的，我们将测试Notch3是否可以在骨骼中下调并逆转骨骼表型 Notch3tm1.1can小鼠；目的2）确定可以针对Notch3tm1.1can突变。我们将确定 notch3tm1.1 ecan突变是否可以专门下调和Notch3TM1.1can骨骼 Notch36691-Taatga反义寡核苷酸的给药反转。和R33阶段目标 3）验证Notch3突变诱导的多能（IPS）细胞中的ASO方法。在R33阶段，我们打算通过创建突变体IPS细胞系来研究疗效，以证明Notch3 ASO在人类细胞中的实用性在下调Notch3突变等位基因中的Notch3 ASO。拟议工作的目标是发展治疗毁灭性Notch3相关疾病的骨骼表现的特定反义技术，治疗骨骼遗传疾病的第一步。