Mechanisms and Treatment of Hajdu Cheney Syndrome

Hajdu Cheney 综合症的机制和治疗

• 批准号: 10655476
• 负责人: Ernesto Canalis
• 金额: $ 36.08万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10655476
• 关键词: Acro-Osteolysis; Affect; Alleles; Antisense Oligonucleotides; Antisense Technology; Behavior; Bone Resorption; Bone remodeling; Cell Differentiation process; Cell Line; Cells; Clinical; Cytolysis; Dependence; Development; Disease; Disease model; Distal; ES01; Effectiveness; Enhancers; Event; Exhibits; Exons; Fracture; Genes; Genetic Diseases; Goals; Hajdu-Cheney Syndrome; Heterozygote; Human; In Vitro; Intervention; Investigation; Knowledge; Lead; Ligands; Modeling; Mus; Mutant Strains Mice; Mutation; Myelogenous; NF-kappa B; Nuclear; Osteoblasts; Osteoclasts; Osteopenia; Other Genetics; Phalanx; Phenotype; Play; Positioning Attribute; Proteins; Regulation; Research; Role; Secondary to; Skeleton; Syndrome; TRANCE protein; Testing; Therapeutic Intervention; Ubiquitination; Work; bone; bone loss; craniofacial; gain of function; human disease; human model; in vivo; induced pluripotent stem cell; mouse model; mutant; notch protein; novel; novel therapeutic intervention; osteoclastogenesis; overexpression; premature; prevent; receptor expression; skeletal; skeletal disorder; tomography

PROJECT SUMMARY/ABSTRACT Notch receptors play a critical role in cell fate decisions and in the regulation of bone remodeling, either directly or through the induction of their target genes, namely Hairy Enhancer of Split (Hes) and Hes-related with YRPW motif (Hey). Hajdu Cheney Syndrome (HCS) is a devastating disease characterized by developmental abnormalities, acroosteolysis and bone loss with fractures. HCS is associated with mutations in exon 34 of NOTCH2 upstream of the PEST domain leading to NOTCH2 stabilization and gain-of-function. We created a mouse model of HCS (Notch2tm1.1Ecan) that presents with osteopenia due to enhanced osteoclastogenesis and bone resorption. These events are secondary to an increase in receptor activator of nuclear factor Kappa B ligand (RANKL) by cells of the osteoblast lineage, and to direct effects of NOTCH2 in cells of the myeloid lineage. In this lineage, the expression of HES1 is induced by NOTCH2 and the inactivation of Hes1 in the osteoclast lineage reverses the in vitro and in vivo phenotype of HCS mutants. Moreover, HES1 induces osteoclastogenesis directly and as a result causes osteopenia in vivo. This reveals a previously unrecognized function of HES1 in osteoclast differentiation and function that will be explored as part of the proposed research. An additional goal of the proposed work is to develop ways to correct the skeletal manifestations of HCS by targeting the mutation with Notch2 antisense oligonucleotides (ASO), a strategy that would be applicable to other genetic disorders of the skeleton. Our specific aims are: Aim 1) To determine the role of HES1 in osteoclastogenesis. Our goals are to induce and inactivate Hes1 specifically in cells of the osteoclast lineage to determine its contribution to osteoclast differentiation and bone remodeling as determined by microcomputed tomography and histomorphometry; Aim 2) To establish that the Notch2tm1.1Ecan mutation can be targeted. We will determine whether the Notch2tm1.1Ecan mutation can be downregulated specifically and the Notch2tm1.1Ecan skeletal phenotype ameliorated by the administration of antisense oligonucleotides targeting the Notch26955C>T mutation; and Aim 3) To validate the mechanisms of the HCS phenotype and ASO approach in NOTCH2 mutant-induced pluripotent (iPS) cells. To this end, we created NOTCH2HCS mutant iPS cell lines to study the impact of the mutation on osteoclastogenesis and the efficacy of ASOs in downregulating NOTCH2 mutant alleles. The goals of the proposed work are to understand the mechanisms and develop specific antisense technology to treat the skeletal manifestations of a devastating NOTCH2-associated disease.

项目概要/摘要 Notch受体在细胞命运决定和骨重塑调节中发挥着关键作用 直接或通过其靶基因的诱导，即Hairy Improver of Split (Hes)和Hes相关 带有 YRPW 主题（嘿）。 Hajdu Cheney 综合症 (HCS) 是一种毁灭性的疾病，其特征是 发育异常、肢端骨质溶解和骨折引起的骨质流失。 HCS 与突变有关 PEST 结构域上游 NOTCH2 的外显子 34 导致 NOTCH2 稳定和功能获得。我们 创建了 HCS 小鼠模型 (Notch2tm1.1Ecan)，该模型由于增强的 破骨细胞生成和骨吸收。这些事件是继发于受体激活剂增加的 成骨细胞谱系细胞的核因子 Kappa B 配体 (RANKL)，以及 NOTCH2 的直接作用 骨髓谱系的细胞。在这个谱系中，HES1 的表达是由 NOTCH2 诱导的， 破骨细胞谱系中 Hes1 的失活可逆转 HCS 突变体的体外和体内表型。 此外，HES1直接诱导破骨细胞生成，从而导致体内骨质减少。这揭示了一个 HES1 在破骨细胞分化和功能中以前未被认识的功能，将作为其中的一部分进行探索 拟议的研究。拟议工作的另一个目标是开发纠正骨骼的方法 通过使用 Notch2 反义寡核苷酸 (ASO) 靶向突变来治疗 HCS 的表现，该策略 将适用于骨骼的其他遗传疾病。我们的具体目标是： 目标 1) 确定 HES1 在破骨细胞生成中的作用。我们的目标是在以下细胞中特异性诱导 Hes1 并使之失活 破骨细胞谱系以确定其对破骨细胞分化和骨重塑的贡献 通过显微计算机断层扫描和组织形态计量学；目标 2) 确定 Notch2tm1.1Ecan 突变可以 成为目标。我们将确定Notch2tm1.1Ecan突变是否可以特异性下调，以及 Notch2tm1.1Ecan 骨骼表型通过施用靶向的反义寡核苷酸得到改善 Notch26955C>T 突变；目标 3) 验证 HCS 表型和 ASO 方法的机制 NOTCH2 突变诱导的多能 (iPS) 细胞。为此，我们创建了 NOTCH2HCS 突变 iPS 细胞系 研究突变对破骨细胞生成的影响以及 ASO 下调的功效 NOTCH2 突变等位基因。拟议工作的目标是了解机制并开发 特定反义技术可治疗破坏性 NOTCH2 相关的骨骼表现 疾病。

项目成果

NOTCH2 promotes osteoclast maturation and metabolism and modulates the transcriptome profile during osteoclastogenesis.

• DOI: 10.1016/j.jbc.2023.105613
• 发表时间: 2024-02
• 期刊: JOURNAL OF BIOLOGICAL CHEMISTRY
• 影响因子: 4.8
• 作者: Canalis, Ernesto;Schilling, Lauren;Yu, Jungeun;Denker, Emily
• 通讯作者: Denker, Emily

NOTCH2 sensitizes the chondrocyte to the inflammatory response of tumor necrosis factor α.

• DOI: 10.1016/j.jbc.2023.105372
• 发表时间: 2023-12
• 期刊: JOURNAL OF BIOLOGICAL CHEMISTRY
• 影响因子: 4.8
• 作者: Canalis, Ernesto;Yu, Jungeun;Singh, Vijender;Mocarska, Magda;Schilling, Lauren
• 通讯作者: Schilling, Lauren