FUNCTIONAL CHARACTERIZATION OF NOVEL DETERMINANTS OF HOLOPROSENCEPHALY (HPE)

前脑无裂畸形 (HPE) 的新决定因素的功能特征

基本信息

批准号：
10596128
负责人：
Ernesto Guccione
金额：
$ 36.34万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-03-05 至 2026-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10596128
关键词：
Accounting Affect Anterior Automobile Driving Biological Process Brain Cause of Death Cells Cephalic Cessation of life Chromatin Clinical Collaborations Congenital Abnormality Craniofacial Abnormalities Critical Pathways Data Defect Development Developmental Process Diagnostic Disease Embryo Embryonic Development Embryonic Structures Epigenetic Process Erinaceidae Event Eye Failure Family Fingers Follow-Up Studies Forebrain Development Gene Expression Genes Genetic Genetic Determinism Genetic Transcription Goals Growth Head Healthcare Systems Heterozygote Holoprosencephaly Human In Vitro Individual Infant Knowledge Link MAP Kinase Gene Mediating Microcephaly Mining Molecular Molecular Diagnosis Morbidity - disease rate Mus Mutate Mutation NOTCH3 gene Nature Newborn Infant Oncogene Deregulation Pathogenicity Pathway interactions Patients Pattern Phenotype Primitive Streaks Prosencephalon Proteins Regenerative Medicine Reporting Research Project Grants Risk Role SHH gene Science Shapes Signal Pathway Signal Transduction Specific qualifier value Structural defect Structure System Testing Therapeutic Transcriptional Regulation United States National Institutes of Health Variant Work brain malformation cohort congenital anomaly defined contribution diagnostic screening disability embryonic stem cell epigenomics genetic testing genetic variant improved in vivo malformation member mouse genetics novel prevent progenitor programs stem targeted sequencing transcription factor transcriptomics

Accounting Affect Anterior Automobile Driving Biological Process Brain Cause of Death Cells Cephalic Cessation of life Chromatin Clinical Collaborations Congenital Abnormality Craniofacial Abnormalities Critical Pathways Data Defect Development Developmental Process Diagnostic Disease Embryo Embryonic Development Embryonic Structures Epigenetic Process Erinaceidae Event Eye Failure Family Fingers Follow-Up Studies Forebrain Development Gene Expression Genes Genetic Genetic Determinism Genetic Transcription Goals Growth Head Healthcare Systems Heterozygote Holoprosencephaly Human In Vitro Individual Infant Knowledge Link MAP Kinase Gene Mediating Microcephaly Mining Molecular Molecular Diagnosis Morbidity - disease rate Mus Mutate Mutation NOTCH3 gene Nature Newborn Infant Oncogene Deregulation Pathogenicity Pathway interactions Patients Pattern Phenotype Primitive Streaks Prosencephalon Proteins Regenerative Medicine Reporting Research Project Grants Risk Role SHH gene Science Shapes Signal Pathway Signal Transduction Specific qualifier value Structural defect Structure System Testing Therapeutic Transcriptional Regulation United States National Institutes of Health Variant Work brain malformation cohort congenital anomaly defined contribution diagnostic screening disability embryonic stem cell epigenomics genetic testing genetic variant improved in vivo malformation member mouse genetics novel prevent progenitor programs stem targeted sequencing transcription factor transcriptomics

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项目摘要

SUMMARY Congenital defects are a leading cause of morbidity worldwide, accounting for the deaths of 330,000 new- born every year. Brain malformations appear to be the most common congenital anomalies and are a major cause of death and lifelong disability. In the majority of cases the cause remains uncertain, due to the complexity and the multi-genic origin of these anomalies. Genes encoding Transcription Factors (TFs) and epigenetic regulators have become relevant candidates given the central role of these proteins in integrating signalling cascades and orchestrating multiple biological processes. Deficiency in their function may disturb entire transcriptional programs, involving several genes and molecular pathways. Here we combine mouse genetics and epigenomic approaches to uncover the role of PRDM15, a previously unsuspected disease-associated epigenetic regulator, in congenital brain malformations. Moreover, by functionally characterizing PRDM15 downstream effectors (e.g. NOTCH and WNT/PCP pathways) we uncover hitherto underappreciated genes mutated in patients with brain malformations (i.e. HPE and microcephaly). Preliminary data: We have characterized the function of PRDM15 in regulating the mouse naïve ESC state. We have then expanded our findings to demonstrate that, in vivo, PRDM15 depletion leads to: 1) embryonic lethality at E12.5-E14.5; 2) patterning defects affecting Anterior/Posterior patterning and forebrain development. In particular we have observed a failure to properly form the Axial Mesendoderm (AME), an embryonic structure necessary for proper anterior specification; 3) Finally, in collaboration with the groups of M.Muenke (NIH) and F.Hildebrandt (Harvard), we have identified heterozygous and homozygous mutations in PRDM15 linked to Holoprocensephaly (HPE) and microcephaly, respectively and mutations in over 100 PRDM15-regulated genes (~20% likely to be damaging) in a large cohort of HPE patients (132 trios and 188 singletons). In AIM1 we propose to define the molecular basis of PRDM15 function, specifically to understand how it regulates, at the level of chromatin, the transcriptional program driving forebrain development. Next, in AIM2 we will establish a causative link between human PRDM15 mutations, HPE-associated genetic variants identified as PRDM15-transcriptional targets, and the associated spectrum of brain malformations (ranging from HPE, to Microcephaly). The significance of these studies is that PRDM15 is a so far uncharacterized critical regulator of embryonic development and knowledge of the downstream regulated pathways will be useful to the field of regenerative medicine and will have diagnostic and clinical implications for patients with holoprosencephaly and microcephaly. The clinical Impact of these studies is that given the multigenic origin of HPE, targeted sequencing of PRDM15, and its key downstream targets, can potentially be added to routine genetic testing in families at risk of carrying other HPE-causing mutations (e.g. SHH, ZIC2, TGIF).

概括先天性缺陷是全球发病率的主要原因，占33万新的死亡每年出生。脑畸形似乎是最常见的先天性异常，并且是主要的死亡原因和终生残疾。在大多数情况下，由于复杂性，原因仍然不确定以及这些异常的多生成起源。编码转录因子（TFS）和表观遗传的基因鉴于这些蛋白质在整合信号传导中的核心作用，监管机构已成为相关的候选者级联和编排多个生物学过程。功能不足可能会打扰整个转录程序涉及多个基因和分子途径。在这里，我们结合了小鼠遗传学和表观基因组学方法，以发现PRDM15的作用，以前先天性脑畸形中无刺相关的表观遗传调节剂。而且，在功能上表征PRDM15下游效应（例如Notch和Wnt/PCP途径）我们发现迄今为止，在脑畸形患者（即HPE和小头畸形）中突变的基因不足。初步数据：我们已经表征了PRDM15在确定小鼠幼稚的ESC状态方面的功能。然后，我们扩展了发现，以证明，在体内，PRDM15部署会导致：1）胚胎 E12.5-E14.5的致死性； 2）影响前/后构图和前脑发育的图案缺陷。特别是我们观察到未能正确形成轴向胚层（AME），这是一种胚胎结构适当的前规范所必需的； 3）最后，与M.Muenke（NIH）和 F. -Hildebrandt（哈佛），我们已经确定了与PRDM15相关的杂合和纯合突变全异脑（HPE）和小头畸形，以及超过100个PRDM15调节基因的突变（〜20％的HPE患者（132例三重奏和188个单例）可能会损害20％）。在AIM1中，我们建议定义PRDM15函数的分子基础，特别是为了了解它如何在染色质水平上调节转录程序驱动前脑发育。接下来，在AIM2中将在人类PRDM15突变之间建立因果关系，与HPE相关的遗传变异鉴定作为PRDM15转录靶标以及脑畸形的相关范围（从HPE到范围小头畸形）。这些研究的重要性是，prdm15是迄今为止胚胎的未表征的关键调节剂下游规范途径的发展和知识将对再生领域有用药物，将对全脑和小头畸形患者具有诊断和临床意义。这些研究的临床影响是，鉴于HPE的多基因起源，有针对性的测序 PRDM15及其主要下游目标可能会添加到有风险的家庭的常规基因测试中携带其他引起HPE的突变（例如SHH，ZIC2，TGIF）。