Homeotic hotspot in the human genome for eye and brain disease

人类眼部和脑部疾病基因组中的同源异型热点

基本信息

批准号：
10416324
负责人：
Thomas M. Glaser
金额：
$ 39.26万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-01 至 2027-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10416324
关键词：
3-Dimensional 9q21 Affect Alleles Anatomy Anophthalmos Anterior Apoptosis Architecture Bilateral Biological Assay Blindness Brain Brain Diseases CRISPR/Cas technology Cell Differentiation process Cells Cerebellar Ataxia Cerebellum Childhood Choroid Plexus Epithelium Chromatin Chromosome Fragile Sites Chromosomes Clinical Clustered Regularly Interspaced Short Palindromic Repeats Coloboma Constitutional Cornea Cre driver DNA DNA Binding Defect Development Disease Drosophila genus Ectoderm Electroporation Embryonic Development Enhancers Epithelial Evolution Exhibits Exons Eye Eye diseases Female Genetic Transcription Genome Gonadal structure Heterogeneity Hi-C Human Human Genetics Human Genome In Vitro Inheritance Patterns Iris Kinetics Lens Placodes Life Light Link Meiosis Methods Microphthalmos Modeling Molecular Mosaicism Mus Mutation Neural tube Nucleotides Optic vesicle Organ Pathogenesis Pathogenicity Patients Pattern Phenotype Pigments Pluripotent Stem Cells Positioning Attribute Prevalence Purkinje Cells Retina Retinal Photoreceptors Retinitis Pigmentosa Rod Running Series Signal Transduction Site Specific qualifier value Specificity Spinocerebellar Ataxias Structure of retinal pigment epithelium Surface Syndrome System Techniques Testing Testis Tissues Transcriptional Regulation Transgenes Visual Fields X Inactivation arm brain malformation cell type embryo tissue gain of function histogenesis in utero induced pluripotent stem cell male malformation mouse model nerve stem cell novel optic cup photoreceptor degeneration prenatal proband promoter relating to nervous system retinal neuron sex sex determination single-cell RNA sequencing stem cells transcription factor

3-Dimensional 9q21 Affect Alleles Anatomy Anophthalmos Anterior Apoptosis Architecture Bilateral Biological Assay Blindness Brain Brain Diseases CRISPR/Cas technology Cell Differentiation process Cells Cerebellar Ataxia Cerebellum Childhood Choroid Plexus Epithelium Chromatin Chromosome Fragile Sites Chromosomes Clinical Clustered Regularly Interspaced Short Palindromic Repeats Coloboma Constitutional Cornea Cre driver DNA DNA Binding Defect Development Disease Drosophila genus Ectoderm Electroporation Embryonic Development Enhancers Epithelial Evolution Exhibits Exons Eye Eye diseases Female Genetic Transcription Genome Gonadal structure Heterogeneity Hi-C Human Human Genetics Human Genome In Vitro Inheritance Patterns Iris Kinetics Lens Placodes Life Light Link Meiosis Methods Microphthalmos Modeling Molecular Mosaicism Mus Mutation Neural tube Nucleotides Optic vesicle Organ Pathogenesis Pathogenicity Patients Pattern Phenotype Pigments Pluripotent Stem Cells Positioning Attribute Prevalence Purkinje Cells Retina Retinal Photoreceptors Retinitis Pigmentosa Rod Running Series Signal Transduction Site Specific qualifier value Specificity Spinocerebellar Ataxias Structure of retinal pigment epithelium Surface Syndrome System Techniques Testing Testis Tissues Transcriptional Regulation Transgenes Visual Fields X Inactivation arm brain malformation cell type embryo tissue gain of function histogenesis in utero induced pluripotent stem cell male malformation mouse model nerve stem cell novel optic cup photoreceptor degeneration prenatal proband promoter relating to nervous system retinal neuron sex sex determination single-cell RNA sequencing stem cells transcription factor

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项目摘要

PROJECT SUMMARY Classical homeotic mutations (e.g. Drosophila Ubx) disrupt embryonic development, transforming one tissue type into another. We have discovered the molecular basis of four human X-linked disorders affecting the eye or brain – BASR syndrome, foveal dysgenesis, retinitis pigmentosa and spinocerebellar ataxia, which have a homeotic or degenerative basis. Each disorder is caused by insertion of a large autosomal DNA segment at the same Xq27 palindromic site near SOX3, which encodes a potent trans- cription factor homologous to SRY (testis determinant). The Xq insertions are predicted to disrupt chromatin architecture, activating SOX3 ectopically in tissues defined by newly juxtaposed enhancers, and altering cell fate (homeosis) via a gain-of-function (GOF). We propose that SOX3 changes retinal pigment epithelia (RPE) into neuroretina in BASR, reprograms cerebellar Purkinje cells in SCAX5, and triggers photoreceptor degeneration in RP24. Using a novel palinsert PCR assay, we defined the breakpoints and candidate enhancers for each insertion. We also identified >10 further Xq27 disorders affecting the eye, brain or other organs – including unsolved cases with a likely similar mechanism. We will define new Xq27 palindrome insertions and test our hypothesis for disease pathogenesis at chromatin and developmental levels, using [1] patient-derived iPSCs, 3D chromatin interaction assays (Hi-C), in vitro differentiation, serial scRNA-seq profiles; and [2] (homol informative mouse transgenes, including the binary CRISPR/Cas9 Hprt HoP-In ogy promoted integration) GOF system we pioneered – with Sox3HA expression activated in RPE, rods, Purkinje cells, or other tissues via established Cre drivers, in a constitutional (XY) or mosaic (XX) pattern, and in a sustained or Dox-inducible manner.

项目摘要经典同源突变（例如果蝇UBX）破坏胚胎发育，将一种组织类型转化为另一种组织。我们发现了四种影响眼睛或大脑的人X连锁疾病 - BASR综合征，凹foveal 发育不全，色素性视网膜炎和脊椎发子共济失调，具有同义或退化基础。每个疾病都是由插入大型常染色体DNA引起的在Sox3附近的同一XQ27 palindromic位点处的分段，该位点编码潜在的反式需要（确定睾丸）。预测XQ插入为了破坏染色质架构，在新的组织中生动地激活SOX3 并置的增强子，并通过功能获得（GOF）改变细胞命运（主管）。我们建议SOX3将视网膜色素上皮（RPE）变成Basr的Neuroretina，在SCAX5中重新编程小脑Purkinje细胞，并触发光感受器变性在RP24中。使用新型的Palinsert PCR分析，我们定义了断点和候选者每个插入的增强剂。我们还确定了> 10进一步影响的XQ27疾病眼睛，大脑或其他器官 - 包括可能具有相似机制的未解决病例。我们将定义新的XQ27腔插入并检验我们的疾病假设使用[1]患者衍生的IPSC，在染色质和发育水平的发病机理， 3D染色质相互作用测定（HI-C），体外分化，串行SCRNA-SEQ轮廓；和[2] （同性恋）内容丰富的鼠标翻译，包括二进制CRISPR/CAS9 HPRT HOPIN OGY促进的集成）GOF系统，我们进行了启动 - Sox3Ha表达通过已建立的CRE驱动器在RPE，杆，Purkinje细胞或其他组织中激活宪法（XY）或马赛克（XX）模式，以及持续或DOX诱导的方式。