Synaptic function of Chromosome 21- encoded microRNAs

21 号染色体编码的 microRNA 的突触功能

• 批准号: 9312335
• 负责人: Heather McGowan
• 金额: --
• 依托单位: RBHS-ROBERT WOOD JOHNSON MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2017-07-02
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9312335
• 关键词: Affect; Biochemical; Biological Assay; Biological Process; Brain; Calcium; Cell Line; Cells; Chromosomes, Human, Pair 21; Code; Cognition Disorders; Collection; Data; Defect; Dendritic Spines; Development; Diploidy; Down Syndrome; Down-Regulation; Electrophysiology (science); Fragile X Syndrome; Functional disorder; Gene Expression Regulation; Genetic; Genetic Transcription; Human; Human Chromosomes; Image; Immunohistochemistry; Impaired cognition; Impairment; Inhibitory Synapse; Intellectual functioning disability; Knowledge; Luciferases; Maintenance; Mediating; Messenger RNA; Methodology; Methods; Methyl-CpG-Binding Protein 2; MicroRNAs; Modeling; Modification; Molecular; Morphology; Mutate; Nervous system structure; Neurodevelopmental Disorder; Neurons; Pathway interactions; Patients; Process; Proteins; Regulation; Reporter; Research; Resolution; Rett Syndrome; Role; Site; Synapses; Synaptic Transmission; Techniques; Technology; Testing; Translating; Translational Repression; Untranslated RNA; Western Blotting; base; cognitive disability; expectation; induced pluripotent stem cell; innovation; insight; knock-down; nervous system disorder; neuropsychiatric disorder; novel; overexpression; public health relevance; synaptic function; synaptogenesis

DESCRIPTION (provided by applicant): The formation and maintenance of appropriate and functional synaptic connections is a highly regulated process, with misregulation resulting in disordered cognition. Given the emerging information about gene expression regulation by non-coding RNAs, synaptic functions are likely to be regulated at least in part by non- coding RNAs, including microRNAs (miRNAs). Several miRNAs have been implicated in spinogenesis, dendritic arborization, and synaptogenesis. Thus, over- or under-expression of miRNAs in the brain could conceivably contribute to synaptic dysfunction resulting in neurological or neuropsychiatric disorders. Human chromosome 21 (HSA21) codes for 5 known miRNAs, and Trisomy 21 (TS21, i.e. Down syndrome) is the most common genetic form of intellectual disability. Thus, TS21 provides a unique model to study the effect of miRNA overexpression on the formation and functionality of synapses. The objective of this project is to elucidate the role of HSA21 miRNAs in synaptic dysfunction in human neurons generated from TS21 patients, which may be implicated in the cognitive disability in TS21 by testing the hypothesis that overexpression of HSA21 miRNAs leads to dysfunction in synaptic transmission that causes cognitive impairment in TS21 patients. Furthermore, there is preliminary evidence that suggests these miRNAs may affect synaptic integrity via a methyl CpG binding protein 2 (MeCP2) dependent pathway. Utilizing the innovative induced pluripotent stem (iPS) cell and induced neuronal (iN) cell technologies, it is possible to study the effects of HSA21 miRNAs on the synapses of human neurons. Following overexpression of HSA21 miRNAs in control iNs, synaptic function will be assessed by morphological and functional analyses, including electrophysiology and Calcium imaging. MeCP2 will be confirmed as a target of these miRNAs by dual luciferase assay, and its role in the miRNA-mediated modification of synapses will be tested via knockdown and rescue. Furthermore, patient-specific iNs will be used to elucidate whether the overexpression of HSA21 miRNAs causes synaptic defects in TS21. After verifying the expression levels of HSA21 miRNAs in TS21 iNs via qPCR and correlating them with the level of MeCP2 (determined by Western Blot), we will morphologically and functionally characterize the synapse for comparison with control iNs. We will then establish a cause-effect relationship between HSA21 miRNA overexpression and synaptic defects using Tough Decoys to antagonize the miRNAs and "rescue" the synaptic function of TS21-iN cells, as well as the expression level of MeCP2. The proposed research is innovative, because we will use interdisciplinary analytical methodologies and combine the newly developed iN cell and iPS cell technologies to examine the functions of HSA21 miRNAs in the nervous system, which will broaden our knowledge of their biological functions, as well as provide insight into mechanistic and molecular bases for the treatment of TS21.

DESCRIPTION (provided by applicant): The formation and maintenance of appropriate and functional synaptic connections is a highly regulated process, with misregulation resulting in disordered cognition. Given the emerging information about gene expression regulation by non-coding RNAs, synaptic functions are likely to be regulated at least in part by non- coding RNAs, including microRNAs (miRNAs). Several miRNAs have been implicated in spinogenesis, dendritic arborization, and synaptogenesis. Thus, over- or under-expression of miRNAs in the brain could conceivably contribute to synaptic dysfunction resulting in neurological or neuropsychiatric disorders. Human chromosome 21 (HSA21) codes for 5 known miRNAs, and Trisomy 21 (TS21, i.e. Down syndrome) is the most common genetic form of intellectual disability. Thus, TS21 provides a unique model to study the effect of miRNA overexpression on the formation and functionality of synapses. The objective of this project is to elucidate the role of HSA21 miRNAs in synaptic dysfunction in human neurons generated from TS21 patients, which may be implicated in the cognitive disability in TS21 by testing the hypothesis that overexpression of HSA21 miRNAs leads to dysfunction in synaptic transmission that causes cognitive impairment in TS21 patients. Furthermore, there is preliminary evidence that suggests these miRNAs may affect synaptic integrity via a methyl CpG binding protein 2 (MeCP2) dependent pathway. Utilizing the innovative induced pluripotent stem (iPS) cell and induced neuronal (iN) cell technologies, it is possible to study the effects of HSA21 miRNAs on the synapses of human neurons. Following overexpression of HSA21 miRNAs in control iNs, synaptic function will be assessed by morphological and functional analyses, including electrophysiology and Calcium imaging. MeCP2 will be confirmed as a target of these miRNAs by dual luciferase assay, and its role in the miRNA-mediated modification of synapses will be tested via knockdown and rescue. Furthermore, patient-specific iNs will be used to elucidate whether the overexpression of HSA21 miRNAs causes synaptic defects in TS21. After verifying the expression levels of HSA21 miRNAs in TS21 iNs via qPCR and correlating them with the level of MeCP2 (determined by Western Blot), we will morphologically and functionally characterize the synapse for comparison with control iNs. We will then establish a cause-effect relationship between HSA21 miRNA overexpression and synaptic defects using Tough Decoys to antagonize the miRNAs and "rescue" the synaptic function of TS21-iN cells, as well as the expression level of MeCP2. The proposed research is innovative, because we will use interdisciplinary analytical methodologies and combine the newly developed iN cell and iPS cell technologies to examine the functions of HSA21 miRNAs in the nervous system, which will broaden our knowledge of their biological functions, as well as provide insight into mechanistic and molecular bases for the treatment of TS21.

项目成果

Bridging the Gap between DNA Methylation, DNA Methylation Readers, and Neurodevelopmental Disorders.

弥合 DNA 甲基化、DNA 甲基化读取器和神经发育障碍之间的差距。

• DOI: 10.1523/jneurosci.1130-16.2016
• 发表时间: 2016
• 期刊: The Journal of neuroscience : the official journal of the Society for Neuroscience
• 作者: McGowan,Heather

Regulatory functions and pathological relevance of the MECP2 3'UTR in the central nervous system.

• DOI: 10.1186/s13619-015-0023-x
• 发表时间: 2015
• 期刊: Cell regeneration (London, England)
• 作者: McGowan H;Pang ZP
• 通讯作者: Pang ZP