Conditional Genetics Rescue of Angelman Syndrome

天使综合症的条件遗传学拯救

基本信息

批准号：
8860218
负责人：
MATTHEW P ANDERSON
金额：
$ 20.36万
依托单位：
BETH ISRAEL DEACONESS MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-06-15 至 2016-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8860218
关键词：
15q Adult Age Alleles Angelman Syndrome Autistic Disorder Bacterial Artificial Chromosomes Behavior Behavioral Binding Brain Cell Nucleus Chimeric Proteins Chromosomes, Human, Pair 15 Cognitive Conceptions Cytoplasm Defect Development Disease Dose Embryo Estrogens Event Exons Eye Functional disorder Future Gene Dosage Gene Expression Genes Genetic Goals Health Heat shock proteins ILK gene Individual Inheritance Patterns Inherited Isodicentric Chromosome Knock-out Laughter Length Location Maps Modeling Motor Mus Mutate Neurons Nuclear Receptors Phenotype Physiological Proteins RNA Splicing Smiling Social Behavior Social Interaction Synapses Tamoxifen Testing Time Transgenes Transgenic Mice Transgenic Organisms Ultrasonics base imprint insight motor deficit mouse model multicatalytic endopeptidase complex neurobehavioral neuronal circuitry neurophysiology neuropsychiatry novel postnatal protein degradation recombinase social communication therapy development trait ubiquitin-protein ligase vector vocalization

项目摘要

DESCRIPTION (provided by applicant): Angelman syndrome results when maternally-inherited 15q11-13 is deleted or imprinted 15q11-13 gene Ube3a is mutated. Autism results when maternal 15q11-13 is triplicated. The parental inheritance patterns are explained by Ube3a the only 15q11-13 gene expressed solely from the maternal allele in neurons. The countervalent 15q disorders display some contrasting behavioral changes. Angelman syndrome presents with intellectual and motor deficits, but also a hypersocial demeanor, while 15q triplication displays impaired social behavior. Interestingly, in additional to the other previousl characterized deficits, we recently established the Angelman syndrome mice display increased social interaction and ultrasonic vocalization. By contrast, tripling Ube3a gene dosage reduced social interaction and vocalization providing a model of autism. The results establish a Goldilock effect for Ube3a gene dosage and social behavior. Reciprocal gene dose-dependent effects also argue some Ube3a effects could arise from ongoing regulatory rather than remote developmental defects. Ube3a acts as an E3 ubiquitin protein ligase to promote protein degradation and as a nuclear receptor co-activator. To test the hypothesis that Ube3a deficiency in Angelman syndrome causes deficits in behavior and circuit function via ongoing regulatory rather than developmental defects, we will test for reversal of the behavioral and circuit deficits when Ube3a is genetically reactivated in adulthood. To achieve this goal, we generated mice carrying a single extra copy of Ube3a-ON transgene that is silenced by a loxP-flanked STOP cassette. The Ube3a-ON transgene is inactive until the loxP-flanked STOP cassette is deleted by Cre recombinase. Crossing Ube3a-ON transgenic to Cre-ER" fusion protein transgenic mice enables tamoxifen- induced Ube3a gene expression. ERTM is mutated, responding to tamoxifen but not estrogen. Cre recombinase is tethered to heat shock proteins in the cytoplasm by ERTM. Tamoxifen penetrates the CNS and binds ERTM, permitting Cre to enter the nucleus and delete the loxP-flanked STOP cassette. If adult rescue is not achieved, we will perform temporal mapping to postnatal or embryonic developmental periods. We also created an Ube3a-OFF transgene where loxP sequences flank Ube3a exon 2 so tamoxifen causes Cre-ERTM to inactivate Ube3a. Crossing Ube3a-OFF/Cre-ERTM into the Angelman model, we will determine if inactivating Ube3a in adulthood or at specific developmental times recreates the disorder. The result will establish the feasibility of rescuing Angelman syndrome and provide insights into the pathophysiological basis and treatment options for the disorder.

描述（由申请人提供）：当含有15q11-13的15q11-13的15q11-13基因UBE3A的15q11-13的15q11-13时，Angelman综合征结果被突变。自闭症产生15q11-13的三次三次三份。 ube3a解释了父母的遗传模式是仅从神经元中母体等位基因表达的唯一15q11-13基因。反价15Q疾病显示出一些对比的行为变化。 Angelman综合征表现出智力和运动缺陷，但也表现出了超社会的举止，而15q三分之一的人会显示社会行为受损。有趣的是，除了其他先前的缺陷外，我们最近建立了Angelman综合征小鼠展示的社交互动和超声波发声。相比之下，UBE3A基因剂量的三倍减少了社会互动和发声，从而提供了自闭症模型。结果为UBE3A基因剂量和社会行为树立了金色效果。相互基因剂量依赖性效应也表明，某些UBE3A效应可能是由于正在进行的监管而不是远程发育缺陷引起的。 UBE3A充当E3泛素蛋白连接酶，可促进蛋白质降解和核受体共激活剂。为了检验Angelman综合征中UBE3A缺乏的假设，通过持续的调节性而不是发育缺陷导致行为和电路功能的缺陷，我们将测试当Ube3a在偶然中遗传重新激活的行为和电路缺陷逆转。为了实现这一目标，我们生成了携带单个Ube3a-on Transgene的额外副本的小鼠，该副本被Loxp-Flank Planks Stop Cassette保持沉默。 UBE3A-ON转基因不活跃，直到Loxp-Flank的停止盒子被CRE重组酶删除为止。将ube3a-On转基因交叉至CRE-ER“融合蛋白转基因小鼠可实现卧氧昔芬诱导的UBE3A基因表达。ERTM被突变，对硫昔芬的反应，但不反应雌激素。CRE重物组织酶在cre cre cre the creTm and creTm and thamoxy and thamoxy and thamoxy intms and thamoxemoxs and thamoxefen and thamoxefen and thamoxefen and cross and thamoxefen and thamoxifen and thamoxifen。核并删除loxp的停止盒，如果没有实现成人的救援，我们将进行时间映射到产后或胚胎发育时期。我们将确定成年后的UBE3A是否会重现该疾病。