Modeling esophageal/respiratory birth defects in human pluripotent stem cell (PSC)-derived fetal tissues
Modeling esophageal/respiratory birth defects in human pluripotent stem cell (PSC)-derived fetal tissues
批准号:
10174986
负责人:
James M Wells
金额:
$32.3万
依托单位国家:
美国
项目类别:
财政年份:
2017
资助国家:
美国
项目状态:
已结题
起止时间:
2017-08-15 至 2022-05-31
关键词:
AchalasiaAffectAutologousBarrett EsophagusBiochemicalBiological AssayBiological ModelsCell LineChIP-seqCollaborationsColonComplexCongenital AbnormalityDefectDevelopmentDiseaseDominant-Negative MutationDorsalEmbryoEndodermEnteric Nervous SystemEosinophilic EsophagitisEpithelialEsophageal AtresiaEsophageal TissueEsophagusFailureFetal TissuesFoodGastroesophageal reflux diseaseGenesGenetic TranscriptionGerm LayersHMG-Box DomainsHumanHuman CharacteristicsImpairmentLeadMalignant neoplasm of esophagusMegaesophagusModelingMolecularMorphogenesisMorphologyMovementMusMuscleMutationOperative Surgical ProceduresOral cavityOrganoidsPathologyPathway interactionsPatientsPhysiologicalPluripotent Stem CellsPrimitive foregut structureProcessRegulator GenesReporterResortRespiratory SystemRoleSignal TransductionSmooth MuscleStomachStratified EpitheliumSurgeonTissue EngineeringTissuesTracheaTracheoesophageal FistulaTranscriptional ActivationTubebasebeta catenincell motilitycell typeconstrictionembryo tissuehuman pluripotent stem cellhuman tissueinduced pluripotent stem cellloss of functionmolecular modelingmotility disordernerve supplynervous system developmentnovelprotein Bprotein protein interactionreconstructionrespiratorysegregationstem cell based approachtissue reconstructiontranscription factortranscriptome sequencingvertebrate embryos
中文摘要
摘要:在人类PSC来源的胚胎组织中建立EA/TEF模型。
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在脊椎动物胚胎的发育过程中,一种常见的前肠管状结构可促进食道的生长和发育。
呼吸道疾病包括一系列复杂的分子生物学和形态生物学过程。
前肠管形成于下段食道,下腹段段形成于下呼吸道,如果不能做到这一点,就会出现这种情况。
导致气管食道出生缺陷,如食道闭锁和气管食道瘘(EA/TEF)。
正如我们在项目2中讨论的那样,关于WNT和BMP如何发出信号以促进人类呼吸暂停的命运,还有很多事情是未知的。
转录因子Nkx2.1的激活。与之形成对比的是,人们对促食道因子知之甚少。
而人类基因组研究也证明,转录因子Sox2的HMG-box基因也参与了人类的分离过程。
食道和呼吸道疾病的血统,然而,无论Sox2是促进食道疾病的命运还是其他行为。
目前尚不清楚这主要是为了抑制胃背侧的呼吸诱导通路。我们的假设是,这两种途径都适用。
他们的机制也参与了正常的食道发育。
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在人类中,大多数导致Ea/TEF的基因仍未确定。然而,一些杂合的基因突变发生在SOX2中。
这会导致EEA和TEF的异常,这与Sox2杂合性缺失的小鼠形成鲜明对比,这两种情况都很正常。
然而,从小鼠胚胎的正常前肠和内胚层中完全丢失Sox2蛋白会导致食道发育不全。
SOX2基因也在食道的肠神经系统疾病(ENS)的发育过程中表达出来。
患有EEA的患者可能会有运动性和缺陷,但我们可以假设一些与EEA相关的基因可能不会影响神经。
发展。然而,一项关于EA相关基因突变如何不同地影响血管上皮细胞和/或血管的研究报告。
在任何物种中都从未完成过食道修复,更不用说人类了。我们将提出几种全新的食道修复技术--PSC--
基于研究Sox2基因和其他与EA相关的基因如何影响人类食道规范的方法。
上皮细胞的形态发生、功能和功能神经的支配,使用的是来自食道的人类多能干细胞。
有机化合物与肠道和神经系统有关。
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在这个项目中,我们的目标是进一步确定食道安全规范的基本机制。
人类的发展首先是专注于SOX2这一关键的食道致癌因素。我们假设SOX2是
这两种行为都是为了抑制人类的呼吸道疾病血统,并通过一种未知的基因来促进食道疾病的命运。
监管监管网络。我们将继续使用一个由PSC派生的更具人性化的未来模式,并将其与SOX2的收益和亏损相结合。
功能障碍是为了识别一个被SOX2基因抑制的呼吸系统GRN基因和一个被SOX2基因抑制的食道癌GRN基因。
家属。相反,我们将无法确定Nkx2.1是否会抑制食道癌的命运。我们将不会利用这一优势。
人类基因SOX2和SOX2基因的可扩充性和文化的可扩充性需要进一步确定人类基因SOX2和SOX2基因的直接转录和转录靶点。
Nkx2.1正在使用核糖核酸-seq和芯片-seq。然后我们将继续调查潜在的TEF的疾病和机制。
此外,EA表示,这些突变不是由Sox2基因突变引起的。我们不会生成包含基于患者的基因突变的PSC品系。
Sox2和他们调查了这些因素是如何影响食道系统和呼吸系统血统的形成的。
确定SOX2基因突变对Wnt基因和BMP信号转导系统的影响,并确定Sox2基因是否通过直接作用于蛋白质-蛋白质而起作用。
与效应蛋白的相互作用包括b-连环蛋白/Tcf和Smads。最后,我们将不会研究EA是如何发挥作用的。
突变对不同类型的食道上皮细胞有不同的影响;;是上皮细胞,是平滑的上皮细胞。
和失弛缓症。考虑到一些患有胃肠失弛缓症的患者与运动障碍相关,包括失弛缓症。
我们将不会调查Sox2基因突变是否也会导致赤字增加。我们将不会使用。
IPSC的这些线是从EEA/TEF中派生出来的,患者在第一个和第二个项目中被确定是什么,以建立一个新的分子生物学缺陷的模型。
这种出生缺陷的根本原因是使用了我们最新的人类PSC衍生的有机化合物模型。
英文摘要
Summary: Modeling EA/TEF In Human PSC-Derived Embryonic Tissues
During development of the vertebrate embryo, a common foregut tube gives rise to the esophagus and
respiratory tract and this involves an array of complex molecular and morphological processes. The dorsal
foregut tube forms the esophagus and the ventral domain forms the respiratory tract, and failure to do so can
result in tracheaesophageal birth defects such as esophageal atresia and tracheoesophageal fistula (EA/TEF).
As discussed in project 2, much is known about how Wnt and BMP signaling promote a respiratory fate by
activation of the transcription factor Nkx2.1. In contrast, little is known about pro-esophageal factors. Mouse
and human studies demonstrate that the HMG-box transcription factor Sox2 is involved in segregation of the
esophageal and respiratory lineages, however whether Sox2 promotes an esophageal fate or acts
predominantly to repress respiratory-inducing pathways the dorsal foregut is unclear. We hypothesize that both
mechanisms are involved in normal esophageal development.
In humans, most genes that cause EA/TEF remain unidentified. However, heterozygous mutations in SOX2
can cause of EA and TEF, which is in contrast to mice with heterozygous loss of Sox2, which are normal.
Complete loss of Sox2 from the foregut endoderm of mouse embryos results in esophageal agenesis, however
Sox2 is also expressed during development of the enteric nervous system (ENS) of the esophagus. Given that
patients with EA can have motility defects, we hypothesize some EA-associated genes may affect ENS
development. However, a study of how EA-associated mutations differentially affect the epithelium and/or ENS
of the esophagus has never been done in any species, let alone humans. We propose several novel PSC-
based approaches to study how Sox2 and other EA-associated genes impact Human esophagus specification,
epithelial morphogenesis, and functional innervation using human pluripotent stem cell-derived esophageal
organoids with an enteric nervous system.
In this project we aim to identify the mechanisms underlying esophageal specification and
development in humans by first focusing on the key esophageal factor Sox2. We hypothesize that SOX2
acts both to repress the respiratory lineage, and promote an esophageal fate via an unidentified gene
regulatory network. We will use a human PSC-derived foregut model in combination with SOX2 gain- and loss-
of-function to identify a respiratory GRN that is repressed by SOX2 and an esophageal GRN that is SOX2-
dependant. Conversely we will determine if NKX2.1 represses the esophageal fate. We will take advantage of
the expandable nature of human foregut cultures to identify direct transcriptional targets of human SOX2 and
NKX2.1 using RNA-seq and ChIP-seq. We will then investigate the disease mechanisms underlying TEF
and EA that are caused by Sox2 mutations. We will generate PSC lines harboring patient-based mutations in
SOX2 and investigate how these impact the formation of the esophageal and respiratory lineages. We will
identify the impact of SOX2 mutations on Wnt and BMP signaling and if Sox2 acts by direct protein-protein
interactions with the effector proteins b-catenin/TCF and Smads. Lastly we will investigate how EA
mutations differentially effect the different cell types of the esophagus;; the epithelial, smooth muscle
and ENS. Given that some patients with EA have associated motility disorders including achalasia 3,
constrictions 4 and megaesophagus 5, we will investigate if Sox2 mutations also have ENS deficits. We will use
iPSC lines derived from EA/TEF patients identified in projects 1 and 2 to model the molecular deficits
underlying this birth defect using our human PSC-derived organoid model.
期刊论文(0)
专著(0)
科研奖励(0)
会议论文
Project-3: Modeling EA/TEF in human organoids
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批准号:10458162
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项目类别:
-
资助金额:$39.75万
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财政年份:2017
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负责人:James M Wells
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依托单位:
Project-3: Modeling EA/TEF in human organoids
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批准号:10647838
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项目类别:
-
资助金额:$39.75万
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财政年份:2017
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负责人:James M Wells
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依托单位:
Human Endocrine Cell Development
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批准号:8814212
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项目类别:
-
资助金额:$45.5万
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财政年份:2012
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负责人:James M Wells
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依托单位:
Generating Human Intestinal Organoids with an ENS.
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批准号:8415736
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项目类别:
-
资助金额:$34.43万
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财政年份:2012
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负责人:James M Wells
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依托单位:
Generating Human Intestinal Organoids with an ENS.
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批准号:8516139
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项目类别:
-
资助金额:$32.82万
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财政年份:2012
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负责人:James M Wells
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依托单位:
Generating Human Intestinal Organoids with an ENS.
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批准号:8665593
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项目类别:
-
资助金额:$7.65万
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财政年份:2012
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负责人:James M Wells
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依托单位:
Mammalian Foregut and Liver Development
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批准号:8204678
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项目类别:
-
资助金额:$36.09万
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财政年份:2009
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负责人:James M Wells
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依托单位:
Mammalian Foregut and Liver Development
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批准号:7895193
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项目类别:
-
资助金额:$10.0万
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财政年份:2009
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负责人:James M Wells
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依托单位:
Mammalian Foregut and Liver Development
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批准号:7753860
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项目类别:
-
资助金额:$40.22万
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财政年份:2009
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负责人:James M Wells
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依托单位:
Mammalian Foregut and Liver Development
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批准号:7582986
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项目类别:
-
资助金额:$39.44万
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财政年份:2009
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负责人:James M Wells
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依托单位:
Mammalian Foregut and Liver Development
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批准号:8018155
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项目类别:
-
资助金额:$36.09万
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财政年份:2009
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负责人:James M Wells
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依托单位:
Stem Cell/Organoid and Genome Editing Core
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批准号:10620719
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项目类别:
-
资助金额:$14.99万
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财政年份:2007
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负责人:James M Wells
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依托单位:
Pluripotent Stem Cell and Organoid Core
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批准号:9312950
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项目类别:
-
资助金额:$14.34万
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财政年份:2007
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负责人:James M Wells
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依托单位:
Pluripotent Stem Cell and Organoid Core
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批准号:9535292
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项目类别:
-
资助金额:$13.95万
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财政年份:2007
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负责人:James M Wells
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依托单位:
Stem Cell/Organoid and Genome Editing Core
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批准号:10442029
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项目类别:
-
资助金额:$14.99万
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财政年份:2007
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负责人:James M Wells
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依托单位:
Mechanisms of endoderm specification along the A-P axis.
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批准号:7092306
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项目类别:
-
资助金额:$27.54万
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财政年份:2006
-
负责人:James M Wells
-
依托单位:
Mechanisms of endoderm specification along the A-P axis
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批准号:7410109
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项目类别:
-
资助金额:$27.67万
-
财政年份:2006
-
负责人:James M Wells
-
依托单位:
Mechanisms of endoderm specification along the A-P axis.
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批准号:7800390
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项目类别:
-
资助金额:$27.4万
-
财政年份:2006
-
负责人:James M Wells
-
依托单位:
Mechanisms of endoderm specification along the A-P axis
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批准号:7615114
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项目类别:
-
资助金额:$27.67万
-
财政年份:2006
-
负责人:James M Wells
-
依托单位:
Mechanisms of endoderm specification along the A-P axis
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批准号:7222756
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项目类别:
-
资助金额:$27.67万
-
财政年份:2006
-
负责人:James M Wells
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依托单位:
海外基金