Co-culture of probiotic bacteria for growth factor delivery in minigut organoids
Co-culture of probiotic bacteria for growth factor delivery in minigut organoids
批准号:
9789051
负责人:
Thomas J Mansell
金额:
$17.63万
依托单位:
依托单位国家:
美国
项目类别:
财政年份:
2018
资助国家:
美国
项目状态:
已结题
起止时间:
2018-09-20 至 2020-07-31
关键词:
3-DimensionalAnimal ExperimentsAnimal ModelBacteriaBehaviorBiological ModelsBiosensorButyratesCell Culture TechniquesCell Differentiation processCell LineCell divisionCell physiologyCell secretionCellsChloride IonChloridesChronic DiseaseCoculture TechniquesCommunicationDetectionDevelopmentDiabetes MellitusDifferentiation AntigensEngineered ProbioticsEngineeringEnterocytesEnteroendocrine CellEnvironmentEpithelial CellsEscherichia coliFluorescence MicroscopyGene ExpressionGenesGeneticGnotobioticGoalsGoldGrowthGrowth FactorHarvestHeartHormonalImageImageryInflammatory Bowel DiseasesIntestinal SecretionsIntestinesKnowledgeLabelMetabolic syndromeModelingMolecularMonitorOrganoidsOutcomeOutputPathway interactionsPeptidesPhenotypePhysiologyProbioticsProductionProtein SecretionProteinsRegulationResourcesSerotoninSignal TransductionStem cellsTechnologyTestingTherapeuticTimeTime Studybasecell growthcell typecommensal bacteriacostcytokinedesignexperimental studygut bacteriagut microbiotahost-microbe interactionshuman diseaseimprovedintestinal cryptintestinal villimicrobialpathogenpathogenic bacteriaphysical chemical interactionpublic health relevanceresponsesmall moleculethree dimensional cell culturetranslational impact
中文摘要
项目摘要
肠道细菌和肠道细胞之间的相互作用已经涉及许多人类
包括炎症性肠病、糖尿病、代谢综合征等在内的疾病。
目前,gnotobiotic动物模型是测试和理解这些的金标准。
相互作用,但动物实验是昂贵的,gnotibiotic殖民地需要大量的
资源我们提出,培养的肠道类器官或“小肠”可以用作
细菌可培养的中间环境和细菌培养的效果
和分泌对哺乳动物基因表达的影响可以在相对较短的时间内监测,
降低成本。我们的方法是共培养工程益生菌,
并积极影响3D培养中肠上皮细胞的发育。
我们建议通过以下具体目标开发这项技术:
目的1:建立小肠细胞分化的真实的实时表型表征
全细胞生物传感器来检测。 我们将开发全细胞生物传感器,
分泌物,如氯化物和血清素使用工程益生菌E。大肠杆菌的
这一目标背后的基本原理是,这些分子的分泌是适当的
肠干细胞分化为不同的细胞类型。荧光生物传感器
输出将以真实的实时操作,这将允许分泌表型的动态可视化,
荧光显微镜
目的2:通过分泌小分子和生长因子来改变小肠的生长,
肠道细菌。我们将设计益生菌,
并对干细胞在三维培养中的增殖有积极影响。这一目标的基本原理是
丁酸的分泌,一种关键的代谢产物,是肠道中自然微生物驱动的,
微生物丁酸生产对肠细胞的影响是深远的。此外,我们希望研究
细菌分泌的生长因子R-spondin-R11在小肠中对生长的积极调节
流明
总之,这些目标的成功实施将形成一种强大的共同发展文化
该平台用于优化3D类器官培养物和研究宿主微生物相互作用。
英文摘要
PROJECT SUMMARY
Interactions between gut bacteria and intestinal cells have been implicated in many human
diseases including inflammatory bowel disease, diabetes,metabolic syndrome, and many more.
Currently, gnotobiotic animal models are the gold standard for testing and understanding these
interactions, but animal experiments are costly and gnotibiotic colonies require large amounts of
resources. We propose that cultured intestinal organoids or "miniguts" can be used as an
intermediate environment in whichbacteria can be cultured and the effect of bacterial culture
and secretion on mammalian gene expression can be monitored at relatively shorter times and
lower costs. Our approach is to co-culture engineered probiotic bacteria to accurately monitor
and actively influence the development of intestinal epithelial cells in 3D culture.
We propose to develop this technology via the following Specific Aims:
Aim 1: Establish real-time phenotypic characterization of cell differentiation in miniguts
by engineered whole-cell biosensors. We will develop whole cell biosensors for intestinal
secretions such as chloride and serotonin using engineered probiotic E. coli bacteria. The
rationale behind this aim is that secretion of these molecules is an indication of proper
differentiation of intestinal stem cells into distinct cell types. The biosensor with fluorescent
output will operate in real-time which will allow dynamic visualization of secretion phenotypes by
fluorescence microscopy.
Aim 2: Alter minigut growth by secretion of small molecules and growth factors from
commensal bacteria. We will engineer probiotic bacteria to deliver molecules that negatively
and positively influence the proliferation of stem cells in 3D culture. The rationale of this aim is
that secretion of butyrate, a key metabolite, is naturally microbially-driven in the gut and the
effect of microbial butyrate production on gut cells is profound. In addition, we wish to study
positive regulation of growth by bacterial secretion the growth factor R-spondin-1 in the minigut
lumen.
Taken together, successful implementation of these aims will develop a robust co-culture
platform for the optimization of 3D organoid cultures and the study of host-microbe interactions.
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会议论文
Location-specific In Vivo Sensing and Imaging of Butyrate in the GI Tract
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批准号:10433447
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项目类别:
-
资助金额:$21.3万
-
财政年份:2022
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负责人:Thomas J Mansell
-
依托单位:
Location-specific In Vivo Sensing and Imaging of Butyrate in the GI Tract
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批准号:10611459
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项目类别:
-
资助金额:$17.41万
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财政年份:2022
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负责人:Thomas J Mansell
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依托单位:
Creating a niche for engineered live biotherapeutics
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批准号:10276995
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项目类别:
-
资助金额:$29.13万
-
财政年份:2021
-
负责人:Thomas J Mansell
-
依托单位:
Creating a niche for engineered live biotherapeutics
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批准号:10427447
-
项目类别:
-
资助金额:$29.07万
-
财政年份:2021
-
负责人:Thomas J Mansell
-
依托单位:
Creating a niche for engineered live biotherapeutics
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批准号:10622534
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项目类别:
-
资助金额:$29.0万
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财政年份:2021
-
负责人:Thomas J Mansell
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依托单位:
海外基金