| 项目编号: | 1703219
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| 项目名称: | A Robust Human Blood-Brain Barrier Model Generated from Induced Pluripotent Stem Cells |
| 作者: | Eric Shusta
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| 承担单位: | University of Wisconsin-Madison
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| 批准年: | 2017
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| 开始日期: | 2017-07-01
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| 结束日期: | 2020-06-30
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| 资助金额: | 400000
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| 资助来源: | US-NSF
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| 项目类别: | Standard Grant
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| 国家: | US
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| 语种: | 英语
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| 特色学科分类: | Engineering - Chemical, Bioengineering, Environmental, and Transport Systems
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| 英文关键词: | bbb model
; shear stress
; small molecule
; project
; scaled-out
; human induced pluripotent stem cell
; model
; resultant bbb endothelial cell
; mature bbb cell
; bbb permeability
; brain drug uptake
; brain endothelial cell
; ipsc-derived bbb endothelial cell
; cartilage cell
; impermeable brain vasculature
; ipsc
; stem cell engineering
; multi-cellular differentiation
; panel
; blood-brain barrier
; bbb endothelial cell
; heart cell
; other stem cell product
; bbb cell purification
; blood brain barrier
; bone cell
; chemotherapeutic
; large-scale
|
| 英文摘要: | PI: Shusta, Eric V.
Proposal: 1703219
Millions of Americans are afflicted with neurological illnesses such as Alzheimer's disease, Parkinson's disease, and cerebral AIDS. However, very few new treatments have resulted, despite significant advances in the development of both small molecule pharmaceuticals and biopharmaceuticals (gene and protein medicines). In large part, the paucity of new therapies is due to a failure to overcome the impermeable brain vasculature, also known as the blood-brain barrier (BBB). Thus, accurate prediction of human BBB permeability prior to clinical trials is one of the most significant challenges in neuropharmaceutical development. A potential solution is the engineering of a cell-based, in vitro model that can be used prior to clinical drug administration to mimic human BBB permeability characteristics observed in vivo. To this end, the research team recently created a BBB model based on brain endothelial cells differentiated from human induced pluripotent stem cells (iPSCs). The resultant BBB endothelial cells possess characteristics that potentially have the transformative capacity for predicting brain drug uptake. The iPSC differentiation process would need to be facile, scalable and robust for broad dissemination to industrial and academic researchers. A major challenge to the ability to scale up is the need for flow-based shear stress, which is not possible at large scale. To address this challenge, this project hypothesizes that the effects of shear stress can be mimicked by employing small molecules that can trigger key shear-induced mechanotransduction pathways, thereby enabling large-scale BBB differentiation. Finally, the fully mature BBB cells will be cryopreserved, scaled-out and benchmarked using a panel of chemotherapeutics with known in vivo BBB permeability. The project is designed to train one full-time graduate student and one undergraduate researcher in this interdisciplinary space of stem cell engineering to prepare them for careers in industry and academia. Stem cell-based BBB modeling will also be disseminated to rural Wisconsin high school students through hands-on experimental modules as part of the UW-Madison Morgridge Research Institute Summer Science Camp.
The project focuses on developing an in vitro model of the blood brain barrier (BBB) that is scalable, chemically defined, and improves upon existing alternatives for in vitro BBB models based on functionally-relevant metrics, including transendothelial electrical resistance (TEER) measurements and the expression of drug transporters. To this end, the project builds on the research team's recently created BBB model based on brain endothelial cells differentiated from human induced pluripotent stem cells (iPSCs). The resultant BBB endothelial cells possess well-developed tight junctions, characteristic transporter expression, and could ultimately possess the transformative capacity for a priori prediction of brain drug uptake. The iPSC differentiation process would need to be facile, scalable and robust for broad dissemination to industrial and academic researchers. In its current embodiment, the BBB model requires a fairly complex combination of undefined medium and matrix, in addition to multi-cellular differentiation that requires BBB cell purification prior to use. To address these challenges, the project plans to completely define the BBB differentiation protocol using an innovative combination of temporal signaling pathway activation and a defined matrix, resulting in a pure population of BBB endothelial cells. Importantly, flow-based shear stress will be applied to further mature the iPSC-derived BBB endothelial cells towards an in vivo-like BBB phenotype that may be therefore more predictive of in vivo drug transport properties; but it is difficult, if not impossible, to apply shear stress at large scale. As a paradigm shift, it is hypothesized that the effects of shear stress can be mimicked by instead employing small molecules that can agonize key shear-induced mechanotransduction pathways, thereby enabling large-scale BBB differentiation. Such proof-of-principle demonstration would motivate using small molecules to substitute for mechanical forces in the differentiation protocols of other stem cell products, such as heart cells, bone cells and cartilage cells, and facilitate their practical large-scale production. Finally, the fully mature BBB cells will be cryopreserved, scaled-out and benchmarked using a panel of chemotherapeutics with known in vivo BBB permeability. In summary, the team proposes to: 1) develop a fully defined protocol for the differentiation of iPSCs to BMECs capitalizing on the sequential induction of Wnt and retinoic acid (RA) signaling; 2) use BMECs to develop BBB-mimics in transwell plates and enhance the BBB phenotype through delivery of small molecules mimicking shear stress effects; and 3) validate the BBB model by measuring the permeability of a panel of chemotherapeutics and comparing these values against known in vivo permeabilities. |
| 资源类型: | 项目
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| 标识符: | http://119.78.100.158/handle/2HF3EXSE/89768
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| Appears in Collections: | 全球变化的国际研究计划
科学计划与规划
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| Recommended Citation: | |
Eric Shusta. A Robust Human Blood-Brain Barrier Model Generated from Induced Pluripotent Stem Cells. 2017-01-01.
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