Computational and experimental modeling of cell function in response to 3D oxygen transport in vitro.

细胞功能响应体外 3D 氧运输的计算和实验模型。

基本信息

批准号：
9895842
负责人：
Ian S Kinstlinger
金额：
$ 2.9万
依托单位：
RICE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-04-16 至 2020-11-15
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9895842
关键词：
3-Dimensional Animal Model Architecture Area Behavior Biochemical Biocompatible Materials Biological Assay Biology Blood Blood Vessels Cardiac Output Cell Culture Techniques Cell Death Cell Density Cell Proliferation Cell Survival Cell model Cell physiology Cells Cellular Morphology Characteristics Computer Models Consumption Convection Coupled Cues Diffusion Engineering Environment Equation Evaluation Experimental Models Exposure to Gel Gene Expression Gene Expression Profile Goals HepG2 Hepatic Hepatocyte Heterogeneity Hydrogels Hypoxia Image Impairment In Vitro Incubated Jordan Knowledge Length Link Liver Maps Measurement Measures Metabolic Metabolism Modeling Organ Oxygen Pattern Perfusion Phenotype Physiological Play Population Positioning Attribute Protocols documentation Radial Regenerative Medicine Reproducibility Research Resources Role Structure Supporting Cell Techniques Therapeutic Time Tissue Engineering Tissue Model Tissues To specify Training Validation Variant Vision base biofabrication cell behavior cell type density design design and construction experimental study human tissue imaging Segmentation in vivo insight novel oxygen transport predictive modeling response spatiotemporal three dimensional cell culture three-dimensional modeling transcriptome sequencing

项目摘要

Abstract To advance regenerative medicine towards recapitulating the structures and functions of human tissues at physi- ologic length scales and with relevant cell densities, strategies must be developed to meet their unrelenting metabolic demands. In order to effectively and efﬁciently oxygenate functional engineered tissues, we must understand how changes in oxygen tension modulate cell viability, proliferation, and phenotype. Decades of studying cell cultures incubated under low oxygen levels have unveiled some aspects of the hypoxic response and many key in- sights into its mechanism. Separately, an astonishing array of biomaterials have been developed which can support cells in 3D environments and can recapitulate native cell morphologies and functions to a much greater extent than 2D culture. However, as emerging areas such as regenerative medicine have sought to incorporate cells within these materials, new questions have emerged regarding the roles played by oxygen transport and hypoxia in directing the density and function of the cell populations. Currently, we lack a comprehensive framework to describe and pre- dict how cell populations will alter their densities and functions over time in the presence of spatiotemporally heterogeneous oxygen gradients. We need to extend our knowledge of cellular responses to hypoxia into 3D and we need to proﬁle how tissue-speciﬁc cell functions are impacted by local oxygen cues. In this proposal, I and a sup- porting team of experts in biomaterials, computational modeling, and liver biology will unify computational models of hypoxic response with engineered model tissues to link oxygen transport with tissue function in 3D. Our ﬁndings will be incorporated into an experimentally validated model capable of predicting how cell popula- tions change in density and function in response to speciﬁed oxygen gradients. Cellular responses to hypoxia will be parameterized by cell-speciﬁc response functions and integrated with oxygen transport equations in an agent-based computational model. We will ﬁt parameters using advanced volumetric imaging and image segmentation along with biochemical assays to map cellular markers of viability, proliferation, hypoxia, and phenotype within 3D hydrogels containing HepG2 liver cells, a well-deﬁned model cell type from a highly metabolic tissue. I hypothesize that our closed-loop computational and experimental workﬂow will yield a scalable model of cell behavior at the tissue level which captures previously unstudied functional responses to hypoxia. Finally, to broadly proﬁle the phe- notypic landscape of cells growing in the presence of oxygen gradients, we will use RNA sequencing to map spatial zonation of cell phenotypes along axial and radial oxygen gradients in perfused hydrogels. Controlled encapsulation of cells within hydrogels of reproducible architecture will enable us to evaluate these spatial patterns in gene expres- sion with a degree of experimental control and reproducibility beyond the capabilities of in vivo approaches. Taken together, these studies will provide fundamental insights into how cells respond to local oxygen gradients in 3D environments. Analysis of the spatial heterogeneity introduced by oxygen gradients is also expected to inspire new paradigms for engineering zones of cell function within tissues.

抽象的推动再生医学在物理层面重述人体组织的结构和功能逻辑长度尺度和相关的细胞密度，必须制定策略以满足其无情的代谢为了有效地为功能性工程组织供氧，我们必须了解这些需求。氧张力的变化如何调节细胞活力、增殖和表型数十年的细胞研究。在低氧水平下培养的培养物揭示了缺氧反应的某些方面和许多关键因素另外，人们还开发了一系列令人惊叹的生物材料来支持这一机制。 3D 环境中的细胞，可以在更大程度上重现原生细胞形态和功能然而，随着再生医学等新兴领域试图将细胞纳入其中。材料中，出现了关于氧运输和缺氧在引导细胞中所起的作用的新问题。目前，我们缺乏一个全面的框架来描述和预测细胞群的密度和功能。决定细胞群在存在时空的情况下如何随时间改变其密度和功能我们需要将细胞对缺氧反应的知识扩展到 3D 和我们需要分析局部氧信号如何影响组织特异性细胞功能。生物材料、计算模型和肝脏生物学专家的移植团队将统一计算缺氧反应模型与工程模型组织将氧运输与组织功能联系起来 3D.我们的发现将被纳入一个经过实验验证的模型中，该模型能够预测细胞数量如何变化。细胞对缺氧的反应将导致密度和功能的变化。由细胞特定的响应函数参数化，并与基于代理的氧传输方程集成我们将使用先进的体积成像和图像分割来拟合参数。生化检测可绘制 3D 水凝胶内活力、增殖、缺氧和表型的细胞标记物含有 HepG2 肝细胞，这是一种来自高度代谢组织的明确模型细胞类型。闭环计算和实验工作流程将产生组织中细胞行为的可扩展模型水平捕获了先前未研究的缺氧功能反应。由于细胞在氧气梯度存在下生长的情况不典型，我们将使用 RNA 测序来绘制空间图受控封装中细胞表型沿轴向和径向氧梯度的分区。水凝胶内细胞的可重复结构将使我们能够评估基因表达中的这些空间模式具有超出体内方法能力的一定程度的实验控制和再现性。总之，这些研究将为细胞如何响应局部氧梯度提供基本见解。对氧梯度引入的空间异质性的分析也有望得到启发。组织内细胞功能工程区域的新范例。