Investigating the influence of biomimetic cues in cardiovascular system formation

组织结构及其力学特性是心脏和血管发育和功能的重要调节因素。我的研究计划由相互关联的项目组成，重点是建立组织结构和生物力学力量、新血管成熟和来自干细胞的心脏细胞之间的相关性。在目标1中，我们将开发一个功能性的“芯片上的微血管系统”，以研究基质组成以及血管内皮细胞和血管周围细胞-细胞相互作用对血管通透性的影响，在一个考虑血流的动态平台上。需要创新的方法来生成功能性微血管，并以准确和可控的方式重现其生理微环境。我们将通过使用脂肪来源的微血管来实现这一点，这些微血管可以自我组装成血管系统，并拥有形成功能网络所需的所有细胞类型。这一平台将首次允许在存在流动的情况下在体外产生复杂的、有功能的微血管；这代表着对现有策略的重大进步，并解决了这些应用在人类细胞来源方面的一个重要缺口。在目标2中，我们将通过对血管细胞(血管内皮细胞和平滑肌细胞)施加循环拉伸来确定牵引力在特定血管动脉特性形成中的影响，并研究它是否更好地导致动脉特性的指定。我们将评估特定动脉基因的表达，涉及血管周围募集、细胞面积、增殖和细胞-细胞相互作用的生长因子。我们研究的创新来自于研究循环伸展作为动脉身份正确分配的成熟线索，并代表了潜在的新的概念进展。由于心脏的发育依赖于血管的形成，在目标3中，我们将研究血管的存在是否会促进干细胞来源的心脏细胞的成熟。我们将通过确定向干细胞来源的心脏细胞添加血管或内皮细胞是否会促进它们的成熟来检验这一假设。细胞成熟度将通过评估分子特征、结构和细胞组织、功能特性和电生理来确定。这些将以不同发育阶段的人类心肌细胞为基准。这是一个具有极高潜在影响的领域，在那里获得人类成体样心肌细胞将是世界上第一次。总而言之，这些研究将扩大我们对心血管成熟和功能机制的理解，并将指导我们努力在体外培养复杂的、具有血管功能的人类心脏组织(我们的长期目标)。

Tissue architecture and its mechanical properties are important modulators of the development and function of the heart and blood vessels. My research program consists of inter-related projects that are focused on establishing the relevance of tissue architecture and biomechanical forces, in the maturation of new blood vessels and of heart cells-derived from stem cells. In Objective 1, we will develop a functional “microvasculature-on-a-chip” to study effects of matrix composition and the modulation of endothelial and perivascular cell-cell interactions in vascular permeability in a dynamic platform that accounts for flow. Innovative approaches are needed for generating functional microvasculatures and recapitulating their physiological microenvironments in an accurate and controllable fashion. We will achieve this by use of adipose-derived microvessels that can self-assemble into a vasculature and have all the cell types necessary to form a functional network. This platform will allow for the first time, the generation of complex, functional microvasculatures in vitro in the presence of flow; which represents a significant advancement to existing strategies and solves an important gap in terms of human cell source for these applications. In Objective 2 we will define the influence of traction forces in the formation of specific vessel arterial identity by applying cyclic stretch to vascular cells (endothelial and smooth muscle cells) and investigating if it would preferably lead to the specification of arterial identity. We will assess the expression of specific arterial genes, growth factors involved in perivascular recruitment, cell area, proliferation and cell-cell interaction. The innovation in our studies comes from investigating cyclic stretch as a maturation cue for the proper assignment of arterial identity and represent a potential novel conceptual advance. Since the development of the heart is dependent on the formation of blood vessels, in Objective 3, we will investigate if the presence of blood vessels improves the maturation of the heart cells derived from stem cells. We will test this hypothesis by determining whether the addition of blood vessels or endothelial cells to stem cell-derived heart cells will enhance their maturation. Cell maturation will be determined by assessing the molecular signature, structural and cellular organization, functional properties and electrophysiology. These will be benchmarked against human cardiomyocytes from different stages of development. This is an area of extremely high potential impact where obtaining human adult-like heart cells in culture would be a world first. Collectively, these studies will expand our understanding of the mechanisms of cardiovascular maturation and function and will guide our efforts towards generating complex, functional vascularized human heart tissues in vitro (our long-term goal).