Mimicking, Exploiting, and Understanding Biology's Heterogeneity in 4D

项目总结 我们的身体由不断变化的组织和器官组成。从… 形态发生和动态平衡对疾病的进展，这些变化都与 定义人类生活的健康和不健康的过程。我在华盛顿大学的研究项目 正在开发强大且独特强大的多学科方法，以模拟、利用和量化 这些变化，特别是当它们在时间和3D空间(即4D)中演变时。在我们实验室的头五年 存在，我们已经：(1)开发了一套合成细胞培养平台，其生化和 生物物理特性可以使用细胞兼容的光化学在4D中进行可逆调制，并且具有 利用这些平台调节增殖、迁移、分化和细胞内信号转导。 和亚细胞分辨率；(2)引入了一种基于光降解材料的方法来产生第一个 细胞载水凝胶生物材料中的内皮化3D血管网络几乎跨越所有尺寸的 天然人类血管系统(包括毛细血管)；(3)报告了第一个用于传授的模块化框架 具有对多种环境线索/生物标志物的精确降解响应的生物材料 用户可编程的布尔逻辑；以及(4)建立了第一批工具，用于在空间和时间上进行解析 蛋白质组学“，能够可视化和量化用户体内和体外产生的蛋白质- 在4D中定义区域。目前的提议扩大了我们集团在每个领域的能力，为 通过从根本上转变的知识获得新的治疗靶点和治疗疾病的方法 基本的细胞生理学。在本项目中，我们将：(1)开发我们的4D可调生物材料来总结和探索 体外心血管发育信号，检查精确的空间和时间 信号蛋白的呈现最终导致有组织的分化；(2)使用合成毛细管来 检查药物作用和耐药性，筛选治疗药物，调查微血管闭塞，血栓形成， 以及许多血液病(如镰状细胞性贫血、球形红细胞增多症)中发生的重塑改变； 开发和部署水凝胶纳米颗粒，对癌症表现出基于逻辑的降解反应-呈现 以前所未有的特异性为肿瘤提供小分子化疗药物的生物标志物；以及(4) 扩展我们的4D蛋白质组策略，以允许在生活中进行光学和生理定义的蛋白质组映射 组织和模式生物。关键是，我们正在开发和实施的方法是细胞、组织、 和疾病不可知性，使人们在产卵时能够更好地理解各种生物过程 疾病诊断、治疗和预防进步的基础。

PROJECT SUMMARY Our bodies consist of an exquisite collection of tissues and organs that undergo constant change. From morphogenesis and homeostasis to the progression of disease, these changes are associated with both the healthy and unhealthy processes that define human life. My research program at the University of Washington is developing robust and uniquely powerful multidisciplinary methodologies to mimic, exploit, and quantify these changes, particularly as they evolve in both time and 3D space (i.e., 4D). In our lab’s first five years of existence, we have: (1) developed a suite of synthetic cell-culture platforms whose biochemical and biophysical properties can be reversibly modulated in 4D using cytocompatible photochemistries, and have utilized these platforms to regulate proliferation, migration, differentiation, and intracellular signaling at single- and sub-cellular resolutions; (2) introduced a photodegradable material-based approach to generate the first endothelialized 3D vascular networks within cell-laden hydrogel biomaterials that span nearly all size scales of native human vasculature (including capillaries); (3) reported the first modular framework for imparting biomaterials with precise degradative responsiveness to multiple environmental cues/biomarkers following user-programmable Boolean logic; and (4) established the first tools for “spatiotemporally resolved proteomics”, enabling visualization and quantification of proteins produced in vitro and in vivo within user- defined regions in 4D. The present proposal expands our group’s capabilities in each of these areas, paving the way to new therapeutic targets and treatments of disease through a fundamentally transformed knowledge of basic cell physiology. In this project, we will: (1) exploit our 4D-tunable biomaterials to recapitulate and probe cardiovascular developmental signaling in vitro, examining the manner in which precise spatial and temporal presentation of signaling proteins culminates in orchestrated differentiation; (2) employ synthetic capillaries to examine drug action and resistance, screen therapeutics, and investigate microvascular occlusion, thrombosis, and altered remodeling that occurs in many hematologic diseases (e.g., sickle cell anemia, spherocytosis); (3) develop and deploy hydrogel nanoparticles exhibiting logic-based degradative response to cancer-presented biomarkers to deliver small molecule chemotherapeutics to tumors with unprecedented specificity; and (4) extend our 4D proteomic strategies to permit optically and physiologically defined proteomic mapping in living tissue and model organisms. Critically, the methods that we are developing and implementing are cell-, tissue-, and disease-agnostic, enabling enhanced understanding of a wide variety of biological processes while laying the foundation for advances in disease diagnosis, treatment, and prevention.