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

点击翻译按钮获取中文摘要

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.