A major goal of systems biology has been to comprehend how molecular circuitry governs information processing and decision-making in multicellular communities. A growing and largely descriptive single-cell atlas of tissues and pathologies has begun providing rich insight into the composition and spatial organization of microenvironments, yet it remains a challenge to understand cause-and-e?ect relationships from such data. How does the signaling state of one cell a?ect that of its neighbors? This simple question is complicated by reaction/di?usion transport in tissue, feedback loops based on cellular response to signaling, and dynamic cell migration. Despite this complexity, understanding principles of multi-scale intercellular communication promises to be a key component in designing cellular- and signaling-based therapies. Unfortunately, it has been di?cult to directly parse signal propagation in tissue because technological gaps have limited our ability to manipulate and monitor cell behavior in situ within native disease microenvironments.# Our proposal addresses these questions by leveraging recent advances in in vivo imaging, nanotechnology, and synthetic biology to generate a framework for image-guided manipulation, real-time monitoring, and systems-level analysis of signal propagation within microenvironmental niches. As an initial application, we will use this framework to understand how myeloid polarization signaling in?uences the tumor microenvironment in metastatic ovarian cancer. We focus in particular on monocyte-derived dendritic cells and macrophages, since they are highly implicated in drug resistance, they are therapeutically manipulated via targeted drugs or adoptive cell therapies, and it remains unclear how their signaling across the spectrum of pro- and anti-in?ammatory states can spread to neighboring cells over space and in time. Although this project will yield fundamental insights into myeloid signaling propagation, we also aim to extend image- guided genetic reprogramming to translationally-relevant modalities with potential therapeutic application. The novel integration of technologies to achieve these goals promises to be ?exible and useful for diverse biological applications where myeloid cells play a role, in cancer and beyond.#
The goal of this proposal is to develop a new paradigm for studying how spatially-de?ned molecular signaling circuits integrate across diverse multicellular communities, based on image-guided manipulation and real- time monitoring of in situ single-cell behaviors. We initially apply our framework to study how myeloid polarization propagates across the tumor microenvironment, with the goal of understanding fundamental principles of multi-scale signaling networks and new therapeutic strategies to exploit them. We anticipate our image-guided approach will bridge microscopy with synthetic biology in a transformative manner that is ?exible, practical, and highly translatable across technology platforms and biological applications. !