Project Summary/Abstract Hypertensive vascular disease is a leading global risk factor for morbidity and mortality, affecting over 116 million people. It is characterized by alterations to extracellular matrix (ECM) composition, biomechanical properties, and aberrant molecular signaling leading to increased blood pressure and vessel stiffening. Extensive work has focused on developing improved pharmacological treatments, tissue engineered blood vessels, and vascularizing engineered volumetric tissue, but often overlook the interdependence between cellular signaling and biomechanical forces leading to disease. Understanding the relationship between ECM biomechanics and receptor signaling, and developing tools to manipulate it, are essential for creating a healthy, mature vascular tissue while avoiding pathological changes. Here I propose that incorporation of spatially defined ECM composition and cellular alignment into a vascular-inspired 3D bioprinted tissue scaffold will produce an engineered small artery that physiologically controls vascular tone and facilitates investigation into how ECM composition and altered receptor trafficking impair vascular reactivity and promote a hypertensive phenotype. I will utilize two novel platforms: FRESH 3D bioprinting to directly fabricate perfusable vasculature from ECM proteins, and a fluorescence-based nanomechanical biosensor (NMBS) for mapping in vivo tissue strain and vascular smooth muscle contractility to Aim 1: Develop a collagen-based 3D bioprinted vascular microfluidic platform integrating controlled fluid flow and endothelialization to replicate vascular ECM biomechanics and endothelial barrier function; Aim 2: Directly pattern ECM structure and cellular organization using FRESH printing in a layer-by-layer manner to recapitulate resistance artery vascular smooth muscle cell and endothelial cell function; and Aim 3:Investigate how pathologic changes in the ECM alters receptor trafficking and impairs vascular reactivity using novel bioinks to replicate a healthy and hypertensive vessel ECM composition and material properties. This proposal seeks to enhance our knowledge of the interplay between biomechanical forces biochemical signaling during vascular development and hypertensive disease progression. The microfluidics (K99) and engineered vascular tissues (R00) created will have wide utility for drug screening and disease modeling. The career development plan, under the guidance of co-mentors Drs. Feinberg and Kleyman, and my advisory committee, will provide advanced training in microfluidics, biomechanical analysis, and vascular biology/disease modeling. The mentored phase capitalizes on an inter-disciplinary mentoring team and substantial research and professional development resources at Carnegie Mellon University, the University of Pittsburgh, and the Vascular Medicine Institute. This K99/R00, combined with my prior expertise in 3D bioprinting, advanced fluorescence microscopy, quantitative image analysis, and cellular/molecular biology, will facilitate my transition to an independent career focused on how ECM composition and structure alter receptor signaling to drive tissue maturation and disease progression.

Project Narrative Vascular disease resulting from hypertension is a leading health problem in the US; yet, it remains unknown how hypertensive changes in the composition of vessel extracellular matrix (ECM) and mechanical properties regulate receptor trafficking and vascular reactivity. There is a critical need to develop an engineering platform capable of patterning ECM structure, composition, and cellular alignment in 3D to build a perfusable vascular network with native biomechanical properties to investigate the interplay between biomechanical forces and biochemical signaling in hypertensive vascular disease. This proposal will use FRESH 3D bioprinting and a newly developed biomechanical strain sensor to 3D bioprint a functional resistance artery and create an ECM- based vascular microfluidic platform to improve our understanding of hypertensive disease progression.