Novel Techniques for Vascular Investigation

National Heart, Lung, And Blood Institute

Investigators

Linked publications, trials & patents

Abstract

Over the last year, weve continued to both fold in existing vascular physiology techniques and to leverage the newly optimized methodology for latex casting of organ-based vasculature into several projects. These techniques were utilized for a recent manuscript highlighting the impact of elastin insufficiency on pulmonary vasculature with and without minoxidil treatment (Knutsen et al, Front Cardiovasc Med.) and are showcased in two new manuscripts in progress looking at the impact of chronic vascular stiffness on aging tissue beds. The ability to visualize both the proximal and distal vasculature simultaneously and in 3D allowed us to see complex changes to the caliber, density, and tortuosity of the vasculature that would have been difficult to appreciate by histology or more traditional CT angiogram. In addition to this technique, were using advanced in-vivo ultrasound techniques to evaluate both vascular function and cardiac changes in the vasculature of aged elastin haploinsufficient mice. We grew our ex vivo vascular phenotyping methods by extending our vessel myography to incorporate pulsatile flow. We then interrogated the integrity of the vascular matrix proteins by the addition of stressors during ex-vivo cyclic hemodynamic simulations thereby yielding information regarding matrix assembly and structure quality. This method was underlined in our recent manuscript focused on the elastin crosslinking protein Lysyl Oxidase (Tsang et al, Int J Mol Sci.). Collaborative efforts have yielded a Cell Rep Methods paper describing a novel method combining multi-modality imaging to generate high-information 3D analysis of intracranial vasculature (Rosenblum et al, Cell Rep Methods) and work is underway applying this new multi-modality method to a collaborative project evaluating the impact of Notch3 insufficiency on intracranial vascular function and brain outcomes. We are also continuing to extend our collaboration with the Zhuang lab exploring Hif2-Alpha and SDHB vascular malformations. Over the next year, we intend to dive more deeply into methods which will enable small vessel visualization in end organs such as light sheet microscopy and to increasingly incorporate AI technologies for interrogation of the the high quantities of data collected with these techniques. Studies underway so far have generated high resolution images which will enable vascular density quantification, enabling us to answer questions about the potential for chronic hypoperfusion in our vascular models.