CAREER: Direct Measurement of Intranuclear Strain and Gene Expression in Single Cells in Vivo

该项目将极大地促进对结构复杂和分层生物组织中细胞核物理调节的理解。细胞核是细胞中的中心膜结合细胞器，其功能是控制对正常生理至关重要的基本细胞活动。更广泛地说，研究活动可能影响组织工程，再生医学和相关领域，通过建立研究核菌株和单细胞基因表达机制的能力，包括研究组织和生物材料中的稀有细胞群。该教育计划旨在提供一种互动和解决问题的学习方法。教育活动强调机械生物学的本科和研究生课程的发展，并扩大获奖者所在机构与儿童博物馆之间的合作关系，以促进从小学习科学和工程。这个教师早期职业发展（Career）奖的研究目标是应用先进的显微镜，图像配准和分子生物学方法来研究细胞核的生物力学和决定基因表达物理调节的基本机制。在该奖项下进行的研究将对受机械力干扰的天然组织中嵌入的单个细胞的细胞核进行成像，并确定细胞骨架蛋白对核内形状、变形和活动的影响。一个多尺度和翻译的方法也将研究正常和转基因小鼠的细胞核，以研究机械力如何调节基因在体内的表达。从研究活动中获得的知识将揭示亚细胞菌株在疾病和损伤中是如何改变的，研究将建立新的分析工具，用于模型系统中细胞核的无创、体内和功能生物力学评估。

This project will significantly advance the understanding of the physical regulation of the cell nucleus in structurally complex and hierarchical biological tissues. The nucleus is a central membrane-bound organelle in the cell that functions to control fundamental cellular activities that are critical to normal physiology. More broadly, the research activities may impact tissue engineering, regenerative medicine, and related fields, by establishing the ability to study mechanisms of nuclear strain and gene expression in single cells, including studies of rare cell populations within tissues and biomaterials. The education plan is designed to provide an interactive and problem-solving approach toward learning. The education activities emphasize undergraduate and graduate course development in mechanobiology, and broaden a partnership between the awardee's institution and a children's museum to promote learning in science and engineering from an early age.The research objective of this Faculty Early Career Development (CAREER) award is to apply advanced microscopy, image registration, and molecular biology methods to study the biomechanics of the cell nucleus and basic mechanisms that determine the physical regulation of gene expression. Studies conducted under this award will image the nucleus of single cells embedded in native tissue that is perturbed by mechanical force, and determine the influence of cytoskeleton proteins on intranuclear shape, deformation, and activity. A multiscale and translational approach will also study the nucleus in normal and genetically modified mice to investigate how mechanical force regulates gene expression in vivo. The knowledge gained from the research activities will reveal how subcellular strain is altered in disease and injury, and studies will establish new analysis tools for the noninvasive, in vivo, and functional biomechanical assessment of nuclei in model systems.