CAREER: Sculpting light in biological tissue: an ultrasound-mediated traveling light source for spatiotemporally precise in vivo gene editing

生物光子学的一个基本挑战是，由于散射和吸收，光在体内的穿透性很差，因此需要进行侵入性程序，如光纤植入和外科组织切除，以提供用于深层组织成像和光激活治疗的光。为了解决这一根本挑战，这个职业项目寻求通过非侵入性超声波在人体内的任何深度或位置按需产生光线，从而实现身体中具有用户定义图案的“光雕刻”。研究小组将使用这种方法在深层组织中进行空间和时间上的精确基因编辑。这一职业项目的科学成果将使调查员及其小组能够更广泛地参与各种教育活动。这些教育活动包括K-12外展和教师培训，为大学前代表不足的少数族裔(URM)学生提供长期指导，以及为服务不足的高中和历史上黑人学院和大学(HBCU)的学生提供动手实验室实习。研究人员的首要职业目标是弥合知识的鸿沟，消除物理科学和生命科学之间的障碍。为了实现这一目标，这一职业项目旨在开发一种在工程超声场下可循环交付和可充电的光源，从而在三维生物组织中实现按需“光雕刻”，从而解决生物光子学的基本挑战，即将光传输到100微米以上的深度。该项目的主要假设是，血液循环中的移动机械发光纳米颗粒(MLNPs)可以作为能量继电器，通过循环MLNPs(青色环)和组织穿透性FUS(聚焦超声)传输局部光发射的需求，以进行基因编辑。该研究计划安排在三个目标下：第一个目标是开发一种稀土掺杂黄长石发光材料MLNPs调色板，其发射波长可调，以响应不同的超声波频率。第二个目标是基于模拟的超声声压场、局部血流动力学和血液循环中MLNPs的光物理性质，从机理上了解MLNPs的超声压力、发射波长和陷阱深度之间的关系，从而获得用户定义的任何生物组织中的光发射时空模式。第三个目标是用超声波在活体内进行“光雕刻”，在活体小鼠中进行时空精确的基因编辑，使用可切换的Cas9和生物发光报告来评估成功。如果成功，这项技术可以广泛扩展到任何需要身体深处光源的应用，包括活体荧光显微镜、脑深部光遗传学和光动力疗法来治疗癌症和病毒感染。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

A fundamental challenge in biophotonics is the poor penetration of light deep inside the body due to scattering and absorption, thereby necessitating invasive procedures, such as optical fiber implantation and surgical tissue removal, to deliver light for deep-tissue imaging and light-activated therapies. To address this fundamental challenge, this CAREER project seeks to produce light on-demand at any depth or location inside the body via noninvasive ultrasound, thus realizing “light sculpting” in the body with user-defined patterns. The investigator’s group will use this method for spatially and temporally precise gene editing in deep tissue. The scientific outcomes of this CAREER project will enable the broadening participation of the Investigator and his group in a multitude of education activities. These education activities include K-12 outreach and teacher training, long-term mentorship to pre-college underrepresented minority (URM) students, and hands-on lab internship to students from underserved high schools and historically Black colleges and universities (HBCUs). The overarching career goal of the investigator is to bridge the gap in knowledge and eliminate the barrier between the physical and life sciences. Towards this goal, this CAREER project aims to develop a circulation deliverable and rechargeable light source under an engineered ultrasound field, thereby leading to on demand “light sculpting” in three-dimensional biological tissue and thus address the fundamental challenge of biophotonics of delivering light to a depth of more than 100 microns. The project’s main hypothesis is that traveling mechanoluminescent nanoparticles (MLNPs) in blood circulation can act as an energy relay that transports demand generation of localized light emission by circulating MLNPs (cyan circles) and tissue-penetrant FUS (focused ultrasound) for gene editing. The research plan is organized under three objectives: the FIRST objective is to develop a palette of rare-earth doped melilite phosphor MLNPs with tunable emission wavelengths in response to different ultrasound frequencies. With mechanistic understanding of the relationship between ultrasound pressure, emission wavelength and trap depth of MLNPs, the SECOND objective is to obtain user-defined spatiotemporal pattern of light emission in any biological tissue based on the simulated ultrasound pressure field, local hemodynamics, and the photophysical properties of blood-circulating MLNPs. The THIRD objective is to perform in vivo "light sculpting" with ultrasound for spatiotemporally precise gene editing in live mice, using photo switchable Cas9 and a bioluminescent reporter to assess success. If successful, this technology can be widely extended to any application requiring a light source deep in the body, including in vivo fluorescence microscopy, deep-brain optogenetics, and photodynamic therapies to treat cancer and viral infections.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.