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

职业：在生物组织中塑造光：超声波介导的行进光源，用于时空精确的体内基因编辑

• 批准号: 2045120
• 负责人: Guosong Hong
• 金额: $ 50万
• 依托单位: Stanford University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-10-01 至 2026-09-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2045120&HistoricalAwards=false
• 关键词: CAREER; Sculpting; light; biological; tissue

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.

生物光子学的一个基本挑战是，由于散射和吸收，光在体内深处的穿透性较差，因此需要侵入性手术，例如光纤植入和手术组织切除，以传输用于深层组织成像和光激活治疗的光。为了应对这一根本挑战，这个CAREER项目寻求通过无创超声波在体内任何深度或位置按需产生光，从而以用户定义的模式实现体内的“光雕刻”。研究人员的小组将使用这种方法在深层组织中进行空间和时间上精确的基因编辑。该职业项目的科学成果将使研究者及其团队能够更广泛地参与多种教育活动。这些教育活动包括 K-12 外展和教师培训、对大学前代表性不足的少数族裔 (URM) 学生的长期指导，以及为来自服务不足的高中和传统黑人学院和大学 (HBCU) 的学生提供实验室实践实习。研究者的首要职业目标是弥合知识差距并消除物理科学和生命科学之间的障碍。为了实现这一目标，该CAREER项目旨在开发一种工程超声场下的循环可传输和可充电光源，从而实现三维生物组织中的按需“光雕刻”，从而解决生物光子学将光传输到100微米以上深度的根本挑战。 该项目的主要假设是，血液循环中移动的机械发光纳米颗粒（MLNP）可以充当能量中继，通过循环 MLNP（青色圆圈）和组织渗透性 FUS（聚焦超声）来传输局部光发射的需求，以进行基因编辑。 该研究计划分为三个目标：第一个目标是开发一系列稀土掺杂黄长石磷光体 MLNP，其发射波长可调，可响应不同的超声波频率。通过对超声压力、发射波长和 MLNP 陷阱深度之间关系的机械理解，第二个目标是基于模拟超声压力场、局部血流动力学和血液循环 MLNP 的光物理特性，获得任何生物组织中用户定义的光发射时空模式。第三个目标是利用超声波进行体内“光雕刻”，以在活体小鼠中进行时空精确的基因编辑，并使用可光切换的 Cas9 和生物发光报告基因来评估成功情况。 如果成功，这项技术可以广泛扩展到任何需要体内深处光源的应用，包括体内荧光显微镜、深部脑光遗传学以及治疗癌症和病毒感染的光动力疗法。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。

项目成果

A palette of rechargeable mechanoluminescent fluids produced by a biomineral-inspired suppressed dissolution approach

由生物矿物启发的抑制溶解方法产生的可充电机械发光液体调色板

• 发表时间: 2022
• 期刊: Journal of the American Chemical Society
• 影响因子: 15
• 作者: Fan Yang, Xiang Wu