CAREER: Rational Design of Phase-Changing Nanomaterials for Spatiotemporal Protein Delivery

非技术摘要本项目将开发超声敏感的纳米材料，可以被声学激活，以高精度将其他不可渗透的生物分子输送到细胞中。这是通过合理设计的氟相变纳米粒子（PCN），它经历液-气相变响应声刺激。虽然多氯化萘作为超声成像剂取得了成功，但其转化为药物输送装置受到货物装载不良和生物学结果多变的阻碍。在先前开创性工作的基础上，该项目将通过了解颗粒组成和组织力学对其声学激活的相互依赖性，为多氯化萘的合理设计开辟新的机会，并优化生物货物的装载和交付。这些目标将通过三个目标来实现：（1）制备生物启发的多氯化萘，并确定物理和化学性质如何影响其超声激活;（2）证明蛋白质成功分散到氟多氯化萘的液体内部，并评估超声下的递送;（3）将基于生物物理细胞的实验与膜力学的计算模型配对，以推进更全面的PCN机制。介导的细胞透化。 这项工作的成功将使PCNs的设计与良好控制的声学生物物理学，产生新的生化工具，可用于探测和操纵复杂的组织微环境中的细胞具有高精度。该项目的研究结果将与推广活动相结合，将生物材料设计和生物物理学的原则纳入K-12教育。教育目标包括：（1）检查将高中生与本科工程设计团队整合在STEM教育中的招聘和坚持的效果;（2）开发和实施超声奥运会计划，为K-12学生提供变革性的实践经验纳米技术和生物声学。利用综合研究和推广计划，通过严格和常规评估的支持，该项目将推进新的生物纳米技术，并提高新兴STEM领域的学生保留率。 技术摘要本CAREER奖的目标是开发一个框架的合理设计的氟相变纳米粒子（PCNs），可以通过超声蒸发，以提供跨膜传递的生物分子进入细胞的空间和时间精度。虽然多氯化萘作为超声造影剂已经取得了成功，但它们转化为临床上有用的运载工具还有待实现。这是两个关键知识差距的结果，包括：（1）缺乏定义明确的结构-活性关系，定义PCN物理化学性质如何调节其相变行为;（2）对局部组织力学如何影响PCN生物物理学的机械理解不完整。使用模板驱动的肽组装，该项目将制备可编程的PCN，以阐明颗粒表面张力和大小对其声学激活的影响，以及确定允许蛋白质货物装载到含氟液体颗粒核心中并从其中递送的条件。同时，该项目将确定颗粒-细胞-基质相互作用如何协同工作，以控制汽化过程中细胞表面多氯化萘的生物物理学。为了在新兴生物技术领域培养和培养新一代的多元化科学家，例如这里所探讨的那些科学家，该项目将制定和实施一项推广计划，将代表性不足的K-12学生纳入STEM研究。活动将包括将服务不足的高中学生纳入本科工程设计团队，并创建一个超声奥运会研讨会，让6-12年级的教师参与，并将生物工程和声学概念融入课堂。通过将拟议的研究计划与外展活动联系起来，该项目将有助于培养一支有竞争力的STEM人才队伍，能够应对生物材料设计、分子生物物理学和生物成像方面的各种挑战。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Non-Technical AbstractThis project will develop ultrasound-sensitive nanomaterials that can be acoustically activated to deliver otherwise-impermeable biomolecules into cells with high precision. This is achieved through the rational design of fluorous phase-changing nanoparticles (PCNs), which undergo liquid-gas phase transitions in response to acoustic stimuli. While PCNs have enjoyed success as ultrasound imaging agents, their translation into drug delivery devices has been impeded by poor cargo loading and variable biologic outcomes. Building upon prior seminal work, this project will open new opportunities in the rational design of PCNs by understanding the interdependence of particle composition and tissue mechanics on their acoustic activation, and optimize the loading and delivery of biologic cargo. These objectives will be accomplished through three aims: (1) Prepare bio-inspired PCNs and identify how physical and chemical properties impacts their ultrasound activation; (2) Demonstrate successful dispersion of proteins into the liquid interior of fluorous PCNs and assess delivery under ultrasound; (3) Pair biophysical cell-based experiments with computational models of membrane mechanics to advance a more comprehensive mechanism for PCN-mediated cell permeabilization. Success of this work will enable the design of PCNs with well controlled acoustic biophysics, yielding new biochemical tools that can be used to probe and manipulate cells within complex tissue microenvironments with high precision. Research findings from this project will be integrated with outreach activities to incorporate principles of biomaterial design and biophysics into K-12 education. Educational objectives include: (1) Examine the effect of integrating high-school students with undergraduate engineering design teams on recruitment and persistence in STEM education; and (2) Develop and implement an Ultrasound Olympics program that will provide transformative hands-on experiences to K-12 students in nanotechnology and bioacoustics. Leveraging an integrated research and outreach program, supported through rigorous and routine assessment, this project will advance new bio-nanotechnologies and improve student retention in emerging STEM fields. Technical AbstractThe goal of this CAREER award is to develop a framework for the rational design of fluorous phase-changing nanoparticles (PCNs) that can be vaporized by ultrasound to afford transmembrane delivery of biomolecules into cells with spatial and temporal precision. Although PCNs have enjoyed success as ultrasound contrast agents, their translation into clinically useful delivery vehicles has yet to be realized. This is a consequence of two key gaps in knowledge that include: (1) a lack of well-defined structure-activity relationships defining how PCN physicochemical properties regulate their phase-changing behavior; and (2) an incomplete mechanistic understanding of how local tissue mechanics impacts PCN biophysics. Using template-driven peptide assembly, this project will prepare programmable PCNs to elucidate the influence of particle surface tension and size on their acoustic activation, as well as identify conditions that allow protein cargo to be loaded into, and delivered from, the fluorous liquid particle core. In parallel, this project will identify how particle-cell-matrix interactions work in concert to control the biophysics of PCNs at the cell surface during vaporization. To engage and train a new generation of diverse scientists in emerging biotechnologies, such as those explored here, this project will develop and implement an outreach program to integrate underrepresented K-12 students in STEM research. Activities will include incorporating underserved high-school students into undergraduate engineering design teams, and the creation of an Ultrasound Olympics workshop to engage grade 6-12 teachers and integrate bioengineering and acoustics concepts into the classroom. By linking the proposed research program with outreach activities, this project will contribute to the development of a competitive STEM workforce capable of tackling diverse challenges in biomaterials design, molecular biophysics and bioimaging.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.