EAGER: Three-Dimensional Printing of Functional Nanobots for Precision Gene Delivery

EAGER：用于精确基因传递的功能纳米机器人的三维打印

• 批准号: 1937653
• 负责人: Shaochen Chen
• 金额: $ 29.98万
• 依托单位: University of California-San Diego
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1937653&HistoricalAwards=false
• 关键词: EAGER; Three; Dimensional; Printing; Functional

Over 100,000 human diseases are caused by genetic alterations in the genome, and only a very small portion of these diseases can be cured. Gene editing represents a pivotal development in disease therapeutics as a powerful tool to correct defects and mutations within the genome. In particular, Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) Cas9 represents a paradigm shift in the ability to make precise, targeted genomic change. Recently, a few approaches have been developed for intracellular delivery of CRISPR/Cas9 complexes. While these approaches have some degree of success, it remains extremely challenging to achieve highly effective and efficient intracellular CRISPR/Cas9 delivery. This EArly-concept Grants for Exploratory Research (EAGER) grant supports research to design, manufacture, and test nanobots or nanoscale robots that can precisely target and deliver CRISPR/Cas9 to diseased cells and release the gene-editing agencies in a controlled fashion. The three-dimensional nanoscale printing method for fabricating the nanobots involves multi-materials printing and could be a powerful tool for scalable nanomanufacturing of functional nanoscale machines for a variety of applications. The nanobots could revolutionize gene or drug delivery to repair genetic disorder of many human diseases, which would have a strong impact on human health. The project offers exciting interdisciplinary training that integrates content from manufacturing to biomaterials to nanomachines to therapeutics for a diverse group of graduate and undergraduate students. Nanoscale printing and nanobots are excellent tools for laboratory demonstrations to attract high school students and teachers, and women and underrepresented minority researchers to science and engineering fields.This project aims to investigate the nanomanufacturing processing of a novel nanobot system for targeted gene or drug delivery at the single cell level. The collaborative research team designs the nanobot using biocompatible materials and uses a nanoscale 3D printing system to fabricate it. The nanobot consists of a magnetic nanomotor and a biodegradable nano-cargo. The nanomotor, which is typically 200 nm round and 400 nm long, is 3D printed by embedding iron oxide magnetic nanoparticles in hydrogel. The nano-cargo, which is of similar dimensions, is also 3D printed by encapsulating CRISPR/Cas9 in a biodegradable hydrogel, so that CRISPR/Cas9 can be released through biodegradation once inside the cell. Fundamental research focuses on investigating the effects of material composition and properties, and nanomanufacturing processing parameters on the nanobot performance. The team also tests the efficacy of the nanobot to deliver CRISPR/Cas9 into cancer cells for tumor suppression. The scalability of the nanomanufacturing process is demonstrated through the reproducible fabrication of an array of nanobots.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.

超过10万种人类疾病是由基因组中的基因改变引起的，这些疾病中只有很小一部分可以治愈。基因编辑代表着疾病治疗学的一个关键发展，它是纠正基因组中缺陷和突变的强大工具。特别是，簇状规则间隔短回文重复序列(CRISPR)Cas9代表了做出精确的、有针对性的基因组变化的能力的范式转变。近年来，CRISPR/Cas9复合体的细胞内给药方法已有较多进展。虽然这些方法在一定程度上取得了成功，但要实现高效和高效的CRISPR/Cas9细胞内递送仍然具有极大的挑战性。这项早期概念探索性研究拨款(AGERGE)支持设计、制造和测试纳米机器人或纳米机器人的研究，这些机器人可以精确地定位并将CRISPR/Cas9运送到患病细胞，并以受控的方式释放基因编辑机构。用于制造纳米机器人的三维纳米打印方法涉及多材料打印，并可能成为用于各种应用的可伸缩纳米制造功能纳米机器的强大工具。纳米机器人可能会彻底改变基因或药物的输送，以修复许多人类疾病的遗传障碍，这将对人类健康产生重大影响。该项目提供令人兴奋的跨学科培训，为不同的研究生和本科生群体提供从制造到生物材料、纳米机械到治疗学的内容。纳米打印和纳米机器人是吸引高中生和教师、女性和少数族裔研究人员进入科学和工程领域的优秀实验室演示工具。本项目旨在研究一种新型纳米机器人系统的纳米制造过程，以在单细胞水平上靶向基因或药物输送。合作研究团队使用生物兼容材料设计了纳米机器人，并使用纳米级3D打印系统来制造它。该纳米机器人由一个磁性纳米马达和一个可生物降解的纳米货物组成。这种纳米电机通常为200 nm圆，400 nm长，通过在水凝胶中嵌入氧化铁磁性纳米颗粒进行3D打印。尺寸相似的纳米货物也可以通过将CRISPR/Cas9包裹在可生物降解的水凝胶中进行3D打印，这样CRISPR/Cas9一旦进入细胞内就可以通过生物降解释放出来。基础研究侧重于研究材料组成和性能以及纳米制造工艺参数对纳米机器人性能的影响。该团队还测试了纳米机器人将CRISPR/Cas9导入癌细胞抑制肿瘤的效果。纳米制造过程的可扩展性通过一系列纳米机器人的可重复制造得到证明。该奖项反映了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。

项目成果

Controlled Growth Factor Release in 3D‐Printed Hydrogels

• DOI: 10.1002/adhm.201900977
• 发表时间: 2019-11
• 期刊: Advanced Healthcare Materials
• 影响因子: 10
• 作者: Pengrui Wang;D. Berry;A. Moran;F. He;Trevor Tam;Luwen Chen;Shaochen Chen

Bioprinting of dual ECM scaffolds encapsulating limbal stem/progenitor cells in active and quiescent statuses.

• DOI: 10.1088/1758-5090/ac1992
• 发表时间: 2021-08-13
• 期刊: Biofabrication
• 影响因子: 9
• 作者: Zhong Z;Balayan A;Tian J;Xiang Y;Hwang HH;Wu X;Deng X;Schimelman J;Sun Y;Ma C;Dos Santos A;You S;Tang M;Yao E;Shi X;Steinmetz NF;Deng SX;Chen S
• 通讯作者: Chen S

3D Printing of a Biocompatible Double Network Elastomer with Digital Control of Mechanical Properties

• DOI: 10.1002/adfm.201910391
• 发表时间: 2020-02
• 期刊: Advanced Functional Materials
• 影响因子: 19
• 作者: Pengrui Wang;D. Berry;Zhaoqiang Song;Wisarut Kiratitanaporn;Jacob Schimelman;A. Moran;F. He;B. Xi;S. Cai;Shaochen Chen

High throughput direct 3D bioprinting in multiwell plates

• DOI: 10.1088/1758-5090/ab89ca
• 发表时间: 2021-04-01
• 期刊: BIOFABRICATION
• 影响因子: 9
• 作者: Hwang, Henry H.;You, Shangting;Chen, Shaochen
• 通讯作者: Chen, Shaochen

Rapid 3D bioprinting of a multicellular model recapitulating pterygium microenvironment.

• DOI: 10.1016/j.biomaterials.2022.121391
• 发表时间: 2022-03
• 期刊: Biomaterials
• 影响因子: 14