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100，000种人类疾病是由基因组中的遗传改变引起的，这些疾病中只有很小一部分可以治愈。基因编辑代表了疾病治疗学的关键发展，是纠正基因组内缺陷和突变的有力工具。特别是，重复的规则间隔短回文重复序列（CRISPR）Cas9代表了进行精确的靶向基因组改变的能力的范式转变。最近，已经开发了一些用于CRISPR/Cas9复合物的细胞内递送的方法。虽然这些方法取得了一定程度的成功，但实现高效和高效的细胞内CRISPR/Cas9递送仍然极具挑战性。EARLY概念的探索性研究赠款（EAGER）资助支持设计，制造和测试纳米机器人或纳米级机器人的研究，这些机器人可以精确地将CRISPR/Cas9靶向并递送到患病细胞，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级印刷方法涉及多材料印刷，并且可以是用于各种应用的功能性纳米级机器的可扩展纳米制造的强大工具。纳米机器人可以彻底改变基因或药物输送，以修复许多人类疾病的遗传紊乱，这将对人类健康产生重大影响。该项目提供了令人兴奋的跨学科培训，为不同的研究生和本科生群体整合了从制造到生物材料到纳米机器到治疗学的内容。纳米打印和纳米机器人是吸引高中生和教师、女性和少数民族研究人员进入科学和工程领域的实验室演示的绝佳工具。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，该系统用于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相容性材料设计纳米机器人，并使用纳米级3D打印系统制造它。纳米机器人由磁性纳米颗粒和可生物降解的纳米货物组成。纳米颗粒通常为200 nm圆形和400 nm长，通过将氧化铁磁性纳米颗粒嵌入水凝胶中进行3D打印。具有类似尺寸的纳米货物也是通过将CRISPR/Cas9封装在可生物降解的水凝胶中进行3D打印的，因此CRISPR/Cas9一旦进入细胞内就可以通过生物降解释放。 基础研究的重点是研究材料的组成和性质，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递送到癌细胞中以抑制肿瘤的功效。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

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