Programmable DNA Nanostructures as Biomedical and Structural Scaffolds

可编程 DNA 纳米结构作为生物医学和结构支架

基本信息

批准号：
10711302
负责人：
Arun Richard Chandrasekaran
金额：
$ 38.56万
依托单位：
STATE UNIVERSITY OF NEW YORK AT ALBANY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-01 至 2028-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10711302
关键词：
3-Dimensional Address Adopted Antibodies Area Award Binding Biodistribution Biological Biological Availability Biosensing Techniques Cell Culture Techniques Cells Chemicals Crystallization Crystallography DNA Data Development Disease Drug Carriers Drug Delivery Systems Drug Screening Face Freezing Health Immune response Ligands Methods Minor Groove Modification Myotonic dystrophy type 1 Nanostructures Nanotechnology National Institute of General Medical Sciences Oligonucleotides Peptides Pharmaceutical Preparations Polycyclic Compounds Positioning Attribute Proteins Radiation induced damage Research Research Personnel Resolution Roentgen Rays Scaffolding Protein Structure Systemic disease Toxic effect Use of New Techniques Validation Work X-Ray Crystallography bioimaging body system design design and construction drug candidate fluorophore improved mouse model nanoparticle practical application pre-clinical scaffold screening self assembly technology platform x-ray free-electron laser

项目摘要

PROJECT ABSTRACT/SUMMARY DNA nanotechnology offers near-atomic control for building structures, with precise positioning of guest molecules such as antibodies, fluorophores and ligands that make them potentially useful in a number of biological applications such as biosensing, drug delivery, cell modulation and bioimaging. However, there are still many challenges that need to be addressed for DNA nanotechnology to reach its full potential for practical applications. In this proposal, we focus on two main areas of development in DNA nanotechnology to address these challenges: (1) Creating a robust, multifunctional drug delivery platform for treating multisystemic diseases, and (2) designing 3D DNA crystals as scaffolds for X-ray structure determination and characterization of such 3D lattices using the new technique of Serial Femtosecond X-ray Crystallography (SFX). For drug delivery, we will develop DNA polyhedra as drug carriers for delivering a new class of modified polycyclic compounds (MPCs) to multiple organ systems and enhancing drug candidate screening using myotonic dystrophy type 1 (DM1) as a testbed disease. Our work will provide quantifiable loading of these minor groove binding drugs and thorough validation of drug delivery efficiency from cell culture to preclinical DM1 mouse models, establishing cell internalization, lack of toxicity and immune response, cell- and disease-specific targeting, bioavailability and biodistribution of the drug-loaded DNA nanostructures. For developing DNA nanostructures as structural scaffolds, we will design and construct DNA motifs that assemble into 3D DNA crystals with different cavity sizes that allow hosting guests of different sizes ranging from nanoparticles to proteins. We will improve resolution of the crystals by programming crystal contacts and incorporating chemical modifications and demonstrate macromolecular scaffolding of proteins using triplex forming oligonucleotides (TFOs) as tethers and peptides using PNA linkers. We will develop methods to grow microcrystals of these DNA motifs for structural analysis using SFX, where diffraction data is collected using high intensity X-ray free-electron lasers, that eliminate the need for large single crystals, freezing, and radiation damage associated with traditional crystallography. This proposed research extends beyond a single disease or health issue, making this work well-suited for the R35 Maximizing Investigators’ Research Award (MIRA) at the NIGMS. In the long-term, I envision that our modular, platform technology using DNA nanostructures can be adopted by other labs for different disease treatments and drug screening (drug delivery) and to obtain crystallographic information of hard-to-crystallize molecules (macromolecular scaffolds).

项目摘要/总结 DNA 纳米技术为建筑结构提供近原子控制，并精确定位宾客抗体、荧光团和配体等分子使其在许多领域具有潜在用途然而，还有生物传感、药物输送、细胞调节和生物成像等生物学应用。 DNA 纳米技术要充分发挥其实际应用潜力，仍需解决许多挑战在本提案中，我们重点关注 DNA 纳米技术的两个主要发展领域，以解决这一问题。这些挑战：（1）创建一个强大的多功能药物输送平台来治疗多系统疾病， (2) 设计 3D DNA 晶体作为支架，用于 X 射线结构测定和表征使用串行飞秒 X 射线晶体学 (SFX) 新技术的 3D 晶格。对于药物输送，我们将开发 DNA 多面体作为药物载体，用于输送新型修饰多环化合物化合物（MPC）对多个器官系统的影响，并使用肌强直增强候选药物筛选 1 型营养不良 (DM1) 作为一种试验疾病，我们的工作将提供这些小沟的可量化负荷。从细胞培养物到临床前 DM1 小鼠的药物输送效率的结合和彻底验证模型，建立细胞内化，缺乏毒性和免疫反应，细胞和疾病特异性载药DNA纳米结构的靶向性、生物利用度和生物分布。为了开发 DNA 纳米结构作为结构支架，我们将设计和构建 DNA 基序组装成具有不同腔体尺寸的 3D DNA 晶体，可容纳不同尺寸的客人，从我们将通过编程晶体接触来提高晶体的分辨率。结合化学修饰并使用三链体展示蛋白质的大分子支架我们将开发使用 PNA 连接体形成寡核苷酸 (TFO) 作为系链和肽的方法。使用 SFX 对这些 DNA 基序的微晶体进行结构分析，其中使用高光谱仪收集衍射数据高强度 X 射线自由电子激光器，无需大型单晶、冷冻和辐射与传统晶体学相关的损伤。这项拟议的研究超越了单一疾病或健康问题，使这项工作非常适合 NIGMS 的 R35 最大化研究者研究奖 (MIRA) 从长远来看，我预计我们的其他实验室可以采用使用 DNA 纳米结构的模块化平台技术来治疗不同的疾病治疗和药物筛选（药物输送）并获得难结晶的晶体学信息分子（高分子支架）。