A Bio-Inspired Artificial Transcription Factor for Regeneration of Functional Hair Cells

用于功能性毛细胞再生的仿生人工转录因子

基本信息

批准号：
9920703
负责人：
KELVIN YUI-HANG KWAN
金额：
$ 32.85万
依托单位：
RUTGERS, THE STATE UNIV OF N.J.
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-05-01 至 2023-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9920703
关键词：
Address Adult Affect Aging American Aminoglycosides Auditory Binding Biological Biomimetics Cell Differentiation process Cell physiology Cells Chromatin Chromatin Structure Clinical Cochlea Coupling DNA Binding DNA Binding Domain DNA Sequence Data Development Disease Embryo Endogenous Factors Ensure Epigenetic Process Equilibrium Exposure to Fibrinogen GFI1 gene Gene Delivery Gene Expression Generations Genes Genetic Transcription Goals Hair Cells Hearing Histone Acetylation Histone Deacetylation Human In Vitro Individual Inner Hair Cells Labyrinth Learning Libraries Loudness Magnetism Malignant Neoplasms Mediating Methods Mission Modeling Molecular Morphology Mus Natural regeneration Neurosciences Nuclear Organ Peptides Pluripotent Stem Cells Promoter Regions Proteins Public Health Publications Recovery Regenerative Medicine Replacement Therapy Research Personnel Sensorineural Hearing Loss Sensory Sensory Hair Social Functioning Social isolation Specificity Structure Supporting Cell Technology Tertiary Protein Structure Testing Therapeutic Effect Thromboplastin Transactivation Translating United States National Institutes of Health Viral Viral Vector Work base biomaterial compatibility cell type clinical translation deafness design disability early childhood genetic manipulation genome-wide hair cell regeneration hearing impairment hearing restoration innovation innovative technologies nanoGold nanomedicine nanoparticle neurosensory neurotransmission ototoxicity overexpression prevent regenerative therapy side effect small molecule social skills sound spatiotemporal stem cell biology stem cell differentiation stem cell fate stem cells tool transcription factor transcription factor S-II transcriptome transdifferentiation

项目摘要

PROJECT SUMMARY Hearing loss during early childhood significantly affects learning and acquisition of social skills, while hearing loss in adults can often result in social isolation and inability to perform many routine social functions. A leading cause of sensorineural hearing loss is the loss of sensory hair cells of the inner ear. A lifetime exposure to aminoglycoside and loud sounds will result in an estimated 15% of adult Americans (~36 million) having some form of hearing loss. A promising approach to mitigate hearing loss and deafness is a cell replacement therapy by transdifferentiating supporting cells into hair cells. Unfortunately, current approaches for transdifferentiation rely on viral delivery may be unsafe and impractical for clinical translation. Therefore, there is a critical need to develop alternative platforms to regulate gene expression and induce transdifferentiation in an efficient, non-viral manner suitable for hearing restoration. To this end, our long-term goal is to develop NanoScript, an innovative, tunable nanoparticle-based artificial transcription factor platform capable of effectively regulating gene expression in a non-viral manner. Using NanoScript, we will transdifferentiate supporting cells into functional hair cells. NanoScript consists of a nanoparticle functionalized with specific small molecules and peptides that are designed to mimic the individual domains of natural transcription factor (TF) proteins. TFs are endogenous, multi-domain proteins that orchestrate many cellular functions including differentiation. Since NanoScript is a functional replica of TF proteins, it can replace virally-delivered TFs for regenerative medicine-based applications. The overall objective of this proposal is to design three NanoScripts that mimic three TFs essential for hair cell differentiation (Gfi1, Pou4f3, and Atoh1; GPA). We will test whether GPA-NanoScript binds to the same DNA sequence and activate gene expression in vitro. Next we will determine if addition of epigenetic modulators to GPA-NanoScript will bind to the same targets as the TF proteins, locally alter the chromatin structure and enhance gene expression. Finally, we will use cochlear explants to determine whether GPA-NanoScript promotes transdifferentiation of supporting cells into hair cells. Generation of nascent hair cells using an ex vivo model will serve as a springboard to test NanoScript technology for regenerative medicine. It will also establish NanoScript as an effective and non-viral tool for researchers to generate functional cells via direct reprogramming.

项目概要儿童早期的听力损失显着影响社交技能的学习和获得，而听力成年人的丧失往往会导致社会孤立和无法履行许多日常社会职能。领先的感音神经性听力损失的原因是内耳感觉毛细胞的损失。一生接触据估计，氨基糖苷类药物和响亮的声音将导致 15% 的美国成年人（约 3600 万）患有某些疾病。听力损失的形式。细胞替代疗法是减轻听力损失和耳聋的一种有前景的方法通过将支持细胞转分化为毛细胞。不幸的是，目前的转分化方法依赖病毒传递对于临床转化来说可能不安全且不切实际。因此，迫切需要开发替代平台来调节基因表达并以有效的非病毒方式诱导转分化适合听力恢复的方式。为此，我们的长期目标是开发 NanoScript，一种创新的、可调节的基于纳米粒子的人工能够以非病毒方式有效调控基因表达的转录因子平台。使用 NanoScript，我们将支持细胞转分化为功能性毛细胞。 NanoScript 由具有特定小分子和肽功能的纳米颗粒，旨在模仿个体天然转录因子 (TF) 蛋白的结构域。 TF 是内源性多结构域蛋白，可协调许多细胞功能，包括分化。由于 NanoScript 是 TF 蛋白的功能复制品，因此它可以取代病毒传递的转录因子，用于基于再生医学的应用。本提案的总体目标的目标是设计三个 NanoScript，模拟毛细胞分化所必需的三个 TF（Gfi1、Pou4f3 和 Atoh1；平均绩点）。我们将测试 GPA-NanoScript 是否与相同的 DNA 序列结合并激活基因表达体外。接下来我们将确定在 GPA-NanoScript 中添加表观遗传调节剂是否会与相同的目标结合作为 TF 蛋白，局部改变染色质结构并增强基因表达。最后，我们将使用耳蜗外植体以确定 GPA-NanoScript 是否促进支持细胞转分化为毛细胞。使用离体模型生成新生毛细胞将作为测试 NanoScript 的跳板再生医学技术。它还将使 NanoScript 成为一种有效的非病毒工具研究人员通过直接重编程产生功能细胞。