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The function of LIN28B and follistatin in supporting cell reprogramming and hair cell regeneration in the murine cochlea

LIN28B 和卵泡抑素在支持小鼠耳蜗细胞重编程和毛细胞再生中的功能

基本信息

批准号：
10513325
负责人：
ANGELIKA DOETZLHOFER
金额：
$ 48.48万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-12-01 至 2026-11-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10513325
关键词：
Ablation Activins Address Adolescent Adult Alleles Attenuated Auditory Biological Assay CRISPR/Cas technology Cell Proliferation Cell Reprogramming Cells Cochlea Cre lox recombination system Cues Data Diphtheria Toxin Disease Doxycycline Ear Fluorescent in Situ Hybridization Follistatin Gene Expression Profile Generations Genes Hair Cells Hearing Human In Vitro Knock-out Labyrinth LoxP-flanked allele Mediating Mitotic Mus Natural regeneration Neonatal Organoids Persons Publishing RNA-Binding Proteins Regenerative capacity Regenerative response Role Signal Transduction Supporting Cell Testing Therapeutic Transforming Growth Factor Beta 2 Transgenes Transgenic Mice Transgenic Organisms Trauma Up-Regulation antagonist candidate selection cell dedifferentiation cell determination cell injury cochlear development deafness experimental study gene function gene network hair cell regeneration hearing impairment hearing restoration in vitro Model in vivo in vivo regeneration inducible Cre insight mature animal mouse genetics mouse model neonatal mice notch protein novel overexpression paralogous gene pluripotency progenitor programs regenerative repair strategy response single molecule single-cell RNA sequencing sound tool transcriptome

项目摘要

Project summary Our proposed study aims to address the function of LIN28B and follistatin in supporting cell reprogramming and hair cell regeneration in the murine cochlea. Loss of auditory hair cells (HCs) due to disease or trauma is permanent and is a leading cause for hearing impairments and deafness in humans. Immature auditory supporting cells (SCs) do regenerate HCs in response to damage but their ability to regenerate lost HCs rapidly declines as SCs undergo maturation and little to no HC regeneration is observed in adult animals. We recently uncovered that the RNA binding protein LIN28B and its paralog LIN28A control the regenerative capacity of cochlear SCs in neonatal cochlear organoids and explants. Whether LIN28A/B has a similar role in cochlear HC regeneration in vivo has yet to be tested. Furthermore, our recent in vitro studies suggest that LIN28B promotes HC regeneration through reprogramming SCs into progenitor-like cells and that such state transition can be further enhanced by the co-activation of the Activin antagonist follistatin. However, whether SCs truly activate a transitional progenitor-like state during HC regeneration and if so, how LIN28B and FST may influence such state transition are still unresolved. In our proposed study we will use mouse genetic tools to address whether LIN28A/B regulates spontaneous cochlear HC regeneration in the immature cochlea in vivo (aim1). Furthermore, we will use single cell RNA sequencing and single molecule FISH to determine whether LIN28B reprograms cochlear SCs into progenitor-like cells during HC regeneration (aim2). Moreover, to establish how LIN28B and FST enhance SC reprogramming and subsequent HC regeneration we will manipulate the function of potential LIN28B and FST effector genes using lentiviral overexpression and CRISPR-Cas9-mediated knockout strategies in cochlear organoids (aim3). Finally, we will determine whether co-expression of LIN28B and FST with Atoh1 enables SCs to regenerate cochlear HCs in the mature cochlea in vivo (aim4).

项目总结