Uncovering mechanisms and progenitors responsible for limb regeneration

揭示负责肢体再生的机制和祖细胞

基本信息

批准号：
9750512
负责人：
Nicholas Leigh
金额：
$ 6.37万
依托单位：
HARVARD UNIVERSITY
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：
2018-07-16 至 2021-07-15
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9750512
关键词：
Affect Ambystoma American Amputation Biological Assay Blood coagulation Cartilage Cell Differentiation process Cell Proliferation Cell Survival Cell physiology Cells Complement Factor D Data Data Set Dermis Development Diabetes Mellitus Disease EGF gene EGF-Like Domain Event Gene Expression Profile Genes Genetic Transcription Goals Heterogeneity Human Immunofluorescence Immunologic In Situ Hybridization Infection Injections Kinetics LIF gene Lead Length Limb Bud Limb structure Location Mammalian Cell Mediating Messenger RNA Molecular Molecular Biology Morphology Mus Muscle Natural regeneration Organism Pain Patients Phenotype Plasmids Prosthesis Psychological Impact RNA RNA Sequences Regenerative response Role Salamander Schwann Cells Stem cells Structure Testing Tissues Transcript Trauma Work appendage base blastema design experimental study gene discovery genetic signature in vivo insight knock-down limb amputation limb regeneration loss of function molecular marker mutant novel progenitor recruit regenerative response single-cell RNA sequencing standard of care stem tool transcriptome von Willebrand Factor

项目摘要

PROJECT SUMMARY Limb loss affects nearly 2 million Americans and comes with complications such as infection, pain, and blood clots, in addition to serious psychological impact. Currently, the standard of care following limb loss is the use of a prosthetic. The ideal response following limb loss would be the regeneration of an exact replicate of the lost limb. This is not the response humans make, but other species such salamanders have the ability to perfectly regenerate lost appendages. Therefore, understanding the remarkable ability of salamanders to regenerate lost limbs may elucidate mechanisms to unlock the regenerative capacity of humans. Following limb amputation salamanders form a structure called the blastema. The blastema is a pool of tissue specific progenitor cells that are responsible for regenerating the lost limb. Recent gains in molecular biology has allowed for discovery of genes that are enriched and specific to the regenerating limb. We have recently identified von Willebrand Factor D and EGF domains (VWDE) as a highly blastema enriched and specific gene. We hypothesize that the blastema contains a pool of transcriptionally distinct progenitor cells and pan- blastemal genes, such as VWDE, support this niche. Our first aim will evaluate the role and functional domains of VWDE in blastema cell function and mammalian cell differentiation. Our second aim will work to uncover other progenitor cells (i.e. dermis, cartilage, Schwann cells) using single cell RNA-sequencing to elucidate the cellular heterogeneity of the blastema. These studies will provide essential information on a novel gene and work to uncover different progenitor cells required for limb regeneration in salamanders. These gains in understanding of salamander limb regeneration will put us closer to unlocking the ability to promote regeneration in humans.

项目概要近 200 万美国人面临肢体损失，并伴有感染、疼痛和出血等并发症血栓，除了严重的心理影响。目前，肢体丧失后的护理标准是使用假肢的。肢体丧失后的理想反应是再生出肢体的精确复制品失去肢体。这不是人类的反应，但火蜥蜴等其他物种有能力完美再生丢失的附肢。因此，了解蝾螈的非凡能力再生失去的肢体可能会阐明释放人类再生能力的机制。下列的截肢蝾螈形成一种称为胚基的结构。胚基是一组特定的组织负责再生失去的肢体的祖细胞。分子生物学的最新进展允许发现丰富且特定于再生肢体的基因。我们最近有鉴定出冯·维勒布兰德因子 D 和 EGF 结构域 (VWDE) 作为高度胚基富集和特异性的基因。我们假设胚基包含一组转录上不同的祖细胞和泛细胞胚芽基因，例如 VWDE，支持这一生态位。我们的首要目标是评估角色和职能领域 VWDE 在胚基细胞功能和哺乳动物细胞分化中的作用。我们的第二个目标将努力揭示其他祖细胞（即真皮、软骨、雪旺细胞）使用单细胞 RNA 测序来阐明胚基的细胞异质性。这些研究将提供有关新基因的重要信息致力于发现蝾螈肢体再生所需的不同祖细胞。这些收获在对蝾螈肢体再生的了解将使我们更接近释放促进能力人类的再生。