Characterization of neural crest stem cells in human hair follicles

描述（由申请人提供）：我们的跨学科研究团队非常适合研究成体干细胞中的microRNA（miRNA）。PI'实验室研究皮肤成体干细胞及其在皮肤稳态和黑素细胞发育中的作用;而共同PI正在领导一个计算生物学实验室，专注于miRNA的功能分析。PI和co-PI尚未合作或共同发表。拟议的研究将利用两种PI的优势，协同作用将促进成体干细胞和黑素细胞发育中的miRNA研究。 尽管有新的证据表明miRNA在干细胞自我更新和分化中起重要作用，但它们在干细胞中的作用刚刚开始被理解，并且它们在神经嵴干细胞（NCSC）和黑素细胞发育中的功能还有待探索。在本修订提案中，我们计划通过深度测序方法探索NCSC向黑素细胞分化过程中miRNA的表达，然后研究调节选定的miRNA表达是否可以提高NCSC向黑素细胞分化的效率。 miRNA研究没有在母基金中提出。新提案将通过解决NCSC分化过程中miRNA的功能为母基金增加一个新的维度，这可能使我们能够发现成体干细胞自我更新和分化的潜在新机制。此外，我们不会将我们的研究局限于使用已建立的微阵列平台来研究已知的miRNA和mRNA，而是使用深度测序方法来探索成体干细胞和黑素细胞中miRNA的表达和3'非翻译区（UTR）中的miRNA结合位点。超高通量测序方法将使我们能够量化特定已知miRNA的绝对拷贝数并发现新的miRNA。近年来研究表明，3 'UTR的长度可能影响miRNA的生物学功能，我们也将探讨miRNA和UTR在干细胞分化过程中的相互作用。结合miRNA和总RNA的深度测序将深入了解miRNA如何在干细胞自我更新和分化中发挥作用。我们希望我们的研究将产生关于miRNA在成体干细胞自我更新和分化以及黑素细胞发育中的功能的重要信息。 公共卫生相关性：这些拟议的研究将拓宽我们对microRNA在人类神经嵴干细胞（hNCSC）和色素细胞发育中的功能的基础知识，并提高我们选择性地从hNCSC中产生分化细胞类型用于再生医学的能力。

DESCRIPTION (provided by applicant): Our interdisciplinary research team is uniquely suited to study microRNA (miRNA) in adult stem cells. The PI' laboratory studies skin adult stem cells and their roles in skin homeostasis and melanocyte development; whereas the co-PI is leading a computational biology lab with focus on the functional analysis of miRNAs. The PI and co-PI have yet collaborated or published together. The proposed studies will utilize the strength of both PIs and the synergy will facilitate the miRNA research in adult stem cells and melanocyte development. Despite emerging evidences that miRNAs play an important role in stem cell self- renewal and differentiation, their role in stem cells has just begun to be understood and their function in neural crest stem cells (NCSCs) and melanocyte development has yet to be explored. In this revision proposal, we plan to explore the expression of miRNA during differentiation of NCSCs to melanocytes by deep sequencing method and then study whether modulation of selected miRNA expression may increase the efficiency of NCSC differentiation to melanocytes. miRNA studies were not proposed in the parent grant. The new proposal will add a new dimension to the parent grant by addressing the functions of miRNAs during NCSC differentiation which may allow us to discover potential novel mechanisms underlying adult stem cell self-renewal and differentiation. In addition, we will not limit our research using established microarray platforms to study known miRNA and mRNA, but explore the expression of miRNA and miRNA binding sites in the 3' untranslated region (UTR) in adult stem cells and melanocytes using deep sequencing methods. The ultra high-throughput sequencing method will allow us to quantify the absolute copy number of specific known miRNAs and to discover novel miRNAs. It has been suggested recently that 3'UTR length may influence the biological function of miRNAs, and we will also interrogate the interaction between miRNA and UTR during stem cell differentiation. Combining deep sequencing of both miRNA and total RNA will gain insight on how miRNAs function in stem cell self-renewal and differentiation. We expect our research will generate essential information on the function of miRNA in adult stem cells self-renewal and differentiation as well as melanocyte development. PUBLIC HEALTH RELEVANCE: These proposed studies will broad our fundamental knowledge about the function of microRNA in human neural crest stem cells (hNCSCs) and pigmented cell development, and increase our ability to selectively produce differentiated cell types from hNCSCs for regenerative medicine.