Legumain dependent gene signatures in stromal progenitors

LGMN has been shown to regulate cell fate decission of stromal progenitors of the bone. To understabnd the underlying mechanisms and gene signatures that are responsive to elevated or diminished levels of LGMN we performed RNA-seq analysis on stable cell lines with diminished (LGMN knockdown) or elevated (LGMN overexpression) levels of LGMN at the progenitor level. Genes responsive to LGMN levels show invers dynamics during osteoblast and adipocyte differentiation. Overall design: Stable cell lines were generated from human stromal progenitor cells of bone marrow origin (BM-hMSC-TERT4 cells) with overexpression or knockdown of the asparaginyl endopeptidase (AEP) legumain (LGMN). RNA-seq was performed on 3 replicates of cells with LGMN overexpression (T4_LGMN) and control (T4_pBabe) as well as LGMN knockdown (T4_shLGMN) and control (T4_shCtrl).

已有研究证实，LGMN可调控骨髓基质祖细胞的细胞命运决定。为阐明LGMN表达水平异常升高或降低时的潜在调控机制及响应性基因特征，本研究针对祖细胞水平下LGMN表达下调（LGMN敲低，LGMN knockdown）或上调（LGMN过表达，LGMN overexpression）的稳定细胞系开展了RNA测序（RNA-seq）分析。受LGMN水平调控的基因在成骨细胞与脂肪细胞分化进程中呈现相反的表达动态。

总体实验设计：本研究从骨髓来源的人基质祖细胞（BM-hMSC-TERT4细胞）中构建了天冬酰胺内肽酶（asparaginyl endopeptidase, AEP）legumain（即LGMN）过表达或敲低的稳定细胞系。分别对LGMN过表达组（T4_LGMN）、对应对照组（T4_pBabe），以及LGMN敲低组（T4_shLGMN）、对应对照组（T4_shCtrl）的细胞各设置3次生物学重复，进行RNA-seq测序。