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A microRNA signature associated with chondrogenic lineage commitment

机译：与软骨生成谱系承诺相关的microRNA标记

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Generating appropriate cartilage for clinical applications to heal skeletal tissue loss is a major health concern. In this regard, cell-based approaches offer a potential therapeutic strategy for cartilage repair, although little is known about the precise mechanism of chondrogenesis. Unrestricted somatic stem cell (USSC) is considered as a suitable candidate because of its potential for differentiating into multiple cell types. Recent studies show that microRNAs (miRNAs) are involved in several biological processes including development and differentiation. To identify the chondro-specific miRNA signature, miRNA patterns of USSCs and differentiated chondrocytes were investigated using microarrays and validation by qPCR. Prior to these analyses, chondrogenic commitment of differentiated USSCs was verified by immunocytochemistry, specific staining and evaluation of some main chondrogenic marker genes. Various in silico explorations (for both putative targets and signalling pathways) and empirical analyses (miRNA transfections followed by qPCR of some chondrogenic indicators) were carried out to support our results. Transient modulation of multiple chondro-miRs (such as mir-630, mir-624 and mir-376) with chondrocyte targets (such as TGFbR, MAP3K, collagens, SMADs and cadherins) as mediators of chondrogenic signalling pathways including cell-cell interactions, TGF-beta, and MAPK signalling suggests a mechanism for genetic induction of chondrogenic differentiation. In conclusion, this research reveals more details about the allocation of USSCs into the chondrocytes through identification of miRNA signature which modulates targets and pathways required for chondrogenic lineage and could provide guidelines for future clinical treatments and anti-miRNA therapies.
机译：产生适当的软骨以用于临床应用以治愈骨骼组织丢失是主要的健康问题。在这方面，基于细胞的方法为软骨修复提供了一种潜在的治疗策略，尽管对软骨形成的确切机制知之甚少。无限制的体干细胞（USSC）被认为是合适的候选者，因为它具有分化为多种细胞类型的潜力。最近的研究表明，microRNA（miRNA）参与了几个生物学过程，包括发育和分化。为了鉴定软骨特异性的miRNA标记，使用微阵列研究了USSC和分化的软骨细胞的miRNA模式，并通过qPCR进行了验证。在进行这些分析之前，已通过免疫细胞化学，特异性染色和一些主要软骨生成标记基因的评估来验证分化的USSC的软骨生成承诺。进行了各种计算机模拟探索（针对既定的靶标和信号传导途径）和经验分析（miRNA转染，然后对某些软骨形成指标进行qPCR）以支持我们的结果。用软骨细胞靶标（例如TGFbR，MAP3K，胶原蛋白，SMAD和钙黏着​​蛋白）作为软骨形成信号通路（包括细胞间相互作用）的介质，瞬时调节多个软骨miR（例如mir-630，mir-624和mir-376）， TGF-beta和MAPK信号提示了软骨诱导分化的遗传诱导机制。总之，这项研究通过鉴定miRNA标记揭示了USSCs在软骨细胞中的分配的更多细节，miRNA标记可调节软骨生成谱系所需的靶标和途径，并可为将来的临床治疗和抗miRNA治疗提供指导。

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