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Single-cell RNA sequencing reveals cellular and molecular heterogeneity in fibrocartilaginous enthesis formation

ABSTRACT: Single-cell RNA sequencing reveals cellular and molecular heterogeneity in fibrocartilaginous enthesis formation

PROVIDER: PRJNA928314 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR23239881.fastq.gz	Fastqsanger.gz
	SRR23239882.fastq.gz	Fastqsanger.gz
	SRR23239883.fastq.gz	Fastqsanger.gz
	SRR23239884.fastq.gz	Fastqsanger.gz
	SRR23239885.fastq.gz	Fastqsanger.gz

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Single-cell RNA sequencing reveals cellular and molecular heterogeneity in fibrocartilaginous enthesis formation

Project description:The attachment site of the rotator cuff (RC) is a classic fibrocartilaginous enthesis, which is the junction between bone and tendon with typical characteristics of a fibrocartilage transition zone. Enthesis development has historically been studied with lineage tracing of individual genes selected a priori, which does not allow for the determination of single-cell landscapes yielding mature cell types and tissues. Here, in together with open source GSE182997 datasets (3 sample) provided by Fang et al, we applied Single-cell RNA sequencing (scRNA-seq) to delineate the comprehensive postnatal RC enthesis growth and the temporal atlas from as early as postnatal day 1 up to postnatal week 8. And we furtherly performed single cell spatial transcriptomic sequencing on postnatal day 1 mice enthesis, in order to deconvoluted bone-tendon junction (BTJ) chondrocytes onto spatial spots. In summary, we deciphered the cellular heterogeneity and the molecular dynamics during fibrocartilage differentiation. Combined with current spatial transcriptomic data, our results provide a transcriptional resource that will support future investigations of enthesis development at the mechanistic level and may shed light on the strategies for enhanced RC healing outcomes.

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Single-cell RNA-seq of the tendon enthesis cells

Project description:Our studies used scRNA-seq analysis to get a full map of transcriptional profiles of enthesis cells and also a seprate Gli1-lineage enthesis stem cells. By harvesting enthesis cells from different development stages for scRNA-seq analysis, we revealed enthesis cell heterogeneity and identified six cell sub-populations using scRNA-seq. We infered cell differentiation trajectories for enthesis stem cells differentiating into mineralizing chondrocytes. A gene regulatory network analysis combined fluorescent in situ hybridization were then used to identify a number of transcription factors coordinating tenogenesis, chondrogenesis, and osteogenesis to form an enthesis with spatially graded mineralization.To further define the enthesis stem cell population, enthesis Gli1-lineage cells were isolated and their transcriptomes were characterized at single cell resolution. These specific Gli1-responsive cells had a linear trajectory and a capacity of chondrogenesis and osteogenesis. The full characterization of transcriptional landscape of tendon enthesis stem cells demonstrates a promising therapeutic strategies using this cell source for enthesis regeneration.

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Single-cell RNA sequencing reveals cellular and molecular heterogeneity in fibrocartilaginous enthesis formation.

Project description:The attachment site of the rotator cuff (RC) is a classic fibrocartilaginous enthesis, which is the junction between bone and tendon with typical characteristics of a fibrocartilage transition zone. Enthesis development has historically been studied with lineage tracing of individual genes selected a priori, which does not allow for the determination of single-cell landscapes yielding mature cell types and tissues. Here, in together with open-source GSE182997 datasets (three samples) provided by Fang et al., we applied Single-cell RNA sequencing (scRNA-seq) to delineate the comprehensive postnatal RC enthesis growth and the temporal atlas from as early as postnatal day 1 up to postnatal week 8. And, we furtherly performed single-cell spatial transcriptomic sequencing on postnatal day 1 mouse enthesis, in order to deconvolute bone-tendon junction (BTJ) chondrocytes onto spatial spots. In summary, we deciphered the cellular heterogeneity and the molecular dynamics during fibrocartilage differentiation. Combined with current spatial transcriptomic data, our results provide a transcriptional resource that will support future investigations of enthesis development at the mechanistic level and may shed light on the strategies for enhanced RC healing outcomes.

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