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Transcriptional profiling of mESC-derived tendon and fibrocartilage cell fate switch [scRNA-seq]

ABSTRACT: Transcriptional profiling of mESC-derived tendon and fibrocartilage cell fate switch [scRNA-seq]

PROVIDER: PRJNA646530 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR12232800.fastq.gz	Fastqsanger.gz
	SRR12232801.fastq.gz	Fastqsanger.gz
	SRR12232802.fastq.gz	Fastqsanger.gz
	SRR12232803.fastq.gz	Fastqsanger.gz
	SRR12232804.fastq.gz	Fastqsanger.gz

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Transcriptional profiling of mESC-derived tendon and fibrocartilage cell fate switch [scRNA-seq]

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Transcriptional profiling of mESC-derived tendon and fibrocartilage cell fate switch.

Project description:The transcriptional regulators underlying induction and differentiation of dense connective tissues such as tendon and related fibrocartilaginous tissues (meniscus and annulus fibrosus) remain largely unknown. Using an iterative approach informed by developmental cues and single cell RNA sequencing (scRNA-seq), we establish directed differentiation models to generate tendon and fibrocartilage cells from mouse embryonic stem cells (mESCs) by activation of TGFβ and hedgehog pathways, achieving 90% induction efficiency. Transcriptional signatures of the mESC-derived cells recapitulate embryonic tendon and fibrocartilage signatures from the mouse tail. scRNA-seq further identify retinoic acid signaling as a critical regulator of cell fate switch between TGFβ-induced tendon and fibrocartilage lineages. Trajectory analysis by RNA sequencing define transcriptional modules underlying tendon and fibrocartilage fate induction and identify molecules associated with lineage-specific differentiation. Finally, we successfully generate 3-dimensional engineered tissues using these differentiation protocols and show activation of mechanotransduction markers with dynamic tensile loading. These findings provide a serum-free approach to generate tendon and fibrocartilage cells and tissues at high efficiency for modeling development and disease.

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Transcriptomic trajectory of the tendon and fibrocartilage fate switch

Project description:This SuperSeries is composed of the SubSeries listed below.

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A Conserved Transcription Factor Regulatory Program Promotes Tendon Fate

Project description:Tendons, which transmit force from muscles to bones, are highly prone to injury. Understanding the mechanisms driving tendon fate would impact efforts to improve tendon healing, yet this knowledge is limited. To find direct regulators of tendon progenitor emergence, we performed a zebrafish high-throughput chemical screen. We established Forskolin as a tenogenic inducer across vertebrates, functioning through Creb1a, which is required and sufficient for tendon fate. Putative enhancers containing cAMP response elements (CRE) in human, mouse and fish, drove specific expression in zebrafish cranial and fin tendons. Analysis of these genomic regions identified motifs for Ebf/EBF transcription factors. Mutation of CRE or Ebf/EBF motifs significantly disrupted enhancer activity and specificity in tendons. Zebrafish ebf1a/ebf3a mutants displayed defects in tendon formation. Notably, Creb1a/CREB1 and Ebf1a/Ebf3a/EBF1 overexpression facilitated tenogenic induction in zebrafish and human pluripotent stem cells. Together, our work reveals functional conservation of two novel transcription factors in promoting tendon fate.

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