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Delineating the heterogeneity of matrix-directed differentiation towards soft and stiff tissue lineages via single-cell profiling.

ABSTRACT: Delineating the heterogeneity of matrix-directed differentiation towards soft and stiff tissue lineages via single-cell profiling.

PROVIDER: PRJNA702075 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR13719762.fastq.gz	Fastqsanger.gz
	SRR13719763.fastq.gz	Fastqsanger.gz
	SRR13719764.fastq.gz	Fastqsanger.gz
	SRR13719765.fastq.gz	Fastqsanger.gz
	SRR13719766.fastq.gz	Fastqsanger.gz

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Delineating the heterogeneity of matrix-directed differentiation towards soft and stiff tissue lineages via single-cell profiling.

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Delineating the heterogeneity of matrix-directed differentiation toward soft and stiff tissue lineages via single-cell profiling.

Project description:Mesenchymal stromal/stem cells (MSCs) form a heterogeneous population of multipotent progenitors that contribute to tissue regeneration and homeostasis. MSCs assess extracellular elasticity by probing resistance to applied forces via adhesion, cytoskeletal, and nuclear mechanotransducers that direct differentiation toward soft or stiff tissue lineages. Even under controlled culture conditions, MSC differentiation exhibits substantial cell-to-cell variation that remains poorly characterized. By single-cell transcriptional profiling of nonconditioned, matrix-conditioned, and early differentiating cells, we identified distinct MSC subpopulations with distinct mechanosensitivities, differentiation capacities, and cell cycling. We show that soft matrices support adipogenesis of multipotent cells and early endochondral ossification of nonadipogenic cells, whereas intramembranous ossification and preosteoblast proliferation are directed by stiff matrices. Using diffusion pseudotime mapping, we outline hierarchical matrix-directed differentiation and perform whole-genome screening of mechanoresponsive genes. Specifically, top-ranked tropomyosin-1 is highly sensitive to stiffness cues both at RNA and protein levels, and changes in TPM1 expression determine the differentiation toward soft versus stiff tissue lineage. Consistent with actin stress fiber stabilization, tropomyosin-1 overexpression maintains YAP1 nuclear localization, activates YAP1 target genes, and directs osteogenic differentiation. Knockdown of tropomyosin-1 reversed YAP1 nuclear localization consistent with relaxation of cellular contractility, suppressed osteogenesis, activated early endochondral ossification genes after 3 d of culture in induction medium, and facilitated adipogenic differentiation after 1 wk. Our results delineate cell-to-cell variation of matrix-directed MSC differentiation and highlight tropomyosin-mediated matrix sensing.

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Next-generation sequencing facilitates quantitative analysis of rat BMSCs in soft or stiff SA hydrogel

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