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The use of ectopic volar fibroblasts to modify skin identity I

ABSTRACT: The use of ectopic volar fibroblasts to modify skin identity I

PROVIDER: PRJNA970379 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR24474300_1.fastq.gz	Fastqsanger.gz
	SRR24474300_2.fastq.gz	Fastqsanger.gz
	SRR24474400_1.fastq.gz	Fastqsanger.gz
	SRR24474400_2.fastq.gz	Fastqsanger.gz
	SRR24474301_1.fastq.gz	Fastqsanger.gz

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The use of ectopic volar fibroblasts to modify skin identity III

Project description:The modification of skin identity, such as the conversion of residual limb/stump (non-volar) skin of amputees to pressure-resistant palmoplantar skin to enhance prosthesis use and minimize skin breakdown, has tremendous clinical potential. Recognizing the capacity of fibroblasts to modify keratinocyte differentiation, we hypothesized that volar fibroblast injections might induce ectopic volar characteristics. First, we observed pressure-responsive volar fibroblast resulting in increased limb development after pressure treatment, then we confirmed this in using a bioprinted skin construct, and later in a clinical trial that showed increased volar measures in non-volar skin after volar fibroblast injections. Bulk and single cell RNA seq demonstrate gene ontology categories of cornified envelope, keratinization and morphogenic pathways. Thus, the long-term engraftment of volar fibroblasts creates a platform for the therapeutic development

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The use of ectopic volar fibroblasts to modify skin identity

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The use of ectopic volar fibroblasts to modify skin identity

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

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In vitro gene signature of cultured volar or nonvolar keratinocytes

Project description:How cell and tissue identity persist despite constant cell turnover is an important biologic question with cell therapy implications. While many mechanisms exist, we investigated the controls for site-specific gene expression in skin given its diverse structures and functions. For example, the transcriptome of in vivo palmoplantar (i.e. volar) epidermis is globally unique including Keratin 9 (KRT9). While volar fibroblasts have the capacity to induce KRT9 in non-volar keratinocytes, we demonstrate here that volar keratinocytes continue to express KRT9 in vitro solo-cultures. Despite this, KRT9 expression is lost with volar keratinocyte passaging, in spite of stable hypo-methylation of its promoter. Coincident with KRT9 loss is a gain of the primitive Keratin 7 and a signature of dsRNA sensing, including the dsRNA receptor DDX58. Exogenous dsRNA inhibits KRT9 expression in early passage volar keratinocytes or in vivo footpads of wild-type mice. Loss of DDX58 in passaged volar keratinocytes rescues KRT9 and inhibits KRT7 expression. Additionally, DDX58 null mice are resistant to the ability of dsRNA to inhibit KRT9 expression. These results demonstrate that the sensing of dsRNA is critical for loss of cell specific gene expression; our results have important implications of how dsRNA sensing is important outside of immune pathways. Keratinocytes were expanded from both the sole and the dorsum of the foot and at passage 4, RNA was extracted and sent for microarray analysis

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