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Single-cell sequencing dissects the transcriptional identity of activated fibroblasts and identifies novel persistent distal tubular injury patterns in kidney

ABSTRACT: Single-cell sequencing dissects the transcriptional identity of activated fibroblasts and identifies novel persistent distal tubular injury patterns in kidney

PROVIDER: PRJNA883649 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR21683913.fastq.gz	Fastqsanger.gz
	SRR21683914.fastq.gz	Fastqsanger.gz
	SRR21683915.fastq.gz	Fastqsanger.gz
	SRR21683916.fastq.gz	Fastqsanger.gz
	SRR21683917_1.fastq.gz	Fastqsanger.gz

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Single-cell sequencing dissects the transcriptional identity of activated fibroblasts and identifies novel persistent distal tubular injury patterns in kidney fibrosis.

Project description:Examining kidney fibrosis is crucial for mechanistic understanding and developing targeted strategies against chronic kidney disease (CKD). Persistent fibroblast activation and tubular epithelial cell (TEC) injury are key CKD contributors. However, cellular and transcriptional landscapes of CKD and specific activated kidney fibroblast clusters remain elusive. Here, we analyzed single cell transcriptomic profiles of two clinically relevant kidney fibrosis models which induced robust kidney parenchymal remodeling. We dissected the molecular and cellular landscapes of kidney stroma and newly identified three distinctive fibroblast clusters with "secretory", "contractile" and "vascular" transcriptional enrichments. Also, both injuries generated failed repair TECs (frTECs) characterized by decline of mature epithelial markers and elevation of stromal and injury markers. Notably, frTECs shared transcriptional identity with distal nephron segments of the embryonic kidney. Moreover, we identified that both models exhibited robust and previously unrecognized distal spatial pattern of TEC injury, outlined by persistent elevation of renal TEC injury markers including Krt8 and Vcam1, while the surviving proximal tubules (PTs) showed restored transcriptional signature. We also found that long-term kidney injuries activated a prominent nephrogenic signature, including Sox4 and Hox gene elevation, which prevailed in the distal tubular segments. Our findings might advance understanding of and targeted intervention in fibrotic kidney disease.

| S-EPMC10764314 | biostudies-literature

Single-cell sequencing dissects the transcriptional identity of activated fibroblasts and identifies novel persistent distal tubular injury patterns in kidney

Project description:Mouse E18.5 wildtype kidney single-cell reference.

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Modeling hereditary distal renal tubular acidosis using patient-derived iPSCs with WDR72 mutations

Project description:Distal renal tubular acidosis (dRTA) is a rare genetic disorder characterized by impaired urine acidification. The WDR72 gene has recently been implicated in dRTA, though its precise functional mechanisms remain unclear. In this study, we generated induced pluripotent stem cell (iPSC) from a patient with WDR72 mutations to model the disease. Upon differentiation into renal tubular epithelial cells, the wild-type and mutant cells (variants c.1777A>G and c.2522T>A) were analyzed for RNA expression, protein localization, and H+-ATPase trafficking. The results indicated no significant changes in these parameters, with the mutant protein retaining its role in acid-base homeostasis. Transcriptomic analysis revealed the top 10 differentially expressed genes (DEGs) were predominantly associated with the collagen-containing extracellular matrix, suggesting a potential link between WDR72 function and extracellular matrix composition. These findings highlight the value of iPSC-derived cellular models in advancing personalized research on dRTA.

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Modeling hereditary distal renal tubular acidosis using patient-derived iPSCs with WDR72 mutations

Project description:Modeling hereditary distal renal tubular acidosis using patient-derived iPSCs with WDR72 mutations

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Cloning and variation of adult renal tubular progenitors

Project description:Kidney damage involves the progressive and inexorable destruction of tubular and glomerular system. However, it is known that the patients survive AKI often recover renal structure and function. Correspondingly, previous studies demonstrated tubular regeneration in mice after massive kidney injury and linked mouse Sox9+ renal progenitor cells to this process. Here we show that renal progenitor cells can be cloned from renal needle biopsy sample of CKD patients. Progenitor cells can readily assembly into “kidney organoids” expressing proximal/distal tubular cell markers in 3D culture.

2022-04-22 | GSE119426 | GEO

Cloning and transplantation of adult renal tubular progenitors

Project description:Kidney damage involves the progressive and inexorable destruction of tubular and glomerular system. However, it is known that the patients survive AKI often recover renal structure and function. Correspondingly, previous studies demonstrated tubular regeneration in mice after massive kidney injury and linked mouse Sox9+ renal progenitor cells to this process. Here we show that progenitor cells can be cloned from mouse medulla and cortex. Clones can be grown from a single cell and indefinitely passaged. Progenitor cells derived from renal medulla can readily assembly into “kidney organoids” expressing proximal/distal tubular cell markers in 3D culture.

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Quantitative proteinomics on WWP2 overexpressed human tubular epithelial cells HK-2

Project description:WWP2 is a E3 ligase which regulates multiple cellular events, such as stress response, differentiation. WWP2 played physiological role in the distal tubules of the kidneys. Whether WWP2 regulated pathophysiological rol in the proximal tubules is unclear. In the present study, we stably overexpressed WWP2 using lenti-viral infection in the human tubular epithelial cells, HK-2, and performed proteinomics analysis to explore WWP2's potential substrates.

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2022-06-28 | GSE199929 | GEO

Tig1 regulates proximo-distal identity during salamander limb regeneration

Project description:Salamander limb regeneration is an accurate process which gives rise exclusively to the missing structures, irrespective of the amputation level. This suggests that cells in the stump have an awareness of their spatial location, a property termed ‘positional identity’. Little is known about how positional identity is encoded, in salamanders or other biological systems. Through single-cell RNAseq analysis, we identified Tig1/RARRES1 as a potential determinant of proximal identity. Tig1 encodes a conserved cell surface molecule, is regulated by retinoic acid and exhibits a graded expression along the proximo-distal axis of the limb. Its overexpression leads to regeneration defects in the distal elements and elicits proximal displacement of blastema cells, while its neutralisation blocks proximo-distal cell surface interactions. Critically, Tig1 reprogrammes distal cells to a proximal identity, upregulating Prod1 and inhibiting HoxA13 and distal transcriptional networks. Thus, Tig1 is a central cell surface determinant of proximal identity in the salamander limb.

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Electric Tubular Anastomosis in Rectal Cancer

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