Generation of Distal Renal Segments Involves a Unique Population of Aqp2+ Progenitor Cells

Gao, Chao; Chen, Lihe; Chen, Enuo; Tsilosani, Akaki; Xia, Yang; Zhang, Wenzheng

doi:10.1681/asn.2021030399

Cited by 14 publications

(37 citation statements)

References 35 publications

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“…Ward clustering uncovered three AQP2-positive clusters ( Figure S2Bb ) that mapped to distinct tubular epithelium. The two AQP2-positive clusters with highest intensity (clusters 1 and 2 Figure S2Bc ) mapped predominantly to collecting duct cells and putative distal epithelial cells (verified by morphology), which could be consistent with APQ2-positive progenitor cells described previously 31 . The AQP2-positive with low intensity mapped to the distal tubular epithelium, suggesting a subpopulation of cells in the distal nephron with a low level of AQP2 ( Figure S2Ad and S2Bc ).…”

Section: Resultssupporting

confidence: 87%

“…VTEA (verified by morphology), which could be consistent with APQ2-positive progenitor cells described previously 31 . The AQP2-positive with low intensity mapped to the distal tubular epithelium, suggesting a subpopulation of cells in the distal nephron with a low level of AQP2 (Figure S2Ad and S2Bc).…”

Section: Building An Extensible Framework For An Integrated Tissue Cy...supporting

confidence: 89%

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Integrated cytometry with machine learning applied to high-content imaging of human kidney tissue for in-situ cell classification and neighborhood analysis

Winfree

McNutt

Khochare

et al. 2021

Preprint

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The human kidney is a complex organ with various cell types that are intricately organized to perform key physiological functions and maintain homeostasis. New imaging modalities such as mesoscale and highly multiplexed fluorescence microscopy are increasingly applied to human kidney tissue to create single cell resolution datasets that are both spatially large and multi-dimensional. These single cell resolution high-content imaging datasets have a great potential to uncover the complex spatial organization and cellular make-up of the human kidney. Tissue cytometry is a novel approach used for quantitative analysis of imaging data, but the scale and complexity of such datasets pose unique challenges for processing and analysis. We have developed the Volumetric Tissue Exploration and Analysis (VTEA) software, a unique tool that integrates image processing, segmentation and interactive cytometry analysis into a single framework on desktop computers. Supported by an extensible and open-source framework, VTEA's integrated pipeline now includes enhanced analytical tools, such as machine learning, data visualization, and neighborhood analyses for hyperdimensional large-scale imaging datasets. These novel capabilities enable the analysis of mesoscale two and three-dimensional multiplexed human kidney imaging datasets (such as CODEX and 3D confocal multiplexed fluorescence imaging). We demonstrate the utility of this approach in identifying cell subtypes in the kidney based on labels, spatial association and their microenvironment or neighborhood membership. VTEA provides integrated and intuitive approach to decipher the cellular and spatial complexity of the human kidney and complement other transcriptomics and epigenetic efforts to define the landscape of kidney cell types.

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Section: Resultssupporting

confidence: 87%

Section: Building An Extensible Framework For An Integrated Tissue Cy...supporting

confidence: 89%

Integrated cytometry with machine learning applied to high-content imaging of human kidney tissue for in-situ cell classification and neighborhood analysis

Winfree

McNutt

Khochare

et al. 2021

Preprint

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“…Adult Aqp2-Expressing Cells Regenerate DCT2/CNT/ CD Cells during Renal Maintenance Embryonic and neonate APs, which are characterized by expressing Aqp2 and V-ATPase subunits B1 and B2, have the capacity for self-renewal, clonogenicity, and multipotency. 18 During development, embryonic Aqp2 1 B1B2 1 cells (APs) emerge at embryonic day 15.5 (E15.5) and differentiate, in a stepwise manner, into five or more cell types to form molecularly distinct DCT2, CNT1, CNT2, and CD segments. This is accomplished by either loss of Aqp2 or B1B2 and acquisition of NCC (DCT2 marker) at E15.5, CAII (another IC marker) at E16.5, or AE1 and Pendrin (markers for a-IC and b-IC, respectively) at postnatal day 1.…”

Section: Resultsmentioning

confidence: 99%

“…Costaining of RFP/AE1, RFP/NCC, or B1/AE1, using rabbit antibodies, was performed according to our previously published sequential staining protocol. 14,18 Briefly, slides were first incubated with one primary antibody overnight at 4 C, followed by hybridization with a respective secondary antibody. After blocking with 2% normal donkey serum/1% BSA/0.05% Tween 20 in PBS, sections were then probed with another primary antibody overnight at 4 C. Finally, sections were incubated with another matched secondary antibody and mounted using ProLong Gold antifade mounting media (Invitrogen).…”

Section: Immunofluorescence Studiesmentioning

confidence: 99%

Aqp2+ Progenitor Cells Maintain and Repair Distal Renal Segments

Gao

Zhang

Chen

et al. 2022

JASN

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Background: Adult progenitor cells presumably have clonogenicity, self-renewal and multipotentiality, and can regenerate cells under various conditions. Definitive evidence demonstrating the existence of such progenitor cells in adult mammalian kidneys is lacking. Methods: We performed in vivo lineage tracing and thymidine analog labeling, using adult Tamoxifen-inducible (Aqp2ECE/+ RFP/+, Aqp2ECE/+ Brainbow/+ and Aqp2ECE/+ Brainbow/Brainbow) and WT mice. The Tamoxifen-inducible mice were analyzed between 1 to 300 days post-induction. Alternatively, WT and Tamoxifen-induced mice were subjected to unilateral ureteral obstruction and thymidine analog labeling and analyzed 2 to 14 days postsurgery. Multiple cell-specific markers were used for high-resolution immunofluorescence confocal microscopy to identify the cell types derived from Aqp2+ cells. Results: Like their embryonic counterparts, adult cells expressing Aqp2 and V-ATPase subunits B1 and B2 (Aqp2+ B1B2+) are the potential Aqp2+ progenitor cells (AP). Adult AP rarely divide to generate daughter cells either maintaining the property of the AP (self-renewal) or differentiating into DCT2/CNT/CD cells (multipotentiality), forming single-cell-derived multiple-cell clones (clonogenicity) during tissue maintenance. They selectively and continuously regenerate DCT2/CNT/CD cells in response to injury resulting from ureteral ligation. AP proliferation demonstrated direct correlation with Notch activation and inversely correlated with development of kidney fibrosis. Derivation of both intercalated and DCT2 cells was found to be cell-division-dependent and -independent manners, most likely through AP differentiation that requires cell division and through direct conversion of AP and/or regular principal cells without cell division, respectively. Conclusion: Our study demonstrates that Aqp2+ B1B2+ cells behave as adult AP to maintain and repair DCT2/CNT1/CNT2/CD segments.

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“…Though speculative, based on the urine cell signature’s bias toward distal nephron segments, those progenitor-like cells – if not dedifferentiated distal TEC – may also be a separate entity of distal/medullary/papillary tip origin, like the recently characterized distal multipotent Aqp2+ progenitor cells in mice(54), and therefore harder to detect via biopsy.…”

Section: Discussionmentioning

confidence: 99%

Urinary single-cell sequencing captures intrarenal injury and repair processes in human acute kidney injury

Klocke

Kim

Skopnik

et al. 2022

Preprint

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Acute kidney injury (AKI) is a major health issue, the outcome of which depends primarily on damage and reparative processes of tubular epithelial cells (TEC). Mechanisms underlying AKI remain incompletely understood, specific therapies are lacking and monitoring the course of AKI in clinical routine is confined to measuring urine output and plasma levels of filtration markers.Here we demonstrate feasibility and potential of a novel approach to assess the cellular and molecular dynamics of AKI by establishing a robust urine-to-single cell RNA sequencing (scRNAseq) pipeline for excreted kidney cells via flow cytometry sorting. We analyzed 42,608 single cell transcriptomes of 40 urine samples from 32 AKI patients and compared our data with reference material from human AKI post-mortem biopsies and published mouse data. We demonstrate that TEC transcriptomes mirror intrarenal pathology and reflect distinct injury and repair processes, including oxidative stress, inflammation, and tissue rearrangement. We also describe an AKI-specific abundant urinary excretion of progenitorlike cells.In conclusion, single cell transcriptomics of kidney cells excreted in urine provides non-invasive, unprecedented insight into cellular processes underlying AKI, thereby opening novel opportunities for target identification, AKI sub-categorization and monitoring of natural disease course and interventions.Graphical abstract

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Generation of Distal Renal Segments Involves a Unique Population of Aqp2+ Progenitor Cells

Abstract: See article for abstract.

Cited by 14 publications

References 35 publications

Integrated cytometry with machine learning applied to high-content imaging of human kidney tissue for in-situ cell classification and neighborhood analysis

Integrated cytometry with machine learning applied to high-content imaging of human kidney tissue for in-situ cell classification and neighborhood analysis

Aqp2+ Progenitor Cells Maintain and Repair Distal Renal Segments

Urinary single-cell sequencing captures intrarenal injury and repair processes in human acute kidney injury

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