scholarly journals Cell profiling of mouse acute kidney injury reveals conserved cellular responses to injury

2020 ◽  
Vol 117 (27) ◽  
pp. 15874-15883 ◽  
Author(s):  
Yuhei Kirita ◽  
Haojia Wu ◽  
Kohei Uchimura ◽  
Parker C. Wilson ◽  
Benjamin D. Humphreys
Keyword(s):  
Acute Kidney Injury ◽  
Proximal Tubule ◽  
Communication Networks ◽  
Rat Kidney ◽  
Kidney Injury ◽  
Human Kidney ◽  
Cellular Responses ◽  
Acute Injury ◽  
Proximal Tubule Cell ◽  
Tubule Cell

After acute kidney injury (AKI), patients either recover or alternatively develop fibrosis and chronic kidney disease. Interactions between injured epithelia, stroma, and inflammatory cells determine whether kidneys repair or undergo fibrosis, but the molecular events that drive these processes are poorly understood. Here, we use single nucleus RNA sequencing of a mouse model of AKI to characterize cell states during repair from acute injury. We identify a distinct proinflammatory and profibrotic proximal tubule cell state that fails to repair. Deconvolution of bulk RNA-seq datasets indicates that this failed-repair proximal tubule cell (FR-PTC) state can be detected in other models of kidney injury, increasing during aging in rat kidney and over time in human kidney allografts. We also describe dynamic intercellular communication networks and discern transcriptional pathways driving successful vs. failed repair. Our study provides a detailed description of cellular responses after injury and suggests that the FR-PTC state may represent a therapeutic target to improve repair.

scholarly journals Cell profiling of mouse acute kidney injury reveals conserved cellular responses to injury

2020 ◽  
Author(s):  
Yuhei Kirita ◽  
Haojia Wu ◽  
Kohei Uchimura ◽  
Parker C. Wilson ◽  
Benjamin D. Humphreys
Keyword(s):  
Acute Kidney Injury ◽  
Proximal Tubule ◽  
Rna Sequencing ◽  
Therapeutic Target ◽  
Kidney Injury ◽  
Cellular Responses ◽  
Acute Injury ◽  
Proximal Tubule Cell ◽  
Tubule Cell ◽  
Single Nucleus

AbstractAfter acute kidney injury (AKI), patients either recover or alternatively develop fibrosis and chronic kidney disease. Interactions between injured epithelia, stroma and inflammatory cells determine whether kidneys repair or undergo fibrosis, but the molecular events that drive these processes are poorly understood. Here, we use single nucleus RNA sequencing of a mouse model of AKI to characterize cell states during repair from acute injury. We identify a distinct proinflammatory and profibrotic proximal tubule cell state that fails to repair. Deconvolution of bulk RNA-seq datasets indicates that this “failed-repair proximal tubule cell” or FR-PTC, state can be detected in other models of kidney injury, increasing in the aging rat kidney and over time in human kidney allografts. We also describe dynamic intercellular communication networks and discern transcriptional pathways driving successful vs. failed repair. Our study provides a detailed description of cellular responses after injury and suggests that the FR-PTC state may represent a therapeutic target to improve repair.Significance StatementSingle nucleus RNA sequencing revealed gene expression changes during repair after acute kidney injury. We describe a small population of proximal tubule cells that fail to repair (FR-PTC). Since this subpopulation expresses abundant pro-inflammatory and profibrotic genes, it may represent a new therapeutic target to improve repair and reduce fibrosis after AKI.

scholarly journals Ataxia Telangiectasia and Rad3-Related Activation by DNA Damage Mitigates Maladaptive Repair after Acute Kidney Injury

Nephron ◽  
2021 ◽  
pp. 1-4
Author(s):  
Joseph V. Bonventre
Keyword(s):  
Dna Damage ◽  
Acute Kidney Injury ◽  
Proximal Tubule ◽  
Ataxia Telangiectasia ◽  
Kidney Injury ◽  
Human Kidney ◽  
Acute Injury ◽  
Proximal Tubule Cell ◽  
Cell Renewal ◽  
Healthy State

DNA damage is an important consequence of injury to the proximal tubule. The proximal tubule cell responds to this damage by mounting a DNA damage response (DDR). Two protein kinases, ataxia-telangiectasia mutated (ATM) or ataxia telangiectasia and Rad3-related (ATR), play an important role in this DDR. If efficient, the DDR can lead to repair of the DNA, cell renewal, and return to a healthy state. In many cases, however, especially in the setting of baseline kidney injury, there is incomplete repair. In human chronic kidney disease (CKD) and in human kidney organoids exposed to acute injury, there is increased evidence of DNA damage and activation of ATR. This review focuses on 3 aspects of the DNA damage and response to it: (1) DNA damage and the DDR precipitated by acute injury; (2) protection afforded by the DDR kinase, ATR, in multiple mouse models of acute kidney injury; and (3) downstream effects of genetic inhibition of ATR in the proximal tubule, leading to maladaptive repair, fibrosis, and CKD.

scholarly journals Insulin-like growth factor binding protein 7 and tissue inhibitor of metalloproteinases-2: differential expression and secretion in human kidney tubule cells

2017 ◽  
Vol 312 (2) ◽  
pp. F284-F296 ◽  
Author(s):  
David R. Emlet ◽  
Nuria Pastor-Soler ◽  
Allison Marciszyn ◽  
Xiaoyan Wen ◽  
Hernando Gomez ◽  
...  
Keyword(s):  
Cell Culture ◽  
Acute Kidney Injury ◽  
Kidney Injury ◽  
Human Kidney ◽  
Distal Tubule ◽  
Cell Types ◽  
Tissue Inhibitor Of Metalloproteinases ◽  
Tubule Cell ◽  
Tubule Cells

We have characterized the expression and secretion of the acute kidney injury (AKI) biomarkers insulin-like growth factor binding protein 7 (IGFBP7) and tissue inhibitor of metalloproteinases-2 (TIMP-2) in human kidney epithelial cells in primary cell culture and tissue. We established cell culture model systems of primary kidney cells of proximal and distal tubule origin and observed that both proteins are indeed expressed and secreted in both tubule cell types in vitro. However, TIMP-2 is both expressed and secreted preferentially by cells of distal tubule origin, while IGFBP7 is equally expressed across tubule cell types yet preferentially secreted by cells of proximal tubule origin. In human kidney tissue, strong staining of IGFBP7 was seen in the luminal brush-border region of a subset of proximal tubule cells, and TIMP-2 stained intracellularly in distal tubules. Additionally, while some tubular colocalization of both biomarkers was identified with the injury markers kidney injury molecule-1 and neutrophil gelatinase-associated lipocalin, both biomarkers could also be seen alone, suggesting the possibility for differential mechanistic and/or temporal profiles of regulation of these early AKI biomarkers from known markers of injury. Last, an in vitro model of ischemia-reperfusion demonstrated enhancement of secretion of both markers early after reperfusion. This work provides a rationale for further investigation of these markers for their potential role in the pathogenesis of acute kidney injury.

scholarly journals THE POLARITY OF THE PROXIMAL TUBULE CELL IN RAT KIDNEY

1972 ◽  
Vol 54 (2) ◽  
pp. 232-245 ◽  
Author(s):  
Hans-G Heidrich ◽  
Rolf Kinne ◽  
Eva Kinne-Saffran ◽  
Kurt Hannig
Keyword(s):  
Proximal Tubule ◽  
Plasma Membranes ◽  
Specific Activity ◽  
Rat Kidney ◽  
High Specific Activity ◽  
Kidney Cortex ◽  
Proximal Tubule Cell ◽  
Surface Charges ◽  
Tubule Cell ◽  
Rat Kidney Cortex

Two different membrane fractions were obtained from a brush-border fraction of rat kidney cortex by using their different electrical surface charges in preparative free-flow electrophoresis. One membrane fraction contained only morphologically intact microvilli and was characterized by a high specific activity of alkaline phosphatase. The other fraction morphologically resembled classical plasma membranes by possessing junctional complexes and a high Na-K-ATPase activity The contamination of the isolated membrane fractions by other cell organelles was extremely low These two fractions represent the apical (luminal) and the basal (interstitial) area of the renal proximal tubule cell membrane and clearly demonstrate the polarity of this cell.

Epidermal growth factor receptor regulation in rat kidney: two models of renal growth

1989 ◽  
Vol 257 (6) ◽  
pp. F1059-F1064 ◽  
Author(s):  
M. T. Behrens ◽  
A. L. Corbin ◽  
M. K. Hise
Keyword(s):  
Folic Acid ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Proximal Tubule ◽  
Rat Kidney ◽  
Kidney Cortex ◽  
Proximal Tubule Cell ◽  
Tubule Cell ◽  
Rat Kidney Cortex ◽  
Epidermal Growth

The binding of 125I-labeled epidermal growth factor (EGF) to plasma membranes prepared from rat kidney cortex was studied following unilateral nephrectomy, a model of proximal tubule cell hypertrophy, and following the administration of folic acid, a model of proximal tubule cell hyperplasia. Binding of 125I-EGF was a linear function of basolateral membrane protein content and time of incubation. Specific binding to luminal brush-border membranes was not evident in these studies. Neither insulin nor insulin-like growth factor I could displace EGF binding, indicating that binding was specific. Scatchard analysis revealed a single binding site. The KD in sham-operated animals 48 h after surgery was 11.2 +/- 1.4 nM, whereas Bmax averaged 95.2 +/- 4.1 fmol/mg protein (n = 3). Similar values were obtained in nephrectomized animals. The Bmax of folic acid-untreated animals averaged 212.5 +/- 6.9 fmol/mg 48 h after administration, whereas that of vehicle-injected controls averaged 85.4 +/- 9.2 (n = 3, P less than 0.001). Differences in binding were not related to changes in affinity, ligand degradation by the preparations, or receptor binding of endogenous EGF. These data indicate that regeneration following folic acid administration is associated with an upregulation of proximal nephron EGF receptors that may play an important role in the mitogenic response.

Expression of MT-3 mRNA in human kidney, proximal tubule cell cultures, and renal cell carcinoma

1997 ◽  
Vol 92 (2) ◽  
pp. 149-160 ◽  
Author(s):  
John G Hoey ◽  
Scott H Garrett ◽  
Mary Ann Sens ◽  
John H Todd ◽  
Donald A Sens
Keyword(s):  
Renal Cell Carcinoma ◽  
Proximal Tubule ◽  
Cell Carcinoma ◽  
Cell Cultures ◽  
Renal Cell ◽  
Human Kidney ◽  
Proximal Tubule Cell ◽  
Tubule Cell ◽  
Kidney Proximal Tubule

Megalin in acute kidney injury: foe and friend

2014 ◽  
Vol 306 (2) ◽  
pp. F147-F154 ◽  
Author(s):  
Ravikiran Mahadevappa ◽  
Rikke Nielsen ◽  
Erik Ilsø Christensen ◽  
Henrik Birn
Keyword(s):  
Acute Kidney Injury ◽  
Animal Models ◽  
Proximal Tubule ◽  
Essential Role ◽  
Kidney Injury ◽  
Current Evidence ◽  
Renal Protection ◽  
Tubule Cell ◽  
Cellular Protection ◽  
And Function

The kidney proximal tubule is a key target in many forms of acute kidney injury (AKI). The multiligand receptor megalin is responsible for the normal proximal tubule uptake of filtered molecules, including nephrotoxins, cytokines, and markers of AKI. By mediating the uptake of nephrotoxins, megalin plays an essential role in the development of some types of AKI. However, megalin also mediates the tubular uptake of molecules implicated in the protection against AKI, and changes in megalin expression have been demonstrated in AKI in animal models. Thus, modulation of megalin expression in response to AKI may be an important part of the tubule cell adaption to cellular protection and regeneration and should be further investigated as a potential target of intervention. This review explores current evidence linking megalin expression and function to the development, diagnosis, and progression of AKI as well as renal protection against AKI.

scholarly journals Modeling acute kidney injury in kidney organoids with cisplatin

2019 ◽  
Author(s):  
Jenny L. M. Digby ◽  
Aneta Przepiorski ◽  
Alan J. Davidson ◽  
Veronika Sander
Keyword(s):  
Dna Damage ◽  
Acute Kidney Injury ◽  
Proximal Tubule ◽  
Kidney Injury ◽  
Induced Pluripotent Stem Cell ◽  
Human Kidney ◽  
Distal Tubule ◽  
Dependent Manner ◽  
Kidney Organoids

ABSTRACTAcute kidney injury (AKI) remains a major global healthcare problem and there is a need to develop human-based models to study AKI in vitro. Towards this goal, we have characterized induced pluripotent stem cell-derived human kidney organoids and their response to cisplatin, a chemotherapeutic drug that induces AKI and preferentially damages the proximal tubule. We found that a single treatment with 50 µM cisplatin induces HAVCR1 and CXCL8 expression, DNA damage (γH2AX) and cell death in the organoids in a dose-dependent manner but greatly impairs organoid viability. DNA damage was not specific to the proximal tubule but also affected the distal tubule and interstitial populations. This lack of specificity correlated with low expression of the proximal tubule-specific SLC22A2/OCT2 transporter for cisplatin. To improve viability, we developed a repeated low-dose regimen of 4x 5 µM cisplatin over 7 days and found this causing less toxicity while still inducing a robust AKI response that included secretion of known AKI biomarkers and inflammatory cytokines. This work validates the use of human kidney organoids to model aspects of AKI in vitro, with the potential to identify new AKI biomarkers and develop better therapies.

Tissue Culture Models of Acute Kidney Injury: From Tubule Cells to Human Kidney Organoids

2022 ◽  
pp. ASN.2021050693
Author(s):  
Julie Bejoy ◽  
Eddie Qian ◽  
Lauren Woodard
Keyword(s):  
Acute Kidney Injury ◽  
Tissue Culture ◽  
Cell Lines ◽  
Renal Cell ◽  
Kidney Injury ◽  
Human Kidney ◽  
Cell Types ◽  
Tubule Cell ◽  
Culture Models ◽  
Kidney Organoids

Acute kidney injury (AKI) affects approximately 13.3 million people around the world each year, causing chronic kidney disease and/or mortality. The mammalian kidney cannot generate new nephrons after postnatal renal damage and regenerative therapies for AKI are not available. Human kidney tissue culture systems can complement animal models of AKI and/or address some of their limitations. Donor-derived somatic cells, such as renal tubule epithelial cells or cell lines (RPTEC/hTERT, ciPTEC, HK-2, Nki-2, and CIHP-1), have been used for decades to permit drug toxicity screening and studies into potential AKI mechanisms. However, tubule cell lines do not fully recapitulate tubular epithelial cell properties in situ when grown under classic tissue culture conditions. Improving tissue culture models of AKI would increase our understanding of the mechanisms, leading to new therapeutics. Human pluripotent stem cells (hPSCs) can be differentiated into kidney organoids and various renal cell types. Injury to human kidney organoids results in renal cell type crosstalk and upregulation of kidney injury biomarkers that are difficult to induce in primary tubule cell cultures. However, current protocols produce kidney organoids that aren't mature and contain off-target cell types. Promising bioengineering techniques, such as bioprinting and "kidney-on-a17 chip" methods, as applied to kidney nephrotoxicity modeling advantages and limitations are discussed. This review explores the mechanisms and detection of AKI in tissue culture, with an emphasis on bioengineered approaches such as human kidney organoid models.

scholarly journals Presence of an extensive clathrin coat on the apical plasmalemma of the rat kidney proximal tubule cell.

1984 ◽  
Vol 98 (5) ◽  
pp. 1630-1636 ◽  
Author(s):  
J S Rodman ◽  
D Kerjaschki ◽  
E Merisko ◽  
M G Farquhar
Keyword(s):  
Proximal Tubule ◽  
Brush Border ◽  
Specific Binding ◽  
Rat Kidney ◽  
Light Chains ◽  
Kidney Cortex ◽  
Immunoperoxidase Staining ◽  
Proximal Tubule Cell ◽  
Tubule Cell ◽  
Kidney Proximal Tubule

The nature of the cytoplasmic coat present on the apical invaginations of the kidney proximal tubule cell was investigated by immuneoverlay and immunocytochemistry of renal brush borders with anticlathrin antibodies. When kidney cortex was prepared for electron microscopy using methods that enhance visualization of clathrin coats, the apical invaginations at the base of the brush border microvilli were seen to be backed by a nearly continuous coating which resembles but is more extensive than the lattice-like clathrin coats found around brain coated vesicles. When isolated brush border fractions were prepared under conditions that preserve the coats, separated by SDS PAGE, and transferred to nitrocellulose, the presence of clathrin heavy and light chains was detected by immuneoverlay using two different affinity-purified anticlathrin IgGs--one that we prepared, which detects only the clathrin light chains, and the other, prepared by Louvard et al. ( Louvard , D., C. Morris, G. Warren, K. Stanley, F. Winkler , and H. Reggio , 1983, EMBO [Eur. Mol. Biol. Organ.] J., 2:1655-1664), which detects both the heavy and light chains. As viewed by light microscopy (immunofluorescence or immunoperoxidase), staining with both anticlathrins was concentrated at the base of the proximal tubule microvilli. Immunoelectron microscopic localizations carried out on brush border fractions (using peroxidase and gold conjugates) demonstrated specific binding of anticlathrin IgGs to the lattice-like cytoplasmic coat. When brush border fractions were reacted with monoclonal antibodies prepared against gp330 and maltase, proteins that serve as markers for the membrane of the apical invaginations and microvilli, respectively ( Kerjaschki , D., L. Noronha - Blob , B. Sacktor , and M. G. Farquhar , 1984, J. Cell Biol., 98:1505-1513), the two proteins retained their restrictive distribution in the brush border. The findings demonstrate (a) that the cytoplasmic coat of the proximal tubule intermicrovillar apical invaginations is composed of clathrin heavy and light chains, and (b) that the differential distribution of proteins in these two brush border microdomains is maintained in appropriately prepared brush border fractions.

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