CCN2 Aggravates the Immediate Oxidative Stress–DNA Damage Response following Renal Ischemia–Reperfusion Injury

Valentijn, Floris A.; Knoppert, Sebastiaan N.; Pissas, Georgios; Rodrigues-Díez, Raúl R.; Márquez‐Expósito, Laura; Broekhuizen, Roel; Mokrý, Michal; Kester, Lennart; Falke, Lucas L.; Goldschmeding, Roel; Ruíz-Ortega, Marta; Eleftheriadis, Theodoros; Nguyen, Tri Q.

doi:10.3390/antiox10122020

Cited by 24 publications

(24 citation statements)

References 53 publications

(60 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Interestingly, CCN2 and CCN5 were elevated in the senescent fibrotic fibroblast clusters as well as the in the sorted tdTom + SnC. The CCN matricellular proteins have been implicated in induction of senescence in a variety of contexts (Joon-Il Jun & Lau, 2011; Joon-II Jun & Lau, 2017; Valentijn et al, 2021). They are also regulatory targets of the YAP/TAZ pathway, a mechanosensing pathway recently highlighted for its potential role in fibrosis (Dwivedi et al, 2020; Mascharak et al, 2022; Zhou et al, 2022).…”

Section: Discussionmentioning

confidence: 99%

Transfer learning of an in vivo-derived senescence signature identifies conserved and tissue-specific senescence across species and diverse pathologies

Cherry

Andorko

Krishnan

et al. 2022

Preprint

View full text Add to dashboard Cite

Senescent cells (SnCs) contribute to normal tissue development and repair but accumulate with aging where they are implicated in a number of pathologies and diseases. Despite their pathological role and therapeutic interest, SnC phenotype and function in vivo remains unclear due to the challenges in identifying and isolating these rare cells. Here, we developed an in vivo-derived senescence gene expression signature using a model of the foreign body response (FBR) fibrosis in a p16Ink4a-reporter mouse, a cell cycle inhibitor commonly used to identify SnCs. We identified stromal cells (CD45-CD31-CD29+) as the primary p16Ink4a expressing cell type in the FBR and collected the cells to produce a SnC transcriptomic signature with bulk RNA sequencing. To computationally identify SnCs in bulk and single-cell data sets across species and tissues, we used this signature with transfer learning to generate a SnC signature score (SenSig). We found senescent pericyte and cartilage-like fibroblasts in newly collected single cell RNAseq (scRNASeq) data sets of murine and human FBR suggesting populations associated with angiogenesis and secretion of fibrotic extracellular matrix, respectively. Application of the senescence signature to human scRNAseq data sets from idiopathic pulmonary fibrosis (IPF) and the basal cell carcinoma microenvironment identified both conserved and tissue-specific SnC phenotypes, including epithelial-derived basaloid and endothelial cells. In a wound healing model, ligand-receptor signaling prediction identified putative interactions between SnC SASP and myeloid cells that were validated by immunofluorescent staining and in vitro coculture of SnCs and macrophages. Collectively, we have found that our SenSig transfer learning strategy from an in vivo signature outperforms in vitro-derived signatures and identifies conserved and tissue-specific SnCs and their SASP, independent of p16Ink4a expression, and may be broadly applied to elucidate SnC identity and function in vivo.

show abstract

Section: Discussionmentioning

confidence: 99%

Transfer learning of an in vivo-derived senescence signature identifies conserved and tissue-specific senescence across species and diverse pathologies

Cherry

Andorko

Krishnan

et al. 2022

Preprint

View full text Add to dashboard Cite

show abstract

“…These EMT-induced changes are mediated by the activation of MAPK-ERK cascade [ 55 , 56 ], ERGF, JAK-STAT3, and NF-κB signaling pathways [ 21 ]. In addition, stimulation with CCN2(IV) also induces cell growth arrest of cultured tubular cells [ 21 ] and activation of senescent-related mechanisms [ 11 ]. Our findings show that CCN2(IV) preferentially binds to tubular epithelial cells in the mouse kidney.…”

Section: Discussionmentioning

confidence: 99%

“…One of these cellular responses is cell growth, in which CCN2 participates in the regulation of proliferation, apoptosis, and migration [ 8 ]. In addition, CCN2 can also promote phenotype changes, as described in T lymphocytes differentiation towards Th17 subtype [ 9 ], in the induction of cellular senescence [ 10 , 11 ], and in the processes of epithelial/endothelial to mesenchymal transitions (EMT/EndMT, respectively) [ 12 , 13 ]. CCN2 is also involved in angiogenesis, cancer, inflammation, and fibrogenesis [ 14 , 15 , 16 , 17 , 18 ].…”

Section: Introductionmentioning

confidence: 99%

“…Preclinical studies have shown that inhibition of endogenous CCN2 by antisense oligonucleotides or gene silencing slows disease progression in experimental diabetic nephropathy, unilateral ureteral obstruction, and nephrectomized transgenic mice overexpressing TGF-β1 [ 19 , 30 , 31 , 32 ]. More recently, studies in CCN2 conditional deficient mice have demonstrated the role of CCN2 in renal fibrosis, cell growth arrest, oxidative stress, and senescence [ 11 , 21 ].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

CCN2 Binds to Tubular Epithelial Cells in the Kidney

et al. 2022

Self Cite

View full text Add to dashboard Cite

Cellular communication network-2 (CCN2), also called connective tissue growth factor (CTGF), is considered a fibrotic biomarker and has been suggested as a potential therapeutic target for kidney pathologies. CCN2 is a matricellular protein with four distinct structural modules that can exert a dual function as a matricellular protein and as a growth factor. Previous experiments using surface plasmon resonance and cultured renal cells have demonstrated that the C-terminal module of CCN2 (CCN2(IV)) interacts with the epidermal growth factor receptor (EGFR). Moreover, CCN2(IV) activates proinflammatory and profibrotic responses in the mouse kidney. The aim of this paper was to locate the in vivo cellular CCN2/EGFR binding sites in the kidney. To this aim, the C-terminal module CCN2(IV) was labeled with a fluorophore (Cy5), and two different administration routes were employed. Both intraperitoneal and direct intra-renal injection of Cy5-CCN2(IV) in mice demonstrated that CCN2(IV) preferentially binds to the tubular epithelial cells, while no signal was detected in glomeruli. Moreover, co-localization of Cy5-CCN2(IV) binding and activated EGFR was found in tubules. In cultured tubular epithelial cells, live-cell confocal microscopy experiments showed that EGFR gene silencing blocked Cy5-CCN2(IV) binding to tubuloepithelial cells. These data clearly show the existence of CCN2/EGFR binding sites in the kidney, mainly in tubular epithelial cells. In conclusion, these studies show that circulating CCN2(IV) can directly bind and activate tubular cells, supporting the role of CCN2 as a growth factor involved in kidney damage progression.

show abstract

“…Endogenous salbutamolβ treatment exacerbates sepsis-related and nephrotoxic AKI by activating the ROS/DNA damage/P53 apoptotic pathway [11]. CCN2 exacerbates ischemic AKI by increasing the expression of γH2AX and p-P53 through oxidative DNA damage [40].The degree of DNA damage is inversely correlated with the estimated glomerular filtration rate (eGFR) [41]. Therefore, activation of the DDR signaling pathway may represent a common mechanism that leads to AKI.…”

Section: Discussionmentioning

confidence: 99%

Inhibition of PLK3 Attenuates Tubular Epithelial Cell Apoptosis after Renal Ischemia–Reperfusion Injury by Blocking the ATM/P53-Mediated DNA Damage Response

Deng

Wei

Xie

et al. 2022

Oxidative Medicine and Cellular Longevity

View full text Add to dashboard Cite

Objective. Renal ischemia–reperfusion (I/R) injury is a major cause of acute kidney injury (AKI) in transplanted kidneys. This study was aimed at exploring the role of PLK3 (polo-like kinase 3) in renal I/R injury, focusing on its relationship with oxidative stress-induced DNA damage and renal tubular epithelial cell (TEC) apoptosis. Methods. TRAP-seq data from the development dataset GSE52004 and the validation dataset GSE121191 were analyzed using GEO2R. PLK3 overexpression plasmids and targeted silencing siRNAs were used in a model of hypoxia/reoxygenation (H/R) injury, and rAAV-9-PLK3-KD were administered to C57BL/6J mice exposed to I/R injury. The ATM-specific inhibitor KU-60019 was used to block the DNA damage response (DDR). Western blotting was performed to measure DDR- and apoptosis-associated protein expression. Cell viability was measured by CCK-8 reagent, and apoptosis was examined by flow cytometry and TUNEL assay. Furthermore, the fluorescent probes H2DCFH-DA and DHE were used to measure ROS production in vitro. The MDA level and SOD activity were measured to assess oxidative stress in vivo. KIM-1 staining and Scr and BUN were used to evaluate kidney injury. Results. The mRNA and protein levels of PLK3 were markedly increased in the H/R injury and I/R injury models. GO terms showed that PLK3 was mainly involved in oxidative stress and DNA damage after renal I/R injury. Overexpression of PLK3 decreased cell viability and increased apoptosis. In contrast, targeted silencing of PLK3 expression decreased the Bax/Bcl-2 ratio by decreasing P53 phosphorylation, thereby reducing TEC apoptosis. Furthermore, KU-60019 reduced PLK3 activation and DDR-induced apoptosis, while overexpression of PLK3 reversed the mitigating effect of KU-60019 on TEC apoptosis. Similarly, rAAV-9-PLK3 KD mice exhibited a lower rate of TEC apoptosis and milder renal damage after I/R injury. Conclusion. We demonstrate for the first time that PLK3 is involved in oxidative stress-induced DNA damage and TEC apoptosis in renal I/R injury. Inhibition of PLK3 attenuates TEC apoptosis after I/R injury by blocking the ATM/P53-mediated DDR. Therefore, PLK3 may serve as a potential therapeutic target for ischemic AKI.

show abstract

CCN2 Aggravates the Immediate Oxidative Stress–DNA Damage Response following Renal Ischemia–Reperfusion Injury

Cited by 24 publications

References 53 publications

Transfer learning of an in vivo-derived senescence signature identifies conserved and tissue-specific senescence across species and diverse pathologies

Transfer learning of an in vivo-derived senescence signature identifies conserved and tissue-specific senescence across species and diverse pathologies

CCN2 Binds to Tubular Epithelial Cells in the Kidney

Inhibition of PLK3 Attenuates Tubular Epithelial Cell Apoptosis after Renal Ischemia–Reperfusion Injury by Blocking the ATM/P53-Mediated DNA Damage Response

Contact Info

Product

Resources

About