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Shroom3 regulates epithelial differentiation during tubular repair after ischemia reperfusion injury

2021· article· en· W3169959569 on OpenAlexafffund
Joanna Cunanan, Anna Li, Hadiseh Khalili, Ashmeet Hunjan, Ahsan U. Khan, Kristina Cunanan, Thomas A. Drysdale, Darren Bridgewater

Bibliographic record

VenueThe FASEB Journal · 2021
Typearticle
Languageen
FieldBiochemistry, Genetics and Molecular Biology
TopicRenal and related cancers
Canadian institutionsWestern UniversityMcMaster University
FundersNatural Sciences and Engineering Research Council of CanadaCanadian Institutes of Health ResearchKidney Foundation of Canada
KeywordsPathologyCell biologyKidneyVimentinEpitheliumNephronActinBiologyMesenchymal stem cellRenal ischemiaIschemiaMyosinCellular differentiationImmunohistochemistryReperfusion injuryMedicineInternal medicineEndocrinology

Abstract

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Shroom3 is an actin‐binding protein that modulates actomyosin dynamics to alter epithelial cell morphology. In adult kidneys, Shroom3 is apically localized in tubular epithelial cells of cortical nephron segments, such as S1 of proximal tubules, distal tubules, and collecting ducts. Genomic variants in Shroom3 are strongly associated with poor renal function and poor outcomes in kidney transplant recipients. After a kidney insult such as ischemia reperfusion injury from renal or cardiac surgeries, the damaged renal epithelium undergoes cell morphology changes to regenerate normal renal epithelial structure. This repair process includes actomyosin remodelling, allowing damaged epithelial cells to undergo de‐differentiation into a mesenchymal cell type, proliferate and re‐differentiate to tubular epithelial cells. In this study, our objective is to determine whether Shroom3 plays a role in renal epithelial repair after an ischemic kidney insult. We performed bilateral renal ischemia/reperfusion (I/R) on 3‐month‐old Shroom3 heterozygous mutant mice ( Shrm3 +/‐ )and wild type (WT) mice. Compared to WT at 10 days post‐I/R, most of the epithelial cells in cortical tubules of Shrm3 +/‐ kidneys had F‐actin and phospho‐Myosin Light Chain‐2 (pMLC‐2) sporadically distributed in the cytoplasm rather than being apically localized. In these cortical tubular cells, immunohistochemistry (IHC) demonstrated that mesenchymal marker Vimentin was highly expressed while epithelial markers N‐cadherin and E‐cadherin were significantly lower than in WT. These findings suggest that Shroom3 regulates tubular repair in an actin dependent manner that facilitates proper epithelial re‐differentiation. The inability to complete tubular epithelial re‐differentiation would result in a worsened kidney histopathology. Analysis of Shrm3 +/‐ kidneys compared to WT at 10 days post‐I/R showed higher levels of Kidney Injury Molecule‐1 in the apical surface of cortical tubular cells as shown by IHC, a 2.5‐fold increase in cell proliferation shown by KI‐67 ( Shrm3 +/‐ =16.5±0.8 vs. WT=6.6±1.8), a 4.4‐fold increase in apoptosis shown by Caspase‐3 ( Shrm3 +/‐ =1.4±0.09 vs. WT=0.32±0.04), a 2.4‐fold increase in inflammation levels shown by F4/80 ( Shrm3 +/‐ =64.5±2.2 vs. WT=26.4±2.5), and a 4.3‐fold increase in fibrosis levels shown by Picrosirius red ( Shrm3 +/‐ =10.3±0.73 vs. WT=2.4±0.15). These histopathological changes translated to severe impairments in kidney function: Shrm3 +/‐ mice had a 4.0‐fold increase in serum creatinine levels at 24 hours post‐I/R and did not return to baseline levels until after 7 days, while WT mice had a 2.0‐fold increase after 24 hours and returned to baseline by 48 hours. Shrm3 +/‐ mice also had higher mortality rates over a 10‐day period ( Shrm3 +/‐ =34.6% vs. WT=14.3%). Taken together, our findings demonstrate that Shroom3 plays key roles in renal epithelium repair after ischemia reperfusion injury likely by regulating actin cytoskeleton remodeling during the repair phase and thus modulating epithelial re‐differentiation. Our results could help explain why patients with genetic anomalies in Shroom3 show worse renal outcomes after an ischemic kidney insult.

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How this classification was reachedexpand

Full frame machine prediction

Teacher imitation

Not calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.

metaresearch head score (Codex)0.000
metaresearch head score (Gemma)0.000
Version: metacan-v3-hybrid-931329e0061cValidation status: machine_predicted_unvalidated
Candidate categoriesnone
Consensus categoriesnone
DomainCandidate signal: none · Consensus signal: none
Study designCandidate signal: Bench or experimental · Consensus signal: Bench or experimental
GenreCandidate signal: Empirical · Consensus signal: Empirical
Teacher disagreement score0.001
Threshold uncertainty score0.004

Distilled classifier scores by category (both heads)

CategoryCodexGemma
Metaresearch0.0000.000
Meta-epidemiology (narrow)0.0000.000
Meta-epidemiology (broad)0.0000.000
Bibliometrics0.0000.000
Science and technology studies0.0000.000
Scholarly communication0.0000.000
Open science0.0000.000
Research integrity0.0000.001
Insufficient payload (model declined to judge)0.0010.000

Machine scores (provisional)

The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.

Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.

Opus teacher head0.003
GPT teacher head0.202
Teacher spread0.199 · how far apart the two teachers sit on this one work
Validation statusscore_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from it

Classification

machine, unvalidated

Machine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.

The models applied no category: nothing in the taxonomy fit this work.
Study designBench or experimental
Domainnot available
GenreEmpirical

How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".

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Citations0
Published2021
Admission routes2
Has abstractyes

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Same venueThe FASEB JournalSame topicRenal and related cancersFrench-language works237,207