MP49-04 MATRIX METALLOPROTEINASE-9 CONTROLS SECRETION OF NERVE GROWTH FACTOR FROM BLADDER CELLS IN VITRO
Bibliographic record
Abstract
You have accessJournal of UrologyCME1 May 2022MP49-04 MATRIX METALLOPROTEINASE-9 CONTROLS SECRETION OF NERVE GROWTH FACTOR FROM BLADDER CELLS IN VITRO Aya Hajj, Aalya Hamouda, Stephanie Sirmakesyan, Philippe Cammisotto, and Lysanne Campeau Aya HajjAya Hajj More articles by this author , Aalya HamoudaAalya Hamouda More articles by this author , Stephanie SirmakesyanStephanie Sirmakesyan More articles by this author , Philippe CammisottoPhilippe Cammisotto More articles by this author , and Lysanne CampeauLysanne Campeau More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002624.04AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Urine of female patients with overactive bladder syndrome (OAB) is characterized by low levels of the neurotrophin NGF with stable levels of its precursor proNGF. The resulting imbalance between NGF and proNGF may originate from an elevated activity of the proteolytic enzyme matrix metalloproteinase-9 (MMP-9) observed in the same samples. On the other hand, an inhibitor of the proinflammatory p75NTR receptor, THX-B, restored NGF levels to normal in a murine model of diabetic voiding dysfunction (Mossa et al 2020). The aim of the present work is to: 1) determine if NGF and MMP-9 are synthesized by cells of the bladder; 2) assess the role of MMP-9 in the synthesis and release of NGF by the same cells; and 3) examine how NGF secretion and MMP-9 might be affected by THX-B. METHODS: Rat bladders were digested by collagenase type IV to generate primary cultures of urothelial and smooth muscle cells. The expression of NGF, proNGF and MMP-9 were assessed by RT-qPCR and by immunoblotting. Cellular localisation of proteins was obtained by immunohistochemistry. Knock-down of MMP-9 was achieved using Crispr-Cas9. Levels of NGF and proNGF were measured by ELISA and enzyme activities by specific enzymatic kits. RESULTS: Urothelial and smooth muscle cells were found to contain and release significant amounts of NGF, proNGF and MMP-9 as revealed by RT-qPCR, immunoblotting and microscopy. The knock-down of MMP-9 using Crispr-Cas9 resulted in the disappearance of MMP-9 mRNA, proteins and enzymatic activity, in both cell types. In the MMP-9 knock-down cells, levels of secreted NGF were multiplied by a factor 4 to 9 while proNGF concentrations were not affected. On the other hand, incubation of urothelial cells with the p75 antagonist THX-B (5 µg/mL) for 24 hours increased NGF secretion and concomitantly decrease the activity of MMP-9. MMP-7, the enzyme converting proNGF to NGF, displayed an increased activity as well. On the other hand, THX-B had no effect on smooth muscle cells. Levels of mRNA were not affected by THX-B in both cell types. Pathways associated to p75NTR, namely erk, jnk, p38MAPK and cyclic AMP were also unchanged by treatment with THX-B. CONCLUSIONS: Bladder cells express, synthesize and release NGF, proNGF and MMP-9. The latter is central in the control of NGF secretion. THX-B targets urothelial cell to enhance NGF secretion by downregulation of MMP-9 and increased in MMP-7 activities, which could explain the improvement of diabetic voiding dysfunction in vivo. Source of Funding: Canadian Urology Association © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e854 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Aya Hajj More articles by this author Aalya Hamouda More articles by this author Stephanie Sirmakesyan More articles by this author Philippe Cammisotto More articles by this author Lysanne Campeau More articles by this author Expand All Advertisement PDF downloadLoading ...
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot 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.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.001 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.001 |
| Insufficient payload (model declined to judge) | 0.021 | 0.005 |
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.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
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".