Abstract 5815: Therapeutic Targeting of the Calmodulin-Cyclin E Interaction in Vascular Smooth Muscle Cells
Bibliographic record
Abstract
Background: Cell cycle regulation in vascular smooth muscle cells (VSMC) is a putative target in proliferative vascular diseases. Having shown that increases in intracellular [Ca 2+ ] are tightly coordinated and critical for VSMC proliferation, we also discovered that binding of Ca 2+ /calmodulin (CaM) to cyclin E/cdk2 is necessary for Ca 2+ -sensitive G 1 -to-S phase progression in VSMC. To test the importance of this, we developed a synthetic C aM- b inding sequence (CBS) that blocked CaM-cyclin E interactions, prevented activation of cdk2, and abrogated Ca 2+ -sensitive G1-to-S transitions in VSMC. Of note, binding of CaM to another target protein, calcineurin, was not altered by CBS. Objective: To evaluate the physiological relevance of targeting CaM-cyclin E interactions in human VSMC in vitro and in a mouse model of neointima formation in vivo. Methods & Results: Human aortic SMC (HA-SMC) were treated with the CBS peptide conjugated to the viral TAT transduction domain to increase bioavailability. Peptide delivery of TAT-CBS was confirmed by immunochemistry. TAT-CBS caused a dose-dependent anti-proliferative effect in HA-SMC, as well as in human aortic endothelial cells and mouse embryonic fibroblasts (MEF) as measured by 3 H-thymidine incorporation. Importantly, these effects were lost in MEF from cyclin E1/E2 double knockout mice, further supporting target specificity for CaM/cyclin E. Moreover, TAT-CBS treatment was not cytotoxic, as it did not increase either LDH or caspase-3 release compared to controls. In vivo application of TAT-CBS in pluronic gel to the outside of injured mouse carotid arteries significantly decreased neointima formation and medial thickening without affecting re-endothelialization as compared to animals treated with vehicle-only or negative-control peptide. Summary: Inhibition of CaM binding to cyclin E/cdk2 with the TAT-CBS peptide significantly decreases HA-SMC proliferation in a dose-dependent manner in vitro. Moreover, application of TAT-CBS-containing gel to injured carotid arteries inhibits VSMC proliferation without affecting re-endothelialization. These data support TAT-CBS as a novel therapy for restenosis, and the general approach of targeting Ca 2+ -sensitive cell cycle control in VSMC.
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How this classification was reachedexpand
Full frame distilled prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.
Codex and Gemma teacher scores by category
| 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 | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.000 | 0.000 |
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.
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 teacher head, 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".