Project Title

Synthesis of 11-[2-arylmethylene)hydrazono]-PBD Derivatives and Evaluation of Their Effects on CB2-Mediated Smooth Muscle Cell Trans-Differentiation to an Osteogenic Phenotype

Authors' Affiliations

Marilyn Hagar, Department of Chemistry, College of Arts and Sciences, East Tennessee State University, Johnson City, TN Dr. Abbas Shilabin, Department of Chemistry, College of Arts and Sciences, East Tennessee State University, Johnson City, TN Dr. Douglas Thewke, Department of Biomedical Sciences, Quillen College of Medicine, East Tennessee State University, Johnson City, TN.

Location

Culp Room 210

Start Date

4-6-2022 11:00 AM

End Date

4-6-2022 11:15 AM

Faculty Sponsor’s Department

Chemistry

Name of Project's Faculty Sponsor

Abbas Shilabin

Additional Sponsors

Abbas Shilabin, Douglas Thewke, Dane Scott

Classification of First Author

Undergraduate Student

Competition Type

Non-Competitive

Type

Boland Symposium

Project's Category

Biological and Chemical Sciences

Abstract or Artist's Statement

Atherosclerotic disease is characterized by the formation of lipid-ladden plaques in artery walls. During later stages of disease, these plaques become calcified by mechanisms involving the trans-differentiation of vascular smooth muscle cells (VSMC) to osteoblast-like cells. Although vascular calcification was thought to be a passive mechanism, evidence shows that this process is heavily modulated by various cell signaling mechanisms, including CB2 endocannabinoid receptors. Previous studies have shown that known CB2 antagonists accelerate VSMCs trans-differentiation to an osteoblast-like phenotype, indicating that this receptor serves an anti-calcification signal. The goal of this investigation is to determine if a series of 11-[2-arylmethylene)hydrazono]-PBD derivatives with established CB2 binding affinity function as CB2 antagonists or agonists in a cell culture model of VSMC osteoblastic trans-differentiation. MOVAS cells were grown in standard media or osteogenic media (to induce trans-differentiation) supplemented with and without the various PBD derivatives. Following the treatment period, the extent of osteoblast-like activity was evaluated by alizarin red staining for calcium deposition. To quantify the staining present, the dye was extracted using cetylpyridinium chloride hydrate solution and then analyzed via UV-Vis spectroscopy at 570 nm. The ability of the derivatives to modulation of osteoblastic transdifferentiation of MOVAS cells was further evaluated by performing Western blot analysis for expression of Runx2, an essential transactivator of osteoblast differentiation. Results of this work determined that some of the PBD derivatives increased the calcification compared to the control, indicating that they likely act as CB2 receptor antagonists, while others decreased calcification compared to the control, indicating that they likely act as CB2 receptor agonists. Not only do these results characterize the interactions of these compounds with CB2 receptors, they demonstrate that these PBD derivatives have biological activity. These results also further implicate CB2 receptors as a regulator of VSMC cell calcification, which could lead to novel drug therapies for the treatment of atherosclerotic plaques.

This document is currently not available here.

Share

COinS
 
Apr 6th, 11:00 AM Apr 6th, 11:15 AM

Synthesis of 11-[2-arylmethylene)hydrazono]-PBD Derivatives and Evaluation of Their Effects on CB2-Mediated Smooth Muscle Cell Trans-Differentiation to an Osteogenic Phenotype

Culp Room 210

Atherosclerotic disease is characterized by the formation of lipid-ladden plaques in artery walls. During later stages of disease, these plaques become calcified by mechanisms involving the trans-differentiation of vascular smooth muscle cells (VSMC) to osteoblast-like cells. Although vascular calcification was thought to be a passive mechanism, evidence shows that this process is heavily modulated by various cell signaling mechanisms, including CB2 endocannabinoid receptors. Previous studies have shown that known CB2 antagonists accelerate VSMCs trans-differentiation to an osteoblast-like phenotype, indicating that this receptor serves an anti-calcification signal. The goal of this investigation is to determine if a series of 11-[2-arylmethylene)hydrazono]-PBD derivatives with established CB2 binding affinity function as CB2 antagonists or agonists in a cell culture model of VSMC osteoblastic trans-differentiation. MOVAS cells were grown in standard media or osteogenic media (to induce trans-differentiation) supplemented with and without the various PBD derivatives. Following the treatment period, the extent of osteoblast-like activity was evaluated by alizarin red staining for calcium deposition. To quantify the staining present, the dye was extracted using cetylpyridinium chloride hydrate solution and then analyzed via UV-Vis spectroscopy at 570 nm. The ability of the derivatives to modulation of osteoblastic transdifferentiation of MOVAS cells was further evaluated by performing Western blot analysis for expression of Runx2, an essential transactivator of osteoblast differentiation. Results of this work determined that some of the PBD derivatives increased the calcification compared to the control, indicating that they likely act as CB2 receptor antagonists, while others decreased calcification compared to the control, indicating that they likely act as CB2 receptor agonists. Not only do these results characterize the interactions of these compounds with CB2 receptors, they demonstrate that these PBD derivatives have biological activity. These results also further implicate CB2 receptors as a regulator of VSMC cell calcification, which could lead to novel drug therapies for the treatment of atherosclerotic plaques.