Description
- Abstract:
- Objective: Hindered drug delivery due to the presence of the blood-tumor barrier (BTB) is one of the limiting factors in effective glioblastoma therapy. We intend to investigate regulatory mediators involved in the brain endothelial integrity and vascular permeability in glioblastoma, using a small molecule 6-bromoindirubin acetoxime (BIA) as a model compound. Methods: RNA-sequencing data from cultured human cerebral microvascular endothelial (hCMEC/D3) cells treated with BIA or vehicle control were analyzed and genes were prioritized based on fold-change and known functions. Expression levels of the lead candidates, including the junctional protein VE-cadherin (CDH5) and transcription factors ETV1 and ERG, were assessed by RT-qPCR, Western blot and immunofluorescence on cultured hCMEC/D3 and primary human brain microvascular endothelial (HBMEC) cells, as well as in vivo treated murine glioblastoma xenograft tissue samples. The specificity and enhancement of drug penetration at the BTB following BIA treatment in vivo were validated by TRITC-dextran and sodium fluorescein tracers. Barrier formation of hCMEC/D3 cells upon ETV1 or ERG inhibition was tested using the electric cell-substrate impedance sensing (ECIS) assay. A human phospho- kinase array was used to screen for possible kinase targets involved in BIA’s mechanism of action. Results: BIA-induced reduction of VE-cadherin expression was consistently observed. ETV1 and ERG expression levels were downregulated after BIA treatment. siRNA of ETV1 seemed to partially inhibit endothelial barrier formation. BIA appeared to inactivate a broad spectrum of kinases, including Yes, p38α, STAT2 and STAT5α/STAT5b. Conclusions: BIA reduces endothelial permeability at the BTB likely via modulation of kinases (i.e. p38α) and transcription factors (i.e. ETV1) that control VE-cadherin expression.
- Notes:
- Thesis (Sc. M.)--Brown University, 2023
Citation
Xu, Zheyun,
"Modulating Endothelial Permeability to Improve Drug Delivery in Glioblastoma"
(2023).
Biotechnology, Biology and Medicine Theses and Dissertations.
Brown Digital Repository. Brown University Library.
https://repository.library.brown.edu/studio/item/bdr:nxh39k24/
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Biology and Medicine Theses and Dissertations
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