High-Frequency Irreversible Electroporation (H-FIRE) Induced Blood–Brain Barrier Disruption Is Mediated by Cytoskeletal Remodeling and Changes in Tight Junction Protein Regulation

Biomedicines

卷:10

Melvin F. Lorenzo¹  Sabrina N. Campelo¹  Rafael V. Davalos¹  John L. Robertson²  Scott S. Verbridge²  Irving C. Allen²  Fang-Chi Hsu³  Brittanie R. Partridge⁴  Jonathan Hinckley⁴  John H. Rossmeisl⁴  Yukitaka Kani⁴ 

[1] Department of Biomedical Engineering and Mechanics, Virginia Tech, Blacksburg, VA 24061, USA;

[2] Department of Biomedical Sciences and Pathobiology, Virginia Tech, Blacksburg, VA 24061, USA;

[3] Department of Biostatistics and Data Sciences, Division of Public Health Sciences, Wake Forest University School of Medicine, Winston-Salem, NC 27101, USA;

[4] Department of Small Animal Clinical Sciences, Virginia Tech, Blacksburg, VA 24061, USA;

关键词: high-frequency irreversible electroporation (H-FIRE);    blood–brain barrier;    glioma;    intracranial drug delivery;   

DOI  :  10.3390/biomedicines10061384

来源: DOAJ

【 摘 要 】

Glioblastoma is the deadliest malignant brain tumor. Its location behind the blood–brain barrier (BBB) presents a therapeutic challenge by preventing effective delivery of most chemotherapeutics. H-FIRE is a novel tumor ablation method that transiently disrupts the BBB through currently unknown mechanisms. We hypothesized that H-FIRE mediated BBB disruption (BBBD) occurs via cytoskeletal remodeling and alterations in tight junction (TJ) protein regulation. Intracranial H-FIRE was delivered to Fischer rats prior to sacrifice at 1-, 24-, 48-, 72-, and 96 h post-treatment. Cytoskeletal proteins and native and ubiquitinated TJ proteins (TJP) were evaluated using immunoprecipitation, Western blotting, and gene-expression arrays on treated and sham control brain lysates. Cytoskeletal and TJ protein expression were further evaluated with immunofluorescent microscopy. A decrease in the F/G-actin ratio, decreased TJP concentrations, and increased ubiquitination of TJP were observed 1–48 h post-H-FIRE compared to sham controls. By 72–96 h, cytoskeletal and TJP expression recovered to pretreatment levels, temporally corresponding with increased claudin-5 and zonula occludens-1 gene expression. Ingenuity pathway analysis revealed significant dysregulation of claudin genes, centered around claudin-6 in H-FIRE treated rats. In conclusion, H-FIRE is capable of permeating the BBB in a spatiotemporal manner via cytoskeletal-mediated TJP modulation. This minimally invasive technology presents with applications for localized and long-lived enhanced intracranial drug delivery.

【 授权许可】

Unknown