Date of Award
2026-05-01
Degree Name
Master of Science
Department
Biological Sciences
Advisor(s)
Charlotte C. Vines
Abstract
C-C chemokine receptor 7 (CCR7) is a G-protein-coupled receptor (GPCR) that is important for immune cell trafficking to and within secondary lymphoid organs, upon stimulation by the ligands CCL19 and CCL21. CCR7 activation by CCL19 promotes T-ALL migration across the blood-brain barrier (BBB) into the Central Nervous System (CNS), where the leukemic T-cells escape standard chemotherapies. Our first study explored the use of the CCL19 antagonist CCL198-83 to inhibit CCL19/CCR7-mediated functions. We identified a targetable cell-surface receptor, CCR7, that can be inhibited to prevent β2-integrin-mediated T-ALL invasion of the CNS and that may serve as a platform for pharmacological inhibition of T-ALL entry into the CNS. Additionally, we have designed a 3D-printed MFD, printed with a newly characterized biocompatible BioMed Clear Resin V1, that allows an in vitro human BBB model that closely recapitulates the physiological state, including: (1) shear stress of 10-20 dyne/cm2; which alone promotes cell adhesion, (2) the presence of human brain microvascular endothelial cells, primary human brain pericytes, and human astrocytes encapsulated in (3) a collagen type IV/ fibronectin extracellular matrix hydrogel. Finally, since GPCRs are regulated by G-protein-coupled receptor kinases (GRKs), we focused on uncovering the role of GRK6 in regulating CCR7-mediated signaling and cellular behavior. Using an orthogonal approach to target CCR7 signaling via CCL19, we used a CRISPR/Cas9 lentiviral system to disrupt GRK6 in T cells. We have generated and validated, by Western blot analysis, T cells with reduced GRK6 expression. Our findings have demonstrated that GRK6 differentially regulates CCL21/CCR7 signaling in a ligand-dependent manner, limiting CCR7-mediated chemotaxis and promoting calcium mobilization in a normal T cell environment. By biasing pharmacological platforms to target inhibition of CCL19 signaling and block CNS migration, these GRK6-modulated regulatory events can be exploited in drug development studies to bias signaling towards a more "favorable" effect.
Language
en
Provenance
Received from ProQuest
Copyright Date
2026-05
File Size
144 p.
File Format
application/pdf
Rights Holder
Cesar Ivan Cardona
Recommended Citation
Cardona, Cesar Ivan, "Development of a Physiologically Relevant 3D In Vitro Model of the Human Blood-Brain Barrier to Study the Mechanisms of CCR7-Induced T-All Invasion of the Central Nervous System" (2026). Open Access Theses & Dissertations. 4644.
https://scholarworks.utep.edu/open_etd/4644