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Design of a Gene Transfer Vector to Deliver a Stabilized Anti‐EGFR RNA Aptamer to the Glioblastoma Microenvironment
Author(s) -
Parikh Sachin,
Hicks Martin J
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.911.5
Subject(s) - epidermal growth factor receptor , microvesicles , cancer research , rna , receptor tyrosine kinase , glioma , biology , angiogenesis , aptamer , brain tumor , genetic enhancement , tyrosine kinase , gene , microrna , receptor , signal transduction , medicine , microbiology and biotechnology , pathology , biochemistry
Glioblastoma multiforme (GBM) is an incurable and aggressive type of brain tumor. It is the most common central nervous system (CNS) malignancy with a median survival of only 14 months. The epidermal growth factor receptor (EGFR) is a type of tyrosine kinase receptor (TKR) dysregulated in about 60% of GBM tumors. EGFR amplification and over‐expression leads to angiogenesis and uncontrolled growth and proliferation of GBM. Although a great deal is known about the biology exhibited by EGFR‐activated GBM, the application of therapies against the biologic processes is limited by the blood‐brain barrier, which restricts systemically administered therapies from reaching the brain. We have created an in vivo tissue culture model to develop a novel strategy to bypass these barriers by developing a gene transfer vector to deliver the genetic sequences of a verified anti‐EGFR RNA therapy aptamer that binds with high affinity to EGFR. To stabilize the structure, we have added inactivated hammerhead ribozymes, and to promote extracellular movement of the aptamer, we use an extracellular RNA “exRNA” localization element. Methodologies include RNA isolation, reverse‐transcription and PCR to detect changes in EGFR transcript expression, as well as ELISA to measure changes in the phosphorylated and active form of EGFR. With the revelation that GBM leads to an abundant increase in exosomes and microvesicles, this strategy may have implications for future therapies directed against GBM.