Bispecific RNA Aptamers for Mediating Blood-Brain Barrier Translocation via Insulin Receptors and Precision Targeting of EGFRvIII+ Glioblastoma Cells

Glioblastoma Multiforme (GBM) is an incredibly aggressive and highly deadly form of brain cancer, with an average survival rate of only 14 months. Many forms of current treatment, such as surgical resection and chemotherapy, have damaging side effects and risk chances of recurrence. Thus, there is a dire need for novel treatment strategies. Recently, a development in bispecific aptamers (short single-stranded nucleic acids) enables the usage of insulin receptors (IR) to cross the brain’s blood-brain barrier (BBB) and target cancer cells. Aptamers can carry an anticancer medication and transport it to cancerous cells. Notably, GBM cells are characterized by the Epidermal Growth Factor Receptor Variant III (EGFRvIII), a mutant form of EGFR, is a promising target for aptamers. I hypothesize that these aptamers will bind to the specific binding sites of IR and EGFRvIII to aid drug transportation crossing the BBB and target cancer receptors. This research utilizes computational techniques, including protein and aptamer modeling, molecular docking simulators, and binding site analysis to identify a bispecific aptamer for IR and EGFRvIII. AlphaFold3, a machine learning-based method, and FARFAR2 provide the 3D structure for the receptors and aptamers, respectively. To predict binding interactions, a deep learning-based approach software, ScanNet, can analyze APT1’s successful binding to the IR. Furthermore, APT1 demonstrated selective binding to the EGFRvIII variant without interacting with EGFR, confirming its specificity and targeted affinity. The aptamer A1 was selected as the most appropriate candidate because it will bind to the EGFRvIII cancer form and get transported across the BBB through IR. However, it will not bind to the wt EGFR. Current research will further develop these aptamers and facilitate the system's design, revolutionizing cancer treatment.