Our research shows that while some PTEN-deleted axons can extend past the injury, scar tissue remains a major barrier. To overcome this, we are developing a dual combinatorial approach that uses AAV we are using a dual strategy: we promote regeneration by silencing PTEN, and we clear obstacles by delivering chABC—an enzyme that breaks down scar tissue. Therefore, we aim to utilize AAV-chABC in combination with AAV-driven PTEN deletion to assess whether the scar is a major impediment for growth for PTEN-deleted axons.
AAV-chABC injections were administered into the spinal cords of mice. Specifically, AAVs were injected at the cervical level 5 (C5) in the spinal cords of uninjured mice. Panels A-B illustrate that AAV-GFP (control) injections did not result in detectable chABC (A) or C-4-S immunofluorescence (B), indicating a lack of chABC enzymatic activity. Conversely, panels C-D demonstrate that AAV-chABC resulted in marked chABC expression (C) and C-4-S immunofluorescence (D) (scale bar 100 µm). These results confirm the feasibility of utilizing AAVs to deliver functional chABC into the spinal cord of mice.