AAV Gene Therapy Strategies for Lysosomal Storage Disorders with Central Nervous System Involvement - Gene Delivery and Therapy for Neurological Disorders - page 289

Fig. 8 NHP within MRI-compatible stereotaxic frame and placement of bilateral stainless steel guidance rods.
Snapshots of navigation module used for convection-enhanced delivery. ( a ) Fluoroscopy ( gray scale ) fused
with presurgical MRI ( blue scale ); ( b ) cone-beam CT ( red scale ) can be used for real-time 3D navigation of
sheaths using multiple oblique views; ( c ) axial view

Adjust trajectories as needed to target the desired structures
(Fig. 8 ), and acquire new dataset to confi rm accuracy of adjusted
trajectory and determine depth from dura to the desired target.

11. Load AAV vector into a 1 mL Luer-Lok™ syringe (BD
3096328) and remove any air bubbles. Attach to a 16G
SmartFlow™ neuroventricular cannula (10 ft line length,
NGS-NC-04, MRI Interventions, Inc), and prime it until fl uid
comes out at the tip.
12. Insert cannulas in the frames and lock. Place the syringes in a
PHD2000 Harvard Pump (Harvard Apparatus). Lower the
cannulas to the correct depth for each target structure.
13. Acquire high-resolution CBCT [ 110 ], and fuse with presurgi-
cal MRI to confi rm correct positioning of the cannula
(Fig. 9 ). Start infusion of AAV vector. In our experiments, we
have used the following parameters with no evidence of
adverse effects b :
(a) Thalamus: 150 ʼ L at 2 ʼ L/min—simultaneously into both

thalami
(b) Cerebral lateral ventricle: 300 L/min

14. Upon completion of the injection, wait for 10 min before
withdrawing the cannula.
15. Close the dura with 3-0 vicryl sutures, and scalp with 3-0
monocryl sutures.
16. Move NHP to the recovery room, and monitor until fully
awake. Administer analgesics for 2 days following surgery.

ʼ L at 50 ʼ