A Comprehensive Map of CNS Transduction by Eight Recombinant Adeno-associated Virus Serotypes Upon Cerebrospinal Fluid Administration in Pigs

Abstract

Cerebrospinal fluid administration of recombinant adeno-associated viral (rAAV) vectors has been demonstrated to be effective in delivering therapeutic genes to the central nervous system (CNS) in different disease animal models. However, a quantitative and qualitative analysis of transduction patterns of the most promising rAAV serotypes for brain targeting in large animal models is missing. Here, we characterize distribution, transduction efficiency, and cellular targeting of rAAV serotypes 1, 2, 5, 7, 9, rh.10, rh.39, and rh.43 delivered into the cisterna magna of wild-type pigs. rAAV9 showed the highest transduction efficiency and the widest distribution capability among the vectors tested. Moreover, rAAV9 robustly transduced both glia and neurons, including the motor neurons of the spinal cord. Relevant cell transduction specificity of the glia was observed after rAAV1 and rAAV7 delivery. rAAV7 also displayed a specific tropism to Purkinje cells. Evaluation of biochemical and hematological markers suggested that all rAAV serotypes tested were well tolerated. This study provides a comprehensive CNS transduction map in a useful preclinical large animal model enabling the selection of potentially clinically transferable rAAV serotypes based on disease specificity. Therefore, our data are instrumental for the clinical evaluation of these rAAV vectors in human neurodegenerative diseases.

Development of Intrathecal AAV9 Gene Therapy for Giant Axonal Neuropathy

“…IT delivery has the added benefits of reduced targeting of virus to peripheral organs and avoidance of AAV9-neutralizing antibodies in the blood as compared to systemic delivery. 17 , 18 , 19 , 37 The biodistribution of AAV9 in small and large animals has been validated across multiple labs, 19 , 21 , 38 and here we show that the distribution of gigaxonin expression 1 year post-IT injection of AAV9/JeT-GAN in mice agrees with that data. Further, IT delivery of vector is a minimally invasive route of injection and is scalable to humans.…”

“…Studies of IT delivery of AAV9 in mice and NHPs show a dose response wherein a higher dose correlated to a higher percentage of transduced cells. 20 , 21 , 38 Notably, none of these studies reached a saturating dose of AAV9, and we have shown that our construct design for GAN is unlikely to introduce overexpression-related toxicity. Together, these data support the safety of increasing the dose administered in patients in order to increase the percent of transduced cells in the spinal cord and brain and maximize the therapeutic benefit.…”

Age-Related 1H NMR Characterization of Cerebrospinal Fluid in Newborn and Young Healthy Piglets

“…[ 28 ]. Due to the lack of specific data, every time point can provide important insights, and the 30 days old model has recently been used to describe new techniques [ 17 , 29 ] and characterize new gene therapy patterns [ 30 ].…”

Gene therapy for Mucopolysaccharidoses

“…Intrathecal (IT) or IV administration of rAAV9 is a less invasive systemic delivery method that efficiently transduces genes into multiple tissues, including the CNS. Delivery of various rAAV serotypes to CNS lesions via CSF administration in a large animal has been studied [63–65]. These studies show that rAAV9 has high transduction efficiency in widespread CNS lesions including the spinal cord.…”