Using an In Vivo Mouse Model to Determine the Exclusion Criteria of Preexisting Anti-AAV9 Neutralizing Antibody Titer of Pompe Disease Patients in Clinical Trials.

The efficacy of adeno-associated virus (AAV)-based gene therapy is dependent on effective viral transduction, which might be inhibited by preexisting immunity to AAV acquired from infection or maternal delivery. Anti-AAV neutralizing Abs (NAbs) titer is usually measured by in vitro assay and used for patient enroll; however, this assay could not evaluate NAbs' impacts on AAV pharmacology and potential harm in vivo. Here, we infused a mouse anti-AAV9 monoclonal antibody into Balb/C mice 2 h before receiving 1.2 × 1014 or 3 × 1013 vg/kg of rAAV9-coGAA by tail vein, a drug for our ongoing clinical trials for Pompe disease. The pharmacokinetics, pharmacodynamics, and cellular responses combined with in vitro NAb assay validated the different impacts of preexisting NAbs at different levels in vivo. Sustained GAA expression in the heart, liver, diaphragm, and quadriceps were observed. The presence of high-level NAb, a titer about 1:1000, accelerated vector clearance in blood and completely blocked transduction. The AAV-specific T cell responses tended to increase when the titer of NAb exceeded 1:200. A low-level NAbs, near 1:100, had no effect on transduction in the heart and liver as well as cellular responses, but decreased transduction in muscles slightly. Therefore, we propose to preclude patients with NAb titers > 1:100 from rAAV9-coGAA clinical trials.

[Routing reseach of vas deferens on anatomy and clinical application].

OBJECTIVE: Studying on the routes of vas deferens to provide anatomy basis for surgical operation, especially, reconstruction of long segment loss of vas deferens. METHODS: The routes of vas deferens were observed and anatomic distances along epididymal, infrainguinal, inguinal, retroperitoneal and ampullar segments of vas deferens, the distances from external ring to extremity of vas deferens were measured respectively in 18 formalin fixed adult cadavers. RESULTS: The vas deferens have a large curve from external ring to extremity in its route, draw it out from the external ring. Eliminating this curve will allow to shorten this segment of vas deferens for vasovasostomy by 6.1 - 12.9 (9.31 +/- 1.30) cm. The length of each segment of vas deferens, respectively, is epididymal: 3.2 - 5.6 (4.53 +/- 0.79) cm, infrainguinal: 4.5 - 9.5 (7.31 +/- 1.78) cm, inguinal: 4.4 - 7.5 (5.52 +/- 0.74) cm, retroperitoneal: 12.5 - 19.5 (16.75 +/- 1.87) cm and ampullar: 2.9 - 3.8 (3.63 +/- 0.23) cm. There was no significant differences in segment length and the distances from external ring to extremity of vas deferens between the right and left. CONCLUSION: Reconstruction of long segment loss of vas deferens can be performed by mobilization retroperitoneal vas deferens and draw it out from external ring. There were no significant differences in lengths of vas deferens and the distances from external ring to vassal extremity between the left and right in adults. The surgical operations of vas deferens are closely related each segment of vasa.