476 Pseudotyped Adeno Associated Virus Gene Transfer in Wound Healing Molecular Therapy �������� ��� ���� ���������������� �������� ���� ������© ����������� �!����� ����"� �������� S186 ��������� � ��[.]
APPLICATIONS IN DISEASE hEPO or rAd5-hEPO, respectively, via intraoral cannulation of both submandibular glands A third group (n=3) received 108 particles of rAAV2-hEPO Additionally, a fourth group (n=3) received 1011 genomes of naked CMV-hEPO-SV40polyA linear DNA (hEPODNA) cut from the plasmid used for the rAd5-hEPO construction Hematocrit (Hct) and serum hEPO levels were measured over a 28week period after vector administration Two days prior to administration, mean Hct levels were 59.8±2% (mean ± SD) and hEPO levels were undetectable in all groups Following vector administration, mean Hct and hEPO levels in the rAd5-hEPO-treated group were maximal on week (75±3% and 28±2 mU/ml, respectively) and returned to pre-administration values (60.5±3% and zero, respectively) by week in all but one animal hEPO levels in this particular animal remained >6mU/ml up until its death at week 10 without any effect on Hct levels (~61%) hEPO levels in the group infected with 2.5x109 particles of rAAV2-hEPO increased over the first weeks, remained stable (~24±36 mU/ml) until week 26 and began to decline thereafter (week 28 ; 13±24 mU/ml) Mean Hct levels in this group reached 75±13% by week 4, further increased to 81±14% at week 10, and remained stable until the end of the experiment We measured hEPO in saliva at week 5, and then values were minimal (0.6±1 mU/ml), indicating preferential secretion of hEPO into the bloodstream hEPO and Hct levels were lower for animals in the group infected with 108 particles of rAAV2-hEPO In the mice treated with linear hEPO-DNA, Hct levels remained normal and hEPO levels were undetectable throughout the experiment Our results indicate that stable and long-term production of therapeutic levels of EPO in the serum can be achieved by administering low doses of rAAV2 vector to the salivary glands 475 Delivery of α1-Antitrypsin to the Lung Via Adeno-Associated Virus Vector-Mediated Gene Transfer Is More Efficient by Intrapleural Delivery Rather Than Skeletal Muscle Administration Bishnu P De,1 Robert Merritt,1 Michael Lam,1 Neil R Hackett,1 Ronald G Crystal.1 Weill Medical College of Cornell University, New York, NY, United States α1-antitrypsin (α1AT), a serine proteinase inhibitor synthesized and secreted by the liver, protects the lung from degradation by neutrophil proteases α1AT deficiency is a common autosomal recessive disorder associated with the accelerated development of emphysema if serum α1AT levels are ≤11 μM (570 μg/ml) Several studies have indicated that adeno-associated virus (AAV)-mediated delivery of α1AT is a promising candidate for a gene therapy for this disease Because the change in scale from mice to adult humans will require higher efficiency of gene expression, we hypothesized that intrapleural administration of an AAV vector expressing α1AT may achieve higher levels of α1AT in the lung compared to administration of the vector at a distant organ To evaluate this hypothesis, a serotype AAV vector expressing the α1AT cDNA under the control of cytomegalovirus-chicken β-actin hybrid promoter (AAV2α1AT) was administered to C57Bl/6 mice by various routes A typical batch of the purified vector had