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Intramuscular injection of AAV8 in mice and macaques is associated with substantial hepatic targeting and transgene expression.

Greig JA, Peng H, Ohlstein J, Medina-Jaszek CA, Ahonkhai O, Mentzinger A, Grant RL, Roy S, Chen SJ, Bell P, Tretiakova AP, Wilson JM - PLoS ONE (2014)

Bottom Line: We found that following IM administration of AAV8 vectors in mice, a portion of the vector reached the liver and hepatic gene expression contributed significantly to total expression of secreted transgenes.Hepatic distribution of vector following IM injection was also noted in rhesus macaques.These pre-clinical data on AAV delivery should inform safe and efficient development of future AAV products.

View Article: PubMed Central - PubMed

Affiliation: Gene Therapy Program, Department of Pathology and Laboratory Medicine, Division of Transfusion Medicine, University of Pennsylvania, TRL Suite 2000, 125 South 31st Street, Philadelphia, PA, 19104, United States of America.

ABSTRACT
Intramuscular (IM) administration of adeno-associated viral (AAV) vectors has entered the early stages of clinical development with some success, including the first approved gene therapy product in the West called Glybera. In preparation for broader clinical development of IM AAV vector gene therapy, we conducted detailed pre-clinical studies in mice and macaques evaluating aspects of delivery that could affect performance. We found that following IM administration of AAV8 vectors in mice, a portion of the vector reached the liver and hepatic gene expression contributed significantly to total expression of secreted transgenes. The contribution from liver could be controlled by altering injection volume and by the use of traditional (promoter) and non-traditional (tissue-specific microRNA target sites) expression control elements. Hepatic distribution of vector following IM injection was also noted in rhesus macaques. These pre-clinical data on AAV delivery should inform safe and efficient development of future AAV products.

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Comparison of expression from the liver-specific TBG promoter following IM and IV vector administration in RAG KO mice.Expression on day 28 post-vector administration of 201Ig IA (A), 2.10A mAb (B), VRC01 mAb (C), and PG9 mAb (D) in RAG KO mice. Expression following IM injection of vectors with either the CMV or TBG promoter was compared to expression levels produced following IV injection of the TBG vector. Mice were injected IM with two 15 µl injections into the right and left gastrocnemius muscles. IV injections were performed as a 100 µl injection via the tail vein. Mice were administered with a dose of either 1010 GC or 1011 GC, numbers indicate dose ×1011 GC. ND, not determined. Expression was measured in serum by ELISA and values are expressed as mean ± SEM (n = 3/group). ***p<0.001.
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pone-0112268-g002: Comparison of expression from the liver-specific TBG promoter following IM and IV vector administration in RAG KO mice.Expression on day 28 post-vector administration of 201Ig IA (A), 2.10A mAb (B), VRC01 mAb (C), and PG9 mAb (D) in RAG KO mice. Expression following IM injection of vectors with either the CMV or TBG promoter was compared to expression levels produced following IV injection of the TBG vector. Mice were injected IM with two 15 µl injections into the right and left gastrocnemius muscles. IV injections were performed as a 100 µl injection via the tail vein. Mice were administered with a dose of either 1010 GC or 1011 GC, numbers indicate dose ×1011 GC. ND, not determined. Expression was measured in serum by ELISA and values are expressed as mean ± SEM (n = 3/group). ***p<0.001.

Mentions: AAV8 vectors expressing anti-SIV/SHIV antibodies (in an immunoadhesin format [i.e., 201Ig IA] or a monoclonal antibody format [i.e., 2.10A mAb] [40]) or human anti-HIV antibodies in a monoclonal antibody format (i.e., VRC01 mAb [41], [42] and PG9 mAb [43]) were injected IM as two 15 µl injections at doses of 1010 GC or 1011 GC into RAG KO mice (Figure 2). Expression of these proteins in serum was determined on day 28 post-vector administration and compared to expression from TBG-containing vectors administered intravenously (IV) via the tail vein. Figure 2 summarizes data from these experiments, in which the two doses of vectors (labeled 0.1 and 1) were evaluated in the context of two comparisons: the CMV versus TBG vectors following IM injection and the TBG vector following IM and IV injection.


Intramuscular injection of AAV8 in mice and macaques is associated with substantial hepatic targeting and transgene expression.

Greig JA, Peng H, Ohlstein J, Medina-Jaszek CA, Ahonkhai O, Mentzinger A, Grant RL, Roy S, Chen SJ, Bell P, Tretiakova AP, Wilson JM - PLoS ONE (2014)

Comparison of expression from the liver-specific TBG promoter following IM and IV vector administration in RAG KO mice.Expression on day 28 post-vector administration of 201Ig IA (A), 2.10A mAb (B), VRC01 mAb (C), and PG9 mAb (D) in RAG KO mice. Expression following IM injection of vectors with either the CMV or TBG promoter was compared to expression levels produced following IV injection of the TBG vector. Mice were injected IM with two 15 µl injections into the right and left gastrocnemius muscles. IV injections were performed as a 100 µl injection via the tail vein. Mice were administered with a dose of either 1010 GC or 1011 GC, numbers indicate dose ×1011 GC. ND, not determined. Expression was measured in serum by ELISA and values are expressed as mean ± SEM (n = 3/group). ***p<0.001.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4230988&req=5

pone-0112268-g002: Comparison of expression from the liver-specific TBG promoter following IM and IV vector administration in RAG KO mice.Expression on day 28 post-vector administration of 201Ig IA (A), 2.10A mAb (B), VRC01 mAb (C), and PG9 mAb (D) in RAG KO mice. Expression following IM injection of vectors with either the CMV or TBG promoter was compared to expression levels produced following IV injection of the TBG vector. Mice were injected IM with two 15 µl injections into the right and left gastrocnemius muscles. IV injections were performed as a 100 µl injection via the tail vein. Mice were administered with a dose of either 1010 GC or 1011 GC, numbers indicate dose ×1011 GC. ND, not determined. Expression was measured in serum by ELISA and values are expressed as mean ± SEM (n = 3/group). ***p<0.001.
Mentions: AAV8 vectors expressing anti-SIV/SHIV antibodies (in an immunoadhesin format [i.e., 201Ig IA] or a monoclonal antibody format [i.e., 2.10A mAb] [40]) or human anti-HIV antibodies in a monoclonal antibody format (i.e., VRC01 mAb [41], [42] and PG9 mAb [43]) were injected IM as two 15 µl injections at doses of 1010 GC or 1011 GC into RAG KO mice (Figure 2). Expression of these proteins in serum was determined on day 28 post-vector administration and compared to expression from TBG-containing vectors administered intravenously (IV) via the tail vein. Figure 2 summarizes data from these experiments, in which the two doses of vectors (labeled 0.1 and 1) were evaluated in the context of two comparisons: the CMV versus TBG vectors following IM injection and the TBG vector following IM and IV injection.

Bottom Line: We found that following IM administration of AAV8 vectors in mice, a portion of the vector reached the liver and hepatic gene expression contributed significantly to total expression of secreted transgenes.Hepatic distribution of vector following IM injection was also noted in rhesus macaques.These pre-clinical data on AAV delivery should inform safe and efficient development of future AAV products.

View Article: PubMed Central - PubMed

Affiliation: Gene Therapy Program, Department of Pathology and Laboratory Medicine, Division of Transfusion Medicine, University of Pennsylvania, TRL Suite 2000, 125 South 31st Street, Philadelphia, PA, 19104, United States of America.

ABSTRACT
Intramuscular (IM) administration of adeno-associated viral (AAV) vectors has entered the early stages of clinical development with some success, including the first approved gene therapy product in the West called Glybera. In preparation for broader clinical development of IM AAV vector gene therapy, we conducted detailed pre-clinical studies in mice and macaques evaluating aspects of delivery that could affect performance. We found that following IM administration of AAV8 vectors in mice, a portion of the vector reached the liver and hepatic gene expression contributed significantly to total expression of secreted transgenes. The contribution from liver could be controlled by altering injection volume and by the use of traditional (promoter) and non-traditional (tissue-specific microRNA target sites) expression control elements. Hepatic distribution of vector following IM injection was also noted in rhesus macaques. These pre-clinical data on AAV delivery should inform safe and efficient development of future AAV products.

Show MeSH
Related in: MedlinePlus