AAV Serotype1 Mediates Early Onset of Gene Expression in Mouse Hearts and Results in betterTherapeutic Effect

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AAV Serotype-1 Mediates Early Onset of Gene Expression in Mouse Hearts and Results in better Therapeutic Effect Hua Su University of California, San Francisco Gene Therapy (2006) 13, 1495–1502

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Adeno-associated viral vector (AAV) is attractive for delivery of genes into myocardium. Transgene expression mediated by commonly used AAV serotype 2 in myocardium has a “time lag”. Therapeutic effectiveness for acute myocardial infarction may be reduced by this “time lag”, because irreversible myocardial injury begins immediately after coronary artery occlusion. 4. Different AAV serotypes mediate differential gene expression in different cells and tissues. 5. Earlier transgene expression and better therapeutic effect might be achieved using a different AAV serotype. Background:

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Study the kinetics of gene expression mediated by different serotypes: 1) AAV-Epo was packaged into capsids of AAV serotypes 1 to 5 and injected to mouse hearts. 2) Epo expression was followed by measuring changes in hematocrits and mRNA expression. AAV-LacZ and MLCVEGF (an AAV vector expressing VEGF in cardiac-specific and hypoxia-inducible manner) were packaged into AAV1 and 2, 1) LacZ expression was studied 1 and 14 days after the vector injection by b-gal staining. 2) Hypoxia induction of VEGF expression was analyzed 4 days after vector injections by real-time RT-PCR. 3. Cardiac function and infarct size were studied 2 weeks after gene transfer. Methods

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AAV1 mediates highest transgene expression in mouse hearts among AAV1-5 RNA expression levels in day 1 to 7 measured by real-time RT- PCR. ** indicate that the differences of gene expression mediated by other serotypes are significant from that mediated by serotype 2. ** ** ** ** ** ** ** ** ** ** ** ** **

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LacZ expression was detected in AAV1-LacZ not in AAV2-LacZ injected heart one day after the vector injection AAV1-LacZ AAV2-LacZ

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AAV1 Mediated Robust LacZ Gene Expression around Injection site

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AAV1-MLCVEGF mediated 80 folds Higher VEGF expression in ischemic hearts than normal hearts. AAV2 delivered MLCVEGF did not mediate significant VEGF expression both in normal and ischemic hearts. AAV1 mediated hypoxia-inducible VEGF expression 4 days after vector injection MLC: Cardiac myosin light-chain-2v promoter; H9: nine copies of Epo HRE; ITR: Inverted terminal repeats; PA: SV40 polyadenylation site.

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Hearts received AAV1 mediated VEGF gene transfer have better function and smaller scars Cardiac Function Fibrous Scar

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More capillaries in AAV1-MLCVEGF injected than AAV2-MLCVEGF and HEPES injected hearts 4 days after vector injection AAV1-MLCVEGF AAV2-MLCVEGF HEPES Sections stained with anti-platelet endothelial cell adhesion molecule 1 (PECAM-1) antibody, showing capillary in hearts collected 4 days after vector injections.

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AAV1 mediated VEGF gene transfer induced more neovascular formation than AAV2 (4 weeks after vector injection) MLC: MLCVEGF; CMV: CMVVEGF

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AAV1 mediates transgene expression in myocardium within one day. Hypoxia-responsive element regulated VEGF gene expression packaged in AAV1 capsid is expressed in response to coronary ligation within 4 days of vector injection. 3. The earlier gene expression resulted in better cardiac function and less fibrosis. 4. Improved therapeutic effect observed with AAV1 mediated VEGF gene transfer is likely due to earlier and greater neovascular formation. Conclusions:

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Acknowledgements Yuet Wai Kan Yu Huang Janice Arakawa-Hoyt Alicia Barcena William Grossman Junya Takagawa Jianqin Ye

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	hua.su@ucsf.edu 5 Years ago.



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PowerPoint Slide Presentation on This paper was published on Gene Therapy (2006) 13, 1495–1502. We more PowerPoint Slide Presentation on This paper was published on Gene Therapy (2006) 13, 1495–1502. We studied gene transduciton efficiency mediated by AAV serotypes 1-5 and found AAV serotype 1 is the most effective on for mediating gene transduction in mouse myocardium. less
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