Modification of a Constitutive to Glucose-Responsive Liver-Specific Promoter Resulted in Increased Efficacy of Adeno-Associated Virus Serotype 8-Insulin Gene Therapy of Diabetic Mice.

Cells
Kian Chuan SiaShu Uin Gan

Abstract

We have previously used a hepatotropic adeno-associated viral (AAV) vector with a modified human insulin gene to treat diabetic mice. The HLP (hybrid liver-specific promoter) used was constitutively active and non-responsive to glucose. In this study, we examined the effects of addition of glucose responsive elements (R3G) and incorporation of a 3' albumin enhancer (3'iALB) on insulin expression. In comparison with the original promoter, glucose responsiveness was only observed in the modified promoters in vitro with a 36 h lag time before the peak expression. A 50% decrease in the number of viral particles at 5 × 109 vector genome (vg)/mouse was required by AAV8-R3GHLP-hINSco to reduce the blood sugar level to near normoglycemia when compared to the original AAV8-HLP-hINSco that needed 1 × 1010 vg/mouse. The further inclusion of an 860 base-pairs 3'iALB enhancer component in the 3' untranslated region increased the in vitro gene expression significantly but this increase was not observed when the packaged virus was systemically injected in vivo. The addition of R3G to the HLP promoter in the AAV8-human insulin vector increased the insulin expression and secretion, thereby lowering the required dosage for basal insulin treatmen...Continue Reading

References

Dec 1, 1993·Molecular and Cellular Biology·M J Diaz GuerraM Raymondjean
Mar 1, 2000·Gene Therapy·P M ThuléL S Phillips
Nov 18, 2000·Gene Therapy·P M Thulé, J M Liu
Dec 25, 2002·Transplantation·Tausif Alam, Hans W Sollinger
May 19, 2005·The Journal of Biological Chemistry·Daniel Duran-SandovalBart Staels
Aug 4, 2006·The Journal of Biological Chemistry·Lin MaHoward C Towle
Sep 4, 2008·Molecular and Cellular Biology·Anaïs PerilhouMireille Vasseur-Cognet
May 2, 2009·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Susan J BurkeDonald K Scott
Dec 2, 2010·Molecular Therapy : the Journal of the American Society of Gene Therapy·Jaeseok HanHee-Sook Jun
Apr 2, 2011·Biochimica Et Biophysica Acta·Audrey Poupeau, Catherine Postic
Apr 14, 2011·Self/nonself·Mary S WongNicholas Manolios
Apr 19, 2013·Blood·Federico Mingozzi, Katherine A High
Jan 8, 2016·Diabetology & Metabolic Syndrome·Freddy Goldberg Eliaschewitz, Tânia Barreto
Mar 25, 2017·Human Gene Therapy·Kenneth I Berns, Nicholas Muzyczka
Oct 19, 2017·Diabetic Medicine : a Journal of the British Diabetic Association·D SlatteryP Choudhary
Dec 7, 2018·Journal of Diabetes Investigation·Kai-Jen TienWayne Huey-Herng Sheu

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BETA
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transfection
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