A gene therapy approach for long-term normalization of blood pressure in hypertensive mice by ANP-secreting human skin grafts — PNAS

A gene therapy approach for long-term normalization of blood pressure in hypertensive mice by ANP-secreting human skin grafts
Jean-Philippe Therriena, Soo Mi Kimb, Atsushi Terunumaa, Yan Qinb, Christine L. Tocka, Wolfgang Pfütznerc, Manabu Ohyamaa,1, Jurgen Schnermannb and Jonathan C. Vogela,2
+ Author Affiliations
Copyright ©2010 by the National Academy of Sciences

aDermatology Branch, National Cancer Institute, and
bNational Institute of Digestive and Diabetes and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892; and
cDepartment of Dermatology and Allogology, University of Giessen and Marburg, 35037 Marburg, Germany
↵ 1 Present address: Department of Dermatology, Keio University School of Medicine, 35 Shinanomachi Shinjuku-ku, Tokyo 160-8582, Japan.

Edited by Ira Pastan, National Cancer Institute, National Institutes of Health, Bethesda, MD, and approved November 23, 2009 (received for review August 10, 2009)

Abstract
The use of bioengineered human skin as a bioreactor to deliver therapeutic factors has a number of advantages including accessibility that allows manipulation and monitoring of genetically modified cells. We demonstrate a skin gene therapy approach that can regulate blood pressure and treat systemic hypertension by expressing atrial natriuretic peptide (ANP), a hormone able to decrease blood pressure, in bioengineered human skin equivalents (HSE). Additionally, the expression of a selectable marker gene, multidrug resistance (MDR) type 1, is linked to ANP expression on a bicistronic vector and was coexpressed in the human keratinocytes and fibroblasts of the HSE that were grafted onto immunocompromised mice. Topical treatments of grafted HSE with the antimitotic agent colchicine select for keratinocyte progenitors that express both MDR and ANP. Significant plasma levels of human ANP were detected in mice grafted with HSE expressing ANP from either keratinocytes or fibroblasts, and topical selection of grafted HSE resulted in persistent high levels of ANP expression in vivo. Mice with elevated plasma levels of human ANP showed lower renin levels and, correspondingly, had lower systemic blood pressure than controls. Furthermore, mice with HSE grafts expressing human ANP did not develop elevated blood pressure when fed a high-salt diet. These findings illustrate the potential of this human skin gene therapy approach to deliver therapeutic molecules systemically for long-term treatment of diverse diseases.

keratinocytes
human engineered skin
hypertension
multi-drug resistance gene
retroviral vectors

Footnotes
2To whom correspondences should be addressed at: Dermatology Branch National Cancer Institute, NIH Bldg. 10 / Room 12N260, 10 Center Drive, MSC 1908, Bethesda, MD 20892-1908. E-mail: jonvogel@mail.nih.gov. Author contributions: J.-P.T., C.L.T., J.S., and J.C.V. designed research; J.-P.T., S.M.K., A.T., Y.Q., and C.L.T. performed research; S.M.K., A.T., Y.Q., C.L.T., W.P., M.O., and J.S. contributed new reagents/analytic tools; J.-P.T., S.M.K., J.S., and J.C.V. analyzed data; and J.-P.T. and J.C.V. wrote the paper.
The authors declare no conflict of interest.
This article is a PNAS Direct Submission.
This article contains supporting information online at
www.pnas.org/cgi/content/full/0908882107/DCSupplemental.

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