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 selectablemarker 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.
| Original language | English |
|---|---|
| Pages (from-to) | 1178-1183 |
| Number of pages | 6 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Volume | 107 |
| Issue number | 3 |
| DOIs | |
| Publication status | Published - 2010 Jan 19 |
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Keywords
- Human engineered skin
- Hypertension
- Keratinocytes
- Multi-drug resistance gene
- Retroviral vectors
ASJC Scopus subject areas
- General
Cite this
A gene therapy approach for long-term normalization of blood pressure in hypertensive mice by ANP-secreting human skin grafts. / Therrien, Jean Philippe; Kim, Soo Mi; Terunuma, Atsushi; Qin, Yan; Tock, Christine L.; Pfützner, Wolfgang; Ohyama, Manabu; Schnermann, Jurgen; Vogel, Jonathan C.
In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 107, No. 3, 19.01.2010, p. 1178-1183.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - A gene therapy approach for long-term normalization of blood pressure in hypertensive mice by ANP-secreting human skin grafts
AU - Therrien, Jean Philippe
AU - Kim, Soo Mi
AU - Terunuma, Atsushi
AU - Qin, Yan
AU - Tock, Christine L.
AU - Pfützner, Wolfgang
AU - Ohyama, Manabu
AU - Schnermann, Jurgen
AU - Vogel, Jonathan C.
PY - 2010/1/19
Y1 - 2010/1/19
N2 - 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 selectablemarker 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.
AB - 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 selectablemarker 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.
KW - Human engineered skin
KW - Hypertension
KW - Keratinocytes
KW - Multi-drug resistance gene
KW - Retroviral vectors
UR - http://www.scopus.com/inward/record.url?scp=75749142715&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=75749142715&partnerID=8YFLogxK
U2 - 10.1073/pnas.0908882107
DO - 10.1073/pnas.0908882107
M3 - Article
C2 - 20080656
AN - SCOPUS:75749142715
VL - 107
SP - 1178
EP - 1183
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
SN - 0027-8424
IS - 3
ER -