Combined transductional and transcriptional targeting improves the specificity of transgene expression in vivo

ABSTRACT The promise of gene therapy for health care will not be realized until gene delivery systems are capable of achieving efficient, cell-specific gene delivery _in vivo_. Here we

describe an adenoviral system for achieving cell-specific transgene expression in pulmonary endothelium. The combination of transductional targeting to a pulmonary endothelial marker

(angiotensin-converting enzyme, ACE) and an endothelial-specific promoter (for vascular endothelial growth factor receptor type 1, flt-1) resulted in a synergistic, 300,000-fold improvement

in the selectivity of transgene expression for lung versus the usual site of vector sequestration, the liver. This combined approach should be useful for the design of other gene delivery

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SUPPORT FOR SAFE AND EFFICACIOUS CARDIAC GENE TRANSFER IN SWINE Article Open access 01 December 2024 REFERENCES * Lane, K.B. et al. Heterozygous germline mutations in BMPR2, encoding a

TGF-beta receptor, cause familial primary pulmonary hypertension. The International PPH Consortium. _Nat. Genet._ 26, 81–84 (2000). Article  CAS  PubMed  Google Scholar  * Deng, Z. et al.

Familial primary pulmonary hypertension (gene PPH1) is caused by mutations in the bone morphogenetic protein receptor-II gene. _Am. J. Hum. Genet._ 67, 737–744 (2000). Article  CAS  PubMed 

PubMed Central  Google Scholar  * Moraes, D. & Loscalzo, J. Pulmonary hypertension: newer concepts in diagnosis and management. _Clin. Cardiol._ 20, 676–682 (1997). Article  CAS  PubMed

  Google Scholar  * Russell, W.C. Update on adenovirus and its vectors. _J. Gen. Virol._ 81, 2573–2604 (2000). Article  CAS  PubMed  Google Scholar  * Rodman, D.M. et al. _In vivo_ gene

delivery to the pulmonary circulation in rats: transgene distribution and vascular inflammatory response. _Am. J. Respir. Cell. Mol. Biol._ 16, 640–649 (1997). Article  CAS  PubMed  Google

Scholar  * Huard, J. et al. The route of administration is a major determinant of the transduction efficiency of rat tissues by adenoviral recombinants. _Gene Ther._ 2, 107–115 (1995). CAS 

PubMed  Google Scholar  * Bergelson, J.M. et al. Isolation of a common receptor for Coxsackie B viruses and adenoviruses 2 and 5. _Science_ 275, 1320–1323 (1997). Article  CAS  PubMed 

Google Scholar  * Tomko, R.P., Xu, R. & Philipson, L. HCAR and MCAR: the human and mouse cellular receptors for subgroup C adenoviruses and group B coxsackieviruses. _Proc. Natl. Acad.

Sci. USA_ 94, 3352–3356 (1997). Article  CAS  PubMed  PubMed Central  Google Scholar  * Yee, D. et al. Adenovirus-mediated gene transfer of herpes simplex virus thymidine kinase in an

ascites model of human breast cancer. _Hum. Gene Ther._ 7, 1251–1257 (1996). Article  CAS  PubMed  Google Scholar  * Wickham, T.J. Targeting adenovirus. _Gene Ther._ 7, 110–114 (2000).

Article  CAS  PubMed  Google Scholar  * Douglas, J.T. et al. Targeted gene delivery by tropism-modified adenoviral vectors. _Nat. Biotechnol._ 14, 1574–1578 (1996). Article  CAS  PubMed 

Google Scholar  * Krasnykh, V.N., Douglas, J.T. & van Beusechem, V.W. Genetic targeting of adenoviral vectors. _Mol. Ther._ 1, 391–405 (2000). Article  CAS  PubMed  Google Scholar  *

Reynolds, P.N. et al. A targetable injectable adenoviral vector for selective gene delivery to pulmonary endothelium _in vivo_. _Mol. Ther._ 2, 562–578 (2000). Article  CAS  PubMed  Google

Scholar  * Franke, F.E. et al. CD143 Workshop: angiotensin-I-converting enzyme (CD143) on endothelial cells in normal and in pathological conditions. In _Leukocyte typing VI_. (eds

Kishimoto, T. et al.) 749–751 (Garland Publishing Inc., New York, NY; 1997). Google Scholar  * Danilov, S.M. et al. Pulmonary uptake and tissue selectivity of antibodies to surface

endothelial antigens: key determinants of vascular immunotargeting. _Am. J. Physiol. (Lung Cell. Mol. Physiol.)_ 280, L1335–L1347 (2001). Article  CAS  Google Scholar  * Leissner, P. et al.

Influence of adenoviral fiber mutations on viral encapsidation, infectivity and _in vivo_ tropism. _Gene Ther._ 8, 49–57 (2001). Article  CAS  PubMed  Google Scholar  * Alemany, R. &

Curiel, D.T. CAR binding ablation does not change biodistribution or toxicity of adenoviral vectors. _Gene Ther._ in press (2001). Google Scholar  * Wickham, T.J., Mathias, P., Cheresh, D.A.

& Nemerow, G.R. Integrins alpha v beta 3 and alpha v beta 5 promote adenovirus internalization but not virus attachment. _Cell_ 73, 309–319 (1993). Article  CAS  PubMed  Google Scholar

  * Nettelbeck, D.M., Jerome, V. & Muller, R. Gene therapy: designer promoters for tumour targeting. _Trends Genet._ 16, 174–181 (2000). Article  CAS  PubMed  Google Scholar  * Adachi,

Y. et al. Midkine promoter-based adenoviral vector gene delivery for pediatric solid tumors. _Cancer Res._ 60, 4305–4310 (2000). CAS  PubMed  Google Scholar  * Koeneman, K.S. et al.

Osteocalcin-directed gene therapy for prostate-cancer bone metastasis. _World J. Urol._ 18, 102–110 (2000). Article  CAS  PubMed  Google Scholar  * Yamamoto, M., Alemany, R., Adachi, Y.,

Grizzle, W.E. & Curiel, D.T. Characterization of the cyclooxygenase-2 promoter in an adenoviral vector and its application for the mitigation of toxicity in suicide gene therapy of

gastrointestinal cancers. _Mol. Ther._ 3, 385–394. (2001). Article  CAS  PubMed  Google Scholar  * Nicklin, S.A. et al. Analysis of cell-specific promoters for viral gene therapy targeted at

the vascular endothelium. _Hypertension_ 38, 65–70 (2001). Article  CAS  PubMed  Google Scholar  * Bristol, J.A., Shirley, P., Idamakanti, N., Kaleko, M. & Connelly, S. _In vivo_ dose

threshold effect of adenovirus-mediated factor VIII gene therapy in hemophiliac mice. _Mol. Ther._ 2, 223–232 (2000). Article  CAS  PubMed  Google Scholar  * Tao, N. et al. Sequestration of

adenoviral vector by Kupffer cells leads to a nonlinear dose response of transduction in liver. _Mol. Ther._ 3, 28–35 (2001). Article  CAS  PubMed  Google Scholar  * Reynolds, P.N. et al.

Targeting adenoviral infection with basic fibroblast growth factor enhances gene delivery to vascular endothelial and smooth muscle cells. _Tumor Targeting_ 3, 156–168 (1998). CAS  Google

Scholar  * Wickham, T.J. et al. Targeted adenovirus gene transfer to endothelial and smooth muscle cells by using bispecific antibodies. _J. Virol._ 70, 6831–6838 (1996). CAS  PubMed  PubMed

Central  Google Scholar  * Harari, O.A. et al. Targeting an adenoviral gene vector to cytokine-activated vascular endothelium via E-selectin. _Gene Ther._ 6, 801–807 (1999). Article  CAS 

PubMed  Google Scholar  * Nicklin, S.A., White, S.J., Watkins, S.J., Hawkins, R.E. & Baker, A.H. Selective targeting of gene transfer to vascular endothelial cells by use of peptides

isolated by phage display. _Circulation_ 102, 231–237 (2000). Article  CAS  PubMed  Google Scholar  * Danilov, S.M. et al. Lung is the target organ for a monoclonal antibody to angiotensin-

converting enzyme. _Lab. Invest._ 64, 118–124 (1991). CAS  PubMed  Google Scholar  * Danilov, S. et al. Interaction of mAb to angiotensin-converting enzyme (ACE) with antigen _in vitro_ and

_in vivo_: antibody targeting to the lung induces ACE antigenic modulation. _Int. Immunol._ 6, 1153–1160 (1994). Article  CAS  PubMed  Google Scholar  * Ring, C.J., Harris, J.D., Hurst, H.C.

& Lemoine, N.R. Suicide gene expression induced in tumour cells transduced with recombinant adenoviral, retroviral and plasmid vectors containing the ERBB2 promoter. _Gene Ther._ 3,

1094–1103 (1996). CAS  PubMed  Google Scholar  * Schuster, D.P., Crouch, E.C., Parks, W.C., Johnson, T. & Botney, M.D. Angiotensin converting enzyme expression in primary pulmonary

hypertension. _Am. J. Respir. Crit. Care Med._ 154, 1087–1091 (1996). Article  CAS  PubMed  Google Scholar  * Hirose, S., Hosoda, Y., Furuya, S., Otsuki, T. & Ikeda, E. Expression of

vascular endothelial growth factor and its receptors correlates closely with formation of the plexiform lesion in human pulmonary hypertension. _Pathol. Int._ 50, 472–479 (2000). Article 

CAS  PubMed  Google Scholar  * Orte, C., Polak, J.M., Haworth, S.G., Yacoub, M.H. & Morrell, N.W. Expression of pulmonary vascular angiotensin-converting enzyme in primary and secondary

plexiform pulmonary hypertension. _J. Pathol._ 192, 379–384. (2000). Article  CAS  PubMed  Google Scholar  * He, T.C. et al. A simplified system for generating recombinant adenoviruses.

_Proc. Natl. Acad. Sci. USA_ 95, 2509–2514 (1998). Article  CAS  PubMed  PubMed Central  Google Scholar  * Morishita, K., Johnson, D.E. & Williams, L.T. A novel promoter for vascular

endothelial growth factor receptor (flt-1) that confers endothelial-specific gene expression. _J. Biol. Chem._ 270, 27948–27953 (1995). Article  CAS  PubMed  Google Scholar  * Graham, F.

& Prevec, L. Manipulation of adenovirus vectors. In _Methods in molecular biology 7—gene transfer and expression techniques_. (eds Murray, E.J. & Walker, J.M.) 109–129 (Humana Press,

Clifton, NJ; 1991). Google Scholar  * Raben, D. et al. Enhancement of radiolabeled antibody binding and tumor localization through adenoviral transduction of the human carcinoembryonic

antigen gene. _Gene Ther._ 3, 567–580 (1996). CAS  PubMed  Google Scholar  * Conry, R.M. et al. Immune response to a carcinoembryonic antigen polynucleotide vaccine. _Cancer Res._ 54,

1164–1168 (1994). CAS  PubMed  Google Scholar  Download references ACKNOWLEDGEMENTS This research was supported by grants from the American Heart Association, NIH-NIDDK P30 DK54781, and the

Avon Products Foundation to P.N.R.; NIH-NCI RO1 CA74242 to D.T.C. and British Heart Foundation (PG97/018 and PG99/097) grants to A.H.B. We would also like to thank Masato Yamamoto for

providing the AdCMVLuc virus, and Long Le, Kathee Mercer, and the UAB Image Analysis Core for technical assistance. AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Division of Human Gene

Therapy, Departments of Medicine, Surgery and Pathology, and Gene Therapy Center, University of Alabama at Birmingham, Birmingham, AL, USA Paul N. Reynolds, Lioudmila Kaliberova, Brian G.

Boatman, William E. Grizzle & David T. Curiel * Department of Medicine and Therapeutics, University of Glasgow, Glasgow, UK Stuart A. Nicklin & Andrew H. Baker * Division of

Cardiology, University of Alabama at Birmingham, Birmingham, AL, USA Brian G. Boatman * Department of Anesthesiology, University of Illinois at Chicago, Chicago, IL, USA Irina V.

Balyasnikova & Sergei M. Danilov Authors * Paul N. Reynolds View author publications You can also search for this author inPubMed Google Scholar * Stuart A. Nicklin View author

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G. Boatman View author publications You can also search for this author inPubMed Google Scholar * William E. Grizzle View author publications You can also search for this author inPubMed 

Google Scholar * Irina V. Balyasnikova View author publications You can also search for this author inPubMed Google Scholar * Andrew H. Baker View author publications You can also search for

this author inPubMed Google Scholar * Sergei M. Danilov View author publications You can also search for this author inPubMed Google Scholar * David T. Curiel View author publications You

can also search for this author inPubMed Google Scholar CORRESPONDING AUTHOR Correspondence to Paul N. Reynolds. RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS

ARTICLE Reynolds, P., Nicklin, S., Kaliberova, L. _et al._ Combined transductional and transcriptional targeting improves the specificity of transgene expression _in vivo_. _Nat Biotechnol_

19, 838–842 (2001). https://doi.org/10.1038/nbt0901-838 Download citation * Received: 26 February 2001 * Accepted: 13 July 2001 * Issue Date: September 2001 * DOI:

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