An engineered VEGF-activating zinc finger protein transcription factor improves blood flow and limb salvage in advanced-age mice.
Advances in understanding the relationship between protein structure and DNA binding specificity have made it possible to engineer zinc finger protein (ZFP) transcription factors to specifically activate or repress virtually any gene. To evaluate the potential clinical utility of this approach for peripheral vascular disease, we investigated the ability of an engineered vascular endothelial growth factor (VEGFa)-activating ZFP (MVZ+426b) to induce angiogenesis and rescue hindlimb ischemia in a murine model. Hindlimb ischemia was surgically induced in advanced-age C57/BL6 mice. Adenovirus (Ad) encoding either MVZ+426b or the fluorescent marker dsRed was delivered to the adducter muscle of the ischemic hindlimb, and the effects on blood flow, limb salvage, and vascularization were assessed. Ad-MVZ+426b induced expression of VEGFa at the mRNA and protein levels and stimulated a significant increase in vessel counts in the ischemic limb. This was accompanied by significantly increased blood flow and limb salvage as measured serially for 4 wk. These data demonstrate that activation of the endogenous VEGFa gene by an engineered ZFP can induce angiogenesis in a clinically relevant model and further document the feasibility of designing ZFPs to therapeutically regulate gene expression in vivo.
Yu
Jun
J
Department of Pharmacology, Yale University School of Medicine, New Haven, Connecticut, USA.
Lei
Li
L
Liang
Yuxin
Y
Hinh
Linda
L
Hickey
Reed P
RP
Huang
Yan
Y
Liu
Dinggang
D
Yeh
Jennifer L
JL
Rebar
Edward
E
Case
Casey
C
Spratt
Kaye
K
Sessa
William C
WC
Giordano
Frank J
FJ
D000256:Adenoviridae / Q000235:genetics; D000375:Aging; D000595:Amino Acid Sequence; D000818:Animals; D001783:Blood Flow Velocity; D005240:Feasibility Studies; D005786:Gene Expression Regulation / Q000235:genetics*; D005813:Genes, Synthetic; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000627:therapeutic use*; D006614:Hindlimb / Q000098:blood supply; D007511:Ischemia / Q000628:therapy*; D017078:Laser-Doppler Flowmetry; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D018919:Neovascularization, Physiologic / Q000235:genetics*; D015202:Protein Engineering; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins; D013329:Structure-Activity Relationship; D014157:Transcription Factors / Q000235:genetics / Q000502:physiology*; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis* / Q000235:genetics; D016335:Zinc Fingers / Q000235:genetics / Q000502:physiology*
C509623:MVZplus426b protein; D012333:RNA, Messenger; D011994:Recombinant Proteins; D014157:Transcription Factors; D042461:Vascular Endothelial Growth Factor A; C467480:vascular endothelial growth factor A, mouse