Transduction of peptide analogs of the small heat shock-related protein HSP20 inhibits intimal hyperplasia.
BACKGROUND
Human saphenous vein (HSV) is the autologous conduit of choice for peripheral vascular reconstructions. However, vasospasm can lead to early graft failure. The leading cause of delayed graft failure is intimal hyperplasia.


OBJECTIVE
To develop a proteomic approach to prevent vein-graft spasm and intimal hyperplasia.


METHODS
Biomimetic peptide analogs of the small heat shock-related protein HSP20, containing a protein transduction domain (PTD), a phosphorylated serine, and a sequence of HSP20 surrounding the phosphorylation site (PTD-pHSP20), or a scrambled sequence of the same amino acids surrounding the phosphorylation site (PTD-scHSP20) were synthesized. The peptides were used in muscle bath and organ culture experiments with human saphenous vein (HSV) segments. Cultured smooth muscle cell lines were used to determine the effect of the peptides on proliferation and migration.


RESULTS
In HSV rings precontracted with norepinephrine, PTD-pHSP20 but not PTD-scHSP20 led to relaxation. There was no significant difference in smooth muscle cell proliferation in cells treated with PTD-pHSP20 compared with PTD-scHSP20. Treatment with PTD-pHSP20 significantly inhibited cellular migration compared with PTD-scHSP20. Control, untreated, and PTD-scHSP20-treated saphenous veins had significant increases in intimal thickness after culture. This intimal thickening was completely inhibited by treatment with PTD-pHSP20.


CONCLUSIONS
Protein transduction of biologically active motifs of HSP20 can affect pathologic and physiologic responses of HSV and represents a novel proteomic-based therapeutic approach.


CONCLUSIONS
We have been a part of the genomics era and are now viewing the emergence of "proteomics." The genome is linear and relatively easy to examine; however the proteome is much more complex and dynamic. In essence, the purpose of gene therapy is to manipulate the genome to produce a particular protein. This manuscript describes a new proteomic approach in which the biologically active part of a protein is directly introduced into vascular cells. Peptides were synthesized which contained a total of 24 amino acids, 11 of which represent a protein transduction domain or "carrier" while the other 13 are the biologically active "cargo." These synthetic peptides prevent spasm (contraction) and intimal hyperplasia in segments of human saphenous vein treated ex vivo. Preclinical development is currently underway to develop these molecules as a proteomic-based vein harvest solution to enhance vein-graft patency.
Tessier
Deron J
DJ
Department of Surgery, Mayo Clinic, Scottsdale, AZ, USA.
Komalavilas
Padmini
P
Liu
Bo
B
Kent
Craig K
CK
Thresher
Jeffrey S
JS
Dreiza
Catherine M
CM
Panitch
Alyssa
A
Joshi
Lokesh
L
Furnish
Elizabeth
E
Stone
William
W
Fowl
Richard
R
Brophy
Colleen M
CM
D002478:Cells, Cultured; D006083:Graft Occlusion, Vascular / Q000209:etiology / Q000517:prevention & control*; D050886:HSP20 Heat-Shock Proteins; D006360:Heat-Shock Proteins / Q000235:genetics / Q000502:physiology*; D006801:Humans; D006965:Hyperplasia / Q000150:complications / Q000517:prevention & control; D032389:Myocytes, Smooth Muscle; D009924:Organ Culture Techniques; D010748:Phosphopeptides / Q000235:genetics / Q000502:physiology; D010750:Phosphoproteins / Q000235:genetics / Q000502:physiology*; D012501:Saphenous Vein / Q000503:physiopathology; D013035:Spasm / Q000150:complications / Q000517:prevention & control*; D014161:Transduction, Genetic / Q000379:methods*; D017539:Tunica Intima / Q000473:pathology* / Q000503:physiopathology
D050886:HSP20 Heat-Shock Proteins; C494382:HSPB6 protein, human; D006360:Heat-Shock Proteins; D010748:Phosphopeptides; D010750:Phosphoproteins