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CTHRSSVVC Peptide as a Possible Early Molecular Imaging Target for Atherosclerosis

View Article: PubMed Central - PubMed

ABSTRACT

The purpose of our work was to select phages displaying peptides capable of binding to vascular markers present in human atheroma, and validate their capacity to target the vascular markers in vitro and in low-density lipoprotein receptor knockout (LDLr−/−) mouse model of atherosclerosis. By peptide fingerprinting on human atherosclerotic tissues, we selected and isolated four different peptides sequences, which bind to atherosclerotic lesions and share significant similarity to known human proteins with prominent roles in atherosclerosis. The CTHRSSVVC-phage peptide displayed the strongest reactivity with human carotid atherosclerotic lesions (p < 0.05), when compared to tissues from normal carotid arteries. This peptide sequence shares similarity to a sequence present in the fifth scavenger receptor cysteine-rich (SRCR) domain of CD163, which appeared to bind to CD163, and subsequently, was internalized by macrophages. Moreover, the CTHRSSVVC-phage targets atherosclerotic lesions of a low-density lipoprotein receptor knockout (LDLr−/−) mouse model of atherosclerosis in vivo to High-Fat diet group versus Control group. Tetraazacyclododecane-1,4,7,10-tetraacetic acid-CTHRSSVVC peptide (DOTA-CTHRSSVVC) was synthesized and labeled with 111InCl3 in >95% yield as determined by high performance liquid chromatography (HPLC), to validate the binding of the peptide in atherosclerotic plaque specimens. The results supported our hypothesis that CTHRSSVVC peptide has a remarkable sequence for the development of theranostics approaches in the treatment of atherosclerosis and other diseases.

No MeSH data available.


Related in: MedlinePlus

CTHRSSVVC-phage to Regular (A) and High-Fat diet (B); and insertless phage to regular (C) and High-Fat diet (D); homing specificity to organ target. Representative sections of mice aortic root from Regular (n = 8) and High-Fat diet (n = 13) groups. Hematoxylin–eosin (HE) staining to aorta was performed to all groups. High-Fat diet group presented a strong positively stained area as indicated by the star (*) to CTHRSSVVC-phage homing (B) and negative to insertless Fd-tet phage on the same diet (D). The CTHRSSVVC-phage bound quantification in atheroma of High-Fat diet mice (B) was reported as means ± SEM 45.50, ±S.D 13.97 and all other sections were negative to phage homing. As a reaction control, primary antibody was omitted. Bar, 20 µm in all panels.
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ijms-17-01383-f007: CTHRSSVVC-phage to Regular (A) and High-Fat diet (B); and insertless phage to regular (C) and High-Fat diet (D); homing specificity to organ target. Representative sections of mice aortic root from Regular (n = 8) and High-Fat diet (n = 13) groups. Hematoxylin–eosin (HE) staining to aorta was performed to all groups. High-Fat diet group presented a strong positively stained area as indicated by the star (*) to CTHRSSVVC-phage homing (B) and negative to insertless Fd-tet phage on the same diet (D). The CTHRSSVVC-phage bound quantification in atheroma of High-Fat diet mice (B) was reported as means ± SEM 45.50, ±S.D 13.97 and all other sections were negative to phage homing. As a reaction control, primary antibody was omitted. Bar, 20 µm in all panels.

Mentions: A strong positive phage staining area was observed in the atherosclerotic tissue sections of all LDLr−/− mice fed with a High-Fat diet that received the CTHRSSVVC-phage injection (Figure 7, marked by the star (*)). However, no positivity was detected in the aorta from the LDLr−/− mice fed with a Regular-diet. Although background for phages was observed in aorta and atherosclerotic lesions from LDLr−/− mice fed on both Regular and High-Fat diet that received the insertless phage injection, this staining was not significant. As a reaction control, primary antibody was omitted in adjacent sections, and hematoxylin–eosin (HE) staining was performed in all samples (Figure 7).


CTHRSSVVC Peptide as a Possible Early Molecular Imaging Target for Atherosclerosis
CTHRSSVVC-phage to Regular (A) and High-Fat diet (B); and insertless phage to regular (C) and High-Fat diet (D); homing specificity to organ target. Representative sections of mice aortic root from Regular (n = 8) and High-Fat diet (n = 13) groups. Hematoxylin–eosin (HE) staining to aorta was performed to all groups. High-Fat diet group presented a strong positively stained area as indicated by the star (*) to CTHRSSVVC-phage homing (B) and negative to insertless Fd-tet phage on the same diet (D). The CTHRSSVVC-phage bound quantification in atheroma of High-Fat diet mice (B) was reported as means ± SEM 45.50, ±S.D 13.97 and all other sections were negative to phage homing. As a reaction control, primary antibody was omitted. Bar, 20 µm in all panels.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5037663&req=5

ijms-17-01383-f007: CTHRSSVVC-phage to Regular (A) and High-Fat diet (B); and insertless phage to regular (C) and High-Fat diet (D); homing specificity to organ target. Representative sections of mice aortic root from Regular (n = 8) and High-Fat diet (n = 13) groups. Hematoxylin–eosin (HE) staining to aorta was performed to all groups. High-Fat diet group presented a strong positively stained area as indicated by the star (*) to CTHRSSVVC-phage homing (B) and negative to insertless Fd-tet phage on the same diet (D). The CTHRSSVVC-phage bound quantification in atheroma of High-Fat diet mice (B) was reported as means ± SEM 45.50, ±S.D 13.97 and all other sections were negative to phage homing. As a reaction control, primary antibody was omitted. Bar, 20 µm in all panels.
Mentions: A strong positive phage staining area was observed in the atherosclerotic tissue sections of all LDLr−/− mice fed with a High-Fat diet that received the CTHRSSVVC-phage injection (Figure 7, marked by the star (*)). However, no positivity was detected in the aorta from the LDLr−/− mice fed with a Regular-diet. Although background for phages was observed in aorta and atherosclerotic lesions from LDLr−/− mice fed on both Regular and High-Fat diet that received the insertless phage injection, this staining was not significant. As a reaction control, primary antibody was omitted in adjacent sections, and hematoxylin–eosin (HE) staining was performed in all samples (Figure 7).

View Article: PubMed Central - PubMed

ABSTRACT

The purpose of our work was to select phages displaying peptides capable of binding to vascular markers present in human atheroma, and validate their capacity to target the vascular markers in vitro and in low-density lipoprotein receptor knockout (LDLr−/−) mouse model of atherosclerosis. By peptide fingerprinting on human atherosclerotic tissues, we selected and isolated four different peptides sequences, which bind to atherosclerotic lesions and share significant similarity to known human proteins with prominent roles in atherosclerosis. The CTHRSSVVC-phage peptide displayed the strongest reactivity with human carotid atherosclerotic lesions (p < 0.05), when compared to tissues from normal carotid arteries. This peptide sequence shares similarity to a sequence present in the fifth scavenger receptor cysteine-rich (SRCR) domain of CD163, which appeared to bind to CD163, and subsequently, was internalized by macrophages. Moreover, the CTHRSSVVC-phage targets atherosclerotic lesions of a low-density lipoprotein receptor knockout (LDLr−/−) mouse model of atherosclerosis in vivo to High-Fat diet group versus Control group. Tetraazacyclododecane-1,4,7,10-tetraacetic acid-CTHRSSVVC peptide (DOTA-CTHRSSVVC) was synthesized and labeled with 111InCl3 in >95% yield as determined by high performance liquid chromatography (HPLC), to validate the binding of the peptide in atherosclerotic plaque specimens. The results supported our hypothesis that CTHRSSVVC peptide has a remarkable sequence for the development of theranostics approaches in the treatment of atherosclerosis and other diseases.

No MeSH data available.


Related in: MedlinePlus