14 research outputs found
Development of a Radiolabeled Peptide-Based Probe Targeting MT1-MMP for Breast Cancer Detection
<div><p>Breast cancer is one of the most frequent and aggressive primary tumors among women of all races. Matrix metalloproteinase (MMPs), a family of zinc- and calcium-dependent secreted or membrane anchored endopeptidases, is overexpressed in varieties of diseases including breast cancer. Therefore, noninvasive visualization and quantification of MMP <i>in vivo</i> are of great interest in basic research and clinical application for breast cancer early diagnosis. Herein, we developed a <sup>99m</sup>Tc labeled membrane type I matrix metalloproteinase (MT1-MMP) specific binding peptide, [<sup>99m</sup>Tc]-(HYNIC-AF7p)(tricine)(TPPTS), for <i>in vivo</i> detection of MDA-MB-231 breast tumor by single photon emission computed tomography (SPECT). [<sup>99m</sup>Tc]-(HYNIC-AF7p)(tricine)(TPPTS) demonstrated nice biostability and high MT1-MMP binding affinity <i>in vitro</i> and <i>in vivo</i>. Tumor-to-muscle ratio was found to reach to the highest (4.17±0.49) at 2 hour after intravenously administration of [<sup>99m</sup>Tc]-(HYNIC-AF7P)(tricine)(TPPTS) into MDA-MB-231 tumor bearing mice. Overall, [<sup>99m</sup>Tc]-(HYNIC-AF7P)(tricine)(TPPTS) demonstrated great potential for MT1-MMP targeted detection <i>in vivo</i> and it would be a promising molecular imaging probe that are probably beneficial to breast cancer early diagnoses.</p></div
Cytotoxicity of AF7p analogs on MDA-MB-231 cells.
<p>No cytotoxicity was observed at different concentrations of AF7p, Cy5.5-AF7p and (HYNIC-AF7P)(tricine)(TPPTS).</p
Fluorescent immunohistochemistry for MT1-MMP expression in MDA-MB-435 and A549 tumors.
<p>More MT1-MMP expressed in MDA-MB-435 tumor was observed than that of in A549 tumor. Green: FITC conjugated donkey anti-Rabbit secondary antibody. Blue: DAPI. All signals were adjusted to the same scale. Scale bar equals10 μm.</p
Characterizations.
<p>(A) Chemical structure of [<sup>99m</sup>Tc]-(HYNIC-AF7p)(tricine)(TPPTS). (B) Radio-HPLC purity of [<sup>99m</sup>Tc]-(HYNIC-AF7p)(tricine)(TPPTS).</p
SPECT/CT images of MDA-MB-231 and A549 tumors by targeting MT1-MMP.
<p>(A) Static planar images of MDA-MB-231 tumor-bearing mice model and A549 tumor-bearing mice model after administration of 11.1 MBq/kg of [<sup>99m</sup>Tc](HYNIC-AF7P)(tricine)(TPPTS) at 0.5 and 2 h post injection. Excess amount (200 μg) of free AF7P was injected for blocking test. (B) Quantification of [<sup>99m</sup>Tc](HYNIC-AF7P)(tricine)(TPPTS) and [<sup>99m</sup>Tc](HYNIC-AF7P)(tricine)(TPPTS) (Blocking) in MDA-MB-231 tumor-bearing mice. Quantification of [<sup>99m</sup>Tc](HYNIC-AF7P)(tricine)(TPPTS) in A549 tumor-bearing mice. ROIs are shown as mean %ID/g ± SD.</p
Solution stability data for [<sup>99m</sup>Tc(HYNIC-AF7P)(tricine)(TPPTS)] in saline (A) and in the presence of excess cysteine (B) (1 mg/mL, pH = 7.4).
<p>Solution stability data for [<sup>99m</sup>Tc(HYNIC-AF7P)(tricine)(TPPTS)] in saline (A) and in the presence of excess cysteine (B) (1 mg/mL, pH = 7.4).</p
Biodistribution of [<sup>99m</sup>Tc]-(HYNIC-AF7p)(tricine)(TPPTS) in mice bearing MDA-MB-231 tumor.
<p>Biodistribution of [<sup>99m</sup>Tc]-(HYNIC-AF7p)(tricine)(TPPTS) in mice bearing MDA-MB-231 tumor.</p
Cell labeling.
<p>(A) Chemical structure of Cy5.5 AF7p. (B) Colocalization of Cy5.5-AF7p and MT1-MMP on MDA-MB-231 and A549 cells. Red color is from Cy5.5 for Cy5.5-AF7p distribution on cellular membrane. Green color is from FITC conjugated donkey anti-rabbit secondary antibody for MT1-MMP localization. Blue color is from DAPI for nuclei visualization. Yellow color is merged color from Red and Green, indicating the colocalization of Cy5.5-AF7p and MT1-MMP. Scale bar equals 10 μm. (C). Blocking test for the specificity of Cy5.5-AF7p to MT1-MMP. Little of Cy5.5-AF7p was able to label MT1-MMP after excess amount of AF7p blocking.</p
Tumor to organ ratios of [<sup>99m</sup>Tc]-(HYNIC-AF7p)(tricine)(TPPTS) in B mice bearing MDA-MB-231 tumor.
<p>Tumor to organ ratios of [<sup>99m</sup>Tc]-(HYNIC-AF7p)(tricine)(TPPTS) in B mice bearing MDA-MB-231 tumor.</p
T/NT for <sup>99m</sup>Tc-RGD and <sup>99m</sup>Tc-MIBI in malignant and benign tumors.
<p>(A) The T/NT for <sup>99m</sup>Tc-RGD in breast cancer was significantly higher than that in benign lesions (p<0.001). (B) The T/NT for <sup>99m</sup>Tc-MIBI in breast cancer was significantly higher than that in benign lesions (p<0.001).</p