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Title: A new fluorescence/PET probe for targeting intracellular human telomerase reverse transcriptase (hTERT) using Tat peptide-conjugated IgM

Abstract

Despite an increasing need for methods to visualize intracellular proteins in vivo, the majority of antibody-based imaging methods available can only detect membrane proteins. The human telomerase reverse transcriptase (hTERT) is an intracellular target of great interest because of its high expression in several types of cancer. In this study, we developed a new probe for hTERT using the Tat peptide. An hTERT antibody (IgG or IgM) was conjugated with the Tat peptide, a fluorescence dye and {sup 64}Cu. HT29 (hTERT+) and U2OS (hTERT−) were used to visualize the intracellular hTERT. The hTERT was detected by RT-PCR and western blot. Fluorescence signals for hTERT were obtained by confocal microscopy, live cell imaging, and analyzed by Tissue-FAXS. In nude mice, tumors were visualized using the fluorescence imaging devices Maestro™ and PETBOX. In RT-PCR and western blot, the expression of hTERT was detected in HT29 cells, but not in U2OS cells. Fluorescence signals were clearly observed in HT29 cells and in U2OS cells after 1 h of treatment, but signals were only detected in HT29 cells after 24 h. Confocal microscopy showed that 9.65% of U2OS and 78.54% of HT29 cells had positive hTERT signals. 3D animation images showed that the probe could target intranuclear hTERT in the nucleus.more » In mice models, fluorescence and PET imaging showed that hTERT in HT29 tumors could be efficiently visualized. In summary, we developed a new method to visualize intracellular and intranuclear proteins both in vitro and in vivo. - Highlights: • We developed new probes for imaging hTERT using Tat-conjugated IgM antibodies labeled with a fluorescent dye and radioisotope. • This probes could be used to overcome limitation of conventional antibody imaging system in live cell imaging. • This system could be applicable to monitor intracellular and intranuclear proteins in vitro and in vivo.« less

Authors:
 [1];  [2];  [2];  [2];  [1];  [2];  [2];  [2];  [1];  [2];  [1];  [2];  [2];  [1];  [2];  [1];  [2];  [2];  [2]
  1. Department of Nuclear Medicine, Seoul National University College of Medicine (Korea, Republic of)
  2. (Korea, Republic of)
Publication Date:
OSTI Identifier:
22606173
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biochemical and Biophysical Research Communications; Journal Volume: 477; Journal Issue: 3; Other Information: Copyright (c) 2016 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; ANIMAL TISSUES; ANTIBODIES; BIOMEDICAL RADIOGRAPHY; COPPER 64; DYES; FLUORESCENCE; IMAGES; IN VITRO; IN VIVO; MEMBRANE PROTEINS; MICE; MICROSCOPY; NEOPLASMS; PEPTIDES; POLYMERASE CHAIN REACTION; POSITRON COMPUTED TOMOGRAPHY; PROBES

Citation Formats

Jung, Kyung oh, Biomedical Sciences, Seoul National University College of Medicine, Cancer Research Institute, Seoul National University College of Medicine, Tumor Microenvironment Global Core Research Center, Seoul National University, Youn, Hyewon, E-mail: hwyoun@snu.ac.kr, Cancer Research Institute, Seoul National University College of Medicine, Tumor Microenvironment Global Core Research Center, Seoul National University, Cancer Imaging Center, Seoul National University Hospital, Seoul, Kim, Seung Hoo, Cancer Research Institute, Seoul National University College of Medicine, Kim, Young-Hwa, Biomedical Sciences, Seoul National University College of Medicine, Cancer Research Institute, Seoul National University College of Medicine, Kang, Keon Wook, Cancer Research Institute, Seoul National University College of Medicine, Chung, June-Key, E-mail: jkchung@snu.ac.kr, Biomedical Sciences, Seoul National University College of Medicine, Cancer Research Institute, Seoul National University College of Medicine, and Tumor Microenvironment Global Core Research Center, Seoul National University. A new fluorescence/PET probe for targeting intracellular human telomerase reverse transcriptase (hTERT) using Tat peptide-conjugated IgM. United States: N. p., 2016. Web. doi:10.1016/J.BBRC.2016.06.068.
Jung, Kyung oh, Biomedical Sciences, Seoul National University College of Medicine, Cancer Research Institute, Seoul National University College of Medicine, Tumor Microenvironment Global Core Research Center, Seoul National University, Youn, Hyewon, E-mail: hwyoun@snu.ac.kr, Cancer Research Institute, Seoul National University College of Medicine, Tumor Microenvironment Global Core Research Center, Seoul National University, Cancer Imaging Center, Seoul National University Hospital, Seoul, Kim, Seung Hoo, Cancer Research Institute, Seoul National University College of Medicine, Kim, Young-Hwa, Biomedical Sciences, Seoul National University College of Medicine, Cancer Research Institute, Seoul National University College of Medicine, Kang, Keon Wook, Cancer Research Institute, Seoul National University College of Medicine, Chung, June-Key, E-mail: jkchung@snu.ac.kr, Biomedical Sciences, Seoul National University College of Medicine, Cancer Research Institute, Seoul National University College of Medicine, & Tumor Microenvironment Global Core Research Center, Seoul National University. A new fluorescence/PET probe for targeting intracellular human telomerase reverse transcriptase (hTERT) using Tat peptide-conjugated IgM. United States. doi:10.1016/J.BBRC.2016.06.068.
Jung, Kyung oh, Biomedical Sciences, Seoul National University College of Medicine, Cancer Research Institute, Seoul National University College of Medicine, Tumor Microenvironment Global Core Research Center, Seoul National University, Youn, Hyewon, E-mail: hwyoun@snu.ac.kr, Cancer Research Institute, Seoul National University College of Medicine, Tumor Microenvironment Global Core Research Center, Seoul National University, Cancer Imaging Center, Seoul National University Hospital, Seoul, Kim, Seung Hoo, Cancer Research Institute, Seoul National University College of Medicine, Kim, Young-Hwa, Biomedical Sciences, Seoul National University College of Medicine, Cancer Research Institute, Seoul National University College of Medicine, Kang, Keon Wook, Cancer Research Institute, Seoul National University College of Medicine, Chung, June-Key, E-mail: jkchung@snu.ac.kr, Biomedical Sciences, Seoul National University College of Medicine, Cancer Research Institute, Seoul National University College of Medicine, and Tumor Microenvironment Global Core Research Center, Seoul National University. Fri . "A new fluorescence/PET probe for targeting intracellular human telomerase reverse transcriptase (hTERT) using Tat peptide-conjugated IgM". United States. doi:10.1016/J.BBRC.2016.06.068.
@article{osti_22606173,
title = {A new fluorescence/PET probe for targeting intracellular human telomerase reverse transcriptase (hTERT) using Tat peptide-conjugated IgM},
author = {Jung, Kyung oh and Biomedical Sciences, Seoul National University College of Medicine and Cancer Research Institute, Seoul National University College of Medicine and Tumor Microenvironment Global Core Research Center, Seoul National University and Youn, Hyewon, E-mail: hwyoun@snu.ac.kr and Cancer Research Institute, Seoul National University College of Medicine and Tumor Microenvironment Global Core Research Center, Seoul National University and Cancer Imaging Center, Seoul National University Hospital, Seoul and Kim, Seung Hoo and Cancer Research Institute, Seoul National University College of Medicine and Kim, Young-Hwa and Biomedical Sciences, Seoul National University College of Medicine and Cancer Research Institute, Seoul National University College of Medicine and Kang, Keon Wook and Cancer Research Institute, Seoul National University College of Medicine and Chung, June-Key, E-mail: jkchung@snu.ac.kr and Biomedical Sciences, Seoul National University College of Medicine and Cancer Research Institute, Seoul National University College of Medicine and Tumor Microenvironment Global Core Research Center, Seoul National University},
abstractNote = {Despite an increasing need for methods to visualize intracellular proteins in vivo, the majority of antibody-based imaging methods available can only detect membrane proteins. The human telomerase reverse transcriptase (hTERT) is an intracellular target of great interest because of its high expression in several types of cancer. In this study, we developed a new probe for hTERT using the Tat peptide. An hTERT antibody (IgG or IgM) was conjugated with the Tat peptide, a fluorescence dye and {sup 64}Cu. HT29 (hTERT+) and U2OS (hTERT−) were used to visualize the intracellular hTERT. The hTERT was detected by RT-PCR and western blot. Fluorescence signals for hTERT were obtained by confocal microscopy, live cell imaging, and analyzed by Tissue-FAXS. In nude mice, tumors were visualized using the fluorescence imaging devices Maestro™ and PETBOX. In RT-PCR and western blot, the expression of hTERT was detected in HT29 cells, but not in U2OS cells. Fluorescence signals were clearly observed in HT29 cells and in U2OS cells after 1 h of treatment, but signals were only detected in HT29 cells after 24 h. Confocal microscopy showed that 9.65% of U2OS and 78.54% of HT29 cells had positive hTERT signals. 3D animation images showed that the probe could target intranuclear hTERT in the nucleus. In mice models, fluorescence and PET imaging showed that hTERT in HT29 tumors could be efficiently visualized. In summary, we developed a new method to visualize intracellular and intranuclear proteins both in vitro and in vivo. - Highlights: • We developed new probes for imaging hTERT using Tat-conjugated IgM antibodies labeled with a fluorescent dye and radioisotope. • This probes could be used to overcome limitation of conventional antibody imaging system in live cell imaging. • This system could be applicable to monitor intracellular and intranuclear proteins in vitro and in vivo.},
doi = {10.1016/J.BBRC.2016.06.068},
journal = {Biochemical and Biophysical Research Communications},
number = 3,
volume = 477,
place = {United States},
year = {Fri Aug 26 00:00:00 EDT 2016},
month = {Fri Aug 26 00:00:00 EDT 2016}
}