Efficient detection of human circulating tumor cells without significant production of false-positive cells by a novel conditionally replicating adenovirus

Fuminori Sakurai, Nobuhiro Narii, Kyoko Tomita, Shinsaku Togo, Kazuhisa Takahashi, Mitsuhiro Machitani, Masashi Tachibana, Masaaki Ouchi, Nobuyoshi Katagiri, Yasuo Urata, Toshiyoshi Fujiwara, Hiroyuki Mizuguchi

Research output: Contribution to journalArticle

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Abstract

Circulating tumor cells (CTCs) are promising biomarkers in several cancers, and thus methods and apparatuses for their detection and quantification in the blood have been actively pursued. A novel CTC detection system using a green fluorescence protein (GFP)–expressing conditionally replicating adenovirus (Ad) (rAd-GFP) was recently developed; however, there is concern about the production of false-positive cells (GFP-positive normal blood cells) when using rAd-GFP, particularly at high titers. In addition, CTCs lacking or expressing low levels of coxsackievirus–adenovirus receptor (CAR) cannot be detected by rAd-GFP, because rAd-GFP is constructed based on Ad serotype 5, which recognizes CAR. In order to suppress the production of false-positive cells, sequences perfectly complementary to blood cell–specific microRNA, miR-142-3p, were incorporated into the 3′-untranslated region of the E1B and GFP genes. In addition, the fiber protein was replaced with that of Ad serotype 35, which recognizes human CD46, creating rAdF35-142T-GFP. rAdF35-142T-GFP efficiently labeled not only CAR-positive tumor cells but also CAR-negative tumor cells with GFP. The numbers of false-positive cells were dramatically lower for rAdF35-142T-GFP than for rAd-GFP. CTCs in the blood of cancer patients were detected by rAdF35-142T-GFP with a large reduction in false-positive cells.

Original languageEnglish
Pages (from-to)16001
Number of pages1
JournalMolecular Therapy - Methods and Clinical Development
Volume3
DOIs
Publication statusPublished - Mar 16 2016

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Circulating Neoplastic Cells
Adenoviridae
Fluorescence
Proteins
Neoplasms
3' Untranslated Regions
MicroRNAs

ASJC Scopus subject areas

  • Genetics
  • Molecular Biology
  • Molecular Medicine

Cite this

Efficient detection of human circulating tumor cells without significant production of false-positive cells by a novel conditionally replicating adenovirus. / Sakurai, Fuminori; Narii, Nobuhiro; Tomita, Kyoko; Togo, Shinsaku; Takahashi, Kazuhisa; Machitani, Mitsuhiro; Tachibana, Masashi; Ouchi, Masaaki; Katagiri, Nobuyoshi; Urata, Yasuo; Fujiwara, Toshiyoshi; Mizuguchi, Hiroyuki.

In: Molecular Therapy - Methods and Clinical Development, Vol. 3, 16.03.2016, p. 16001.

Research output: Contribution to journalArticle

Sakurai, Fuminori ; Narii, Nobuhiro ; Tomita, Kyoko ; Togo, Shinsaku ; Takahashi, Kazuhisa ; Machitani, Mitsuhiro ; Tachibana, Masashi ; Ouchi, Masaaki ; Katagiri, Nobuyoshi ; Urata, Yasuo ; Fujiwara, Toshiyoshi ; Mizuguchi, Hiroyuki. / Efficient detection of human circulating tumor cells without significant production of false-positive cells by a novel conditionally replicating adenovirus. In: Molecular Therapy - Methods and Clinical Development. 2016 ; Vol. 3. pp. 16001.
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