A novel method for assessing the renal biopsy specimens using an activatable fluorescent probe

Takuji Iyama,Tsutomu Kanda,Tomoaki Takata,Marie Yamamoto,Takaaki Sugihara,Kentaro Yamada,Yukari Mae,Yasuteru Urano,Masaya Ogawa,Hajime Isomoto,Shintaro Hamada,Satoko Fukuda,Ayami Ida,Sosuke Taniguchi

doi:10.1038/s41598-020-69077-w

Takuji Iyama, Tsutomu Kanda + Show 12 more

Open Access

https://doi.org/10.1038/s41598-020-69077-w

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Journal: Scientific Reports	Publication Date: Jul 21, 2020
Citations: 8	License type: open-access

Affiliation: Tottori University, The University of Tokyo

Abstract

Gamma-glutamyl hydroxymethyl rhodamine green (gGlu-HMRG) is an activatable fluorescent probe that can be activated by γ-glutamyltranspeptidase (GGT). The expression of GGT in the kidney, which is one of the major organs exhibiting enhanced GGT expression, is exclusively localised to the cortex. Here, we aimed to investigate the feasibility of gGlu-HMRG as a probe for the on-site assessment of renal biopsy specimens. gGlu-HMRG fluorescent probe was applied to the renal proximal tubular epithelial cells and cortical collecting duct cells in vitro, mouse kidneys ex vivo, and human biopsy specimens. In addition, the fluorescence intensities in the cortex and the medulla were comparatively evaluated in the biopsy specimens. The fluorescence signal was rapidly detected in the renal proximal tubular epithelial cells, whereas that in the cortical collecting duct cells was not detected. The fluorescence signal was detected in the mouse kidneys ex vivo without markedly affecting the tissue morphology. In the human biopsy specimens, the fluorescence signal in the cortex was significantly distinct from that in the medulla (p < 0.05). Thus, this fluorescent probe can be used to distinctly identify the renal cortex in the biopsy specimens.

Highlights

Gamma-glutamyl hydroxymethyl rhodamine green is an activatable fluorescent probe that can be activated by γ-glutamyltranspeptidase (GGT)
The fluorescence signal was clearly detected in the renal proximal tubule epithelial cells (RPTECs), whereas it could not be detected in the cortical collecting duct cells (Fig. 1A,B)
The fluorescence signal in the RPTECs was stronger when gGlu-HMRG was applied at higher concentration and the intensity gradually increased immediately after the administration of gGlu-HMRG (Fig. 1)

Summary

Introduction

Gamma-glutamyl hydroxymethyl rhodamine green (gGlu-HMRG) is an activatable fluorescent probe that can be activated by γ-glutamyltranspeptidase (GGT). We aimed to investigate the feasibility of gGlu-HMRG as a probe for the on-site assessment of renal biopsy specimens. GGlu-HMRG fluorescent probe was applied to the renal proximal tubular epithelial cells and cortical collecting duct cells in vitro, mouse kidneys ex vivo, and human biopsy specimens. In the human biopsy specimens, the fluorescence signal in the cortex was significantly distinct from that in the medulla (p < 0.05). GGlu-HMRG could detect pancreatic cancers in biopsy specimens obtained from endoscopic ultrasound-guided fine-needle a spiration[5]. The novel gGlu-HMRG-based imaging technique can be applied to evaluate various biopsy specimens other than detecting cancer. We investigated the feasibility of this activatable fluorescent probe for on-site evaluation of renal biopsy specimens

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