American Association for Cancer Research
10780432ccr204518-sup-255819_2_supp_7025875_qqvpbb.xlsx (12.57 kB)

Supplementary Table S5, from A Novel Topical Fluorescent Probe for Detection of Glioblastoma

Download (12.57 kB)
posted on 2023-03-31, 22:46 authored by Yosuke Kitagawa, Shota Tanaka, Mako Kamiya, Yugo Kuriki, Kyoko Yamamoto, Takenori Shimizu, Takahide Nejo, Taijun Hana, Reiko Matsuura, Tsukasa Koike, Erika Yamazawa, Yoshihiro Kushihara, Satoshi Takahashi, Masashi Nomura, Hirokazu Takami, Shunsaku Takayanagi, Akitake Mukasa, Yasuteru Urano, Nobuhito Saito

Real-time PCR original data of lysates from surgical specimens


Translational Research Program


Japan Agency for Medical Research and Development

Grant-in-Aid for Young Scientists

Grant-in-Aid for Sicentific Research

Japan Society for the Promotion of Science

AMED Practical Research for Innovative Cancer Control



Five-aminolevulinic acid (5-ALA) is widely used as an intraoperative fluorescent probe for radical resection of high-grade glioma, and thus aids in extending progression-free survival of patients. However, there exist some cases where 5-ALA fails to fluoresce. In some other cases, it may undergo fluorescence quenching but cannot be orally readministered during surgery. This study aimed to develop a novel hydroxymethyl rhodamine green (HMRG)-based fluorescence labeling system that can be repeatedly administered as a topical spray during surgery for the detection of glioblastoma. We performed a three-stage probe screening using tumor lysates and fresh tumor tissues with our probe library consisting of a variety of HMRG probes with different dipeptides. We then performed proteome and transcript expression analyses to detect candidate enzymes responsible for cleaving the probe. Moreover, in vitro and ex vivo studies using U87 glioblastoma cell line were conducted to validate the findings. The probe screening identified proline-arginine–HMRG (PR-HMRG) as the optimal probe that distinguished tumors from peritumoral tissues. Proteome analysis identified calpain-1 (CAPN1) to be responsible for cleaving the probe. CAPN1 was highly expressed in tumor tissues which reacted to the PR-HMRG probe. Knockdown of this enzyme suppressed fluorescence intensity in U87 glioblastoma cells. In situ assay using a mouse U87 xenograft model demonstrated marked contrast of fluorescence with the probe between the tumor and peritumoral tissues. The novel fluorescent probe PR-HMRG is effective in detecting glioblastoma when applied topically. Further investigations are warranted to assess the efficacy and safety of its clinical use.