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Rapid intraoperative visualization of breast lesions with γ-glutamyl hydroxymethyl rhodamine green.

Ueo H, Shinden Y, Tobo T, Gamachi A, Udo M, Komatsu H, Nambara S, Saito T, Ueda M, Hirata H, Sakimura S, Takano Y, Uchi R, Kurashige J, Akiyoshi S, Iguchi T, Eguchi H, Sugimachi K, Kubota Y, Kai Y, Shibuta K, Kijima Y, Yoshinaka H, Natsugoe S, Mori M, Maehara Y, Sakabe M, Kamiya M, Kakareka JW, Pohida TJ, Choyke PL, Kobayashi H, Ueo H, Urano Y, Mimori K - Sci Rep (2015)

Bottom Line: Cleavage of the probe by GGT generates green fluorescence.We found that fluorescence derived from cleavage of gGlu-HMRG allowed easy discrimination of breast tumors, even those smaller than 1 mm in size, from normal mammary gland tissues, with 92% sensitivity and 94% specificity, within only 5 min after application.We believe this rapid, low-cost method represents a breakthrough in intraoperative margin assessment during breast-conserving surgery.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Surgery, Kyushu University Beppu Hospital, 4546 Tsurumihara, Beppu 874-0838 [2] Department of Surgery and Sciences, Graduate School of Medical Sciences, Kyushu University, Fukuoka 812-8582.

ABSTRACT
We previously developed γ-glutamyl hydroxymethyl rhodamine green (gGlu-HMRG) as a tool to detect viable cancer cells, based on the fact that the enzyme γ-glutamyltranspeptidase (GGT) is overexpressed on membranes of various cancer cells, but is not expressed in normal tissue. Cleavage of the probe by GGT generates green fluorescence. Here, we examined the feasibility of clinical application of gGlu-HMRG during breast-conserving surgery. We found that fluorescence derived from cleavage of gGlu-HMRG allowed easy discrimination of breast tumors, even those smaller than 1 mm in size, from normal mammary gland tissues, with 92% sensitivity and 94% specificity, within only 5 min after application. We believe this rapid, low-cost method represents a breakthrough in intraoperative margin assessment during breast-conserving surgery.

No MeSH data available.


Related in: MedlinePlus

Application of the gGlu-HMRG fluorescence method in surgical margins of BCS specimens.Specimen 1 was diagnosed pathologically as DCIS and specimen 2 was diagnosed as invasive ductal carcinoma (papillotubular). (a) Gross picture. (b) Fluorescence image just before gGlu-HMRG administration. (c) Fluorescence image 5 minutes after gGlu-HMRG administration. Increased brightness was observed in some areas. (d) Red and yellow colors indicate fluorescence-positive areas. Red areas were identified as malignant lesions. Yellow areas were fluorescent positive and did not identified as malignant lesions. (e) HE stained image after formalin fixation. Malignant regions identified from pathological findings are colored green. (f) The area in the red box of (e) is magnified. Red arrows showed malignant lesions in the cross section of surgical margin. (g) Gross picture. Red-circled areas showed malignant lesions diagnosed from pathological findings. (h–j) Red and yellow areas showed fluorescence-positive areas. Red areas were fluorescent positive and identified as malignant lesions. Yellow areas were fluorescent positive and did not identified as malignant lesions. The area in the white box of (h) is magnified in (i) and (j). i) The picture at 5 minutes after gGlu-HMRG adminstration. (j) After analyzing the picture, we found and colored fluorescent positive area as red and yellow. (k) HE-staining image same region as (i) and (j) after formalin fixation. Red arrows indicated extensive intraductal components of invasive ductal carcinoma.
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f2: Application of the gGlu-HMRG fluorescence method in surgical margins of BCS specimens.Specimen 1 was diagnosed pathologically as DCIS and specimen 2 was diagnosed as invasive ductal carcinoma (papillotubular). (a) Gross picture. (b) Fluorescence image just before gGlu-HMRG administration. (c) Fluorescence image 5 minutes after gGlu-HMRG administration. Increased brightness was observed in some areas. (d) Red and yellow colors indicate fluorescence-positive areas. Red areas were identified as malignant lesions. Yellow areas were fluorescent positive and did not identified as malignant lesions. (e) HE stained image after formalin fixation. Malignant regions identified from pathological findings are colored green. (f) The area in the red box of (e) is magnified. Red arrows showed malignant lesions in the cross section of surgical margin. (g) Gross picture. Red-circled areas showed malignant lesions diagnosed from pathological findings. (h–j) Red and yellow areas showed fluorescence-positive areas. Red areas were fluorescent positive and identified as malignant lesions. Yellow areas were fluorescent positive and did not identified as malignant lesions. The area in the white box of (h) is magnified in (i) and (j). i) The picture at 5 minutes after gGlu-HMRG adminstration. (j) After analyzing the picture, we found and colored fluorescent positive area as red and yellow. (k) HE-staining image same region as (i) and (j) after formalin fixation. Red arrows indicated extensive intraductal components of invasive ductal carcinoma.

Mentions: Since these results indicated that the gGlu-HMRG fluorescent method might indeed be suitable for intraoperative pathological margin assessment, we next applied it to examine malignant lesions in the margin of BCS specimens (N = 7 from 5 patients). We compared the fluorescence-positive areas with the malignant lesions independently identified by pathologists. All of the identified malignant lesions showed up as fluorescence-positive areas (Fig. 2 and Supplementary Fig. 4). These results support the validity of this fluorescence-based method as a new clinical tool for pathologists to identify candidate tumorous and proliferative lesions for pathological confirmation. From another point of view, gGlu-HMRG could be useful to identify regions containing no malignancy as fluorescence increase-negative regions (Fig. 2 and Supplementary Fig. 4 and 5), where intraoperative microscopic examination by pathologists would not be required.


Rapid intraoperative visualization of breast lesions with γ-glutamyl hydroxymethyl rhodamine green.

Ueo H, Shinden Y, Tobo T, Gamachi A, Udo M, Komatsu H, Nambara S, Saito T, Ueda M, Hirata H, Sakimura S, Takano Y, Uchi R, Kurashige J, Akiyoshi S, Iguchi T, Eguchi H, Sugimachi K, Kubota Y, Kai Y, Shibuta K, Kijima Y, Yoshinaka H, Natsugoe S, Mori M, Maehara Y, Sakabe M, Kamiya M, Kakareka JW, Pohida TJ, Choyke PL, Kobayashi H, Ueo H, Urano Y, Mimori K - Sci Rep (2015)

Application of the gGlu-HMRG fluorescence method in surgical margins of BCS specimens.Specimen 1 was diagnosed pathologically as DCIS and specimen 2 was diagnosed as invasive ductal carcinoma (papillotubular). (a) Gross picture. (b) Fluorescence image just before gGlu-HMRG administration. (c) Fluorescence image 5 minutes after gGlu-HMRG administration. Increased brightness was observed in some areas. (d) Red and yellow colors indicate fluorescence-positive areas. Red areas were identified as malignant lesions. Yellow areas were fluorescent positive and did not identified as malignant lesions. (e) HE stained image after formalin fixation. Malignant regions identified from pathological findings are colored green. (f) The area in the red box of (e) is magnified. Red arrows showed malignant lesions in the cross section of surgical margin. (g) Gross picture. Red-circled areas showed malignant lesions diagnosed from pathological findings. (h–j) Red and yellow areas showed fluorescence-positive areas. Red areas were fluorescent positive and identified as malignant lesions. Yellow areas were fluorescent positive and did not identified as malignant lesions. The area in the white box of (h) is magnified in (i) and (j). i) The picture at 5 minutes after gGlu-HMRG adminstration. (j) After analyzing the picture, we found and colored fluorescent positive area as red and yellow. (k) HE-staining image same region as (i) and (j) after formalin fixation. Red arrows indicated extensive intraductal components of invasive ductal carcinoma.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4499838&req=5

f2: Application of the gGlu-HMRG fluorescence method in surgical margins of BCS specimens.Specimen 1 was diagnosed pathologically as DCIS and specimen 2 was diagnosed as invasive ductal carcinoma (papillotubular). (a) Gross picture. (b) Fluorescence image just before gGlu-HMRG administration. (c) Fluorescence image 5 minutes after gGlu-HMRG administration. Increased brightness was observed in some areas. (d) Red and yellow colors indicate fluorescence-positive areas. Red areas were identified as malignant lesions. Yellow areas were fluorescent positive and did not identified as malignant lesions. (e) HE stained image after formalin fixation. Malignant regions identified from pathological findings are colored green. (f) The area in the red box of (e) is magnified. Red arrows showed malignant lesions in the cross section of surgical margin. (g) Gross picture. Red-circled areas showed malignant lesions diagnosed from pathological findings. (h–j) Red and yellow areas showed fluorescence-positive areas. Red areas were fluorescent positive and identified as malignant lesions. Yellow areas were fluorescent positive and did not identified as malignant lesions. The area in the white box of (h) is magnified in (i) and (j). i) The picture at 5 minutes after gGlu-HMRG adminstration. (j) After analyzing the picture, we found and colored fluorescent positive area as red and yellow. (k) HE-staining image same region as (i) and (j) after formalin fixation. Red arrows indicated extensive intraductal components of invasive ductal carcinoma.
Mentions: Since these results indicated that the gGlu-HMRG fluorescent method might indeed be suitable for intraoperative pathological margin assessment, we next applied it to examine malignant lesions in the margin of BCS specimens (N = 7 from 5 patients). We compared the fluorescence-positive areas with the malignant lesions independently identified by pathologists. All of the identified malignant lesions showed up as fluorescence-positive areas (Fig. 2 and Supplementary Fig. 4). These results support the validity of this fluorescence-based method as a new clinical tool for pathologists to identify candidate tumorous and proliferative lesions for pathological confirmation. From another point of view, gGlu-HMRG could be useful to identify regions containing no malignancy as fluorescence increase-negative regions (Fig. 2 and Supplementary Fig. 4 and 5), where intraoperative microscopic examination by pathologists would not be required.

Bottom Line: Cleavage of the probe by GGT generates green fluorescence.We found that fluorescence derived from cleavage of gGlu-HMRG allowed easy discrimination of breast tumors, even those smaller than 1 mm in size, from normal mammary gland tissues, with 92% sensitivity and 94% specificity, within only 5 min after application.We believe this rapid, low-cost method represents a breakthrough in intraoperative margin assessment during breast-conserving surgery.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Surgery, Kyushu University Beppu Hospital, 4546 Tsurumihara, Beppu 874-0838 [2] Department of Surgery and Sciences, Graduate School of Medical Sciences, Kyushu University, Fukuoka 812-8582.

ABSTRACT
We previously developed γ-glutamyl hydroxymethyl rhodamine green (gGlu-HMRG) as a tool to detect viable cancer cells, based on the fact that the enzyme γ-glutamyltranspeptidase (GGT) is overexpressed on membranes of various cancer cells, but is not expressed in normal tissue. Cleavage of the probe by GGT generates green fluorescence. Here, we examined the feasibility of clinical application of gGlu-HMRG during breast-conserving surgery. We found that fluorescence derived from cleavage of gGlu-HMRG allowed easy discrimination of breast tumors, even those smaller than 1 mm in size, from normal mammary gland tissues, with 92% sensitivity and 94% specificity, within only 5 min after application. We believe this rapid, low-cost method represents a breakthrough in intraoperative margin assessment during breast-conserving surgery.

No MeSH data available.


Related in: MedlinePlus