FLGS-01. Development of novel topical fluorescent probe for intraoperative rapid detection of glioma

Abstract PURPOSE Fluorescence imaging is an important surgical adjunct in malignant glioma surgery. 5-aminolevulinic acid (5-ALA) has been proven effective for radical tumor resection and extended progression-free survival in a phase III randomized trial and therefore integrated into surgery for malignant glioma. Importantly, however, some limitations still exist in its use, which include false positivity and false negativity as well as inability of re-administration. In this study, we aimed to develop a novel, spray-type fluorescent probe using hydroxymethyl rhodamine green (HMRG) as a fluorescent scaffold. METHODS We have previously established a fluorescent probe library comprised of more than 320 kinds of HMRG probes. They have HMRG as a fluorescent scaffold with various types of dipeptides attached to it. Primary probe screening was performed using the homogenized tumor samples from patients with glioblastoma operated at our institution. Secondary screening followed using the selected probes and fresh tumor samples obtained from patients with glioblastoma operated from 2016 until 2018. Diced electrophoresis gel (DEG) assay, two-dimensional gel electrophoresis followed by a multi-well plate-based fluorometric assay, was performed to identify responsible enzymes for the selected probe. Further experiments with inhibitors, real-time PCR, immunohistochemistry, and western blotting were performed for confirmation. RESULTS Proline-arginine-HMRG (PR-HMRG) was selected as a candidate probe based upon the above two-step screenings. It achieved 79.4% accuracy in receiver operating characteristic curve analysis. Calpain-1 was found to be responsible to cleave PR-HMRG probe by DEG-proteome analysis. Calpain-1 protein was highly expressed in tumor tissues which reacted to PR-HMRG probe. CONCLUSIONS Our innovative screening method was able to find PR-HMRG as a novel fluorescent probe effective for rapid detection of glioblastoma. A preclinical study is planned to assess the efficacy and safety of the selected probe.

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SURG-10. DEVELOPMENT OF NOVEL TOPICAL FLUORESCENT PROBE FOR INTRAOPERATIVE RAPID DETECTION OF GLIOMA

Neuro-Oncology ◽

10.1093/neuonc/noab196.780 ◽

2021 ◽

Vol 23 (Supplement_6) ◽

pp. vi197-vi197

Author(s):

Shota Tanaka ◽

Yosuke Kitagawa ◽

Mako Kamiya ◽

Yugo Kuriki ◽

Kyoko Yamamoto ◽

...

Keyword(s):

Malignant Glioma ◽

Fluorescent Probe ◽

Rapid Detection ◽

Aminolevulinic Acid ◽

Characteristic Curve ◽

Screening Method ◽

Tumor Resection ◽

Phase Iii ◽

Glioma Surgery ◽

Two Dimensional Gel Electrophoresis

Abstract PURPOSE Fluorescence imaging is an important surgical adjunct in malignant glioma surgery. 5-aminolevulinic acid (5-ALA) has been proven effective for radical tumor resection and extended progression-free survival in a phase III randomized trial and therefore integrated into surgery for malignant glioma. Importantly, however, some limitations still exist in its use, which include false positivity and false negativity as well as inability of re-administration. In this study, we aimed to develop a novel, spray-type fluorescent probe using hydroxymethyl rhodamine green (HMRG) as a fluorescent scaffold. METHODS We have previously established a fluorescent probe library comprised of more than 320 kinds of HMRG probes. They have HMRG as a fluorescent scaffold with various types of dipeptides attached to it. Primary probe screening was performed using the homogenized tumor samples from patients with glioblastoma operated at our institution. Secondary screening followed using the selected probes and fresh tumor samples obtained from patients with glioblastoma operated from 2016 until 2018. Diced electrophoresis gel (DEG) assay, two-dimensional gel electrophoresis followed by a multi-well plate-based fluorometric assay, was performed to identify responsible enzymes for the selected probe. Further experiments with inhibitors, real-time PCR, immunohistochemistry, and western blotting were performed for confirmation. RESULTS Proline-arginine-HMRG (PR-HMRG) was selected as a candidate probe based upon the above two-step screenings. It achieved 79.4% accuracy in receiver operating characteristic curve analysis. Calpain-1 was found to be responsible to cleave PR-HMRG probe by DEG-proteome analysis. Calpain-1 protein was expressed at significantly higher level in tumors that were fluoresced by PR-HMRG than in those that were not. CONCLUSIONS Our innovative screening method was able to find PR-HMRG as a novel fluorescent probe effective for rapid detection of glioblastoma. A preclinical study is planned to assess the efficacy and safety of the selected probe.

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Counterbalancing risks and gains from extended resections in malignant glioma surgery: a supplemental analysis from the randomized 5-aminolevulinic acid glioma resection study

Journal of Neurosurgery ◽

10.3171/2010.3.jns097 ◽

2011 ◽

Vol 114 (3) ◽

pp. 613-623 ◽

Cited By ~ 176

Author(s):

Walter Stummer ◽

Jörg-Christian Tonn ◽

Hubertus Maximilian Mehdorn ◽

Ulf Nestler ◽

Kea Franz ◽

...

Keyword(s):

Malignant Glioma ◽

Aminolevulinic Acid ◽

Malignant Gliomas ◽

Neurological Function ◽

Phase Iii ◽

Neurological Deficits ◽

Secondary Outcome ◽

Karnofsky Performance Scale ◽

Glioma Surgery ◽

Repeat Surgery

Object Accumulating data suggest more aggressive surgery in patients with malignant glioma to improve outcome. However, extended surgery may increase morbidity. The randomized Phase III 5-aminolevulinic acid (ALA) study investigated 5-ALA–induced fluorescence as a tool for improving resections. An interim analysis demonstrated more frequent complete resections with longer progression-free survival (PFS). However, marginal differences were found regarding neurological deterioration and the frequency of additional therapies. Presently, the authors focus on the latter aspects in the final study population, and attempt to determine how safety might be affected by cytoreductive surgery. Methods Patients with malignant gliomas were randomized for fluorescence-guided (ALA group) or conventional white light (WL) (WL group) microsurgery. The final intent-to-treat population consisted of 176 patients in the ALA and 173 in the WL group. Primary efficacy variables were contrast-enhancing tumor on early MR imaging and 6-month PFS. Among secondary outcome measures, the National Institutes of Health Stroke Scale (NIH-SS) score and the Karnofsky Performance Scale (KPS) score were used for assessing neurological function. Results More frequent complete resections and improved PFS were confirmed, with higher median residual tumor volumes in the WL group (0.5 vs 0 cm3, p = 0.001). Patients in the ALA group had more frequent deterioration on the NIH-SS at 48 hours. Patients at risk were those with deficits unresponsive to steroids. No differences were found in the KPS score. Regarding outcome, a combined end point of risks and neurological deficits was attempted, which demonstrated results in patients in the ALA group to be superior to those in participants in the WL group. Interestingly, the cumulative incidence of repeat surgery was significantly reduced in ALA patients. When stratified by completeness of resection, patients with incomplete resections were quicker to deteriorate neurologically (p = 0.0036). Conclusions Extended resections performed using a tool such as 5-ALA–derived tumor fluorescence, carries the risk of temporary impairment of neurological function. However, risks are higher in patients with deficits unresponsive to steroids.

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Efficacy of 5-aminolevulinic acid-induced fluorescence detection in malignant glioma surgery

International Congress Series ◽

10.1016/j.ics.2004.03.324 ◽

2004 ◽

Vol 1268 ◽

pp. 1290

Author(s):

Takashi Maruyama ◽

Yoshihiro Muragaki ◽

Masahiko Tanaka ◽

Hiroshi Iseki ◽

Ichiro Sakuma ◽

...

Keyword(s):

Malignant Glioma ◽

Fluorescence Detection ◽

Aminolevulinic Acid ◽

Glioma Surgery

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To what extent will 5-aminolevulinic acid change the face of malignant glioma surgery?

CNS Oncology ◽

10.2217/cns.15.10 ◽

2015 ◽

Vol 4 (4) ◽

pp. 265-272 ◽

Cited By ~ 6

Author(s):

Ricardo Díez Valle ◽

Sonia Tejada Solis

Keyword(s):

Malignant Glioma ◽

Aminolevulinic Acid ◽

Glioma Surgery ◽

The Face

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5-Aminolevulinic Acid Fluorescence-Guided Resection of Intramedullary Ependymoma: Report of 9 Cases

Operative Neurosurgery ◽

10.1227/neu.0b013e31827bc7a3 ◽

2012 ◽

Vol 72 (2) ◽

pp. ons159-ons168 ◽

Cited By ~ 12

Author(s):

Tomoo Inoue ◽

Toshiki Endo ◽

Kenichi Nagamatsu ◽

Mika Watanabe ◽

Teiji Tominaga

Keyword(s):

Spinal Cord ◽

Aminolevulinic Acid ◽

Tumor Resection ◽

Extent Of Resection ◽

Labeling Index ◽

World Health ◽

Histopathological Diagnosis ◽

Glioma Surgery ◽

Ki 67 ◽

Grade Ii

Abstract BACKGROUND: Resection guided by 5-aminolevulinic acid (5-ALA) fluorescence has proved to be useful in intracranial glioma surgery. However, the effects of 5-ALA on spinal cord tumors remain unknown. OBJECTIVE: To evaluate the usefulness of 5-ALA fluorescence-guided resection of intramedullary ependymoma for achieving maximum tumor resection. METHODS: This study included 10 patients who underwent surgical resection of an intramedullary ependymoma. Nine patients were orally administered 5-ALA (20 mg/kg) 2 hours before the induction of anesthesia. 5-ALA fluorescence was visualized with an operating microscope. Tumors were removed in a standardized manner with electro-physiological monitoring. The extent of resection was evaluated on the basis of intra-operative findings and postoperative magnetic resonance imaging. Histopathological diagnosis was established according to World Health Organization 2007 criteria. Cell proliferation was assessed by Ki-67 labeling index. RESULTS: 5-ALA fluorescence was positive in 7 patients (6 grade II and 1 grade III) and negative in 2 patients (grade II). Intraoperative findings were dichotomized: Tumors covered by the cyst were easily separated from the normal parenchyma, whereas tumors without the cyst appeared to be continuous to the spinal cord. In these cases, 5-ALA fluorescence was especially valuable in delineating the ventral and cranial and caudal margins. Ki-67 labeling index was significantly higher in 5-ALA-positive cases compared with 5-ALA-negative cases. All patients improved neurologically or stabilized after surgery. CONCLUSION: 5-ALA fluorescence was useful for detecting tumor margins during surgery for intramedullary ependymoma. When combined with electrophysiological monitoring, fluorescence-guided resection could help to achieve maximum tumor resection safely.

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Observational, retrospective study of the effectiveness of 5-aminolevulinic acid in malignant glioma surgery in Spain (The VISIONA study)

Neurología (English Edition) ◽

10.1016/j.nrleng.2013.05.004 ◽

2014 ◽

Vol 29 (3) ◽

pp. 131-138 ◽

Cited By ~ 3

Author(s):

R. Díez Valle ◽

J. Slof ◽

J. Galván ◽

C. Arza ◽

C. Romariz ◽

...

Keyword(s):

Retrospective Study ◽

Malignant Glioma ◽

Aminolevulinic Acid ◽

Glioma Surgery

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SURG-18. 5-AMINOLEVULINIC ACID FLUORESCENCE (5-ALA) GUIDED RESECTION OF GLIOBLASTOMA, LESSON LEARNED FROM 19 CASES

Neuro-Oncology ◽

10.1093/neuonc/noz175.1018 ◽

2019 ◽

Vol 21 (Supplement_6) ◽

pp. vi243-vi243

Author(s):

Jinmo Cho

Keyword(s):

Aminolevulinic Acid ◽

Tumor Resection ◽

High Grade Glioma ◽

Extent Of Resection ◽

High Grade ◽

Total Resection ◽

Glioma Surgery ◽

Term Survival ◽

Tumor Bed ◽

Key Factor

Abstract BACKGROUND 5-ALA is known as useful tool for high grade glioma resection and the accumulation extent of 5-ALA is known as far beyond gadolinium enhancement. Extent of resection is key factor for favorable outcome and long-term survival for high grade glioma patients and 5-ALA might increase extent of resection. We present our experience of 5-ALA guided glioma surgeries METHODS Total 19 patients were performed 5-ALA guided surgery. They ingested 20mg/kg, four hours before craniotomy. We tried to perform supra-total resection rather than gross total resection according to the tumor consistency and if the tumor located relatively non-eloquent area, we tried to perform lobectomy rather than lesionectomy. After tumor resection, we inspect the tumor bed under 5-ALA fluorescence, and we confirmed the complete loss of fluorescence on the tumor resected bed. We check the MRI within 48 hour after operation and assess the extent of resection RESULTS Among the 19 patients, 15 patients were confirmed glioblastoma and 3 anaplastic astrocytoma and 1 anaplastic oligoastrocytoma. We confirmed all enhancing lesion was completely removed, however, 2 patients show residual non-enhancing lesion in post-operative MRI. Two patients suffered temporary hemiparesis and 2 patients show permanent visual field defect. CONCLUSION 5-ALA is useful tool for glioma surgery. Resection extent could be increased, however, non-enhancing lesion in the high grade gliomas, might be missed under 5-ALA guidance.

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