Fluorescence‐based discrimination of breast cancer cells by direct exposure to 5‐aminolevulinic acid

Morita, Midori; Tanaka, Hideo; Kumamoto, Yoshiaki; Nakamura, Akihiro; Harada, Yoshinori; Ogata, Toru; Sakaguchi, Kôichi; Taguchi, Tetsuya; Takamatsu, Tetsuro

doi:10.1002/cam4.2466

Cited by 18 publications

(9 citation statements)

References 47 publications

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“…This is the first oncological study to metabolically isolate and purify GBM cells representative of infiltrative residual disease, from which candidate therapy targets are likely more clinically relevant and present a methodological approach broadly applicable for solid cancers amenable to 5-ALA-induced metabolic labeling such as melanoma or breast cancer. 16 , 17 We report on the RNAseq data acquired from these different tumor areas and validate a novel potential therapeutic target found on the invasive zone 5-ALA-positive cells.…”

mentioning

confidence: 64%

Metabolism-based isolation of invasive glioblastoma cells with specific gene signatures and tumorigenic potential

Smith

Rowlinson

Estevez-Cebrero

et al. 2020

Neuro-Oncology Advances

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Background Glioblastoma (GBM) is a highly aggressive brain tumor with rapid subclonal diversification, harboring molecular abnormalities that vary temporo-spatially, a contributor to therapy resistance. Fluorescence guided neurosurgical resection utilizes administration of 5-aminolevulinic acid (5ALA) generating individually fluorescent tumor cells within a background population of non-neoplastic cells in the invasive tumor region. The aim of the study was to specifically isolate and interrogate the invasive GBM cell population using a novel 5ALA based method. Methods We have isolated the critical invasive GBM cell population by developing 5ALA-based metabolic fluorescence activated cell sorting. This allows purification and study of invasive cells from GBM without an overwhelming background “normal brain” signal to confound data. The population was studied using RNAseq, rtPCR and immunohistochemistry, with gene targets functionally interrogated on proliferation and migration assays using siRNA knockdown and known drug inhibitors. Results RNAseq analysis identifies specific genes such as SERPINE1 which is highly expressed in invasive GBM cells but at low levels in the surrounding normal brain parenchyma. siRNA knockdown and pharmacological inhibition with specific inhibitors of SERPINE1 reduced the capacity of GBM cells to invade in an in vitro assay. Rodent xenografts of 5ALA positive cells were established and serially transplanted, confirming tumorigenicity of the fluorescent patient derived cells but not the 5ALA negative cells. Conclusions Identification of unique molecular features in the invasive GBM population offer hope for developing more efficacious targeted therapies compared to targeting the tumor core and for isolating tumor sub-populations based upon intrinsic metabolic properties.

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confidence: 64%

Metabolism-based isolation of invasive glioblastoma cells with specific gene signatures and tumorigenic potential

Smith

Rowlinson

Estevez-Cebrero

et al. 2020

Neuro-Oncology Advances

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“…The activity of ABC superfamily G member 2 (ABCG2), which is used to discharge PpIX, also affects the PpIX FI ( Figure 1 ) [ 24 ]. Figure 2 shows the effect of Ko143, a specific inhibitor of ABCG2, on PpIX FI in human breast cancer cells and xenografts [ 26 ]. The combined exposure of cultured MCF-7 and MDA-MB 231 cancer cells to 5-ALA and Ko143 boosts PpIX-FI compared to that in cultured non-cancer MCF10A cells, suggesting an improvement in the accuracy of 5-ALA PDD ( Figure 2 A) [ 26 ].…”

Section: 5-ala Metabolic Pathwaymentioning

confidence: 99%

“…Figure 2 shows the effect of Ko143, a specific inhibitor of ABCG2, on PpIX FI in human breast cancer cells and xenografts [ 26 ]. The combined exposure of cultured MCF-7 and MDA-MB 231 cancer cells to 5-ALA and Ko143 boosts PpIX-FI compared to that in cultured non-cancer MCF10A cells, suggesting an improvement in the accuracy of 5-ALA PDD ( Figure 2 A) [ 26 ]. Increased FI attributed to the combined administration of 5-ALA and Ko143 was also observed in cancer cells acquired by fine needle aspiration in a murine tumor model ( Figure 2 B) [ 26 ].…”

Section: 5-ala Metabolic Pathwaymentioning

confidence: 99%

“…The combined exposure of cultured MCF-7 and MDA-MB 231 cancer cells to 5-ALA and Ko143 boosts PpIX-FI compared to that in cultured non-cancer MCF10A cells, suggesting an improvement in the accuracy of 5-ALA PDD ( Figure 2 A) [ 26 ]. Increased FI attributed to the combined administration of 5-ALA and Ko143 was also observed in cancer cells acquired by fine needle aspiration in a murine tumor model ( Figure 2 B) [ 26 ]. In addition, clinically used ABCG2-interacting kinase inhibitors, such as lapatinib, gefitinib, sunitinib, and PD169316, have also been reported to increase 5-ALA induced PpIX-FI in triple-negative breast cancer cell lines [ 27 ].…”

Section: 5-ala Metabolic Pathwaymentioning

confidence: 99%

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5-Aminolevulinic Acid-Induced Protoporphyrin IX Fluorescence Imaging for Tumor Detection: Recent Advances and Challenges

Harada

Murayama

Takamatsu

et al. 2022

IJMS

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5-Aminolevulinic acid (5-ALA) is a natural amino acid and a precursor of heme and chlorophyll. Exogenously administered 5-ALA is metabolized into protoporphyrin IX (PpIX). PpIX accumulates in cancer cells because of the low activity of ferrochelatase, an enzyme that metabolizes PpIX to heme. High expression of 5-ALA influx transporters, such as peptide transporters 1/2, in cancer cells also enhances PpIX production. Because PpIX radiates red fluorescence when excited with blue/violet light, 5-ALA has been used for the visualization of various tumors. 5-ALA photodynamic diagnosis (PDD) has been shown to improve the tumor removal rate in high-grade gliomas and non-muscular invasive bladder cancers. However, 5-ALA PDD remains a challenge as a diagnostic method because tissue autofluorescence interferes with PpIX signals in cases where tumors emit only weak signals, and non-tumorous lesions, such as inflammatory sites, tend to emit PpIX fluorescence. Here, we review the current outline of 5-ALA PDD and strategies for improving its diagnostic applicability for tumor detection, focusing on optical techniques and 5-ALA metabolic pathways in both viable and necrotic tumor tissues.

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“…Kobuchi et al already investigated this topic on different cancer cells and found an important variation in ABCG2 expression from one cell type to another [ 39 ]. More recently, Morita et al investigated the selective accumulation of PpIX in breast cancer cells from a tumor-bearing mouse model [ 120 ]. In a confocal fluorescence microscopy assay, they found that MCF7 and MDA-MB-231 did highly respond to 5-ALA stimulation 2 h after treatment unlike the normal epithelial MCF10A cells, supporting a selectivity for breast cancer.…”

Section: Concerning Ppix Selective Accumulation In Neoplastic Cellsmentioning

confidence: 99%

A Recap of Heme Metabolism towards Understanding Protoporphyrin IX Selectivity in Cancer Cells

Kiening

Lange

2022

IJMS

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Mitochondria are essential organelles of mammalian cells, often emphasized for their function in energy production, iron metabolism and apoptosis as well as heme synthesis. The heme is an iron-loaded porphyrin behaving as a prosthetic group by its interactions with a wide variety of proteins. These complexes are termed hemoproteins and are usually vital to the whole cell comportment, such as the proteins hemoglobin, myoglobin or cytochromes, but also enzymes such as catalase and peroxidases. The building block of porphyrins is the 5-aminolevulinic acid, whose exogenous administration is able to stimulate the entire heme biosynthesis route. In neoplastic cells, this methodology repeatedly demonstrated an accumulation of the ultimate heme precursor, the fluorescent protoporphyrin IX photosensitizer, rather than in healthy tissues. While manifold players have been proposed, numerous discrepancies between research studies still dispute the mechanisms underlying this selective phenomenon that yet requires intensive investigations. In particular, we wonder what are the respective involvements of enzymes and transporters in protoporphyrin IX accretion. Is this mainly due to a boost in protoporphyrin IX anabolism along with a drop of its catabolism, or are its transporters deregulated? Additionally, can we truly expect to find a universal model to explain this selectivity? In this report, we aim to provide our peers with an overview of the currently known mitochondrial heme metabolism and approaches that could explain, at least partly, the mechanism of protoporphyrin IX selectivity towards cancer cells.

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Fluorescence‐based discrimination of breast cancer cells by direct exposure to 5‐aminolevulinic acid

Cited by 18 publications

References 47 publications

Metabolism-based isolation of invasive glioblastoma cells with specific gene signatures and tumorigenic potential

Metabolism-based isolation of invasive glioblastoma cells with specific gene signatures and tumorigenic potential

5-Aminolevulinic Acid-Induced Protoporphyrin IX Fluorescence Imaging for Tumor Detection: Recent Advances and Challenges

A Recap of Heme Metabolism towards Understanding Protoporphyrin IX Selectivity in Cancer Cells

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