Human High Molecular Weight–Melanoma-Associated Antigen: Utility for Detection of Metastatic Melanoma in Sentinel Lymph Nodes

Goto, Yasufumi; Ferrone, Soldano; Arigami, Takaaki; Kitago, Minoru; Tanemura, Atsushi; Sunami, Eiji; Nguyen, Sandy L.; Turner, Roderick R.; Morton, Donald L.; Hoon, Dave S.B.

doi:10.1158/1078-0432.ccr-07-1842

Cited by 28 publications

(33 citation statements)

References 24 publications

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“…34,35 Goto et al has shown that immunohistochemical detection using HMW-MAAspecific antibodies were more sensitive and specific than the antibodies MART-1, S100p, and HMB-45 which are commonly used for detection of malignant melanoma metastases in sentinel lymph nodes. 36 In this study, we developed a DTIC resistant cell line, FEMX-200DR, from the sensitive cell line FEMX to examine whether induced DTIC resistance also is a determining factor for the effectiveness of 9.2.27PE. The induced DTIC resistance resulted in a higher resistance level to apoptosis in the FEMX-200DR cells, as higher concentrations of the apoptotic inducer STS was needed to obtain IC 50 .…”

Section: Discussionmentioning

confidence: 99%

Anti-Melanoma Activity of the 9.2.27PE Immunotoxin in Dacarbazine Resistant Cells

Risberg

Fodstad²,

Andersson³

2010

Journal of Immunotherapy

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We have earlier shown that the 9.2.27 Pseudomonas Exotoxin A (PE) immunotoxin (IT) efficiently kills melanoma cells through inhibition of protein synthesis followed by some morphologic and biochemical features of apoptosis, a different cell killing mechanism than the one caused by Dacarbazine (DTIC), a chemotherapeutic drug used to treat malignant melanoma. To examine whether induced DTIC resistance also is a determining factor for the effectiveness of 9.2.27PE IT, we developed a DTIC resistant subline, FEMX-200DR, from the DTIC sensitive cell line FEMX. The cell variants were treated with 9.2.27PE, an IT binding to the high molecular weight-melanoma associated antigen (HMW-MAA) expressed on most malignant melanoma cells. The IT was equally effective in killing the FEMX-200DR and the FEMX cells, and the cell death was primarily caused by inhibition of protein synthesis. The DNA repair enzyme and apoptotic marker PARP, a substrate of caspase-3, was inactivated, although we observed only a minor activation of caspase-3 and caspase-8, intracellular proteases involved in apoptosis. In addition to being DTIC resistant, the FEMX-200DR cells were also more resistant to apoptosis than the parent cells as a 3 times higher concentration of the apoptotic inducer Staurosporine was needed to obtain IC50. Furthermore, in early passage malignant melanoma cell lines established from lymph node metastases, the 9.2.27PE caused a time-dependent and dose-dependent decrease in cell viability independent of their DTIC sensitivity. These findings show that the 9.2.27PE IT efficiently can cause cell death in malignant melanoma cells independent of their level of resistance to apoptosis and DTIC.

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Section: Discussionmentioning

confidence: 99%

Anti-Melanoma Activity of the 9.2.27PE Immunotoxin in Dacarbazine Resistant Cells

Risberg

Fodstad²,

Andersson³

2010

Journal of Immunotherapy

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“…Paraffin-embedded tissue microarrays (TMA) for AJCC stage IV melanoma established at JWCI included 262 stage IV metastatic tumors with paired stage III tumor tissues, each in duplicate, from 137 patients (28,29). TMA also contained seven normal tissues with limited or no proliferation (adrenal gland, brain, kidney, and liver), in duplicate.…”

Section: Methodsmentioning

confidence: 99%

“…TMA also contained seven normal tissues with limited or no proliferation (adrenal gland, brain, kidney, and liver), in duplicate. TMA were clinically well-annotated with greater than 5 years follow-up (28,29). Immunohistochemistry (IHC) was performed as previously described (28,29), using the following Abs: mouse anti-human KPC1 Ab (1:50 dilution, #ab57549; Abcam), rabbit anti-human NF-κB1 p50 Ab (1:200 dilution, #sc-114; Santa Cruz Biotechnology), or mouse anti-human p27 Ab (1:100 dilution, #610241; BD Biosciences).…”

Section: Methodsmentioning

confidence: 99%

Epigenetic Regulation of KPC1 Ubiquitin Ligase Affects the NF-κB Pathway in Melanoma

Iida

Ciechanover

Marzese

et al. 2017

Clinical Cancer Research

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Purpose Abnormal activation of the NF-κB pathway induces a more aggressive phenotype of cutaneous melanoma. Understanding the mechanisms involved in melanoma NF-κB activation may identify novel targets for this pathway. KPC1, an E3 ubiquitin ligase, is a regulator of NF-κB pathway. The objective of this study was to investigate the mechanisms regulating KPC1 expression and its clinical impact in melanoma. Experimental Design The clinical impact of KPC1 expression and its epigenetic regulation were assessed in large cohorts of clinically well-annotated melanoma tissues (tissue micro-arrays; n=137, JWCI cohort; n=40) and The Cancer Genome Atlas database (TCGA cohort, n=370). Using melanoma cell lines, we investigated the functional interactions between KPC1 and NF-κB, and the epigenetic regulations of KPC1, including DNA methylation and microRNA expression. Results We verified that KPC1 suppresses melanoma proliferation by processing NF-κB1 p105 into p50, thereby modulating NF-κB-target gene expression. Concordantly, KPC1 expression was down-regulated in AJCC stage IV melanoma compared to early stages (stage I/II p=0.013, stage III p=0.004), and low KPC1 expression was significantly associated with poor overall survival in stage IV melanoma (n=137, Hazard Ratio 1.810, p=0.006). Furthermore, our data showed that high miR-155-5p expression, which is controlled by DNA methylation at its promoter region (TCGA; Pearson’s r −0.455, p<0.001), is significantly associated with KPC1 down-regulation (JWCI; p=0.028, TCGA; p=0.003). Conclusions This study revealed novel epigenetic regulation of KPC1 associated with NF-κB pathway activation, promoting metastatic melanoma progression. These findings suggest the potential utility of KPC1 and its epigenetic regulation as theranostic targets.

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“…mAb were purified from ascitic fluid by sequential precipitation with ammonium sulphate and caprylic acid (24). The purity and activity of mAb preparations were assessed by SDS-PAGE analysis, and by testing with CSPG4-bearing melanoma cells in a binding assay.…”

Section: Methodsmentioning

confidence: 99%

Limited genomic heterogeneity of circulating melanoma cells in advanced stage patients

Ruiz

Luttgen

et al. 2015

Phys. Biol.

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Purpose Circulating melanoma cells (CMCs) constitute a potentially important representation of time-resolved tumor biology in patients. To date, genomic characterization of CMCs has been limited due to the lack of a robust methodology capable of identifying them in a format suitable for downstream characterization. Here, we have developed a methodology to detect intact CMCs that enables phenotypic, morphometric and genomic analysis at the single cell level. Experimental design Blood samples from 40 metastatic melanoma patients and 10 normal blood donors (NBD) were prospectively collected. A panel of 7 chondroitin sulfate proteoglycan 4 (CSPG4)-specific monoclonal antibodies (mAb) was used to immunocytochemically label CMCs. Detection was performed by automated digital fluorescence microscopy and multi-parametric computational analysis. Individual CMCs were captured by micromanipulation for whole genome amplification (WGA) and copy number variation (CNV) analysis. Results Based on CSPG4 expression and nuclear size, 1 to 250 CMCs were detected in 22 (55%) of 40 metastatic melanoma patients (0.5 to 371.5 CMCs/ml). Morphometric analysis revealed that CMCs have a broad spectrum of morphologies and sizes but exhibit a relatively homogeneous nuclear size that was on average 1.5-fold larger than that of surrounding PBMCs. CNV analysis of single CMCs identified deletions of CDKN2A and PTEN, and amplification(s) of TERT, BRAF, KRAS and MDM2. Furthermore, novel chromosomal amplifications in chr12, 17 and 19 were also found. Conclusions Our findings show that CSPG4 expressing CMCs can be found in the majority of advanced melanoma patients. High content analysis of this population may contribute to develop effective therapeutic strategies.

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Human High Molecular Weight–Melanoma-Associated Antigen: Utility for Detection of Metastatic Melanoma in Sentinel Lymph Nodes

Cited by 28 publications

References 24 publications

Anti-Melanoma Activity of the 9.2.27PE Immunotoxin in Dacarbazine Resistant Cells

Anti-Melanoma Activity of the 9.2.27PE Immunotoxin in Dacarbazine Resistant Cells

Epigenetic Regulation of KPC1 Ubiquitin Ligase Affects the NF-κB Pathway in Melanoma

Limited genomic heterogeneity of circulating melanoma cells in advanced stage patients

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