HER2 fluorescent in situ hybridization signal degradation: a 10-year retrospective study

Wu, Jessie M.; Galibois, Andrey-Ann; Seebocus, Neha; Apu, Anupam; Hnatovska, Marta; Wilson, Annie; Shortill, Teresa; Turashvili, Gulisa

doi:10.1007/s10549-020-06048-9

Cited by 3 publications

(3 citation statements)

References 24 publications

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“…Dual-probe HER2 FISH signal degradation over time and under different slide storage conditions should be considered. Storing slides at −80 °C has been shown to allow for signal retention, while at room temperature, signals start to degrade, with CEP17 signal loss at a faster rate [ 37 ]. Additionally, laboratories should participate in quality assurance and proficiency testing programs specifically for HER2 IHC and ISH in EC, which need to be established.…”

Section: Discussionmentioning

confidence: 99%

Implementation of HER2 Testing in Endometrial Cancer, a Summary of Real-World Initial Experience in a Large Tertiary Cancer Center

Plotkin,

Olkhov-Mitsel,

Huang

et al. 2024

Cancers

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HER2-targeted therapies have transformed the management of advanced or recurrent serous endometrial cancer (EC), leading to an increased clinical demand for HER2 testing. Despite its adoption in select academic centers, the global extent of such tumor testing is unclear. In this study, we report on the initial two-year experience of HER2 testing at a major academic center with a reference gynecologic oncology service and biomarker reference laboratory. All patients who underwent HER2 testing based on physician discretion, reflex HER2 testing, and reference laboratory requests were included. From February 2021 to October 2023, HER2 testing was performed on 192 tumor tissue samples from 180 EC patients. Serous carcinoma constituted 52% of samples, reflecting diagnostic challenges and limited therapeutic options for advanced EC. HER2 positivity was found in 28% of all cases and 30% of p53-aberrant cases. An immunohistochemistry (IHC) score of 3+ was found in 15% of samples, while IHC 2+ was found in 45% (13% IHC 2+/ISH+ and 32% IHC 2+/ISH−). The newly identified ‘HER2-low’ category comprised 46% of the samples. Heterogeneity was noted in 42% of HER2-positive cases, with complex patterns in 3%. NGS and HER2 IHC-FISH showed a 24% discordance, attributed to intratumoral heterogeneity, tumor cellularity, a small number of amplified cells, and the HER2/CEP17 ratio near the cut-off. This study offers real-world insights into HER2 testing in EC, highlighting the challenges and underscoring the need for standardized guidelines in specimen handling, proficiency testing, and scoring criteria to enhance patient management and therapeutic decision-making.

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Section: Discussionmentioning

confidence: 99%

Implementation of HER2 Testing in Endometrial Cancer, a Summary of Real-World Initial Experience in a Large Tertiary Cancer Center

Plotkin,

Olkhov-Mitsel,

Huang

et al. 2024

Cancers

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show abstract

“…This technology is expected to be widely applied. It largely facilitates the application of flow cytometry to the research using zebrafish models and provides another option for the detection of specific gene expression levels, especially when failing to label their mRNAs through routinely used in situ hybridization assays due to the cumbersome steps and possible signal degradation (Wu et al 2021 ).…”

Section: Limitations and Prospectsmentioning

confidence: 99%

Zebrafish: a convenient tool for myelopoiesis research

Jing

2023

Cell Regen

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Myelopoiesis is the process in which the mature myeloid cells, including monocytes/macrophages and granulocytes, are developed. Irregular myelopoiesis may cause and deteriorate a variety of hematopoietic malignancies such as leukemia. Myeloid cells and their precursors are difficult to capture in circulation, let alone observe them in real time. For decades, researchers had to face these difficulties, particularly in in-vivo studies. As a unique animal model, zebrafish possesses numerous advantages like body transparency and convenient genetic manipulation, which is very suitable in myelopoiesis research. Here we review current knowledge on the origin and regulation of myeloid development and how zebrafish models were applied in these studies.

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“…It can quantitatively evaluate the copy number and ratio of the HER2 gene and the reference gene of chromosome 17 centromere (CEP17) of BC. However, this method is cumbersome and susceptible to the attenuation of fluorescent dye signals ( 15 , 16 ). Recently, new HER2 detection methods have been proposed ( 17 ), such as a fully automated HER2 gene-protein assay (GPA), next-generation sequencing (NGS), or digital PCR (dPCR), and RNA FISH assay (RCasFISH) ( 18 – 21 ).…”

Section: Introductionmentioning

confidence: 99%

Lentiviral vector–based xenograft tumors as candidate reference materials for detection of HER2-low breast cancer

Wei

Xu²,

Cao³

et al. 2022

Front. Oncol.

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The human epidermal growth factor receptor 2 (HER2) is an important biomarker that plays a pivotal role in therapeutic decision-making for patients with breast cancer (BC). Patients with HER2-low BC can benefit from new HER2 targeted therapy. For ensuring the accurate and reproducible detection of HER2-low cancer, reliable reference materials are required for monitoring the sensitivity and specificity of detection assays. Herein, a lentiviral vector was used to transduce the HER2 gene into MDA-MB-231 cells that exhibited low HER2 density, and the cells were characterized by droplet digital PCR to accurately determine the copy number variation. Then, the formalin-fixed paraffin-embedded (FFPE) samples from xenografts were prepared and evaluated for suitability as candidate reference materials by immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). The FFPE reference materials were selected on the basis of IHC score of 2+ and negative FISH result to meet the requirement for HER2-low BC detection. Furthermore, the FFPE reference materials exhibited typical histological structures that resembled the clinical BC specimens. These novel FFPE reference materials displayed the high stability and homogeneity, and they were produced in high quantity. In summary, we generated high-quality reference materials for internal quality control and proficiency testing in HER2-low detection.

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HER2 fluorescent in situ hybridization signal degradation: a 10-year retrospective study

Cited by 3 publications

References 24 publications

Implementation of HER2 Testing in Endometrial Cancer, a Summary of Real-World Initial Experience in a Large Tertiary Cancer Center

Implementation of HER2 Testing in Endometrial Cancer, a Summary of Real-World Initial Experience in a Large Tertiary Cancer Center

Zebrafish: a convenient tool for myelopoiesis research

Lentiviral vector–based xenograft tumors as candidate reference materials for detection of HER2-low breast cancer

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