MODAPLEX Copy Number High Analysis Kit

Features

• Detect copy number variations using 6 reference and 8 target regions enabling TCGA classification
• Fast results – 4 h from DNA to result
• Tested with gDNA derived from 100 endometrial cancer FFPE samples
• Experience the advantages of a thorough control strategy that ensures a high level of accuracy

The MODAPLEX Copy Number High Analysis Kit (RUO) is a fluorescent PCR-based multiplex assays which detects chromosomal aberrations within the chromosomal locations 17p12 and 19p13.3 in human endometrial cancer (EC) DNA samples. Copy Number Variations (CNVs) within these locations are biomarkers for the EC molecular subtype of copy number high (CNH, or p53 abnormal).

Biomarkers

Target regions (chromosomal location)

Chromosome 19:

19pT1 (Zytoband 19p13.3)

19pT2 (Zytoband 19p13.3)

19pT3 (Zytoband 19p13.3)

19pT4 (Zytoband 19p13.3)

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Chromosome 17:

17pT1 (Zytoband 17p12)

17pT2 (Zytoband 17p12)

17pT3 (Zytoband 17p12)

17pT4 (Zytoband 17p12)

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Reference regions (chromosomal location)

1pRef1 (Zytoband 1p31.3)

1pRef2 (Zytoband 1p31.3)

2pRef1 (Zytoband 2q35)

2pRef2 (Zytoband 2q35)

12pRef1 (Zytoband 12q24.31)

12pRef2 (Zytoband 12q24.31)

Scientific Background

An integrated genomic characterization of endometrial cancer (EC) was performed by The Cancer Genome Atlas Research Network (TCGA), which identified four distinct molecular subgroups [1].:

• MSI (microsatellite instablility)
• POLE mutated (ultramutated)
• copy number low (or NSMP - no specific molecular profile)
• copy number high (CNH)

The molecular stratification of endometrial cancer is of prognostic value as it allows the classification of individual tumors into low-, intermediate- and high risk. Therefore, subtyping of all endometrial cancer pathology specimens, regardless of histological type, is now recommended by several guidelines, including ESMO, the European Society of Gynecological Oncology (ESGO), and the European Society of Pathology (ESP) [2, 3].

Copy number high positive endometrial cancers are characterized by an accumulation of large chromosomal aberrations such as deletions or amplifications (copy number variations). In addition, TP53 mutations occur frequently in this subgroup. Consequently, immunohistochemical staining (IHC) of p53 was established as a surrogate marker for the detection of the high copy number group by two research groups in 2015/2016 and applied in the Proactive Molecular Risk Classifier for Endometrial Cancer (ProMisE) [4, 5]. However, p53 IHC is known to be technically challenging and staining interpretation can vary between diagnostic centers.

The MODAPLEX Copy Number High Analysis Kit allows the identification of the CNH subtype in endometrial carcinomas  with an accuracy comparable to whole genome sequencing. This is achieved by semi-quantifying number of alleles in chromosomal regions, which have been identified as highly unstable in CNH endometrial carcinomas. These regions, located on chromosomes 17 and 19 (17p12 and 19p13.3), have been identified through a comprehensive analysis of next-generation sequencing data of endometrial cancers.  Additionally, several chromosomal stable regions were identified (1p31.3, 2q35, 12q24.31). Together this allows the semi-quantification of the copy numbers of the chromosome 17 and 19 CNH-associated target locations. On top of that, multiple targets are amplified within the target- and reference locations guaranteeing a high level of accuracy and robustness of the product.

1. Kandoth, C. et al. Integrated genomic characterization of endometrial carcinoma. Nature 497, 67–73; 10.1038/nature12113 (2013).
2. Oaknin, A. et al. Endometrial cancer. ESMO Clinical Practice Guideline for diagnosis, treatment and follow-up. Ann Oncol 33, 860–877; 10.1016/j.annonc.2022.05.009 (2022).
3. Concin, N. et al. ESGO/ESTRO/ESP guidelines for the management of patients with endometrial carcinoma. Int J Gynecol Cancer 31, 12–39; 10.1136/ijgc-2020-002230 (2021).
4. Talhouk, A. et al. A clinically applicable molecular-based classification for endometrial cancers. Br J Cancer 113, 299–310; 10.1038/bjc.2015.190 (2015).
5. Stelloo, E. et al. Improved Risk Assessment by Integrating Molecular and Clinicopathological Factors in Early-stage Endometrial Cancer-Combined Analysis of the PORTEC Cohorts. Clin Cancer Res 22, 4215–4224; 10.1158/1078-0432.CCR-15-2878 (2016)