MODAPLEX POLE/POLD1 Mutation Analysis Kit

Features

• Detect and differentiate 16 POLE and 3 POLD1 mutations in one well
• Save valuable FFPE tissue due to low DNA input amount (4 ng)
• Obtain precise insights using a test developed with colorectal and endometrial cancer samples
• Take advantage of an easy and fast workflow with turnaround time of 4h

The MODAPLEX POLE/POLD1 Mutation Analysis Kit is a qualitative and comprehensive PCR-based multiplex assay for the detection of 19 single nucleotide mutations within the exonuclease domains of the POLE and POLD1 genes in human DNA derived from formalin-fixed, paraffin-embedded (FFPE) samples using the MODAPLEX instrument. The assay must be used by qualified and trained personnel in a professional laboratory environment only. Results are intended solely for research use and not for diagnostic procedures.

Biomarkers

POLE: T278M, P286L, P286H, P286R, S297A, S297F, F367S, V411L (G>C), V411L (G>T), H422N, L424V, P436R, M444K, A456P, S459F, A465V‍

POLD1: D316N, C319Y, S478N

Scientific Background

Inactivating mutations in the exonuclease domain of the catalytic subunits of the DNA polymerases epsilon and delta 1 (POLE and POLD1) lead to impaired proofreading during DNA replication and consequently to dramatically increased mutation rates of ≥ 100 mut/Mb [1]. Consequently, pathogenic POLE and POLD1 mutations are now being evaluated as potential predictive biomarkers of response to immune checkpoint inhibitors independent of the MSI status [2,3,4].

The Cancer Genome Atlas Research Network (TCGA) performed an integrated genomic, transcriptomic, and proteomic characterization of endometrial cancer. Four distinct genomic subgroups were identified that also correspond to clinical prognosis, with the novel POLE ultramutated group being of particular interest due its favorable outcomes [5]. This molecular classification has now been incorporated into endometrial carcinoma guidelines, published by the European Society of Gynaecological Oncology (ESGO), the European Society of Pathology (ESP) and European Society for Medical Oncology (ESMO) [6,7].

Detection and differentiation of 19 somatic and rare germline mutations in the polymerase epsilon and polymerase delta-1 exonuclease domains is effectively achievable using the MODAPLEX POLE/POLD1 Mutation Analysis Kit. The test enables researchers to advance promptly in clinical research, as the test is optimized to evaluate POLE and POLD1 mutations that have been described to be:

• primarily associated with ultramutation resulting in a high mutational burden and a very distinct mutational pattern [2, 3, 10, 11, 12]
• found predominantly in colorectal and endometrial cancer, but have also been reported for gastric, breast, ovarian, lung and brain cancers [2, 8, 9, 12]
• associated with a high expression of immune-checkpoint proteins and T-cell markers [3,12,13]

1. Castellucci et al., DNA Polymerase ɛ Deficiency Leading to an Ultramutator Phenotype. The oncologist 22, 497–502 (2017).
2. Bourdais et al., Polymerase proofreading domain mutations. Critical reviews in oncology/hematology 113, 242–248 (2017).
3. Mehnert et al., Immune activation and response to pembrolizumab in POLE-mutant endometrial cancer. The Journal of clinical investigation 126, 2334–2340 (2016).
4. Garmezy et al., Clinical and Molecular Characterization of POLE Mutations as Predictive Biomarkers of Response to Immune Checkpoint Inhibitors in Advanced Cancers. JCO precision oncology 6, (2022).
5. Kandoth et al., Integrated genomic characterization of endometrial carcinoma. Nature 497, 67–73 (2013).
6. Concin et al., ESGO/ESTRO/ESP guidelines for the management of patients with endometrial carcinoma. Int. J. Gynecol. Cancer 31, 12–39 (2021).
7. Oaknin et al., Endometrial cancer. Ann. Oncol. 33, 860–877 (2022).
8. Rayner et al., A panoply of errors. Nat. Rev. Cancer 16, 71–81 (2016).
9. Hoang et al., Polymerase Epsilon Exonuclease Domain Mutations in Ovarian Endometrioid Carcinoma. Int. J. Gynecol. Cancer 25, 1187–1193 (2015).
10. Alexandrov et al., The repertoire of mutational signatures in human cancer. Nature 578, 94–101 (2020).
11. León-Castillo et al., Interpretation of somatic POLE mutations in endometrial carcinoma. The Journal of pathology 250, 323–335 (2020).
12. Ma et al., POLE/POLD1 mutation and tumor immunotherapy. Journal of experimental & clinical cancer research: CR 41 (2022).
13. Top et al., The somatic POLE P286R mutation defines a unique subclass of colorectal cancer featuring hypermutation, representing a potential genomic biomarker for immunotherapy. Oncotarget 7, 68638–68649 (2016).