Validation of the fully automated IdyllaTM system for microsatellite instability detection in Chinese colorectal cancers

doi:10.19401/j.cnki.1007-3639.2022.11.006

Abstract

Abstract:

Background and purpose: Microsatellite instability (MSI) and mismatch repair (MMR) defects have attracted much attention recently as one of the important biomarkers for pan-tumor immunotherapy such as colorectal cancer (CRC). Conventional methods for MSI detection with polymerase chain reaction (PCR) often require non-tumor controls as well as a long turnaround time. The Idylla^TM system as a fully automated quantitative PCR unit with a turnaround time of only 2.5 h should be proved for its reliability among Chinese CRC patients before application. Methods: Formalin-fixed paraffin-embedded (FFPE) tissue samples were collected from 87 CRC patients in Fudan University Shanghai Cancer Center from March 2017 to March 2019 and were subjected to detect MMR protein expression and MSI using immunochemistry (IHC), conventional PCR (Promega MSI analysis) and the Idylla^TM MSI system, respectively. Results: Of the 87 CRC patients enrolled, 56 cases were identified to be deficient MMR (dMMR)/MSI-high (MSI-H) and 31 to be proficient MMR (pMMR)/microsatellite stable (MSS) by both IHC and Promega MSI analysis. However, 4 cases with dMMR/MSI-H were determined to be MSS and one case with pMMR/MSS to be MSI-H using Idylla^TM MSI system, yielding a sensitivity of 92.9% (52/56), specificity of 96.8% (30/31) and an overall concordance of 94.3% (82/87). Then, the above five discordant cases were re-examined using Idylla^TM with resection tissues, as a result, four cases with ≥20% tumor contents changed to be consistent with IHC/Promega MSI assay, however, there was still one inconsistent case with only 5% tumor content. Subsequently, the case was reconfirmed using Idylla^TM with enriched tumor tissue by macrodissection, as expected, the result was demonstrated to be MSI-H, consistent with Promega MSI analysis and IHC. Furthermore, repeatability of the Idylla^TM MSI assay was proved to be reliable in 5 cases with various tumor contents (20%-60%). Conclusion: The Idylla^TM MSI system provides a fast, reliable and fully automated solution to MSI detection in Chinese CRC patients with high sensitivity, specificity and reproducibility, especially for CRC with more than 20% tumor contents.

Key words: Colorectal cancer, Microsatellite instability, Mismatch repair, Idylla^TM, Immunochemistry, Polymerase chain reaction

CLC Number:

WANG Qian, ZHANG Jing, YU Chengli, BAO Longlong, CAI Xu, JIANG Wenhua, HUANG Dan, SHENG Weiqi, ZHU Xiaoli, ZHOU Xiaoyan, BAI Qianming. Validation of the fully automated Idylla^TM system for microsatellite instability detection in Chinese colorectal cancers[J]. China Oncology, 2022, 32(11): 1084-1090.

Figures/Tables 3

Tab. 1

Fig. 1

Tab. 2

References 23

[1]	VASAIKAR S, HUANG C, WANG X J, et al. Proteogenomic analysis of human colon cancer reveals new therapeutic opportunities[J]. Cell, 2019, 177(4): 1035-1049.e19. doi: S0092-8674(19)30292-2 pmid: 31031003
[2]	BOLAND C R, GOEL A. Microsatellite instability in colorectal cancer[J]. Gastroenterology, 2010, 138(6): 2073-2087.e3. doi: 10.1053/j.gastro.2009.12.064 pmid: 20420947
[3]	LYNCH H T, DE LA CHAPELLE A. Hereditary colorectal cancer[J]. N Engl J Med, 2003, 348(10): 919-932. doi: 10.1056/NEJMra012242
[4]	HERMAN J G, UMAR A, POLYAK K, et al. Incidence and functional consequences of hMLH1 promoter hypermethylation in colorectal carcinoma[J]. Proc Natl Acad Sci USA, 1998, 95(12): 6870-6875. doi: 10.1073/pnas.95.12.6870
[5]	CHEN M L, CHEN J Y, HU J, et al. Comparison of microsatellite status detection methods in colorectal carcinoma[J]. Int J Clin Exp Pathol, 2018, 11(3): 1431-1438.
[6]	DUDLEY J C, LIN M T, LE D T, et al. Microsatellite instability as a biomarker for PD-1 blockade[J]. Clin Cancer Res, 2016, 22(4): 813-820. doi: 10.1158/1078-0432.CCR-15-1678 pmid: 26880610
[7]	EVRARD C, TACHON G, RANDRIAN V, et al. Microsatellite instability: diagnosis, heterogeneity, discordance, and clinical impact in colorectal cancer[J]. Cancers (Basel), 2019, 11(10): E1567.
[8]	COHEN R, HAIN E, BUHARD O, et al. Association of primary resistance to immune checkpoint inhibitors in metastatic colorectal cancer with misdiagnosis of microsatellite instability or mismatch repair deficiency status[J]. JAMA Oncol, 2019, 5(4): 551-555. doi: 10.1001/jamaoncol.2018.4942 pmid: 30452494
[9]	UMAR A, BOLAND C R, TERDIMAN J P, et al. Revised Bethesda Guidelines for hereditary nonpolyposis colorectal cancer (Lynch syndrome) and microsatellite instability[J]. J Natl Cancer Inst, 2004, 96(4): 261-268. doi: 10.1093/jnci/djh034 pmid: 14970275
[10]	LINDEMAN N I, CAGLE P T, BEASLEY M B, et al. Molecular testing guideline for selection of lung cancer patients for EGFR and ALK tyrosine kinase inhibitors: guideline from the College of American Pathologists, International Association for the Study of Lung Cancer, and Association for Molecular Pathology[J]. J Thorac Oncol, 2013, 8(7): 823-859. doi: 10.1097/JTO.0b013e318290868f pmid: 23552377
[11]	SAMAISON L, GRALL M, STAROZ F, et al. Microsatellite instability diagnosis using the fully automated Idylla platform: feasibility study of an in-house rapid molecular testing ancillary to immunohistochemistry in pathology laboratories[J]. J Clin Pathol, 2019, 72(12): 830-835. doi: 10.1136/jclinpath-2019-205935 pmid: 31235541
[12]	LEE M, CHUN S M, SUNG C O, et al. Clinical utility of a fully automated microsatellite instability test with minimal hands-on time[J]. J Pathol Transl Med, 2019, 53(6): 386-392. doi: 10.4132/jptm.2019.09.25 pmid: 31606978
[13]	LI X, XU J, LI L, et al. Evaluation of a fully automated idylla test system for microsatellite instability in colorectal cancer[J]. Clin Colorectal Cancer, 2019, 18(4): e316-e323. doi: 10.1016/j.clcc.2019.05.006
[14]	ZWAENEPOEL K, HOLMGAARD DUELUND J, DE WINNE K, et al. Clinical performance of the idylla MSI test for a rapid assessment of the DNA microsatellite status in human colorectal cancer[J]. J Mol Diagn, 2020, 22(3): 386-395. doi: S1525-1578(19)30458-1 pmid: 31881332
[15]	LE D T, URAM J N, WANG H, et al. PD-1 blockade in tumors with mismatch-repair deficiency[J]. N Engl J Med, 2015, 372(26): 2509-2520. doi: 10.1056/NEJMoa1500596
[16]	LE D T, DURHAM J N, SMITH K N, et al. Mismatch repair deficiency predicts response of solid tumors to PD-1 blockade[J]. Science, 2017, 357(6349): 409-413. doi: 10.1126/science.aan6733 pmid: 28596308
[17]	AGUIAR P N Jr, TADOKORO H, FORONES N M, et al. MMR deficiency may lead to a high immunogenicity and then an improvement in anti-PD-1 efficacy for metastatic colorectal cancer[J]. Immunotherapy, 2015, 7(11): 1133-1134. doi: 10.2217/imt.15.84 pmid: 26568256
[18]	ZHAO H, THIENPONT B, YESILYURT B T, et al. Mismatch repair deficiency endows tumors with a unique mutation signature and sensitivity to DNA double-strand breaks[J]. Elife, 2014, 3: e02725. doi: 10.7554/eLife.02725
[19]	UGUEN A, TRONCONE G. A review on the Idylla platform: towards the assessment of actionable genomic alterations in one day[J]. J Clin Pathol, 2018, 71(9): 757-762. doi: 10.1136/jclinpath-2018-205189 pmid: 29903742
[20]	PÉCRIAUX A, FAVRE L, CALDERARO J, et al. Detection of microsatellite instability in a panel of solid tumours with the Idylla MSI Test using extracted DNA[J]. J Clin Pathol, 2021, 74(1): 36-42. doi: 10.1136/jclinpath-2020-206581 pmid: 32513848
[21]	PENG J J, HUANG D, POSTON G, et al. The molecular heterogeneity of sporadic colorectal cancer with different tumor sites in Chinese patients[J]. Oncotarget, 2017, 8(30): 49076-49083. doi: 10.18632/oncotarget.16176 pmid: 28416767
[22]	VELASCO A, TOKAT F, BONDE J, et al. Multi-center real-world comparison of the fully automated Idylla^TM microsatellite instability assay with routine molecular methods and immunohistochemistry on formalin-fixed paraffin-embedded tissue of colorectal cancer[J]. Virchows Arch, 2021, 478(5): 851-863. doi: 10.1007/s00428-020-02962-x
[23]	HAUSE R J, PRITCHARD C C, SHENDURE J, et al. Classification and characterization of microsatellite instability across 18 cancer types[J]. Nat Med, 2016, 22(11): 1342-1350. doi: 10.1038/nm.4191 pmid: 27694933

Case ID	Tumor content	Section	MSI score of each marker							Idylla^TM MSI
Case ID	Tumor content	Section	ACVR2A	BTBD7	DIDO1	MRE11	RYR3	SEC31A	SULF2	Idylla^TM MSI
81	30%	2	0.00	0.00	0.00	0.00	0.00	0.00	0.00	MSS
	30%	3	0.00	0.00	0.00	0.00	0.00	0.00	0.00	MSS
	30%	4	0.00	0.00	0.00	0.00	0.00	0.00	0.00	MSS
44	30%	2	1.00	0.97	0.86	0.93	0.16	0.93	0.37	MSI-H
	30%	3	1.00	0.97	0.86	0.88	0.14	0.97	0.43	MSI-H
	30%	4	1.00	0.97	0.88	0.92	0.18	0.97	0.29	MSI-H
5	40%	2	1.00	0.19	0.87	1.00	0.93	0.01	0.98	MSI-H
	40%	3	1.00	0.21	0.93	1.00	0.94	0.01	0.98	MSI-H
	40%	4	1.00	0.28	0.92	1.00	0.95	0.02	0.99	MSI-H
28	50%	2	0.99	0.63	0.95	0.60	0.19	0.01	0.02	MSI-H
	50%	3	1.00	0.91	0.97	0.61	0.03	0.13	0.01	MSI-H
	50%	4	1.00	0.62	0.99	1.00	0.15	0.00	0.92	MSI-H
39	60%	2	1.00	1.00	0.96	0.99	0.99	0.41	0.98	MSI-H
	60%	3	1.00	1.00	0.97	1.00	0.99	0.55	0.97	MSI-H
	60%	4	1.00	1.00	0.97	1.00	0.99	0.58	0.98	MSI-H