O.V. Shatkovskaya ¹, D.R. Kaidarova ¹, M.G. Orazgaliyeva ¹, E.B. Satbayeva ^1,2, S.O. Ossikbayeva ¹, A.B. Koishibaeva ¹

1. «Kazakh Institute of Oncology and Radiology» JSC, Almaty, the Republic of Kazakhstan;
2. «Almaty oncological center» MSE on REM, Almaty, the Republic of Kazakhstan

DOI: https://www.doi.org/10.52532/2521-6414-2023-2-68-15-21

UDC: 616.24-006:575.224.22

Year: 2023 issure: 68 number: 2 pages: 15-21

Download PDF:

ABSTRACT

Relevance: Currently, molecular diagnosis in NSCLC in Kazakhstan includes detection of EGFR, ALK driver mutations status, and PD-L1-status, but not ROS1, what limits the access of patients with this driver mutation to vital therapy.
The study aimed to optimize the methods of molecular genetic diagnosis of patients with NSCLC by introducing ROS1 testing in the Republic of Kazakhstan.
Methods: The biopsy and surgical material of non-small cell lung cancer (NSCLC) fixed in 10% buffered formalin was studied. After the initial morphological diagnosis of adenocarcinoma, EGFR, and ALK mutation status determination, EGFR, and ALK-negative tumor assays were sent for further determination of ROS1 mutation status. First, we performed immunohistochemistry (IHC) using the Ventana BenchMark Ultra platform using the ROS1 antibody (SP283) and the OptiView DAB Detection Kit imaging system. After that, samples with positive and doubtful IHC results were sent for RT-PCR (reverse transcriptase polymerase chain reaction) to confirm the ROS1 mutation status.
Results: A total of 99 tumor samples from patients with EGFR-negative and ALK-negative lung adenocarcinoma were studied by IHC from January 01 till September 30, 2022. The results of IHC staining were assessed as: 0 (negative) – 59 samples, 1+ (negative) – 25 samples, 2+ (doubtful) – 12 samples, 3+ (positive) – 3 samples. Cases with ≥70% immunostaining were considered positive. Samples with an IHC stain score of 2+ (doubtful), 3+ (positive), and a few samples of 1+ were sent for confirmation by PCR.
Overall, 22 samples were tested using RT-PCR, and results were considered as follows:1 (4%) – positive, 13 (59%) – negative, 8 (37%) -– invalid.
Conclusion: A large proportion of positive and questionable results were obtained when determining ROS1 mutation status using IHC, and a large proportion of invalid results during subsequent RT-PCR testing. Choosing methods for nationwide ROS1 implementation, one should evaluate the economics of the methods to be implemented and compare them with a standard validated FISH method.
Keywords: ROS1 molecular genetic diagnostics, lung cancer (LC), non-small cell lung cancer (NSCLC), immunohistochemistry (IHC), reverse transcriptase polymerase chain reaction (RT-PCR), fluorescence in situ hybridization (FISH).

List of sources used:

1. Кайдарова Д.Р., Шатковская О.В., Онгарбаев Б.Т., Сейсенбаева Г.Т., Ажмагамбетова А.Е., Жылкайдарова А.Ж., Лаврентьева И.К., Саги М.С. Показатели онкологической службы РК, 2021 г. (Статистические и аналитические материалы) / под ред. Д.Р. Кайдаровой. – Алматы, 2022. – 384 с. [Kaidarova D.R., Shatkovskaja O.V., Ongarbaev B.T., Sejsenbaeva G.T., Azhmagambetova A.E., Zhylkaidarova A.Zh., Lavrent’eva I.K., Sagi M.S. Pokazateli onkologicheskoj sluzhby RK, 2021 g. (Statisticheskie i analiticheskie materialy) / pod red. D.R. Kaidarovoy. – Almaty, 2022. – 384 s. (in Russ.)]. https://doi.org/10.52532/1-11-2021-1-384
2. WHO, International Agency for Research on Cancer. Global Cancer Observatory in 2020. Kazakhstan fact sheets. – 2021. https://gco.iarc.fr/today/data/factsheets/populations/398-kazakhstan-fact-sheets.pdf. 29.05.2023.
3. Loo P.S., Thomas S.C., Nicolson M.C., Fyfe M.N., Kerr K.M. Subtyping of undifferentiated non-small cell carcinomas in bronchial biopsy specimens // J. Thor. Oncol. – 2010. – Т. 5. – №. 4. – Р. 442-447. https://doi.org/10.1097/JTO.0b013e3181d40fac
4. National Cancer Institute. Seer Cancer Statistics Review 1975-2017. https://seer.cancer.gov/archive/csr/1975_2017/results_merged/sect_15_lung_bronchus.pdf. 29.05.2023.
5. Lewis D.R., Check D.P., Caporaso N.E., Travis W.D., Devesa S.S. US lung cancer trends by histologic type // Cancer. – 2014. – Т. 120. – №. 18. – Р. 2883-2892. https://doi.org/10.1002/cncr.28749
6. Клинический протокол диагностики и лечения «Рак легкого»: утв. протоколом Объединенной комиссии по качеству медицинских услуг Министерства здравоохранения Республики Казахстан 1 июля 2022 года, №164 [Klinicheskij protokol diagnostiki i lecheniya «Rak legkogo»: utv. protokolom Ob»edinennoj komissii po kachestvu medicinskix uslug Ministerstva zdravooxraneniya Respubliki Kazaxstan 1 ijulya 2019 goda, №164 (in Russ.)]. https://diseases.medelement.com/disease/рак-легкого-кп-рк-2022/17273
7. Stimson L., La Thangue N.B. Biomarkers for predicting clinical responses to HDAC inhibitors // Cancer Let. – 2009. – Т. 280. – №. 2. – Р. 177-183. https://doi.org/10.1016/j.canlet.2009.03.016
8. Kris M.G., Johnson B.E., Berry L.D., Kwiatkowski D.J., Iafrate A.J., Wistuba I.I., Varella-Garcia M., Franklin W.A., Aronson S.L., Su P.S., Shyr Y., Camidge D.R., Sequist L.V., Glisson B.S., Khuri F.R., Garon E.B., Pao W., Rudin C., Schiller J., Haura E.B., Socinski M., Shirai K., Chen H., Giaccone G., Ladanyi M., Kugler K., Minna J.D., Bunn P.A. Using multiplexed assays of oncogenic drivers in lung cancers to select targeted drugs // JAMA. – 2014. – Vol. 311 (19). – Р. 1998. https://doi.org/10.1001/jama.2014.3741
9. Planchard D., Popat S., Kerr K., Novello S., Smit E.F., Faivre-Finn C., Mok T.S., Reck M., Van Schil P.E., Hellmann M.D., Peters S. Metastatic non-small cell lung cancer: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up // Ann. Oncol. – 2018. – Vol. 29. – Р. iv192-iv237. https://doi.org/10.1093/annonc/mdy474
10. National Comprehensive Cancer Network, Inc. NCCN Clinical Practice Guidelines in Oncology (NCCN Guidelines^®) for Non-Small Cell Lung Cancer. – V.3.2023. https://www.nccn.org/professionals/physician_gls/pdf/nscl.pdf. 29.05.2023
11. Об утверждении Комплексного плана по борьбе с онкологическими заболеваниями в Республике Казахстан на 2023-2025 годы. Постановление Правительства РК от 29 июня 2018 года, № 395 [Ob utverzhdenii kompleksnogo plana po bor’be s onkologicheskimi zabolevaniyami v Respublike Kazaxstan na 2018 – 2022 gody. Postanovlenie Pravitel’stva RK ot 29 iyunya 2018 goda, № 395 (in Russ.)]. https://adilet.zan.kz/rus/docs/P1800000395. 29.05.2023
12. Об утверждении комплексного плана по борьбе с онкологическими заболеваниями в Республике Казахстан на 2018 – 2022 годы. Утв. Постановлением Правительства РК от 29 июня 2018 года, № 395 [Ob utverzhdenii kompleksnogo plana po bor’be s onkologicheskimi zabolevaniyami v Respublike Kazaxstan na 2018 – 2022 gody. Utv. Postanovleniem Pravitel’stva RK ot 29 iyunya 2018 goda, № 395 (in Russ.)]. https://adilet.zan.kz/rus/docs/P1800000395. 29.05.2023
13. Lin J. J., Shaw A. T. Recent advances in targeting ROS1 in lung cancer // J. Thor. Oncol. – 2017. – Vol. 12 (11). – Р. 1611-1625. https://doi.org/10.1016/j.jtho.2017.08.002
14. Chin L.P., Soo R.A., Soong R., Ou S.H.I. Targeting ROS1 with anaplastic lymphoma kinase inhibitors: a promising therapeutic strategy for a newly defined molecular subset of non–small-cell lung cancer // J. Thor. Oncol. – 2012. – Vol. 7 (11). – С. 1625-1630. https://doi.org/10.1097/JTO.0b013e31826baf83
15. Tsao M.S., Hirsch F.R., Yatabe Y. IASLC atlas of ALK and ROS1 testing in lung cancer. Aurora (Colorado). – Aurora, CO, USA: International Association for the Study of Lung Cancer, 2016. https://www.iaslc.org/research-education/publications-resources-guidelines/iaslc-atlas-alk-and-ros1-testing-lung-cancer
16. Lindeman N.I., Cagle P.T., Aisner D.L., Arcila M.E., Beasley M.B., Bernicker E.H., Colasacco C., Dacic S., Hirsch F.R., Kerr K., Kwiatkowski D.J., Ladanyi M., Nowak J.A., Sholl L., Temple-Smolkin R., Solomon B., Souter L.H., Thunnissen E., Tsao M.S., Ventura C. B., Wynes M.W., Yatabe Y. Updated molecular testing guideline for the selection of lung cancer patients for treatment with targeted tyrosine kinase inhibitors: guideline from the College of American Pathologists, the International Association for the Study of Lung Cancer, and the Association for Molecular Pathology // Arch. Pathol. Lab. Med. – 2018. – Vol. 142 (3). – Р. 321-346. https://doi.org/10.5858/arpa.2017-0388-CP
17. Naidoo J., Drilon A. Molecular diagnostic testing in non-small cell lung cancer // Am. J. Hematol. Oncol. – 2014. – Vol. 10 (4). – P. 4-11. https://gotoper-com.s3.amazonaws.com/_media/_pdf/AJHO14Sept_01_NSCLC.pdf
18. Программа RUSSCO «Совершенствование молекулярно-генетической диагностики в Российской Федерации». Практическое руководство для врачей. – 54 с. [Programma RUSSCO «Sovershenstvovanie molekulyarno-geneticheskoj diagnostiki v rossijskoj federacii». Prakticheskoe rukovodstvo dlya vrachej. – 54 s. (in Russ.)]. http://cancergenome.ru/materials/guide.pdf. 06.06.2023
19. Roche. Roche launches first in vitro diagnostic IHC test to detect ROS1 protein in cancers. https://diagnostics.roche.com/nl/en/news-listing/2019/roche-launches-first-in-vitro-diagnostic-ihc-test-to-detect-ros1-protein-in-cancers.html. 29.05.2023.
20. Cha Y.J., Lee J.S., Kim H.R., Lim S.M., Cho B.C., Lee C.Y., Shim H.S. Screening of ROS1 rearrangements in lung adenocarcinoma by immunohistochemistry and comparison with ALK rearrangements // PLoS One. – 2014. – Vol. 9 (7). – Р. e103333. https://doi.org/10.1371/journal.pone.0103333
21. Shan, L., Lian, F., Guo, L., Qiu, T., Ling, Y., Ying, J., Lin, D. Detection of ROS1 gene rearrangement in lung adenocarcinoma: comparison of IHC, FISH and real-time RT-PCR // PLoS One. – 2015. – Т. 10. – №. 3. – Р. e0120422. https://doi.org/10.1371/journal.pone.0120422
22. Boyle, T. A., Masago, K., Ellison, K. E., Yatabe, Y., Hirsch, F. R. Ros1 immunohistochemistry among major genotypes of Non–Small-Cell lung cancer // Clin. Lung Cancer. – 2015. – Т. 16. – №. 2. – Р. 106-111. https://doi.org/10.1016/j.cllc.2014.10.003
23. Shaw A.T., Ou S.H.I., Bang Y.J., Camidge D.R., Solomon B.J., Salgia R., Riely G.J., Varella-Garcia M., Shapiro G.I., Costa D.B., Doebele R.C., Le L.P., Zheng Z., Tan W., Stephenson P., Shreeve S.M., Tye L.M., Christensen J.G., Wilner K.D., Clark J.W., Iafrate A.J. Crizotinib in ROS1-rearranged non–small-cell lung cancer // New Engl. J. Med. – 2014. – Vol. 371 (21). – Р. 1963-1971. https://www.nejm.org/doi/full/10.1056/nejmoa1406766
24. Mazières J., Zalcman G., Crinò L., Biondani P., Barlesi F., Filleron T., Dingemans A.C., Léna H., Monnet I., Rothschild S.I., Cappuzzo F., Besse B., Thiberville L., Rouvière D., Dziadziuszko R., Smit E.F., Wolf J., Spirig C., Pecuchet N., Leenders F., Heuckmann J.M., Diebold J., Milia J.D., Thomas R.K., Gautschi O. Crizotinib therapy for advanced lung adenocarcinoma and a ROS1 rearrangement: results from the EUROS1 cohort // J. Clin. Oncol. – 2015. – Vol. 3(9). https://ascopubs.org/doi/10.1200/JCO.2014.58.3302