Регуляторные белки эпителиально-мезенхимального перехода в колоректальном раке: от биологии к клиническому применению

Представлены современные данные о роли регуляторных белков эпителиально-мезенхимального перехода (ЭМП) при колоректальном раке (КРР) в контексте молекулярной стратификации опухолей и персонализированного подбора терапии. Эпителиально-мезенхимальный переход представляет собой ключевой биологический процесс, связанный с инвазией, метастазированием, химиорезистентностью и иммунным уклонением опухоли. Особое внимание уделено характеристике молекулярных подтипов КРР согласно консенсусной классификации (CMS), в частности, CMS4-подтипу с мезенхимальными признаками, для которого характерен высокий уровень ЭМП и неблагоприятный прогноз.

В лекции обсуждаются молекулярные и иммуногистохимические маркеры, отражающие активацию ЭМП, включая CDX2, ZEB1, HTR2B, FRMD6, BMI-1 и ROR1. Рассматриваются особенности экспрессии и сигнальные каскады, с которыми ассоциированы указанные белки, их влияние на опухолевую агрессивность, устойчивость к терапии и перспективы клинического применения. На основании анализа литературных данных рассматриваются возможности использования этих белков в качестве прогностических и потенциальных предиктивных маркеров, а также терапевтических мишеней.

1. Sung H., Ferlay J., Siegel R.L., Laversanne M., Soerjomataram I., Jemal A. et al. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J. Clin. 2021;71(3):209– 249. DOI: 10.3322/caac.21660.

2. Dharwadkar P., Greenan G., Singal A.G., Murphy C.C. Is colorectal cancer in patients younger than 50 years of age the same disease as in older patients? Clin. Gastroenterol. Hepatol. 2021;19(1):192–194.e3. DOI: 10.1016/j.cgh.2019.10.028.

3. Каприн А.Д., Старинский В.В., Шахзадова А.О. Злокачественные новообразования в России в 2023 году (заболеваемость и смертность). М.: Издательский центр МНИОИ им. П.А. Герцена, 2024:276.

4. Argiles G., Tabernero J., Labianca R., Hochhauser D., Salazar R., Iveson T. et al. ESMO Guidelines Committee. Electronic address: clinicalguidelines@esmo.org. Localised colon cancer: ESMO clinical practice guidelines for diagnosis, treatment and follow-up. Ann. Oncol. 2020;31(10):1291–1305. DOI: 10.1016/j.annonc.2020.06.022.

5. Müller M.F., Ibrahim A.E., Arends M.J. Molecular pathological classification of colorectal cancer. Virchows Arch. 2016; 469(2):125–134. DOI: 10.1007/s00428-016-1956-3.

6. Guinney J., Dienstmann R., Wang X., de Reynies A., Schlicker A., Soneson C. et al. The consensus molecular subtypes of colorectal cancer. Nat. Med. 2015;21(11):1350–1356. DOI: 10.1038/nm.3967.

7. Källberg J., Harrison A., March V., Berzina S., Nemazanyy I., Kepp O. et al. Intratumor heterogeneity and cell secretome promote chemotherapy resistance and progression of colorectal cancer. Cell Death Dis. 2023;14(5):306. DOI: 10.1038/s41419-023-05806-z.

8. Deng J., Tian A.L., Pan H., Sauvat A., Leduc M., Liu P. et al. Everolimus and plicamycin specifically target chemoresistant colorectal cancer cells of the CMS4 subtype. Cell Death Dis. 2021;12(11):978. DOI: 10.1038/s41419-021-04270-x.

9. Peters N.A., Constantinides A., Ubink I., van Kuik J., Bloemendal H.J., van Dodewaard J.M. et al. Consensus molecular subtype 4 (CMS4)-targeted therapy in primary colon cancer: a proof-of-concept study. Front. Oncol. 2022;12:969855. DOI: 10.3389/fonc.2022.969855.

10. Chen C., Huang S., Chen C.L., Su S.B., Fang D.D. Isoliquiritigenin inhibits ovarian cancer metastasis by reversing epithelial-to-mesenchymal transition. Molecules. 2019;24(20):3725. DOI: 10.3390/molecules24203725.

11. Пучинская М.В. Эпителиально-мезенхимальный переход в норме и патологии. Архив патологии. 2015;77(1):75–83. DOI: 10.17116/patol201577175-.

12. Peinado H., Olmeda D., Cano A. Snail, Zeb and bHLH factors in tumour progression: an alliance against the epithelial phenotype. Nat. Rev. Cancer. 2007;(6):415–428. DOI: 10.1038/nrc2131.

13. Xu J., Lamouille S., Derynck R. TGF-beta-induced epithelial to mesenchymal transition. Cell Res. 2009;19(2):156–172. DOI: 10.1038/cr.2009.5.

14. Gheldof A., Berx G. Cadherins and epithelial-to-mesenchymal transition. Prog. Mol. Biol. Transl. Sci. 2013;116:317– 336. DOI: 10.1016/B978-0-12-394311-8.00014-5.

15. Tian Q., Xue Y., Zheng W., Sun R., Ji W., Wang X. et al. Overexpression of hypoxia-inducible factor 1α induces migration and invasion through Notch signaling. Int. J. Oncol. 2015;47(2):728–738. DOI: 10.3892/ijo.2015.3056.

16. Zhang B., Zhao R., Wang Q., Zhang Y.J., Yang L., Yuan Z.J. et al. An EMT-related gene signature to predict the prognosis of triple-negative breast cancer. Adv. Ther. 2023;40(10):4339– 4357. DOI: 10.1007/s12325-023-02577-z.

17. Jonckheere S., Adams J., De Groote D., Campbell K., Berx G., Goossens S. et al. Epithelial-mesenchymal transition (EMT) as a therapeutic target. Cells Tissues Organs. 2022;211(2):157– 182. DOI: 10.1159/000512218.

18. Beck F., Stringer E.J. The role of Cdx genes in the gut and in axial development. Biochem. Soc. Trans. 2010;38(2):353– 357. DOI: 10.1042/BST0380353.

19. Young T., Rowland J.E., van de Ven C, Bialecka M., Novoa A., Carapuco M. et al. CDX and HOX genes differentially regulate posterior axial growth in mammalian embryos. Dev. Cell. 2009;17(4):516–526. DOI: 10.1016/j.devcel.2009.08.010.

20. Werling R.W., Yaziji H., Bacchi C.E., Gown A.M. CDX2, a highly sensitive and specific marker of adenocarcinomas of intestinal origin: an immunohistochemical survey of 476 primary and metastatic carcinomas. Am. J. Surg. Pathol. 2003;27(3):303–310. DOI: 10.1097/00000478-200303000-00003.

21. Tomasello G., Barni S., Turati L., Ghidini M., Pezzica E., Passalacqua R. et al. Association of CDX2 expression with survival in early colorectal cancer: a systematic review and meta-analysis. Clin. Colorectal Cancer. 2018;17(2):97–103. DOI: 10.1016/j.clcc.2018.02.001.

22. Baba Y., Nosho K., Shima K., Freed E., Irahara N., Philips J. et al. Relationship of CDX2 loss with molecular features and prognosis in colorectal cancer. Clin. Cancer Res. 2009;15(14):4665– 4673. DOI: 10.1158/1078-0432.CCR-09-0401.

23. Karamitopoulou E., Zlobec I., Panayiotides I., Patsouris E.S., Peros G., Rallis G. et al. Systematic analysis of proteins from different signaling pathways in the tumor center and the invasive front of colorectal cancer. Hum. Pathol. 2011;42(12):1888– 1896. DOI: 10.1016/j.humpath.2010.06.020.

24. Nallanthighal S., Heiserman J.P., Cheon D.J. Collagen type XI alpha 1 (COL11A1): a novel biomarker and a key player in cancer. Cancers (Basel). 2021;13(5):935. DOI: 10.3390/cancers13050935.

25. Wu Y.H., Chou C.Y. Collagen XI alpha 1 chain, a novel therapeutic target for cancer treatment. Front. Oncol. 2022;12:925165. DOI: 10.3389/fonc.2022.925165.

26. Bae J.M., Lee T.H., Cho N.Y., Kim T.Y., Kang G.H. Loss of CDX2 expression is associated with poor prognosis in colorectal cancer patients. World J. Gastroenterol. 2015;21(5):1457– 1467. DOI: 10.3748/wjg.v21.i5.1457.

27. Aasebo K., Dragomir A., Sundstrom M., Mezheyeuski A., Edqvist P.H., Eide G.E. et al. CDX2: a prognostic marker in metastatic colorectal cancer defining a better BRAF mutated and a worse KRAS mutated subgroup. Front. Oncol. 2020;10:8. DOI: 10.3389/fonc.2020.00008.

28. Shigematsu Y., Inamura K., Mise Y., Saiura A., Rehnberg E., Yamamoto N. et al. CDX2 expression is concordant between primary colorectal cancer lesions and corresponding liver metastases independent of chemotherapy: a single-center retrospective study in Japan. Oncotarget. 2018;9(24):17056– 17065. DOI: 10.18632/oncotarget.24708.

29. Ziranu P., Pretta A., Pozzari M., Maccioni A., Badiali M., Fanni D. et al. CDX2 expression correlates with clinical outcomes in MSI-H metastatic colorectal cancer patients receiving immune checkpoint inhibitors. Sci. Rep. 2023;13(1):4397. DOI: 10.1038/s41598-023-31538-3.

30. Dalerba P., Sahoo D., Paik S., Guo X., Yothers G., Song N. et al. CDX2 as a prognostic biomarker in stage II and stage III colon cancer. N. Engl. J. Med. 2016;374(3):211–222. DOI: 10.1056/NEJMoa1506597.

31. Caramel J., Ligier M., Puisieux A. Pleiotropic roles for ZEB1 in cancer. Cancer Res. 2018;78(1):30–35. DOI: 10.1158/0008-5472.CAN-17-2476.

32. Zhang P., Sun Y., Ma L. ZEB1: at the crossroads of epithelial-mesenchymal transition, metastasis and therapy resistance. Cell Cycle. 2015;14(4):481–487. DOI: 10.1080/15384101.2015.1006048.

33. Cho H.J., Oh N., Park J.H., Kim K.S., Kim H.K., Lee E. et al. ZEB1 collaborates with ELK3 to repress E-cadherin expression in triple-negative breast cancer cells. Mol. Cancer Res. 2019;17(11):2257–2266. DOI: 10.1158/1541-7786.MCR-19-0380.

34. Guo Y., Lu X., Chen Y., Clark G., Trent J., Cuatrecasas M. et al. Opposing roles of ZEB1 in the cytoplasm and nucleus control cytoskeletal assembly and YAP1 activity. Cell Rep. 2022;41(1):111452. DOI: 10.1016/j.celrep.2022.111452.

35. Fernandez-De-Los-Reyes I., Gomez-Dorronsoro M., Monreal-Santesteban I., Fernandez-Fernandez A., Fraga M., Azcue P. et al. ZEB1 hypermethylation is associated with better prognosis in patients with colon cancer. Clin. Epigenetics. 2023;15(1):193. DOI: 10.1186/s13148-023-01605-7.

36. Noman M.Z., Janji B., Abdou A., Hasmim M., Terry S., Tan T.Z. et al. The immune checkpoint ligand PD-L1 is upregulated in EMT-activated human breast cancer cells by a mechanism involving ZEB-1 and miR-200. Oncoimmunology. 2017;6(1):e1263412. DOI: 10.1080/2162402X.2016.1263412.

37. Zhang X., Zhang Z., Zhang Q., Zhang Q., Sun P., Xiang R. et al. ZEB1 confers chemotherapeutic resistance to breast cancer by activating ATM. Cell Death Dis. 2018;9(2):57. DOI: 10.1038/s41419-017-0087-3.

38. Wu Z., Zhang L., Xu S., Lin Y., Yin W., Lu J. et al. Predictive and prognostic value of ZEB1 protein expression in breast cancer patients with neoadjuvant chemotherapy. Cancer Cell Int. 2019;19:78. DOI: 10.1186/s12935-019-0793-2.

39. Chen H., Lu W., Huang C., Ding K., Xia D., Wu Y. et al. Prognostic significance of ZEB1 and ZEB2 in digestive cancers: a cohort-based analysis and secondary analysis. Oncotarget. 2017;8(19):31435–31448. DOI: 10.18632/oncotarget.15634.

40. Chen B., Chen B., Zhu Z., Ye W., Zeng J., Liu G. et al. Prognostic value of ZEB-1 in solid tumors: a meta-analysis. BMC Cancer. 2019;19(1):635. DOI: 10.1186/s12885-019-5830-y.

41. Chen D., Yu W., Aitken L., Gunn-Moore F. Willin/FRMD6: a multi-functional neuronal protein associated with Alzheimer’s disease. Cells. 2021;10(11):3024. DOI: 10.3390/cells10113024.

42. Haldrup J., Strand S.H., Cieza-Borrella C., Jakobsson M.E., Riedel M., Norgaard M. et al. FRMD6 has tumor suppressor functions in prostate cancer. Oncogene. 2021;40(4):763–776. DOI: 10.1038/s41388-020-01548-w.

43. Wang T., Guo H., Zhang L., Yu M., Li Q., Zhang J. et al. FERM domain-containing protein FRMD6 activates the mTOR signaling pathway and promotes lung cancer progression. Front. Med. 2023;17(4):714–728. DOI: 10.1007/s11684-022-0959-5.

44. Angus L., Moleirinho S., Herron L., Sinha A., Zhang X., Niestrata M. et al. Willin/FRMD6 expression activates the Hippo signaling pathway kinases in mammals and antagonizes oncogenic YAP. Oncogene. 2012;31(2):238–250. DOI: 10.1038/onc.2011.224.

45. Ma S., Meng Z., Chen R., Guan K.L. The Hippo pathway: biology and pathophysiology. Annu. Rev. Biochem. 2019;88:577– 604. DOI: 10.1146/annurev-biochem-013118-111829.

46. Mokhtari R.B., Ashayeri N., Baghaie L., Sambi M., Satari K., Baluch N. et al. The Hippo pathway effectors YAP/TAZTEAD oncoproteins as emerging therapeutic targets in the tumor microenvironment. Cancers (Basel). 2023;15(13):3468. DOI: 10.3390/cancers15133468.

47. Akrida I., Bravou V., Papadaki H. The deadly cross-talk between Hippo pathway and epithelial-mesenchymal transition (EMT) in cancer. Mol. Biol. Rep. 2022;49(10):10065–10076. DOI: 10.1007/s11033-022-07590-z.

48. Guan C., Chang Z., Gu X., Liu R. MTA2 promotes HCC progression through repressing FRMD6, a key upstream component of Hippo signaling pathway. Biochem. Biophys. Res. Commun. 2019;515(1):112–118. DOI: 10.1016/j.bbrc.2019.05.025.

49. Haldrup J., Strand S.H., Cieza-Borrella C., Jakobsson M.E., Riedel M., Norgaard M. et al. FRMD6 has tumor suppressor functions in prostate cancer. Oncogene. 2021;40(4):763–776. DOI: 10.1038/s41388-020-01548-w.

50. Von Koskull A., Hagstrom J., Haglund C., Kaprio T., Bockelman C. et al. High-tissue FRMD6 expression predicts better outcomes among colorectal cancer patients. Biomarkers. 2024;29(3):127–133. DOI: 10.1080/1354750X.2024.2321916.

51. Wang T., Guo H., Zhang L., Yu M., Li Q., Zhang J. et al. FERM domain-containing protein FRMD6 activates the mTOR signaling pathway and promotes lung cancer progression. Front. Med. 2023;17(4):714-728. DOI: 10.1007/s11684-022-0959-5.

52. Schmuck K., Ullmer C., Engels P., Lübbert H. Cloning and functional characterization of the human 5-HT2B serotonin receptor. FEBS Lett. 1994;342(1):85–90. DOI: 10.1016/0014-5793(94)80590-3.

53. Jia J., Wang M., Xing S., Huang Z., Jiang Y. Crosstalk between enteric serotonergic neurons and colorectal cancer stem cells to initiate colorectal tumorigenesis. Front. Oncol. 2022;12:1054590. DOI: 10.3389/fonc.2022.1054590.

54. Yu H., Qu T., Yang J., Dai Q. Serotonin acts through YAP to promote cell proliferation: mechanism and implication in colorectal cancer progression. Cell Commun. Signal. 2023;21(1):75. DOI: 10.1186/s12964-023-01096-2.

55. Li T., Wei L., Zhang X., Fu B., Zhou Y., Yang M. et al. Serotonin receptor HTR2B facilitates colorectal cancer metastasis via CREB1-ZEB1 axis mediated epithelial-mesenchymal transition. Mol. Cancer Res. 2024;22(6):538–554. DOI: 10.1158/1541-7786.MCR-23-0513.

56. Liu P., Yin Y.L., Wang T., Hou L., Wang X.X., Wang M. et al. Ligand-induced activation of ERK1/2 signaling by constitutively active Gs-coupled 5-HT receptors. Acta Pharmacol. Sin. 2019;40(9):1157–1167. DOI: 10.1038/s41401-018-0204-6.

57. Xu M., Wang S., Wang Y., Wu H., Frank J.A., Zhang Z. et al. Role of p38γ MAPK in regulation of EMT and cancer stem cells. Biochim. Biophys. Acta Mol. Basis Dis. 2018;1864(11):3605– 3617. DOI: 10.1016/j.bbadis.2018.08.024.

58. Tu R.H., Wu S.Z., Huang Z.N., Zhong Q., Ye Y.H., Zheng C.H. et al. Neurotransmitter receptor HTR2B regulates lipid metabolism to inhibit ferroptosis in gastric cancer. Cancer Res. 2023;83(23):3868–3885. DOI: 10.1158/0008-5472.CAN-23-1012.

59. Soll C., Jang J.H., Riener M.O., Moritz W., Wild P.J., Graf R. et al. Serotonin promotes tumor growth in human hepatocellular cancer. Hepatology. 2010;51(4):1244–1254. DOI: 10.1002/hep.23441.

60. Henriksen R., Dizeyi N., Abrahamsson P.A. Expression of serotonin receptors 5-HT1A, 5-HT1B, 5-HT2B and 5-HT4 in ovary and in ovarian tumours. Anticancer Res. 2012;32(4):1361–1366.

61. Jacobs J.J., Kieboom K., Marino S., DePinho R.A., van Lohuizen M. The oncogene and Polycomb-group gene bmi-1 regulates cell proliferation and senescence through the ink4a locus. Nature. 1999;397(6715):164–168. DOI: 10.1038/16476.

62. Xu J., Li L., Shi P., Cui H., Yang L. The crucial roles of Bmi-1 in cancer: implications in pathogenesis, metastasis, drug resistance, and targeted therapies. Int. J. Mol. Sci. 2022;23(15):8231. DOI: 10.3390/ijms23158231.

63. Gray F., Cho H.J., Shukla S., He S., Harris A., Boytsov B. et al. BMI1 regulates PRC1 architecture and activity through homoand hetero-oligomerization. Nat. Commun. 2016;7:13343. DOI: 10.1038/ncomms13343.

64. Li H., Song F., Chen X., Li Y., Fan J., Wu X. et al. Bmi-1 regulates epithelial-to-mesenchymal transition to promote migration and invasion of breast cancer cells. Int. J. Clin. Exp. Pathol. 2014;7(6):3057–3064.

65. Jiang Z.Y., Ma X.M., Luan X.H., Liuyang Z.Y., Hong Y.Y., Dai Y. et al. BMI-1 activates hepatic stellate cells to promote the epithelial-mesenchymal transition of colorectal cancer cells. World J. Gastroenterol. 2023;29(23):3606–3621. DOI: 10.3748/wjg.v29.i23.3606.

66. Paranjape A.N., Balaji S.A., Mandal T., Krushik E.V., Nagaraj P., Mukherjee G. et al. BMI1 regulates self-renewal and epithelial to mesenchymal transition in breast cancer cells through Nanog. BMC Cancer. 2014;14:785. DOI: 10.1186/1471-2407-14-785.

67. Wang M.C., Li C.L., Cui J., Jiao M., Wu T., Jing L.I. et al. BMI-1, a promising therapeutic target for human cancer. Oncol. Lett. 2015;10(2):583–588. DOI: 10.3892/ol.2015.3361.

68. Pourjafar M., Samadi P., Karami M., Najafi R. Assessment of clinicopathological and prognostic relevance of BMI-1 in patients with colorectal cancer: A meta-analysis. Biotechnol. Appl. Biochem. 2021;68(6):1313–1322. DOI: 10.1002/bab.2053.

69. Masiakowski P., Carroll R.D. A novel family of cell surface receptors with tyrosine kinase-like domain. J. Biol. Chem. 1992;267(36):26181–26190.

70. Borcherding N., Kusner D., Liu G.H., Zhang W. ROR1, an embryonic protein with an emerging role in cancer biology. Protein Cell. 2014;5(7):496–502. DOI: 10.1007/s13238-014-0059-7.

71. Neiheisel A., Kaur M., Ma N., Havard P., Shenoy A.K. et al. Wnt pathway modulators in cancer therapeutics: An update on completed and ongoing clinical trials. Int. J. Cancer. 2022;150(5):727–740. DOI: 10.1002/ijc.33811.

72. Cetin M., Odabas G., Douglas L.R., Duriez P.J., Balcik-Ercin P., Yalim-Camci I. et al. ROR1 expression and its functional significance in hepatocellular carcinoma cells. Cells. 2019;8(3):210. DOI: 10.3390/cells8030210.

73. Zhou J.K., Zheng Y.Z., Liu X.S., Gou Q., Ma R., Guo C.L. et al. ROR1 expression as a biomarker for predicting prognosis in patients with colorectal cancer. Oncotarget. 2017;8(20):32864–32872. DOI: 10.18632/oncotarget.15860.

74. Zhang S., Cui B., Lai H., Liu G., Ghia E.M., Widhopf G.F. 2nd et al. Ovarian cancer stem cells express ROR1, which can be targeted for anti-cancer-stem-cell therapy. Proc. Natl. Acad. Sci. USA. 2014;111(48):17266–17271. DOI: 10.1073/pnas.1419599111.

75. Saleh R.R., Antrás J.F., Peinado P., Pérez-Segura P., Pandiella A., Amir E. et al. Prognostic value of receptor tyrosine kinase-like orphan receptor (ROR) family in cancer: A meta-analysis. Cancer Treat. Rev. 2019;77:11–19. DOI: 10.1016/j.ctrv.2019.05.006.

76. Kamrani A., Mehdizadeh A., Ahmadi M., Aghebati-Maleki L., Yousefi M. Therapeutic approaches for targeting receptor tyrosine kinase-like orphan receptor-1 in cancer cells. Expert Opin. Ther. Targets. 2019;23(5):447–456. DOI: 10.1080/14728222.2019.1602608.

77. Zhao Y., Zhang D., Guo Y., Lu B., Zhao Z.J., Xu X. et al. Tyrosine kinase ROR1 as a target for anti-cancer therapies. Front. Oncol. 2021 28;11:680834. DOI: 10.3389/fonc.2021.680834.

78. Lin H., Cheng J., Mu W., Zhou J., Zhu L. et al. Advances in universal CAR-T cell therapy. Front. Immunol. 2021;12:744823. DOI: 10.3389/fimmu.2021.744823.

79. Meng S., Li M., Qin L., Lv J., Wu D., Zheng D. et al. The onco-embryonic antigen ROR1 is a target of chimeric antigen T cells for colorectal cancer. Int. Immunopharmacol. 2023;121:110402. DOI: 10.1016/j.intimp.2023.110402.

80. O’Connor E.S., Greenblatt D.Y., LoConte N.K., Gangnon R.E., Liou J.I., Heise C.P. et al. Adjuvant chemotherapy for stage II colon cancer with poor prognostic features. J. Clin. Oncol. 2011;29(25):3381–3388. DOI: 10.1200/JCO.2010.34.3426.

81. Benson A.B., Venook A.P., Al-Hawary M.M., Cederquist L., Chen Y.J., Ciombor K.K. et al. NCCN Guidelines Insights: Colon Cancer, Version 2.2018. J. Natl. Compr. Canc. Netw. 2018;16(4):359–369. DOI: 10.6004/jnccn.2018.0021.

82. Liu Z., Lu T., Li J., Wang L., Xu K., Dang Q. et al. Clinical significance and inflammatory landscape of a novel recurrence-associated immune signature in Stage II/III colorectal cancer. Front. Immunol. 2021;12:702594. DOI: 10.3389/fimmu.2021.702594.

83. Chen K., Collins G., Wang H., Toh J.W.T. Pathological features and prognostication in colorectal cancer. Curr. Oncol. 2021;28(6):5356–5383. DOI: 10.3390/curroncol28060447.

84. Pagès F., Mlecnik B., Marliot F., Bindea G., Ou F.S., Bifulco C. et al. International validation of the consensus Immunoscore for the classification of colon cancer: a prognostic and accuracy study. Lancet. 2018;391(10135):2128–2139. DOI: 10.1016/S0140-6736(18)30789-X.

85. Malla M., Loree J.M., Kasi P.M., Parikh A.R. Using circulating tumor DNA in colorectal cancer: current and evolving practices. J. Clin. Oncol. 2022;40(24):2846–2857. DOI: 10.1200/JCO.21.02615.