Association of pSTAT3, pSyk with c-Myc, p53, BCL2 proteins expression and survival of patients with diffuse large B-cell lymphoma

Background. Diffuse large B cell lymphoma (DLBCL) is one of the most common non-Hodgkin’s lymphomas. The effectiveness of R-CHOP (rituximab, cyclophosphamide, hydroxydaunorubicin, oncovin, prednisone) therapy was observed in about 60 % of patients, in 20–30 % of cases DLBCL is characterized by a refractory course. Standard clinical prognostic criteria do not accurately determine the prognosis of the disease. In this regard, the problem of developing additional predictors of DLBCL course remains relevant today and is directly related to the molecular pathogenesis of the disease.
The objective: to determine the relationship of pSTAT3, pSyk with c-Myc, p53, BCL2 oncoproteins expression and survival of DLBCL patients.
Materials and methods. Biopsy specimens of lymph nodes and other organs and tissues obtained from 100 patients with newly diagnosed DLBCL were used for the study. All patients received standard first-line immunochemotherapy according to the R-CHOP regimen. Determination of the relative amount of tumor cells expressing pSTAT3, pSyk was carried out using immunohistochemical and morphometric methods. The threshold value of proteins expression was calculated using the ROC analysis: for pSTAT3 it was 68 % of positive tumor cells, for pSyk – 28 %. The relationship of pSTAT3, pSyk with c-Myc, p53, BCL2 expression was determined using the Pearson χ2  test. Overall (OS) and progression-free (PFS) survival were calculated using the Kaplan–Meier method (log-rank test).
Results. The suprathreshold pSTAT3 expression was found to be associated with a high content of c-Myc and p53 transcription factors in tumor cells (p = 0.015 and p = 0.010, respectively). In the group of patients with isolated overexpression of pSTAT3, the 5-year OS and PFS were lower, and the risk of death and disease progression was almost 1.5 times higher than in patients with low protein expression (p = 0.015 and p = 0.011, respectively). When studying the co-expression of pSTAT3 and pSyk, the lowest OS and PFS were recorded in the subjects with a simultaneous high expression of these proteins. With this combination of markers, the probability of an adverse event in patients when calculating OS increased by 2.9 times (p = 0.003; hazard ratio 2.9; 95 % confidence interval 1.43–5.85), and when analyzing PFS – by 2.3 times (p = 0.021; hazard ratio 2.3; 95 % confidence interval 1.14–4.87) compared with other variants of their simultaneous expression.
Conclusion. Overexpression of pSTAT3 is associated with unfavorable biological tumor characteristics and poor patient survival. The combined suprathreshold expression of the pSTAT3 and pSyk proteins is associated with lower OS and PFS values compared to their isolated expression.

1. Bahar T., Chowdhury Z.Z., Rahman Sh. et al. Clinicopathological correlation with outcome of diffuse large B cell lymphoma: experience in a specialized cancer care centre in Bangladesh. J Medicine 2021;22(1):3–6. DOI:10.3329/jom.v22i1.51383.

2. Vaneeva E.V., Rosin V.A., Dyakonov D.A. et al. Assessment of the prognostic value of pSTAT3 expression in diffuse large B-cell lymphoma in a Russian sample of patients. Sibirskiy nauchnyy meditsinskiy zhurnal = Siberian Scientific Medical Journal 2019;39(5):125–33. (In Russ.). DOI:10.15372/SMMJ20190515.

3. Mondello P., Mian M. Frontline treatment of diffuse large B-cell lymphoma: Beyond R-CHOP. Hematol Oncol 2019;37(4):333–44. DOI:10.1002/hon.2613.

4. Swerdlow S.H., Campo E., Harris N.L. et al. WHO classification of tumours of haematopoietic and lymphoid tissues. 4th edn. Lyon, France: IARC, 2017.

5. Roschewski M., Staudt L.M., Wilson W.H. et al. Diffuse large B-cell lymphoma treatment approaches in the molecular era. Nat Rev Clin Oncol 2014;11(1):12–23. DOI:10.1038/nrclinonc.2013.197.

6. Misyurina A.E., Kravchenko S.K., Obukhova T.N. et al. Expression of MYC and BCL2 proteins in patients with diffuse large B-cell lymphoma. Klinicheskaya onkogematologiya = Clinical Oncohematology 2015;1(8):44–53. (In Russ.).

7. Rastorguev S.M., Koroleva D.A., Boulygina E.S. et al. Clinical and prognostic value of molecular markers of diffuse large B-cell lymphoma. Klinicheskaya onkogematologiya = Clinical Oncohematology 2019:12(1): 95–100. (In Russ.). DOI:10.21320/2500-2139-201912-1-95-100.

8. Miao Y., Medeiros L., Xu-Monette Z.Y. et al. Dysregulation of cell survival in diffuse large B cell lymphoma: mechanisms and therapeutic targets. Front Oncol 2019;9:1–17. DOI:10.3389/fonc.2019.00107.

9. Brachet-Botineau M., Polomski M., Neubauer A. et al. Pharmacological inhibition of oncogenic STAT3 and STAT5 signaling in hematopoietic cancers. Cancers 2020;12:240. DOI:10.3390/cancers12010240.

10. Seda V., Mraz M. Eur B-cell receptor signalling and its crosstalk with other pathways in normal and malignant cells. J Haematol 2014;94(3):193–205. DOI:10.1111/ejh.12427.

11. Nikitin E.A. B-cell receptor signaling pathway: mechanisms and inhibitors. Klinicheskaya onkogematologiya = Clinical Oncohematology 2014;7(3): 251–63. (In Russ.).

12. Wossning T., Herzog S., Köhler F. et al. Deregulated Syk inhibits di_erentiation and induces growth factor-independent proliferation of pre-B cells. J Exp Med 2006;203:2829–40. DOI:10.1084/jem.20060967.

13. Liu H., Zhu L., Sun C. et al. Clinical significance and prognostic value of STAT3 expression in patients with diffuse large B-cell lymphoma treated with rituximab-CHOP therapy. Int J Clin Exp Med 2016;9(6):9356–64.

14. Kovrigina A.M., Baykov V.V. Pathological differential diagnosis of primary myelofibrosis. Study guide. Moscow, Saint-Petersburg: Novartis Pharma, 2014. 11 р. (In Russ.).

15. Kolosenko I., Yu Y., Busker S. et al. Identification of novel small molecules that inhibit STAT3-dependent transcription and function. PLoS One 2017;12(6): e0178844. DOI:10.1371/journal.pone.

16. Fei Y., Yu J., Li Y. et al. Plasma soluble PD-L1 and STAT3 predict the prognosis in diffuse large B cell lymphoma patients. J Cancer 2020;11(23):7001–8. DOI:10.7150/jca.47816.

17. Ok C.J., Xu-Monette Z.Y., Tzankov A. et al. STAT3 expression and clinical inplications in de novo diffuse large B cell lymphoma: a report from the International DLBCL RituximabCHOP consortium program. Blood 2013;122:365. DOI:10.1182/blood.V122.21.365.365.