Study On Prognostic Markers Of Diffuse Large B - Cell

Part â…  Immunohistochemical Algorithm only is not Enough for Predicting the Outcome of Patients with Diffuse Large B Cell Lymphoma treated with chemoimmunotherapyObjectives: The clinical and pathological heterogeneity of diffuse large B cell lymphoma(DLBCL) makes prognostic assessment difficult. Gene expression profiling(GEP) technique which can reproducibly divide DLBCL into three groups. However, GEP is expensive and impractical to perform in routine laboratories, algorithms using the method of immunohistochemistry(IHC) were proposed as a surrogate for GEP analysis. But the power of the algorithms was being suspected in chemoimmunotherapy era. It is notable that most of these algorithms have been developed and tested only in Western countries; few reports have confirmed the practicability of their use in Eastern nations. The purpose of this study was to analyze the prognositc value of four algorithms in Chinese DLBCL patients treated with chemoimmunotherapy.Methods: In this study, we retrospectively studied 244 de novo DLBCL who had been diagnosed between February 2006 and January 2014. All of the paraffin-embedded sections were reviewed by two hematopathologists and the diagnoses were based on the World Health Organization(WHO) classification criteria. Among these 244 patients, 141 cases were treated with chemoimmunotherapy which was used for prognostic analysis. Using IHC, we reassessed the prognostic value of the four conventional algorithms and proteins expression in each algorithm in DLBCL patients. Additionally, in order to improve in prognostic value of IHC algorithms, we analyzed MYC and BCL2 gene translocation and protein expression by IHC and and fluorescence in situ hybridization(FISH). Chi-squared and Fisher exact test were used to see consistency among algorithms. Spearman test was used to analyze correlations among different variables. The prognostic value of the four algorithms, protein expression in each of them, and MYC or BCL2 gene aberration was assessed by survival analysis.Results: According to the algorithms applied in this study, 244 cases of de novo DLBCL patients could be further investigated in an IHC process. For the four IHC algorithms(Hans, Choi, Tally and Visco-Young algorithms), 99 and 145, 119 and 125, 69 and 175, 107 and 137 were assigned to GCB and non-GCB, respectively. Neither single protein within algorithms nor the IHC algorithms themselves had strong prognostic power. Using MYC aberration(MA) either on the genetic or protein levels, we established a new algorithm called MA that could divide patients into distinct prognostic groups. Patients of MA had much shorter overall survival(OS) and progression-free survival(PFS) than non-MA(median OS: 25.7 months vs not reached; median PFS: 14.4 months vs not reached; P< 0.0001 for both).Conclusions: Since the low prognostic value of IHC algorithms, using additional prognostic markers not associated with cell of origin may be an alternative way to predict outcome of DLBCL. MA algorithm showed robust prognostic value in predicting outcome of DLBCL. However, since the limited cases used in our study, studies with large samples should be performed to confirm the value of our algorithm and optimize the prognostic system of DLBCL.Partâ…¡ The distinct clinical features and prognosis of the CD10+MUM1+ and CD10-BCL6-MUM1- diffuse large B-cell lymphomaObjectives: Using an immunohistochemistry based method, DLBCL can be classified into germinal centre B-cell(GCB) and non-GCB subtypes by Hans algorithm, which have different biological and pathological features. However, the prognostic role of Hans algorithm was contradictory in the literature. In addition, CD10+MUM1+ and CD10-BCL6-MUM- groups show contradictory immunophenotype with their COO. The purpose of this study was to analyze the clinical features and prognostic value of these groups.Methods: We reviewed the medical records of patients who diagnosed as de novo DLBCL at our hospital between September 2006 and October 2014. A total of 601 cases were included in the study. Among these, 306 cases were treated with rituximab based chemoimmunotherapy and well follow up. IHC was peiformed in 601 patients with antibodies in Hans algorithm. Additional 42 cases in CD10+MUM1+ and CD10-BCL6-MUM- groups were analyszed by IHC with three other IHC algorithms(Choi, Tally and Visco-Young algorithms). We then reassessed the clinical features of special groups with Hans algorithm in 601 DLBCL patients and prognostic value in 306 cases who were treated with chemoimmunotherapy.Results: Patients with GCB subtype had better OS and PFS than non-GCB cases. However, the groups of double positive(CD10+MUM1+, DP) and triple negative(CD10-BCL6-MUM-, TN) showed different clinical characteristics to their original category. In addition, 61.9%(13/21) of the DP group in Hans algorithm were recognized as non-GCB group with Tally algorithm and 52.4%(11/21) of the TN group in Hans algorithm belonged to GCB group in Tally algorithm. Other algorithms showed high consistency with Hans algorithm both in the DP and TN group. The DP group showed similar OS(median OS: both not reached, P=0.3650) and PFS(median PFS: 47.0 vs. 32.7 months, P=0.0878) with those of the non-GCB group. The TN group showed similar OS(median OS: both not reached, P=0.9278) and PFS(median PFS: both not reached, P=0.9420) with those of the GCB group.Conclusions: Recognition of specific entities in Hans algorithm could help us to accurately predict outcome of the patients and choose the best clinical management for them.Part â…¢ MYC or BCL2 copy number aberration is a strong predictor of outcome in patients with diffuse large B-cell lymphoma Objectives: DLBCL is the most common non-Hodgkin lymphomas(NHL). MYC rearrangement concurrent with BCL2 or / and BCL6 rearrangement, also called double hit lymphoma(DHL), constitute a specific cohort of patients with extremely poor outcome. However, most studies on DHL were reported in Western nations. The prognositc value these genes aberrations need to be further investigated the Chinese population.Methods: In this study, we enrolled 246 adult patients with de novo DLBCL that had been diagnosed between February 2006 and January 2014. All of the patients were treated with chemoimmunotherapy or chemotherapy alone. Using IHC and FISH, we investigated MYC, BCL2 and BCL6 gene aberration with DLBCL patients in Chinese population. Cases with additional signals of MYC(MYC break apart probe) along with two aqua signals of centromere 8 per nuclei were considered as MYC CNA(copy number aberration). Cases with additional signals of BCL2 and normal IGH signals(BCL2/IGH probe) and without BCL2 gene break apart(BCL2 gene break apart probe) were considered as BCL2 CNA. Three or four copies of the gene studied was considered a gain, whereas more than four copies was considered an amplification. The association between gene or protein aberrations was analyzed with Chi-squared, Fisher exact and Spearman test. The survival differences of MYC, BCL2 gene aberration were analyzed using the Kaplan-Meier method and were compared by using the log-rank test.Results: Among the 246 DLBCL patients, MYC CNA was detected in 18 cases(7.5%), which was less frequent than MYC rearrangement(12.5%, 30/240). BCL2 CNA was observed in 65(27.1%) cases, which was more common than IGH/BCL2 rearrangement(12.5%, 30/240). Among these, 3.8%(9/240) were identified as having concurrent CNA involving MYC and BCL2(double CNA), which was a little more frequent than classic DHL(2.9%, 7/240). Both MYC CNA and MYC rearrangement were associated with MYC expression. However, only a trend of association were observed between MYC CNA and MYC rearrangement. Similarly, Both BCL2 CNA and BCL2 rearrangement were associated with BCL2 expression. No association was found between BCL2 CNA and BCL2 rearrangement. Patients with MYC or BCL2 CNA had significantly worse OS and PFS than negative cases(MYC CNA: median OS: 15.8 months vs. not reached, P<0.0001; median PFS: 6.1 vs. 29.0 months, P<0.0001; BCL2 CNA: median OS: 24.1 months vs. not reached, P<0.0001; median PFS: 13.2 vs. 74.7 months, P<0.0001). No survival differences were observed between gain and amplification with MYC or BCL2 CNA. BCL2 CNA had much inferior outcome than BCL2 rearrangement or BCL2 expression(median OS: 24.1 months vs. not reached vs. not reached, P=0.0021; median PFS: 13.2 vs. 37.7 vs. 20.7 months, P=0.0008) while MYC CNA predicted similar OS(median OS: 15.8 vs. 18.3 vs. 23.0 months, P=0.1228) and PFS(median PFS: 6.1 vs. 9.7 vs. 11.6 months, P=0.2750) with MYC rearrangement or MYC expression. Patients with both MYC and BCL2 CNA had a similar outcome to classic DHL or protein double expression(MYC and BCL2 or BCL6 coexpression). In multivariate analysis, MYC CNA(except for PFS), BCL2 CNA and double CNA were independent worse prognostic factors for OS and PFS.Conclusions: Patients with MYC or BCL2 CNA constituted a unique group with extremely poor outcome and may require more aggressive treatment regimens.Part IV Epstein-Barr virus positive diffuse large B-cell lymphoma predict poor outcome, regardless of the age Objectives: Epstein-Barr virus(EBV) positive DLBCL is defined as patients older than 50 years which predict poor outcome in most studies. Recent reports showed young patients with sound immune status could also be affected. However, the prognositc value of EBV positive DLBCL patients of the young group is controversial. The purpose of this study was to analyze the clinical feature and prognosis value of EBV positive DLBCL patients in different age groups.Methods: In this study, we reviewed the medical records of 250 patients who diagnosed as de novo DLBCL at our hospital between July 2006 and December 2014. All of the patients were treated with chemoimmunotherapy. Using IHC, we analyzed the pathological features in DLBCL patients. FISH analysis was performed with MYC and IGH/BCL2 translocation probe. Epstein-Barr virus-encoded RNA(EBER) in situ hybridization was carried out using a fluorescein-conjugated EBER oligonucleotide probe and the purified Ig G fraction of a mouse monoclonal anti-fluorescein antibody. The COO was classified according to Hans, Choi, Tally and Visco-Young algorithms. The different clinical and pathological features were analyzed with EBV status in the different age groups. The survival different was performed using the Kaplan-Meier method and were compared by using the log-rank test.Results: The incidence of EBV positivity was 10.4%(26/250). The EBV positive patients shared many unfavorable prognostic characteristics, regardless of elderly or young group. EBV positive patients, both in the elderly and young groups, showed significantly worse OS(median OS, elderly group: 37.0 months vs. not reached, P=0.0337; young group: 36.5 months vs. not reached, P<0.0001) and PFS(median PFS: elderly group: 20.7 months vs. not reached, P<0.0001; young group: 20.5 months vs. not reached, P=0.0010) than negative cases.Conclusions: Since EBV positive DLBCL patients, regardless of age, shared similar poor prognostic features and showed worse outcome than negative cases, we suggest that the age criterion of EBV positive DLBCL, and possibly the name itself, be modified in future.