Bcl-2 Status Research Articles

Efficacy of venetoclax-dexamethasone (VenDex) v pomalidomide-dexamethasone (PomDex) in patients (Pts) with t(11;14)-positive relapsed/refractory multiple myeloma [t(11;14)+ RRMM]: Phase 3 CANOVA study biomarker subgroup analysis.

7510 Background: The randomized Phase 3 CANOVA study’s (NCT03539744) primary analysis showed multiple numerically improved efficacy endpoints with VenDex v PomDex, but the primary endpoint of mPFS by IRC was not statistically significant. Here, we report correlative biomarker analyses from CANOVA. Methods: CANOVA is a randomized, global, open-label Phase 3 study of VenDex v PomDex in pts with t(11;14)+ RRMM and ≥2 prior lines of therapy (LOTs). The primary endpoint was mPFS by IRC. Key secondary endpoints included ORR, ≥VGPR, mOS, and MRD negativity (<10-5). In post hoc mPFS sensitivity analysis, disease progression, death, and start of next LOT were defined as events. BCL2 gene expression by RNAseq (3.7 log2 FPKM prespecified median cutoff for BCL2high v BCL2low) and chr1q abnormalities by whole-exome sequencing (normal, gain, or amp) were assessed centrally from pretreatment, CD138-enriched BM aspirates. Results: mPFS, post hoc mPFS, and mOS in the VenDex arm were similar by BCL2 status, with numerically higher ORR, ≥VGPR, and MRD negativity rates in the BCL2highsubgroup. In contrast, mPFS and mOS within the PomDex arm were numerically longer in the BCL2lowsubgroup, despite equal ORR in both subgroups (Table). Presence of 1q abnormalities was evenly distributed across BCL2high v BCL2low subgroups in the VenDex arm (51% v 45%) but not in the PomDex arm (59% v 31%). Normal 1q and gain(1q) subgroups had numerically improved mPFS, mOS, ORR, ≥VGPR, and MRD negativity with VenDex; pts with amp(1q) had poor outcomes regardless of treatment, albeit the sample size was small. Conclusions: Pts with BCL2high or gain(1q) had numerically improved clinical efficacy with VenDex v PomDex. Clinical benefit was consistent across BCL2 subgroups ( BCL2high or BCL2low) with VenDex but not PomDex. Pts with amp(1q) fared poorly irrespective of treatment arm. Clinical trial information: NCT03539744 . [Table: see text]

Molecular Profiling of High-Grade B-Cell Non-Hodgkin Lymphomas and Its Clinicopathologic Correlation in an Indian Tertiary Cancer Care Center

Abstract Introduction Double-expressor and double-hit lymphomas (DHL) are known to be more aggressive and have poor outcomes with standard chemotherapy regimens. Objectives To assess the incidence of DHL and triple-hit lymphomas (THL) and correlate them with clinicopathologic parameters. Materials and Methods Patients who were diagnosed with high-grade lymphomas from April 2021 to September 2022 were prospectively followed up, and details comprising clinical and pathological parameters, including the immunohistochemistry expression status and gene rearrangements of MYC, BCL2, and BCL6, were recorded. Results The incidence of DHL and THL in our study was 16.43%. The separate incidence of the DHL-BCL2, DHL-BCL6, and THL groups was 16.43, 13.69, and 2.73%, respectively. The germinal center B cell subtype of histology was predominantly seen in DHLs. MYC, BCL2, and BCL6 expressions do not correlate well with translocations of these genes. Conclusion Double protein expression cannot be used for screening to decide which patients should undergo fluorescence in situ hybridization, as this would result in missing 4.5% of DHLs.

Does the Expression of Vascular Endothelial Growth Factor (VEGF) and Bcl-2 Have a Prognostic Significance in Advanced Non-Small Cell Lung Cancer?

Lung cancer is the most common cause of mortality from malignant tumors worldwide. The five-year survival rate for people with advanced stages varies considerably, from 35.4% to 6.9%. The angiogenic potential of bcl2 is not well known, nor is the way in which tumor cells with excessive bcl2 expression affect VEGF production. Hypothetically, given that tumor growth, progression and metastasis are dependent on angiogenesis, the antiapoptotic effect is expected to form a link between these two molecules. The aim of this study was to evaluate the relationship between bcl-2 and VEGF expression, clinicopathological features and survival in 216 patients with advanced NSCLC. Archival tumor tissues were examined by immunohistochemistry for the expression of bcl-2 and VEGF. Immunoreactivity for bcl-2 was observed in 41.4% of NSCLCs, 51% of squamous and 34.8% of adenocarcinomas-expressed Bcl-2. There was an inverse correlation of mononuclear stromal reaction and bcl-2 expression in adenocarcinoma (p < 0.0005). A total of 71.8% NSCLCs were VEGF positive, 56% of squamous and 82.2% of adenocarcinomas. High level of VEGF expression was significantly associated with histology type (p = 0.043), low histology grade (p = 0.014), clinical stage IV (p = 0.018), smoking history (p = 0.008) and EGFR mutations (p = 0.026). There was an inverse correlation in the expression of Bcl-2 and VEGF in NSCLC patients (p = 0.039, r = -0.163). Two-year survival of patients with unresectable NSCLC was 39.3%, and 50% of patients were alive at 17 months. Our results demonstrated no difference in survival for patients in advanced NSCLC grouped by bcl-2 and VEGF status. Additionally, we observed an inverse correlation in the expression of Bcl-2 and VEGF in NSCLC and mononuclear reaction and bcl-2 expression in adenocarcinomas.

Prognostic impact of miR-125b and miR-155b and their relationship with MYC and TP53 in diffuse large B-cell lymphoma: cell-of-origin classification matters.

Tumoral microRNAs, such as miR-125b and miR-155b, are important gene expression regulators with complex pathogenetic mechanisms. However, their role in DLBCL, especially when cell-of-origin classification is considered, are still to be elucidated. In a series of 139 DLBCL cases considering germinal center (GC) versus nonGC subtypes, we investigated miR-125b and miR-155b expression by in situ hibridization and their association with some immunophenotypic presentations, including MYC, BCL2 and TP53 expression, MYC, BCL2 and BCL6 translocation status, as well as clinicopathological features and outcomes. miR-125b detection was positively correlated to the Ki-67 index (P = 0.035) in the nGC. Considering the GC subgroup, the percentage of miR-125b positive cells was also correlated to either MYC and MYC/BCL2 double expression (P = 0.047 and P = 0.049, respectively). When it comes to nGC patients, miR-155b percentage and intensity, as well as Allred score, were positively correlated to disease progression (P = 0.038, P = 0.057 and P = 0.039, respectively). In a multivariate analysis, GC phenotype was a significant independent factor associated with higher OS (P = 0.007) and, considering the nGC group, although not significant, the expression of TP53, miR-125b and miR-155b seems to be potential prognostic biomarkers in these tumors. This study demonstrated different pathways based on cell-of-origin classification and highlighted different clinical outcomes. miR-125b, miR-155b and TP53 expression may also represent potential prognostic factors in nGC-DLBCL.

P1190: OUTCOMES BY BCL2 AND MYC EXPRESSION AND REARRANGEMENTS IN UNTREATED DIFFUSE LARGE B-CELL LYMPHOMA FROM THE POLARIX TRIAL

Background: Overexpression of BCL2 and MYC, and translocation of MYC, BCL2, and BCL6, are associated with poorer outcomes in patients with diffuse large B-cell lymphoma (DLBCL; Horn et al. Blood 2013). We previously reported progression-free survival (PFS) from POLARIX in subgroups of patients with DLBCL receiving Pola-R-CHP or R-CHOP, including patients with double expressor lymphoma (DEL; favoring Pola-R-CHP: hazard ratio [HR] 0.64, 95% confidence interval [CI] 0.42–0.97), and double- or triple-hit lymphoma (DHL/THL; favoring R-CHOP: HR 3.81, 95% CI 0.82–17.64) (Tilly et al. NEJM 2022). Aims: This is a prespecified exploratory analysis, to further analyze immunohistochemistry (IHC) expression status of BCL2 and MYC, and rearrangements (R) of BCL2, BCL6 and MYC as independent prognostic markers. The prognostic impact of DEL, and BCL2 and MYC overexpression within treatment arms was also explored. Methods: BCL2 and MYC protein expression were assessed by IHC and identified as IHC+ (≥50% [BCL2]/≥40% [MYC]) or IHC−; MYC-R, BCL2-R, and BCL6-R were detected by fluorescence in situ hybridization (Tilly et al. NEJM 2022). All patients provided informed consent. Pola-R-CHP is an investigational combination. Exploratory multivariate Cox regression models were adjusted for treatment, stratification factors (International Prognostic Index, bulky disease, geographic region), age >60 years, cell of origin, and biomarker evaluated, as appropriate. Results: The prevalence, univariate HR, and 2-year PFS estimates of BCL2+/MYC+ and BCL2-R/MYC-R/BCL6-R subgroups are presented (Table) except for BCL6-R, because all four PFS events in this subgroup were with Pola-R-CHP. Multivariate analyses results for Pola-R-CHP vs R-CHOP in patients with BCL2+ (HR 0.60, 95% CI 0.43–0.86) and those with MYC+ (HR 0.63, 95% CI 0.45–0.89) were similar to results of univariate analyses. Multivariate analyses of other subgroups were not performed due to the low patient number with BCL2-R/MYC-R/BCL6-R. While patients with DEL treated with Pola-R-CHP had improved PFS vs patients treated with R-CHOP (HR 0.64, 95% CI 0.42–0.97), the prognostic impact of DEL vs non-DEL was identified in the R-CHOP arm (univariate HR 1.53, 95% CI 1.06–2.21; multivariate HR 1.29, 95% CI 0.88–1.91) vs the Pola-R-CHP arm (univariate HR 1.10, 95% CI 0.72–1.69; multivariate HR 1.42, 95% CI 0.89–2.28). Similarly, patients with BCL2+ had inferior PFS vs patients with BCL2– in the R-CHOP arm (univariate HR 1.96, 95% CI 1.31–2.93; multivariate HR 1.74, 95% CI 1.14–2.66); however, no prognostic difference was detected in the Pola-R-CHP arm (univariate HR 1.30, 95% CI 0.85–2.01; multivariate HR 1.56, 95% CI 0.99–2.47). No prognostic impact of MYC+ vs MYC– was detected in either arm (R-CHOP: univariate HR 1.17, 95% CI 0.77–1.77; multivariate HR 1.00, 95% CI 0.65–1.55; Pola-R-CHP: univariate HR 0.83, 95% CI 0.54–1.27; multivariate HR 0.91, 95% CI 0.58–1.42). Image:Summary/Conclusion: Multivariate analyses support the benefit of Pola-R-CHP in patients with BCL2+ and MYC+ DLBCL. The poor prognostic impact associated with DEL, which was mainly driven by BCL2+, appears reduced in Pola-R-CHP- vs R-CHOP-treated patients.

Abstract 2575: Avanbulin, the active moiety of the tumor checkpoint controller lisavanbulin (BAL101553), has anti-lymphoma activity

Abstract Background: Avanbulin (BAL27862) is a microtubule-targeting agent (MTA) that promotes tumor cell death by modulating the spindle assembly checkpoint. Preclinical studies in glioblastoma (GBM) have shown that avanbulin is also active in human cancer cells resistant to other MTAs used in the clinic. Lisavanbulin (BAL101553) is a water-soluble, oral, lysine prodrug of avanbulin, currently in phase 1/2 for patients with recurrent GBM and, in combination with radiotherapy, for newly diagnosed GBM patients. Preclinical studies and clinical signals have also suggested an association between lisavanbulin activity and expression of microtubule plus-end binding protein (EB1). MTAs are active agents for lymphoma patients, as exemplified by the inclusion of vincristine in the R-CHOP regimen, standard treatment for diffuse large B cell lymphoma (DLBCL). Thus, our aim was to evaluate avanbulin, the active moiety of lisavanbulin, for its potential anti-lymphoma activity. Methods: Lymphoma cell lines derived from activated B cell like (ABC) and germinal center B cell type (GCB) DLBCL were exposed to increasing doses of avanbulin; cell proliferation was measured with MTT. Apoptosis induction was performed with annexin V staining on cells incubated in a live cell imaging system (Incucyte) treated with 5, 10, 20 or 40nM avanbulin. Images were taken every 4h for 72h. Cell cycle analysis was performed after 24, 48 and 72h of drug exposure at 20nM. Results: 26 DLBCL cell lines, treated for 72h with increasing avanbulin doses, showed high anti-proliferative activity of the compound, with a median IC50 = 11nM. No differences in terms of IC50 were observed comparing ABC (n=8) to GCB (n=18) DLBCLs, nor based on MYD88, TP53 and BCL2 status. Cell lines with MYC translocation were less sensitive than the ones without translocation. All lymphoma cell lines exhibited high EB1 RNA expression, with no correlation with sensitivity to avanbulin.Live cell imaging demonstrated a strong apoptosis induction in 15 out of 17 cell lines tested at 20 and 40nM. Among the 15 cell lines undergoing apoptosis, 8 cell lines already showed an apoptosis induction within 24h of drug exposure, 6 lines between 24 and 48h and 1 line after 48h exposure. These data confirmed the high activity of avanbulin already seen with the MTT assay.Cell cycle experiments performed at 24, 48 and 72h confirmed the strong cytotoxic effect of avanbulin in 4 DLBCL cell lines, including 2 ABC (OCILY3 and TMD8) and 2 GCB (SUDHL5 and SUDHL16) lines. Sub-G0 accumulation was already present at 24h in all 4 cell lines tested, with an increase over time. Especially at 24h, where we also observed a G2-M arrest in 3 out of 4 cell lines. Conclusions: Our data demonstrate the high cytotoxic anti-lymphoma activity of avanbulin, suggesting a potential activity of its prodrug lisavanbulin in lymphoma patients. Citation Format: Filippo Spriano, Luca Aresu, Luciano Cascione, Alberto J. Arribas, Nicole Forster-Gross, Felix Bachmann, Heidi Lane, Francesco Bertoni. Avanbulin, the active moiety of the tumor checkpoint controller lisavanbulin (BAL101553), has anti-lymphoma activity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2575.

Outcomes by BCL2 and MYC expression and rearrangements in untreated diffuse large B-cell lymphoma (DLBCL) from the POLARIX trial.

7517 Background: Overexpression of BCL2 and MYC, and translocation of MYC, BCL2, and BCL6, are associated with poorer outcomes in patients (pts) with DLBCL (Horn et al. Blood 2013). We previously reported progression-free survival (PFS) from POLARIX in subgroups of pts with DLBCL receiving Pola-R-CHP or R-CHOP, including pts with double expressor lymphoma (DEL; favoring Pola-R-CHP: hazard ratio [HR] 0.64, 95% confidence interval [CI] 0.42–0.97), and double- or triple-hit lymphoma (DHL/THL; favoring R-CHOP: HR 3.81, 95% CI 0.82–17.64) (Tilly et al. NEJM 2022). In this prespecified exploratory analysis, we further analyzed immunohistochemistry (IHC) expression status of BCL2, MYC, and rearrangements (R) of BCL2, BCL6 and MYC as independent prognostic markers. We also explored the prognostic impact of DEL within treatment arms. Methods: BCL2 and MYC protein expression were assessed by IHC and identified as IHC+ (≥50% [BCL2]/≥40% [MYC]) or IHC−; MYC-R, BCL2-R, and BCL6-R were detected by fluorescence in situ hybridization (Tilly et al. NEJM, 2022). Exploratory multivariate Cox regression models were adjusted for treatment, stratification factors (IPI, bulky disease, geographic region), age &gt;60 years, cell of origin, and biomarker evaluated, as appropriate. Results: The prevalence, univariate HR, and 2-year (yr) PFS estimates of BCL2+/MYC+ and BCL2-R/ MYC-R/ BCL6-R subgroups are presented (Table) except for BCL6-R, because all 4 PFS events in this subgroup were with Pola-R-CHP. Multivariate analyses results for Pola-R-CHP vs R-CHOP in pts with BCL2+ (HR 0.60, 95% CI 0.43–0.86) and in pts with MYC+ (HR 0.63, 95% CI 0.45–0.89) were similar to results of univariate analyses. Multivariate analyses of other subgroups were not performed due to the low pt number with BCL2-R/ MYC-R/ BCL6-R. While pts with DEL treated with Pola-R-CHP had improved PFS vs pts treated with R-CHOP (HR 0.64, 0.42–0.97), the prognostic impact of DEL vs non-DEL was more pronounced with R-CHOP (univariate HR 1.53, 95% CI 1.06–2.21; multivariate HR 1.29, 95% CI 0.88–1.91) vs Pola-R-CHP (univariate HR 1.10, 95% CI 0.72–1.69; multivariate HR 1.42, 95% CI 0.89–2.28). Conclusions: Multivariate analyses support the benefit of Pola-R-CHP in pts with BCL2+ and MYC+ DLBCL. The poor prognostic impact associated with DEL appears reduced in Pola-R-CHP- vs R-CHOP-treated pts. Clinical trial information: NCT03274492. [Table: see text]

Patient-reported outcomes provide prognostic information for survival in patients with diffuse large B-cell lymphoma: Analysis of 1239 patients from the GOYA study.

PurposeWe investigated the prognostic value of pretreatment patient‐reported outcomes (PROs) in patients with diffuse large B‐cell lymphoma (DLBCL) receiving obinutuzumab/rituximab plus chemotherapy in the GOYA phase III study.MethodsPatients completed the European Organization for Research and Treatment of Cancer Quality of Life (EORTC QLQ‐C30) and the Functional assessment of chronic illness therapy‐Lymphoma (FACT–Lym) lymphoma subscale (LYMS) during the study. PRO scales with high prognostic value were identified through Cox regression analyses of overall survival (OS) and progression‐free survival (PFS). These scales were evaluated in terms of their additional prognostic value beyond the International Prognostic Index (IPI). A preliminary assessment was performed to evaluate whether the scales provided improved patient‐risk stratification beyond IPI.ResultsOne thousand two hundred and fifty‐nine patients with valid pretreatment PRO scales were included in the analyses, and complete pretreatment data were available for 1239/1414 patients (87.6%). Four PRO scales with high prognostic value were identified: FACT–Lym LYMS and EORTC QLQ‐C30 physical functioning, global health status/quality of life (QoL), and fatigue. All four scales retained significant prognostic value for OS and PFS after IPI adjustment (all p < 0.05). After adjusting for multiple clinical variables (IPI, cell of origin, BCL2 status, and total metabolic tumor volume), all four scales retained significant prognostic value (all p < 0.05) for OS. Only the EORTC QLQ‐C30 physical functioning scale was significant (p < 0.05) for PFS after adjustment for multiple clinical variables.ConclusionsIn this large population of patients with DLBCL, pretreatment PROs provided prognostic information for OS and PFS beyond the well‐established IPI.

Clinicopathological analysis of follicular lymphoma with BCL2, BCL6, and MYC rearrangements.

Most follicular lymphomas (FL) show t(14;18)/IGH-BCL2 translocation, but rearrangement (R) negative cases exist. A series of 140 FL patients with a BCL2, BCL6, and MYC gene status examined by fluorescence in situ hybridization (FISH) were classified into five groups: (a) BCL2-R group (BCL2-R/BCL6-G/MYC-G) (G, germline), 77 cases; (b) BCL2/BCL6 double-R group (BCL2-R/BCL6-R/MYC-G), 16 cases; (c) BCL6-R group (BCL2-G/BCL6-R/MYC-G), 16 cases; (d) MYC-R group (BCL2-R or G/BCL6-R or G/MYC-R), three cases; (e) Triple-G group (BCL2-G/BCL6-G/MYC-G), 28 cases. The BCL6-R group had different clinicopathological characteristics. It showed lower rates of an advanced clinical stage and bone marrow invasion, less disease progression (p = 0.036), and a 'trend' toward a favorable progression-free survival (PFS) (p = 0.06). It also showed higher rates of grade 3A and MUM1-expression, and when analyzing the interfollicular spread pattern of CD20-positive cells, had fewer cases showing the IF3+ pattern (high interfollicular spread). Moreover, cases with BCL6-R and/or BCL6 gain (with cases of BCL2 rearrangement and/or of copy number gain excluded) correlated with favorable PFS (p = 0.014) and less IF3+ pattern (p = 0.007). We demonstrated that BCL6-R FLs showed unique clinicopathological findings, and FISH of BCL2, BCL6, and MYC is useful for FL diagnosis and clinical management.

Differentiated approach in the treatment of diffuse large B-cell lymphoma according to c-MYC and BCL2 status

Background. The treatment results of patients with diffuse large B-cell lymphoma (DLBCL) with c-MYC proto-oncogene and BCL2 protein positivity according to the CHOP-21 ± R scheme remain unsatisfactory, which dictates the need to intensify the therapy regimen. Due to the fact that most authors combine DH (double hit) lymphomas and DE (double expressor) lymphomas for analysis, these data do not allow forming an accurate idea of the independent significance of the phenomenon of activated transcription factor c-MYC and BCL2 protein in patients with DLBCL and the results of treatment specifically in this group. Some researchers believe that the presence of c-MYC gene aberration and BCL2 protein expression are associated with negative clinical characteristics. Nevertheless, the clinical features of this subgroup of DLBCL remain poorly understood.The objective of the study was to analyze the incidence of c-MYC gene aberrations, BCL2 expression in DLBCL patients and to evaluate the effectiveness of intensive R + Hyper-CVAD chemotherapy regimen in this group of patients.Materials and methods. We analyzed the data of 80 patients aged 18–65 years with a newly diagnosed DLBCL for the period from 2018 to 2020. The diagnosis was established by histological and immunohistochemical examination. The tumor stages (II–IV) were assessed according to the Ann Arbor classification; the patient’s somatic status – according to the ECOG scale with a total of no more than 3 points. To detect c-MYC gene aberrations in the tumor tissue, fluorescent in situ hybridization was performed; to determine the BCL2 protein expression and Ki-67 proliferative index – an immunohistochemical study was performed. Patients with identified c-MYC gene aberrations received treatment, which included 8 courses of polychemotherapy according to the R + Hyper-CVAD scheme and high-dose chemotherapy according to the BFR scheme with autologous hematopoietic stem cell transplantation. Patients with DLBCL, in whom c-MYC gene aberration was not detected, were assigned to the group of intermediate and low malignancy. They received a polychemotherapy regimen according to the R-CHOP-21 scheme (up to 8 courses).Results and conclusion. For the first time in Kazakhstan, we studied the incidence of c-MYC proto-oncogene aberrations, BCL2 protein expression and proliferative activity (Ki-67) in DLBCL patients, which made it possible to identify groups of highly aggressive tumor and intermediate, low malignancy, apply personalized polychemotherapy programs and evaluate the immediate treatment efficacy. This approach made it possible to increase treatment effectiveness in the group of patients with highly aggressive DLBCL and to achieve comparable results with therapy in patients with a more favorable prognosis.

Bcl-6-dependent risk stratification by nuclear expression of Peli1 in diffuse large B-cell lymphoma.

Background/Aim: Peli1 is an E3 ubiquitin ligase involving lymphomagenesis by lysine 63 ubiquitination-mediated stabilization of Bcl-6 with in diffuse large B-cell lymphoma (DLBCL). Materials and Methods: We categorized nuclear expression of Peli1 according to Bcl-6 status by immunohistochemistry in DLBCL (n=100), and analyzed clinicopathologic association with prognosis. Results: We established Bcl-6/Peli1 risk model composed of high risk (Bcl-6+/Peli1+ or Bcl-6-/Peli1-; n=64) and low risk (Bcl-6+/Peli1- or Bcl-6-/Peli1+; n=36). High risk group had more frequent non-GCB subtype (83% vs 64%; p=0.033) and Bcl-6-negativity (69% vs 28%; p<0.001) than low risk group. Univariate survival analysis for progression-free survival (PFS) and overall survival (OS) revealed Bcl-6/Peli1 risk group (p=0.026 and p=0.021) and other conventional variables including international prognostic index (IPI), stage, ECOG performance status, number of extranodal sites were significant prognostic factors, along with B symptoms for OS. In multivariate analysis for PFS, Bcl-6/Peli1 risk group (p=0.032; HR=3.29), IPI (p=0.013; HR=3.39) and ECOG PS (p=0.035; HR=3.08) were independent prognostic factors. In multivariate analysis for OS, Bcl-6/Peli1 risk group (p=0.048; HR=7.87) and IPI (p=0.001; HR=12.15) were associated with prognosis. Conclusions: DLBCL had distinctive risk groups according to pairs of nuclear Peli1 and Bcl-6 expression. These results suggest the potential role of Peli1 and Bcl-6 in risk assessment in DLBCL.

Clinicopathologic Characteristics and Genomic Profiling of De Novo CD5 Positive Diffuse Large B-Cell Lymphoma: A Multicenter Retrospective Study in China

De novo CD5 positive diffuse large B cell lymphoma (CD5+ DLBCL) has poor survival in the era of immunochemotherapy. We conducted a multi-center retrospective study to explore the clinicopathologic characteristics, genomic profiling and prognostic elements of 61 CD5+ DLBCL and 60 CD5- DLBCL patients in China. In contrast with CD5- DLBCL, elderly onset, advanced stage, central nervous system (CNS) involvement, as well as MYC/BCL-2 and P53 overexpression were more prevalent in CD5+ DLBCL. In addition, most of CD5+ DLBCL patients were of non-germinal center B-cell-like (non-GCB) and activated B-cell-like (ABC) subtype according to immunohistochemistry and Lymph2Cx assay. Genetic analysis by next generation sequencing (NGS) showed the proportion of MCD subtype in CD5+ DLBCL was higher than that of CD5- DLBCL (50% vs 5%, p = 0.0007). Compared with CD5- cohort, CD5+ DLBCL patients showed poorer 5-year overall survival (OS) (70.9% vs 39.0%, p<0.001), independent of cell-of-origin and MYC/BCL-2, P53 and BCL-6 status. Multivariate analysis showed that age >76 years, advanced stage, CNS involvement and hypoalbuminemia were independent factors associated with poor prognosis in CD5+ DLBCL. In conclusion, CD5+ DLBCL conveyed poor prognosis and showed distinctive clinicopathologic characteristics and predominant genetic features of ABC and MCD subtypes. DisclosuresNo relevant conflicts of interest to declare.

AID is a poor prognostic marker of high-grade B-cell lymphoma with MYC and BCL2 and/or BCL6 rearrangements.

Diffuse large B-cell lymphoma with MYC rearrangement is defined as double/triple-hit lymphoma (DHL/THL) or single-hit lymphoma (SHL) by the inclusion of the BCL2 and BCL6 rearrangements status. DHL/THL is called as "high-grade B-cell lymphoma with MYC and BCL2 and/or BCL6 rearrangements" in the World Health Organization 2017 Classification of Tumors of Hematopoietic and Lymphoid Tissues. To find a prognostic biomarker of DHL/THL, we firstly examined 19 cases (molecular analysis series;10 cases of DHL/THL and 9 cases of SHL) with gene expression profile analysis. The gene expression profile analysis showed that the high expression of AICDA was associated with an adverse prognosis in DHL/THL, but not in SHL. Then, we evaluated immunohistochemical expression of AID, the protein product of AICDA, in 50 cases (molecular analysis series of 19 cases and additional immunohistochemistry series of 31 cases; 12 cases of DHL/THL and 19 cases of SHL) and confirmed that its expression was also associated with an adverse prognosis in DHL/THL. Therefore, AICDA and AID can be a predictor of an adverse clinical outcome in DHL/THL and immunohistochemistry of AID is useful to find DHL/THL-adverse prognosis group.

High Inter- and Intratumoral Variability of Ki67 Labeling Index in Newly Diagnosed Prostate Cancer with High Gleason Scores

Background: The majority of studies investigating the role of Ki67 labeling index (LI) in prostate carcinoma (PC) focused on localized PC treated radically, where Ki67 LI is regarded as a prognostic marker. The relevance of Ki67 in advanced PC remains largely unexplored. While Gleason score is still one of the best indicators of clinical outcomes in PC, differences in progression-free survival and overall survival in patients with high Gleason scores suggest that additional factors are involved in tumor progression. Understanding the underlying mechanisms could help to optimize treatment strategies for an individual patient. Here, we aimed to determine the inter- and intratumoral distribution of Ki67 LI in patients with PC with high Gleason scores and to correlate Ki67 LI with the status of ERG, PTEN, and Bcl-2. Methods: Immunohistochemistry for Ki67, ERG, PTEN, and Bcl-2 was performed on core needle biopsies from 112 patients with newly diagnosed PC Gleason score 8, 9, and 10. Results: Using a cutoff of ≥10%, 17/112 cases (15%) had a homogeneously low and 95/112 cases (85%) a high Ki67 LI. 41% of cases showed intratumoral heterogeneity containing areas with low and high proliferation. There was no association between Ki67 LI and ERG, PTEN, or Bcl-2 status. Conclusions: Our data demonstrate major inter- and intratumoral variability of Ki67 LI in high-grade PC with a surprisingly low Ki67 LI in a subset of cases. Further studies are necessary to explore the molecular basis and potential clinical implications of a paradoxically low proliferation rate in high-grade PC.

Diffuse large B-cell lymphoma genetics - simplifying the subtyping.

Diffuse large B-cell lymphoma genetics - simplifying the subtyping.

CORRELATION OF BCL 2 ONCOPROTEIN EXPRESSION AND ITS PROGNOSTIC SIGNIFICANCE IN COLORECTAL CARCINOMA WITH AGE, SITE GRADE AND STAGE

Background &amp; Objective: Colorectal cancer is the third most prevalent malignancy with high mortality rate, necessitating markers that predict survival and guide the treatment. Previous studies have examined the immunohistochemical expression of Bcl-2, anti apoptotic marker, in colorectal carcinoma, but results have been contradictory.To evaluate the histopathological features of colorectal carcinoma, immunohistochemical expression of Bcl-2 must be analyzed to find out statistical association of Bcl-2 expression with certain prognostic factors histopathologic type, grade and TNM staging and also clinical parameters like age gender, site. Methods: This prospective study was conducted on the colectomy specimens of colorectal carcinoma,over a period of 2 years 6months From may 2018 to November 2020 The tumor morphology and Bcl-2 status were evaluated by immunohistochemistry in each case,with the inclusion criteria of resected intestinal specimens among which only malignant epithelial lesions and exclusion of all benign lesions. Results:The study included 40 cases,with age group of patients 51-60(37.5%) years and male:female ratio of 1.2:1.Bcl-2 positivity was seen in 37.5% of the cases.Weak,moderate,and strong expression of Bcl-2 was seen in 62.5%, 25%, and 12.5% of cases respectively. Even though early stages of colorectal carcinoma showed greater frequency of Bcl-2 expression than advanced stages (25% versus 12.5%), however this association was not statistically significant. Conclusion: The expression of bcl 2 out of 40 resected specimens of colorectal carcinoma correlated with various histological and clinical paramaters .bcl 2 positivity was assessed by semiquantitaive method and its expression was decreased with increase in grade and stage of the tumour and increased with the early stages of colorectal cancer

Histone Deacetylase Inhibitor Depsipeptide (FK228) Induces Apoptosis in Leukemic Cells by Facilitating Mitochondrial Translocation of Bax, Which Is Enhanced by Proteasome Inhibitor Bortezomib.

Histone deacetylase (HDAC) inhibitors are a promising candidate for molecular-targeted therapy for leukemia. In this study, we investigated the mechanisms of cytotoxic effects of depsipeptide (FK228), one of the most effective HDAC inhibitors against leukemia, using human myeloid leukemic cell lines HL-60 and K562. We found that FK228 activated caspase-9 and a subsequent caspase cascade by perturbing the mitochondrial membrane to release cytochrome c, which was almost completely blocked by overexpression of Bcl-2. The mitochondrial damage was caused by the translocation of Bax but not other proapoptotic Bcl-2 family proteins to the mitochondria. FK228 did not affect the interaction between Bax and Bax-adaptor proteins such as 14–3–3 theta and Ku70. FK228-induced apoptosis and mitochondrial translocation of Bax were markedly enhanced by the proteasome inhibitor bortezomib. The synergistic action of FK228 and Bortezomib was at least partly mediated through conformational changes of Bax, which facilitate its translocation to the mitochondria. These results suggest that the combination of HDAC inhibitors and proteasome inhibitors is useful for the treatment of leukemia especially in the context of molecular-targeted therapy. The status of Bcl-2 and Bax may influence the sensitivity of tumors to this combination, and thus, can be a target of further therapeutic intervention.

Results from VERONICA: A randomized, phase II study of second-/third-line venetoclax (VEN) + fulvestrant (F) versus F alone in estrogen receptor (ER)-positive, HER2-negative, locally advanced, or metastatic breast cancer (LA/MBC).

1004 Background: For patients (pts) with ER-positive, HER2-negative MBC, CDK4/6 inhibitors + endocrine therapy (ET) is standard first-line treatment, with single-agent ET considered for second-line. Nevertheless, most pts progress. A novel therapeutic target is the antiapoptotic protein BCL2, which is overexpressed in ̃85% of primary ER-positive breast cancers. VEN is a potent, selective BCL2 inhibitor that has shown promising clinical activity in pts with ER-positive and BCL2-positive MBC who have received prior ET. We report the prespecified primary and updated (for overall survival [OS]) analysis of VERONICA (NCT03584009), a phase II study of VEN + F vs F in ER-positive, HER2-negative LA/MBC. Methods: Pts were ≥18-year-old women with ER-positive, HER2-negative LA/MBC, who received ≤2 prior lines of ET and no prior chemotherapy in the LA/MBC setting and experienced disease recurrence/progression during/after CDK4/6 inhibitor therapy (received ≥8 weeks prior). Pts were randomized 1:1 to VEN (oral; 800 mg daily) + F (intramuscular; 500 mg day 1 and 15 of cycle 1; day 1 of subsequent 28-day cycles) or F, and were treated until disease progression, unacceptable toxicity, withdrawal of consent, death, or predefined study end. Pts were stratified by prior lines of therapy in the LA/MBC setting (1 vs 2) and BCL2 status (high vs low). Primary endpoint was clinical benefit rate (CBR; complete response, partial response, and stable disease ≥24 weeks). Secondary endpoints included progression-free survival (PFS) and OS; safety and exploratory subgroup analyses were also conducted. Results: At primary analysis (cutoff: Aug 5, 2020), 103 pts had been randomized (intention-to-treat [ITT] population). Median age was 58.0 and 59.5 years in the VEN + F and F arms, respectively. CBR was similar between arms (VEN + F: 11.8% [n = 6/51; 95% confidence interval (CI) 4.44–23.87]; F: 13.7% [7/51; 5.70–26.26]; risk difference: -1.96% [95% CI -16.86–12.94]). Median PFS was 2.69 months (95% CI 1.94–3.71) in the VEN + F vs 1.94 months (1.84–3.55) in the F arm (stratified hazard ratio: 0.94 [95% CI 0.61–1.45]). Results for CBR and PFS were similar in the BCL2-high and -low subgroups vs the ITT population. More grade 3–4 adverse events (AEs) were observed in the VEN + F vs F arm (n = 13/50 [26%] vs 6/51 [11.8%]). AEs observed with VEN + F were consistent with their individual safety profiles. At updated analysis (cutoff: Oct 22, 2020), OS data were not mature (35.0% event/pt ratio); median OS was 16.99 months in the VEN + F vs not reached in the F arm (stratified hazard ratio: 2.06 [1.04–4.09]). Conclusions: From the primary analysis, VERONICA did not show an improved CBR or PFS with VEN + F, vs F alone, in pts with endocrine- and CDK4/6 inhibitor-refractory LA/MBC. Biomarker analysis is ongoing. Clinical trial information: NCT03584009 .

Radiologically guided percutaneous core needle biopsy of the spleen: a reliable and safe diagnostic procedure for neoplastic and reactive conditions.

Percutaneous core needle biopsy (CNB) of the spleen is rarely performed, due to concerns about its complications and low diagnostic yield. However, this procedure represents a potentially useful diagnostic tool, especially in patients with splenomegaly and no definitive diagnosis after a clinical and radiological work-up. We report the data on a cohort of 45 radiologically guided percutaneous core needle biopsies of the spleen from 44 patients performed at two centres. Platelet count and prothrombin time were within normal limits in all patients at the time of the procedure. The biopsy was ultrasound-guided in all cases except one, which was guided by computed tomography. An 18G needle was used in 82% of the cases, followed by 16G (10.2%) and 20G (7.8%) needles. The biopsy provided sufficient material for histological examination (including immunohistochemical studies) in 41 cases (91.1%). Haematological malignancies were most commonly diagnosed (52.3%); diffuse large B cell lymphoma (DLBCL) was the most frequent, followed by splenic marginal zone lymphoma (SMZL). For the most recent cases of DLBCL, the CNB provided sufficient material for fluorescence in-situ hybridisation to assess the status of MYC, BCL2 and BCL6. This allowed the identification of a case of high-grade B cell lymphoma with MYC and BCL2 rearrangement. Major complications were not reported; minor complications occurred in three cases (6.7%). Our data demonstrate that radiologically guided percutaneous CNB should be considered as a valid diagnostic tool, as it provides quick and reliable histological diagnoses avoiding the complications and risks of splenectomy.

Abstract OT-28-03: VICKI: A Phase Ib/II, randomized, placebo-controlled, study of venetoclax plus ado-trastuzumab emtansine (T-DM1) in patients (pts) with previously treated HER2-positive locally advanced (LA) or metastatic breast cancer (MBC)

Abstract Background ~15-20% of primary, invasive breast cancer (BC) overexpresses HER2 and, despite survival improvements, there remains an unmet need for further progress. The antibody-drug conjugate T-DM1 is approved for HER2-positive LA and/or MBC that has previously been treated with trastuzumab and a taxane (separately or in combination), and as adjuvant therapy for HER2-positive early BC, where there is residual invasive disease after neoadjuvant taxane and trastuzumab- or HER2-based treatment. Venetoclax (GDC-0199/ABT-199), an oral, selective small-molecule inhibitor of the antiapoptotic protein BCL-2, is approved for treatment of chronic lymphocytic leukemia, small lymphocytic lymphoma, and acute myeloid leukemia. BCL-2 may play a key role in HER2-positive BC, and venetoclax has shown promising activity in estrogen receptor-positive, BCL-2-positive MBC. We describe VICKI (Venetoclax in Combination with Kadcyla), a Phase Ib/II, randomized, double-blind, placebo-controlled, study of venetoclax plus T-DM1 in previously treated HER2-positive LA/MBC (NCT04298918). Trial design The study comprises a Phase Ib stage (dose escalation and expansion cohorts) and a randomized Phase II stage. Phase II will be initiated following identification of the recommended Phase II dose of venetoclax in Phase Ib (400 mg or 800 mg). Pts will be randomized 1:1 to T-DM1 (intravenous 3.6 mg/kg q3w) plus venetoclax or placebo. Randomization will be stratified per BCL-2 status (BCL-2 high vs. low), visceral disease (Yes vs. No), and HER2 immunohistochemistry (IHC) 3+ status (Yes vs. No). Eligibility Adult pts with HER2-positive (IHC 3+ or IHC 2+/in situ hybridization-positive), previously treated, unresectable, histologically or cytologically confirmed invasive LA/MBC are eligible. Pts will have measurable disease per RECIST v1.1 and an Eastern Cooperative Oncology Group performance status of 0 or 1. Pts in Phase II will have BCL-2 expression status by IHC (≥50% of pts BCL-2 high) and will not have received prior treatment with T-DM1, venetoclax, or anti-HER2 drug conjugates. Aims The Phase II co-primary efficacy endpoints will be objective response rate (ORR) and progression-free survival (PFS) per RECIST v1.1 (both investigator-assessed). Secondary and exploratory efficacy endpoints will include duration of response, overall survival, clinical benefit rate, and patient-reported outcomes. Non-efficacy endpoints will be pharmacokinetics, immunogenicity, biomarkers, and safety. Statistical methods In Phase II, the primary efficacy populations will include all randomized pts according to their assigned treatment arm (intention-to-treat). A point estimate and 95% CI for ORR and the difference in ORR between treatment groups will be calculated using the normal approximation to the binomial distribution. PFS will be defined as time from randomization to the first occurrence of disease progression or death from any cause. Kaplan-Meier methodology will be used to estimate median PFS. An interim analysis is planned when ~56 PFS events have occurred. Primary efficacy analysis will occur when 161 pts have had a PFS event. Cox proportional-hazards models, stratified by the stratification factors, will be used to estimate the hazard ratio with 95% CI. Safety will be analyzed per treatment received in pts who received any study treatment (safety population). Accrual Target accrual is ~226-284 pts at 145 sites globally (Phase Ib dose escalation: 6-24 pts; Phase Ib expansion cohorts: ~20-40 pts; Phase II: 220 pts). Accrual is ongoing. Contact information For more information or to refer a patient, email global-roche-genentech-trials@gene.com or call 1-888-662-6728 (USA only). Citation Format: Geoffrey J Lindeman, Erika Hamilton, Ian Krop, Bora Lim, Shanu Modi, Cristina Saura, Rupal Desai, Bradford J Danner, Tharu M Fernando, Shengchun Kong, Fatema A Legrand, Federico Nasroulah. VICKI: A Phase Ib/II, randomized, placebo-controlled, study of venetoclax plus ado-trastuzumab emtansine (T-DM1) in patients (pts) with previously treated HER2-positive locally advanced (LA) or metastatic breast cancer (MBC) [abstract]. In: Proceedings of the 2020 San Antonio Breast Cancer Virtual Symposium; 2020 Dec 8-11; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2021;81(4 Suppl):Abstract nr OT-28-03.