GCB Patients Research Articles

Matched control analysis suggests that R-CHOP followed by (R)-ICE may improve outcome in non-GCB DLBCL compared with R-CHOP

AbstractRituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) is considered the standard-of-care for patients with advanced-stage diffuse large B-cell lymphoma (DLBCL), despite findings that patients with nongerminal center B-cell like (non-GCB) have significantly worse outcome with this regimen. We evaluated the prognostic significance of baseline risk factors, including cell of origin (COO) classified by the Hans algorithm, within an alternative chemoimmunotherapy program. At Memorial Sloan Kettering Cancer Center (MSK), 151 patients with DLBCL received sequential R-CHOP induction and (R)-ICE (rituximab, ifosfamide, carboplatin, and etoposide) consolidation. Outcome analysis based on COO was validated with a propensity score–matched cohort treated with R-CHOP from the Mayo Clinic component of the Molecular Epidemiology Resource (MER). Among the patients with GCB (n = 69) and non-GCB (n = 69) at MSK, event-free survival (EFS) of non-GCB was superior to that of GCB (hazard ratio [HR], 0.53; 95% confidence interval [CI], 0.29-0.98). Overall survival (OS) demonstrated an association in the same direction but was not statistically significant (HR, 0.68; 95% CI, 0.33-1.42). Propensity score–matched patients from MSK (n = 108) demonstrated a small attenuation in the HRs for EFS (HR, 0.57; 95% CI, 0.27-1.18) and OS (HR, 0.76; 95% CI, 0.33-1.79) and were no longer statistically significant. In contrast, the matched MER cohort (n = 108) demonstrated an EFS association (HR, 1.17; 95% CI, 0.70-1.95) and OS association (HR, 1.13; 95% CI, 0.64-2.00) in the opposite direction, but were also not statistically significant. R-CHOP induction and (R)-ICE consolidation may overcome the negative prognostic impact of the non-GCB phenotype, per the Hans algorithm, and can be preferentially selected for this population. This trial was registered at www.ClinicalTrials.gov as #NCT00039195 and #NCT00712582.

THE PROGNOSTIC SIGNIFICANCE OF CD10+ MUM1+ GCB SUBTYPE OF DIFFUSE LARGE B-CELL LYMPHOMAS

PURPOSE - Diffuse Large B-Cell Lymphoma can be subclassified into the prognostically distinct groups of GCB and non-GCB using the immunohistochemical Hans algorithm. However, in real life, patients classified as GCB show differing outcomes even when treated with the same chemotherapy regimen. CD10 is a GCB marker, while MUM1 is a non-GCB marker, but Hans algorithm classifies CD10+ MUM1+ patients as GCB DLBCL. We explored whether CD10+ MUM1+ GCB DLBCL patients had different outcomes compared to rest of the GCB DLBCL patients. METHODS - In this study, we retrospectively analyzed the clinical records of 427 DLBCL patients treated with standard chemo-immunotherapy at Kidwai Memorial Institute of Oncology, Bengaluru from 2013 to 2019. The clinical characteristics, response rate and overall survival of CD10+ MUM1+ GCB group was compared with MUM1-ve GCB group and non-GCB group. RESULTS - Among the patients studied, 187 (43%) patients were of GCB subtype, whereas 240 (57%) patients belonged to the non-GCB subtype. Additionally, 61 (14.3%) patients were of the CD10+ MUM1+ GCB immune-phenotype. No significant differences could be demonstrated in the clinical characteristics of CD10+ MUM1+ GCB and MUM1- GCB patients. Patients with CD10+ MUM1+ GCB subtype exhibited a significantly inferior response rate compared to those with MUM1- GCB subtype (81% versus 92%; p=0.04). Although, both non-GCB and CD10+ MUM1+ GCB subtypes displayed diminished overall survival in comparison to MUM1- GCB patients, (non-GCB = 33 months, CD10+ MUM1+ GCB = 33 months and MUM1-ve GCB = 40.4 months, p = 0.21) it did not reach statistical significance CONCLUSION - CD10+ MUM1+ GCB patients may be prognostically more similar to non-GCB patients, as suggested by the inferior response rates and numerically inferior OS compared to other GCB patients. This needs further confirmation in prospective studies.

Nivolumab and Ibrutinib for Treatment of Patients with Refractory or Relapsed Central Nervous System Lymphoma

Background: Central Nervous System (CNS) lymphoma is a distinct entity of large B-cell lymphoma (LBCL) with a unique genomic profile with similarities to the activated B-cell (ABC) subtype, which may offer potential targets for therapy. These aberrancies commonly include MYD88 mutations, which enhances sensitivity to Bruton's tyrosine kinase (BTK) inhibitors, and translocation or copy number alterations of 9p24, which may associate with sensitivity with programmed death (PD)-targeted therapies. Expression of PD-L1 is common in CNS lymphoma and tumor infiltrating lymphocytes, and the majority of CNS lymphomas have a copy gain of 9p24.1. This suggests an attenuated immune response against CNS lymphoma. Ibrutinib in combination with nivolumab has shown safety and clinical activity in patients with relapsed non-Hodgkin lymphoma or chronic lymphocytic leukemia in a phase 1/2 trial, and therefore we hypothesized that this combination may be active in CNS lymphoma. Methods: We conducted an investigator-initiated, open label, single center, phase 2 clinical trial combining ibrutinib with nivolumab to treat patients with relapsed CNS lymphoma (NCT03770416). Adult patients were eligible if they had CNS lymphoma (primary or secondary) refractory to or relapsed after at least 1 prior CNS directed therapy, with adequate organ and bone marrow function, without prior ibrutinib or PD1 inhibitor therapy, and did not require persistent high dose steroids. The trial had two sequentially enrolled cohorts: Cohort A included ibrutinib 560mg oral daily for one cycle (28-days per cycle), followed by ibrutinib combined with nivolumab 240mg IV every 14 days. Cohort B included ibrutinib and nivolumab during the first cycle. Patients with a partial response or greater after 6 cycles of combined ibrutinib and nivolumab could continue therapy for up to 2 years or until progression of disease or unacceptable toxicity occurs. The primary objective was to determine the best overall response rate (ORR) of ibrutinib and nivolumab. Secondary objectives included the ORR of ibrutinib as a lead in prior to the combination, best complete response (CR) rate, landmark survival outcomes, and the safety of the combination. The trial planned to enroll 20 patients in both cohort A and B, but closed early due to slow accrual, due in part to the COVID19 pandemic. Results: The trial accrued 18 patients from February 2019 to June 2022, with all patients evaluable for response. The median age was 63 years (range 43-88), 33% were >70 years, and 78% were female. 89% had primary CNS lymphoma, 50% had non-GCB, 22% had GCB, and 28% were unable to be classified due to missing components of the Hans algorithm. The median number of prior lines of therapy was 2 (range, 1-4), 55% had refractory disease, 17% had prior stem cell transplant, and 11% had prior CAR T-cell therapy. 10 patients were enrolled to Cohort A (ibrutinib lead in), and 8 patients were enrolled to Cohort B (initial combination therapy). Adverse events included 50% with fatigue, 33% with nausea, and 28% with mucositis. For both cohorts, the best ORR was 77.8% (95% CI: 52~94%) and the best CR rate was 50% (95% CI: 26-74%). In cohort A, the ORR of ibrutinib as a lead in after 1 cycle was 90% (95% CI: 55.5-99.7%), and the CR rate was 60%. In GCB patients (n = 4), the ORR and CR rates were 25%. In non-GCB patients (n =9), the ORR and CR rates were 100% and 56%, respectively. With a median follow up time of 34.6 months, the median overall survival (OS) was 25.4 months (95% CI 12.5-NR), and the OS rate at 12 months was 70%. The median progression free survival (PFS) was 6.6 months (95% CI: 2.9 - NR), and the PFS rate at 18 months was 47%. 4 patients withdrew from protocol therapy in CR, including 2 who proceeded to stem cell transplant consolidation, and 2 who stopped due to fatigue. 3 patients withdrew from protocol therapy in PR, 2 due to mucositis and 1 due to arthralgias. 1 patient died of COVID19 infection with a CR after 4 cycles of therapy. Conclusion: Ibrutinib and Nivolumab resulted in significant clinical activity in patients with refractory/relapsed CNS lymphoma. Our trial is limited by small patient numbers but suggests the potential for targeted therapy combinations in future larger clinical studies.

Linperlisib in Conjunction with Gemcitabine and Oxaliplatin in Relapsed/Refractory Diffuse Large B-Cell Lymphoma: A Multicenter, Single-Arm Phase Ib/II Study

Background: Diffuse large B-cell lymphoma (DLBCL) represents the most common subtype of non-Hodgkin lymphoma (NHL), comprising approximately 30-40% of new NHL cases. Although immunochemotherapy is the current front-line treatment for DLBCL, nearly 40% of patients will face disease relapse or refractoriness to the initial therapy, with a suboptimal prognosis. Gemcitabine in combination with oxaliplatin (GEMOX) offers an established treatment regimen with a manageable side-effect profile, thus providing a viable backbone for innovative treatment combinations. Linperlisib, a selective PI3Kδ inhibitor, has demonstrated its ability to inhibit the growth of PI3Kδ-expressing human tumor cells, as evidenced by both in vitro and in vivo studies. The tolerability, pharmacokinetics, and preliminary efficacy of linperlisib were affirmed in a preceding phase I study, laying the groundwork for its further exploration in patients with relapsed or refractory (r/r) DLBCL. This study aimed to investigate the efficacy and safety of combining linperlisib with GEMOX in treating r/r DLBCL patients. Methods: In this multicenter, single-arm phase Ib/II trial (NCT04500561), histologically confirmed r/r DLBCL patients across six Chinese centers were enrolled and received oral linperlisib (80 mg/day) alongside intravenous GEMOX (gemcitabine 1000 mg/m 2 on day 1, and oxaliplatin 100 mg/m 2 on day 2) every three weeks, with GEMOX administered for a maximum of six cycles. The primary endpoint was the objective response rate (ORR), with secondary outcomes including disease control rate (DCR), time to response (TTR), duration of response (DOR), progression-free survival (PFS), and overall survival (OS). Results: Between September 2020 and June 2021, 39 patients were enrolled and received the study treatment, all of whom were included in the efficacy and safety analysis. By the data cutoff in February 2023, and after a median follow-up of 24.1 months, three patients were still on treatment. The ORR reached 53.8% (95%CI: 37.2, 69.9), and the DCR was recorded at 64.1% (95%CI: 47.2, 78.8) (Figure 1). The median TTR was 1.3 months (range: 1.2-4.0). The median DOR stood at 5.7 months (95%CI: 4.3-9.1), and the median PFS was 5.4 months (95%CI: 1.8-6.7). The median OS had not yet been reached by the cutoff, with the one-year OS rate standing at 65.5% (95%CI: 48.1-78.3). Stratification by subtype revealed an ORR of 66.7% (6/9) among germinal center B-cell like (GCB) DLBCL patients compared to 40% (10/25) among non-GCB DLBCL patients. Median PFS was reported at 3.0 and 3.4 months for GCB and non-GCB DLBCL patients, respectively. The most prevalent grade 3 or higher treatment-related adverse events included decreases in neutrophil count (46.2%), platelet count (41.0%), and white blood cell count (28.2%). Conclusion: The combination of linperlisib with GEMOX exhibits a favorable safety profile and promising clinical efficacy in patients with r/r DLBCL who have received a median of two prior systemic therapies. These findings support the potential role of linperlisib combined with GEMOX as an active and safe therapeutic option for patients with r/r DLBCL, and set a foundation for further research into linperlisib's potential in combination with other treatment regimens.

Beneficial Effect of Microrna-155 in Vincristine Response and Clinical Outcome in Especially GCB Diffuse Large B-Cell Lymphoma

▪R-CHOP is standard treatment for previously untreated diffuse large B-cell lymphoma (DLBCL). Approximately 35-40% of patients relapse following R-CHOP due to drug-specific molecular resistance, and outcome with salvage therapy remain poor. Several clinical trials have tried to improve the R-CHOP regimen by dose adjustments and add on drugs, but with limited clinical benefit, emphasizing that improved understanding of R-CHOP resistance are still relevant and highly needed. Vincristine is essential to the efficacy of R-CHOP, and despite its use for decades, little is known about determinants of resistance in the treatment of DLBCL, which is important when attempting to improve outcome.We hypothesized that miRNAs are involved in the underlying molecular mechanisms by which Vincristine resistance work and that miRNAs hold potential as biomarkers of Vincristine response. By analyses of miRNA microarrays from DLBCL cell lines with different Vincristine sensitivity, we identified lower miR-155 expression in resistant cell lines.The hypothesis was further studied by two different approaches; first, by experimental manipulations of miR-155 expression by lentiviral vectors in DLBCL cell lines, studying the direct impact of miR-155 in Vincristine response, and secondly, by clinical outcome analysis of the prognostic impact of miR-155 expression in two independent DLBCL cohorts.Our results document that miR-155 is directly involved in Vincristine response in DLBCL cells since over-expression of miR-155 increased the Vincristine sensitivity, especially in GCB classified cell lines. Consistently, when miR-155 was downregulated or knocked-out by the CRISPR-Cas9 technology the cells became more resistant.Prognostic impact of miR-155 was evaluated in an in-house cohort of 69 R-CHOP treated patients having miR-155 expression determined by RT-qPCR and in an independent DLBCL meta cohort of 586 patients, who had the miR-155 precursor MIR155HG measured by microarray. Survival analysis was performed overall and according to ABC/GCB subclasses. miR-155 expression did not correlate with clinical outcome after R-CHOP treatment in the entire DLBCL cohort or restricted to ABC classified patients. However, we observed that GCB classified patients with low miR-155 levels had significantly shorter overall and progression-free survival than GCB patients with high miR-155 expression levels (Figure 1). Multivariate Cox regression showed that miR-155 expression added significant and independent prognostic information to the already well-established international prognostic index (IPI) when the analysis was restricted to the GCB subclass.In summary, experimentally we confirmed a direct link between miR-155 and Vincristine response in DLBCL overall and in particular GCB DLBCL. This association is supported by prognostic evaluation in two clinical cohorts treated with R-CHOP, that documents an inferior clinical outcome in GCB classified patients with low miR-155 expression. This suggests that the role of miR-155 in Vincristine response is important enough to affect overall- and progression-free survival especially within the GCB subclass. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Clinical Impact of Somatic Copy Number Alterations in Circulating Tumor DNA from Diverse Lymphoma Subtypes

Introduction:Somatic copy number alterations (SCNAs) constitute a major class of genomic aberrations known to have functional significance across cancers, including prognostic value in lymphoma. For example, diffuse large B-cell lymphoma (DLBCL) patients with tumors harboring SCNAs in MYC, BCL2, or TP53 tend to have inferior outcomes (Quesada, ASH 2016). However, detection of copy number alterations without invasive tissue biopsies remains difficult. We applied Cancer Personalized Profiling by Deep Sequencing (CAPP-Seq), a targeted high-throughput sequencing platform, to noninvasively detect and monitor genome-wide SCNAs in patients with DLBCL, primary mediastinal B-cell lymphoma (PMBCL), and classical Hodgkin lymphoma (cHL).Methods: We profiled pre-treatment blood samples from 209 patients with diverse lymphomas including 168 DLBCL, 20 cHL, and 21 PMBCL, using a custom 314kb gene panel targeting recurrently altered regions in these cancers. Additionally, we sequenced 139 tumor biopsies to use as matched tissue gold standards for SCNA confirmation. In each sample, we interrogated the sequencing read distribution to summarize focal and broad SCNAs directly from plasma. All thresholds were tuned for 95% specificity, which were empirically verified in independent withheld healthy controls. Results were orthogonally validated through tissue karyotyping, fluorescence in situ hybridization (FISH), and whole genome sequencing (WGS) (Fig 1A and Fig 1B).Results: We determined our limit of detection (LOD) for detecting SCNAs in plasma at ~1% circulating tumor fraction with 77% sensitivity; we observed higher sensitivity above 5% circulating tumor fraction (sensitivity = 95%). In our cohort, 70% of patients had over 1% circulating tumor burden with representation from all clinical stages. In DLBCL patients, we detected gains in MYC (12%), BCL2 (24%), BCL6 (19%), and PD-L1 (23%), as well as losses in TP53 (27%) and CDKN2A (19%). SCNAs detected in plasma were 85% concordant with those detected in matched tumor biopsies. Additionally, PD-L1 copy number gains were more frequently detected in both PMBCL (63%, p= 0.026) and cHL (71%, p= 0.012) than in DLBCL (23%), consistent with prior reports (Chapuy B, Blood 2016; Roemer MG, J. Clin. Oncol. 2016).We also demonstrate that noninvasive detection of copy number gains in MYC and BCL2, as well as loss of TP53, are negatively prognostic for survival (p < 0.0001, p= 0.0013, p < 0.0001, respectively) (Fig 1C). Furthermore, we identified associations between DLBCL cell-of-origin classification and specific SCNAs, with non-GCB DLBCL enriched for BCL2, BCL6, and PD-L1 copy number gains (39% vs 17%; 31% vs 15%, and 39% vs 25%, respectively). Circulating tumor burden was not statistically different between GCB and non-GCB patients. TP53 losses were more frequently observed in relapsed/refractory patients compared to treatment naïve patients (22% vs 45%, p= 0.023).We performed a preliminary analysis monitoring disease over time in three patients with clear SCNAs detected in plasma at diagnosis. Across longitudinal blood samples (n= 42), SCNA-based disease detection demonstrated 95% concordance with mutation-based disease detection, with a LOD down to ~0.1% circulating tumor burden. Our results suggest noninvasive SCNA profiling could provide an independent method for detecting residual disease, particularly in tumors characterized by structural alterations. Further validation and expansion of this monitoring method will be explored and presented.Conclusions: Noninvasive identification of genome-wide SCNAs is feasible via targeted next generation sequencing. Our approach allows for both genotyping and monitoring of copy number gains and losses from plasma, without requiring matched germline or tissue specimens. This strategy not only allows for the noninvasive genotyping of tumor SCNAs, but also provides an orthogonal method for monitoring residual disease from liquid biopsies using next generation sequencing.Figure 1: A) Matched tissue FISH staining confirms arm-level PD-L1 amplification (red) detected in plasma. D9Z1 (green) was used as an assay control. B) SCNAs detected in plasma were concordant with karyotype analysis of a matched tumor biopsy. C) Copy number alterations in MYC, BCL2, and TP53 are adversely prognostic for survival in R-CHOP treated DLBCL patients. [Display omitted] DisclosuresHüttmann:Bristol-Myers Squibb, Takeda, Celgene, Roche: Honoraria; Gilead, Amgen: Other: Travel cost. Gaidano:Roche: Consultancy, Honoraria; Janssen: Consultancy, Honoraria; Amgen: Consultancy, Honoraria; Gilead: Consultancy, Honoraria; AbbVie: Consultancy, Honoraria. Westin:Kite Pharma: Membership on an entity's Board of Directors or advisory committees; Apotex: Membership on an entity's Board of Directors or advisory committees; Novartis Pharmaceuticals Corporation: Membership on an entity's Board of Directors or advisory committees; Celgene: Membership on an entity's Board of Directors or advisory committees. Advani:Bayer Healthcare Pharmaceuticals: Research Funding; Gilead: Consultancy; Bristol-Myers Squibb: Consultancy, Research Funding; Celgene: Research Funding; Janssen: Research Funding; Genentech: Research Funding; Seattle Genetics: Research Funding; Nanostring: Consultancy; Spectrum: Consultancy; Millennium: Research Funding; FortySeven: Research Funding; Pharmacyclics: Research Funding; Regeneron: Research Funding; Agensys: Research Funding; Infinity: Research Funding; Juno Therapeutics: Consultancy; Kura: Research Funding; Merck: Research Funding; Pharmacyclics: Consultancy; Cell Medica: Research Funding; Sutro: Consultancy. Diehn:Varian Medical Systems: Research Funding; Roche: Consultancy; Novartis: Consultancy; Quanticel Pharmaceuticals: Consultancy. Alizadeh:Genentech: Consultancy; Celgene: Consultancy; Gilead: Consultancy; Roche: Consultancy; CiberMed: Consultancy.

Adjuvant therapy in the treatment of resected nonmetastatic gallbladder cancer of stage II-IV: A generalized propensity score analysis.

471 Background: The benefit of adjuvant therapy for gallbladder cancer (GBC) remains controversial due to the limited evidence. The current study was aimed to explore the efficacy of adjuvant therapy in patients with resected non-metastatic GBC and to establish a prognosis model to predict the survival benefit of GCB patients with different adjuvant therapies. Methods: Patients with resected non-metastatic GBC of stage II-IV were selected from the Surveillance, Epidemiology, and End Results database and divided into non-radiotherapy and chemotherapy (NCRT) group, chemotherapy (CT) group, and chemoradiotherapy (CRT) group. Generalized propensity score (GPS) and inverse probability of treatment weighting (IPTW) was used to reduce the imbalances between groups. Nomogram was constructed based on Cox proportional hazard model, and the model was validated for discrimination and calibration. Results: Among the 2689 enrolled patients, 1193 (44.4%) patients were classified as stage II, 1371 (51.0%) as stage III, and 125 (4.6%) as stage IV according to the 8th American Joint Commotion Cancer staging manual. Patients in NCRT, CT, and CRT groups were 1703, 444, and 542 respectively. After the IPTW, absolute standardized differences of baseline characteristics were less than 0.1 both in patients with stage II tumors and with stage III-IV tumors. In patients with GBC of stage II, no significant difference in OS was observed between the three treatment groups (P &gt; 0.05). In patients with GBC of stage III-IV, CT group has a superior OS compared with the NCRT group (P &lt; 0.001), and CRT group has a superior OS compared with the CT(P &lt; 0.001) and NCRT (P &lt; 0.001) group. Sensitivity analysis showed consistent results. A nomogram was constructed for patients with stage III-IV tumors to predict the survival benefit of adjuvant therapies. The C-index was 0.673 (95% CI: 0.654-0.692) in the validation using the training set (diagnosed at 2004-2012) and was 0.707 (95% CI: 0.677-0·739) in the internal validation using the validation set (diagnosed at 2013-2015). The calibration curves indicated that the predicted probability closely corresponded to the actual observation OS. Conclusions: Patients with GBC of stage II could not benefit from adjuvant therapy. Patients with GBC of stage III-IV could benefit from chemotherapy and chemoradiotherapy while chemoradiotherapy provide a superior OS. A nomogram was built to predict the survival benefit of different adjuvant therapies in patients with GBC of stage III-IV.

Impact of Cell of Origin (COO) on Long Term Outcomes Post Autologous Hematopoietic Cell Transplant in Patients with Relapsed/ Refractory Chemotherapy Sensitive De-Novo Diffuse Large B-Cell Lymphoma (DLBCL)

Introduction The molecular basis for the clinical heterogeneity observed in DLBCL was initially elucidated with gene expression profiling, identifying three distinct subgroups with prognostic relevance based on COO namely activated B-cell like (ABC), germinal center B-cell like (GCB) and a type 3 subtype. Shortly afterwards the Han's algorithm was developed allowing for widespread applicability and correlation of gene expression with protein expression. GCB profile at diagnosis is generally associated with superior long term outcomes compared to non-GCB in patients treated with conventional chemo-immunotherapy. Patients with relapsed/refractory disease (R/R) after front-line therapy undergo salvage treatment and, upon demonstration of objective response, proceed with high dose therapy and autologous transplantation (auto-HCT). Whether COO by Han's algorithm retains its prognostic significance post auto-HCT in patients with R/R DLBCL is not well established. Methods A retrospective analysis was carried out using the Mayo database across all three sites. It included all patients between 18-75 years of age who were diagnosed with de-novo DLBCL and underwent first auto-HCT for R/R disease. Patients must have received salvage treatment and shown chemotherapy sensitive disease. Patients with transformed DLBCL or other variants and those who underwent auto-HCT in first complete remission for high risk disease were excluded. Patients who had less than a partial response at time of transplantation and still underwent auto-HCT were also excluded. Primary endpoints included overall survival (OS); relapse and relapse-free survival (RFS). Continuous variables were summarized as mean (standard deviation) and median (range). Categorical variables were reported as frequency (percentage). Continuous baseline variables were compared between GCB and non-GCB using Wilcoxon rank sum test and categorical baseline variables were compared with Chi-squared test. Kaplan-Meier method was used to estimate 5 and 10-year rates of freedom from long-term events and draw corresponding survival curves. Multivariate Cox regression models were used to evaluate the impact of cell of origin on long-term survival after adjusting for baseline factors. All tests were two-sided with alpha level set at 0.05 for statistical significance. Results A total of 357 patients underwent auto-HCT between 2005 and 2018. Cell of origin was determined for 284 patients and these were included in the analysis. Median age was 61 (range: 19-78) years, and 64% patients were male. GCB and non-GCB patients did not differ significantly with regards to baseline factors (Table 1). During a median follow up of 1.7 years, 139 patients died, 151 had relapse and 175 either died or had relapse. Five year OS for GCB patients versus (vs) non-GCB was at 44% vs 64% (p=0.004) (Figure 1); relapse was at 67% vs 49% (p=0.012) and RFS was at 28% vs 50% (p=0.003). The difference between GCB and non-GCB groups remained statistically significant in multivariate analysis, with GCB patients 60% more likely to die and 50% more likely to experience relapse compared to non-GCB patients. The only other factor that retained significance in multivariate analysis for both endpoints was response at the time of transplant (Table 2A-C). Conclusions In this analysis, GCB DLBCL results in worse OS and higher relapse vs. non-GCB DLBCL following an auto-HCT. It is unclear if relapse/progression of GCB subtype after front-line therapy alters its natural history into a more aggressive disease. Disclosures Ansell: Trillium: Research Funding; ADC Therapeutics: Research Funding; Affimed: Research Funding; Bristol Myers Squibb: Research Funding; AI Therapeutics: Research Funding; Takeda: Research Funding; Seattle Genetics: Research Funding; Regeneron: Research Funding. Foran:Xencor: Research Funding; Trillium: Research Funding; Takeda: Research Funding; Kura Oncology: Research Funding; Aptose: Research Funding; Aprea: Research Funding; Actinium: Research Funding; Boehringer Ingelheim: Research Funding; Abbvie: Research Funding; BMS: Membership on an entity's Board of Directors or advisory committees; Pfizer: Membership on an entity's Board of Directors or advisory committees; Servier: Membership on an entity's Board of Directors or advisory committees; Novartis: Membership on an entity's Board of Directors or advisory committees; Revolution Medicine: Consultancy; H3Biosciences: Research Funding; Agios: Honoraria, Research Funding. Tun:Celgene: Research Funding; Mundipharma: Research Funding; Curis: Research Funding; TG Therapeutics: Research Funding; Acrotech: Research Funding; DTRM Biopharma: Research Funding; Bristol-Myers Squibb: Research Funding. Kharfan-Dabaja:Daiichi Sankyo: Consultancy; Pharmacyclics: Consultancy.

Inhibitor of DNA Binding 3 Is a Potential Prognostic Biomarker in Non-GCB Diffuse Large B-Cell Lymphoma

Background: Diffuse large B-cell lymphoma (DLBCL) is the most common subtype of non-Hodgkin lymphoma. The majority of patients can be cured under the treatment of R-CHOP, but one third of patients will relapse and have a worse prognosis. To exploring a biomarker which can predict clinical behavior may aid in the implementation of novel therapeutic strategies. Inhibitor of DNA-binding 3 (ID3) is a helix loop-helix transcription factor, which negatively regulates the cell proliferation. ID3 mutations occur in 34-68% of Bukitt's lymphoma (BL) and are assiociated with lymphomagenesis. A variety of nonsense and frameshift mutations in ID3 as well as missense mutations in the region encoding the HLH domain reduces or eliminates ID3 protein function in BL. ID3 mutation is rare in DLBCL. The prognostic value of ID3 protein level remains unclear in DLBCL. Therefore, we conducted a retrospective study to investigate the impact of ID3 expression in DLBCL patients. Methods: One hundred DLBCL patients treated at Fudan University Shanghai Cancer Center with available diagnostic biopsy material were identified. ID3 expression was determined and quantified by immunohistochemistry (IHC) with staining intensity and percentage of positive cells (IRS). The IRS cut off value for ID3 positive is equal or more than 6. Kaplan-Meier and life table method were used to calculate progression free survival (PFS) and overall survival (OS). Curves were compared with the long-rank test in both groups. Correlation between the ID3 expression and clinical variables were tested by Pearson Chi Square test and a two-sided P value of &amp;lt;0.05 was considered to be statistically significant. Result: Seventy-seven (77%) of patients was overexpressed ID3 and 23% patients was shown ID3 negative. Among these pateints, ID3 positive patients had a trend of better outcome without a significance. The PFS and OS were 65% and 68% in ID3 negative group; 80% and 85% in ID3 positive group, respectively. According to cell of origin, the ID3 deficiency were 24% (10/41) in GCB patients and 22% (13/59) in non-GCB patients (p=0.82). the ID3 overexpression was associated with a better OS compared with ID3 negative patients in non-germinal center B-cell lymphoma group (non-GCB) (85% vs 46%, p=0.047). Although the PFS of ID3 positive patients was better than negative group (77% vs 59%, p=0.256), there was no significance. In GCB patients, there was no differences between the ID3 positive group and negative groups in terms of PFS or OS. The basic characteristics were no difference between ID3 positive and negative groups. Conclusion: This is the first report demonstrating that ID3 overexpression is a positive predictor of clinical outcome in non-GCB lymphoma patients. ID3 is a potential novel target to treat lymphoma and further investigation of ID3 need to be done in the near future. (Research supported by National Natural Science Foundation of China (no.81670177, no.81001048) Figure Disclosures No relevant conflicts of interest to declare.

Cell-of-Origin Subtyping of Diffuse Large B-Cell Lymphoma by Using a qPCR-based Gene Expression Assay on Formalin-Fixed Paraffin-Embedded Tissues

The well-established cell-of-origin (COO) algorithm categorizes diffuse large B-cell lymphoma (DLBCL) into activated B-cell-like (ABC) and germinal center B-cell-like (GCB) subgroups through gene expression profiling. We aimed to develop and validate a qPCR-based gene expression assay to determine the COO subgroups of DLBCL with formalin-fixed paraffin-embedded (FFPE) tissue. We first established a DLBCL transcriptome database of 1,016 samples retrieved from three published datasets (GSE10846, GSE22470, and GSE31312). With this database, we identified a qPCR-based 32-gene expression signature (DLBCL-COO assay) that is significantly associated with the COO subgroups. The DLBCL-COO assay was further validated in a cohort of 160 Chinese DLBCL patients. Biopsy samples from DLBCL patients with paired FFPE and fresh frozen tissue were collected to assign COO subtypes based on the immunohistochemistry (IHC) algorithm (Han's algorithm), DLBCL-COO assay, and global gene expression profiling with RNA-seq. For 111 paired FFPE and fresh DLBCL samples, the concordance between the IHC, qPCR, and RNA-seq methods was 77.5% and 91.9%, respectively. The DLBCL-COO assay demonstrated a significantly superior concordance of COO determination with the “gold standard” RNA-seq compared with the IHC assignment with Han's algorithm (P = 0.005). Furthermore, the overall survival of GCB patients defined by the DLBCL-COO assay was significantly superior to that of ABC patients (P = 0.023). This effect was not seen when the tumors were classified by the IHC algorithm. The DLBCL-COO assay provides flexibility and accuracy in DLBCL subtype characterization. These findings demonstrated that the DLBCL-COO assay might serve as a useful tool for guiding prognostic and therapeutic options for DLBCL patients.

THE CORRELATION BETWEEN CELL OF ORIGIN SUBTYPE WITH OVERALL SURVIVAL OF DIFFUSE LARGE B-CELL LYMPHOMA PATIENTS IN KARIADI GENERAL HOSPITAL SEMARANG

Background: DLBCL is the most common type of NHL in the world. DLBCL based on cell of origin is divided into GCB and non-GCB. The diagnosis of DLBCL has not been routinely done to its cell of origin, and there have not been many studies that discuss the DLBCL subtype and the overall survival of the patients, especially in Kariadi General Hospital. This study aims to determine the correlation of DLBCL cell of origin with the 2-year overall survival of DLBCL patients in Kariadi General Hospital. Methods: This is an observational analytic study of 40 DLBCL patients in Kariadi General Hospital from January to August 2017. The data collection including: age of diagnosis, location, stage and 2-year overall survival. Data analysis used chi square test and Kaplan Meier curve. Results: GCB patients had higher 2-year overall survival than non-GCB subtype significantly (p: 0.047), with a 2-year survival rate of GCB subtype was 66.7% and non-GCB subtype was 31.6%. GCB patients tend to have early stage than non-GCB subtype significantly (p:0.028). Conclusion: DLBCL GCB subtype patients had significantly higher 2-year overall survival therefore it has better prognosis than non-GCB subtype.

Quantitative Proteomics in Diffuse Large B-Cell Lymphoma Patients Reveal Novel Overexpressed Proteins and Potentially Druggable Targets in the ABC Subtype

Background: The cell-of-origin (COO) concept, based on gene expression profiling (GEP), dividing diffuse large B-cell lymphoma (DLBCL) patients into germinal center B cell (GCB) or activated B cell (ABC) subtypes, is a well-established subclassification where ABC patients have an inferior survival. The hallmark the ABC-type is constitutive activation of nuclear factor kappa B (NF-κB), often due to mutations in the B-cell receptor (BCR) signaling pathway. This has been the underlying rationale for adding newer drugs, such as bortezomib, ibrutinib or lenalidomide, to R-CHOP for ABC patients. However, none of these combinations studied in phase III trials have shown any clinical benefit. So, the complexity of ABC DLBCL is probably not only explained by genetic alterations and transcriptional changes as gene expression not necessarily correlate with protein expression, and protein action and dynamics are not caught by genomics-based techniques. Instead, using methods to measure global protein expression and interactions could offer new insights into the ABC subtype and possibly aid in the identification of novel drug targets. Aim: To study possible differences in global protein expression between ABC and GCB DLBCL subtypes using quantitative proteomics. Patients: A total of 213 adult DLBCL patients in western Sweden diagnosed between 1/1 2004 and 31/12 2016, were included. All patients received immunochemotherapy (R-CHOP). Primary mediastinal large B-cell lymphoma, primary CNS lymphoma, HIV-related lymphoma and transformed lymphoma were excluded. From archived formalin-fixed, paraffin-embedded (FFPE) tissue sections, from the time of diagnosis, a core biopsy (1 mm diameter) were obtained from each patient sample. Methods: COO was determined using the Hans immunohistochemistry algorithm. For 92 of the 213 patients, COO was also determined using the gene expression Lymph2cx chip: 14% changed subtype group from either non-GCB to GCB (n=8), GCB to ABC (n=4) or GCB to unclassified (n=1). From the FFPE samples a proteomic analysis was performed. In short, peptides were labelled using tandem mass tag (TMT) according to the manufacturer instructions and samples were analysed on an Orbitrap Fusion Tribrid mass spectrometer. The data files were merged for identification and relative quantification using Proteome Discoverer version 1.4.The search used the Human Swissprot Database version August 2016 using Mascot 2.3 as a search engine. The differentially expressed proteins were analysed using STRING version 10.0, for pathway analysis we used the Reactome database resource, and for potentially druggable proteins we used the Human Protein Atlas website which holds protein information of the current FDA approved drugs directed to 672 separate human proteins. Results: In all, 3078 proteins could be identified in all patients and 793 proteins were differentially expressed (p&lt;0.05 adjusted for mass significance according to Benjamin-Hochberg) between ABC and GCB patients. Of these, 410 proteins were overexpressed in the ABC group. Among the most expressed proteins were several well-known ABC-associated proteins (such as IRF4/MUM1, HSP90B1, CCDC50 and STAT3) in addition to a large number of proteins previously not described in ABC DLBCL, e.g. neudesin, BLNK, MPST, BPGM, SUB1, SP140, PCK2, PARP4, SRP54, SRP68, SRP72, TRPV2, IGF2R and FGD2. A majority of the 410 proteins were closely linked with an enrichment p-value &lt; 1x10-16(Fig. 1) and the most enriched pathways were immune system (FDR rate 3.3 x 10-27), interferon signaling (2.9 x 10-7), antigen processing (8.8 x 10-7) and down-modulation of cell surface receptors (4.7 x 10-5). Most interestingly, we also found that 16 proteins overexpressed in the ABC group could be potential drug targets for an FDA approved drug, e.g. high affinity immunoglobulin gamma Fc receptor I, CD47, HDAC2, ELANE and carbonic anhydrase 1. Conclusions: In this large proteomic study we found a number of overexpressed proteins in the ABC subtype, previously not described in DLBCL. Even though functional studies aimed at individual proteins and protein interactions to evaluate potential clinical effect are needed, our findings reveal novel proteins that could be potential druggable targets in ABC DLBCL patients. Figure 1 Disclosures Enblad: Kite/Gilead: Membership on an entity's Board of Directors or advisory committees. Mellqvist:Amgen, Janssen, Oncopeptides, Sanofi, Sandoz, Takeda: Honoraria. Andersson:Abbvie and Janssen: Membership on an entity's Board of Directors or advisory committees; Gilead, Janssen and Roche: Consultancy; Gilead: Research Funding.

Addition of Drug-Response Specific Micro-RNAs to the International Prognostic Index Improves Prognostic Stratification of GCB-DLBCL Patients Treated with R-CHOP

Background: Patients with Diffuse large B-cell lymphoma (DLBCL) in approximately 40% of cases suffer from primary refractory disease and treatment induced immuno-chemotherapy resistance demonstrating that standard provided treatment regimens are not sufficient to cure all patients. Early detection of resistance is of great importance and defining microRNA (miRNA) involvement in resistance could be useful to guide treatment selection and help monitor treatment administration while sparing patients for inefficient, but still toxic therapy. Concept and Aims: With information on drug-response specific miRNAs, we hypothesized that multi-miRNA panels can improve robustness of individual clinical markers and serve as a prognostic classifier predicting disease progression in DLBCL patients. Methods: Fifteen DLBCL cell lines were tested for sensitivity towards rituximab (R), cyclophosphamide (C), doxorubicin (H), and vincristine (O). Cell line specific seeding concentrations was used to ensure exponential growth and each cell line was subjected to 16 concentrations in serial 2-fold dilutions and number of metabolic active cells was evaluated after 48 hours of drug exposure using MTS assay. For each drug, we ranked the cell lines according to their sensitivity and categorized them as sensitive, intermediate responsive, or resistant. Differential miRNA expression analysis between sensitive and resistant cell lines identified 43 miRNAs to be associated with response to compounds of the R-CHOP regimen, by selecting probes with a log fold change larger than 2. Baseline miRNA expression data were obtained for each cell line in untreated condition, and differential miRNA expression analysis identified 43 miRNAs associated to response to R-CHOP. Using the Affymetrix HG-U133+2 platform, expression levels of the miRNA precursors were assessed in 701 diagnostic DLBCL biopsies, and miRNA-panel classifiers were build using multiple Cox regression or random survival forest. Results: Generated prognostic miRNA-panel classifiers were tested for predictive accuracies and were subsequently evaluated by Brier scores and time varying area under the ROC curves (tAUC). Progression-free survival (PFS) was chosen as the outcome, since it is a treatment evaluation parameter as closely as possible to the time of drug exposure and the tested miRNAs were all associated directly to drug specific response. Furthermore, overall survival (OS) was used for verification of findings. Comparison of analyses conducted for the respective cohorts (All DLBCL, ABC, and GCB patients) showed the lowest prediction errors for all models within the GCB subclass with a multivariate Cox miRNA-panel model including miR-146a, miR-155, miR-21, miR-34a, and miR-23a~miR-27a~miR-24-2 cluster performed the best and successfully stratified GCB-DLBCL patients into high- and low-risk of disease progression. In addition, combination of the miRNA-panel and international prognostic index (IPI) substantially increased prognostic performance in GCB classified patients, indicating a prognostic signal from the response-specific miRNAs independent of IPI. In conclusion: We found as proof of concept that adding gene expression data detecting drug-response specific miRNAs to the clinically established IPI improved the prognostic stratification of GCB-DLBCL patients treated with R-CHOP. Disclosures No relevant conflicts of interest to declare.

MYC and BCL2 mRNA Expression As Determined By NGS Predicts Survival in DLBCL in GCB but Not in ABC Subgroup

Introduction: RNA expression profiling using Next Generation Sequencing (NGS) provides important and reproducible information on expression levels of various genes. We studied the expression levels of MYC and BCL2 in patients with diffuse large B-Cell lymphoma (DLBCL) using NGS targeted RNA sequencing. We correlated levels of expression of these genes with outcome. Methods: RNA extracted from 441 FFPE samples with DLBC lymphoma and sequenced targeting 1408 genes. The RNA sequencing is based on hybrid capture and the number of reads ranged from 5 to 10 million. RNA quantification was performed using Cufflinks. The RNA levels were normalized to PAX5 mRNA levels. Of these cases, 380 were subclassified as ABC or GCB using expression profiling. The rest were classified as "undetermined", therefore were not included in further analysis. All patients were treated with R-CHOP. Results: The expression of MYC and BCL2 mRNA was slightly higher in ABC as compared with GCB (P=0.01 and P=0.02, respectively). However, in the GCB subtype, patients with MYC expression above the median showed significantly shorter survival as compared with those below the median (P=0.0007, Log-rank test). In contrast, there was no significant difference in survival using median of MYC expression as cutoff in patients classified as ABC subtype (P=0.38). Using upper 15% cut-off point for BCL2 mRNA expression, GCB patients with high BCL2 expression had significantly shorter survival (P=0.005). In contrast, there was no significant difference in survival between high and low BCL2 expression groups in the ABC subtype (P=0.1). When both MYC and BCL2 are considered, patients with high expression of both BCL2 and MYC (double-RNA expression) had significantly shorter survival as compared with patients with low expression of both MYC and BCL2 (P=0.0009) when both GCB and ABC groups are considered. Patients with high BCL2 expression also had poor survival similar to those double-RNA expression. Considering only patients with GCB, high expressor of both MYC and BCL2 had significantly worse outcome (P=0.0015) as compared with low expressors of both MYC and BCL2, but patients with high BCL2 also had significantly poor outcome as compared to low expressor of both MYC and BCL2 (P=0.0005). In contrast, there was no difference in survival for high or low MYC and BCL2 expressor in the ABC group. Conclusion: The data support the concept that in DLBCL, MYC and BCL2 mRNA expression levels are clinically relevant in GCB, but not ABC subtype. Furthermore, targeted RNA sequencing might provide a reliable and practical objective approach for the subclassification of DLBCL and determining double-RNA expression lymphoma. Figure Disclosures Albitar: Genomic Testing Cooperative: Employment, Equity Ownership. Tam:Takeda: Consultancy; Paragon Genomics: Consultancy. Hsi:Abbvie: Research Funding; Eli Lilly: Research Funding; Cleveland Clinic&amp;Abbvie Biotherapeutics Inc: Patents &amp; Royalties: US8,603,477 B2; Jazz: Consultancy. Piris:Calgene: Membership on an entity's Board of Directors or advisory committees; Gilead: Membership on an entity's Board of Directors or advisory committees, Research Funding; Jansen: Membership on an entity's Board of Directors or advisory committees, Other: Lecture Fees; Nanostring: Membership on an entity's Board of Directors or advisory committees; Kyowa Kirin: Membership on an entity's Board of Directors or advisory committees; Kura: Research Funding; Millenium/Takeda: Membership on an entity's Board of Directors or advisory committees, Other: Lecture Fees, Research Funding. Kantarjian:Immunogen: Research Funding; BMS: Research Funding; Cyclacel: Research Funding; Pfizer: Honoraria, Research Funding; Agios: Honoraria, Research Funding; Ariad: Research Funding; Amgen: Honoraria, Research Funding; Takeda: Honoraria; AbbVie: Honoraria, Research Funding; Astex: Research Funding; Jazz Pharma: Research Funding; Novartis: Research Funding; Actinium: Honoraria, Membership on an entity's Board of Directors or advisory committees; Daiichi-Sankyo: Research Funding.

Characterization of Methylation Patterns in Diffuse Large B Cell Lymphoma By Genome-Wide Methylation Analysis

Background: Diffuse large B cell lymphoma (DLBCL) is the most common type of non-Hodgkin lymphoma worldwide. Although the reference standard for identifying of the cell types is considered of gene expression profiling (GEP). But immunohistochemistry (IHC) is the most common method commercially available. The purpose of this study was to characterize the circulating cell-free DNA (cfDNA) methylation profile in DLBCL and to compare this profile with methylation observed in formalin fixed paraffin-embedded (FFPE) tissues. Additional efforts were made to correlate the observed methylation patterns with prognostic analysis and selected clinical features. Methods: The cfDNA and DNA of FFPE were extracted from 72 patients and 39 patients respectively. We assessed DNA methylation from plasma samples obtained from 29 individuals with GCB DLBCL at the time before treatment along with 43 samples of non-GCB DLBCL as controls. DNA from FFPE tissues were extracted from 11 individuals of GCB DLBCL and 28 individuals with non-GCB DLBCL. DNA methylation was analyzed with the Infinium MethylationEPIC BeadChip that quantitatively measures the methylation levels of more than 850,000 CpG sites across the genome. M values were used for visualization and intuitive interpretation of the results. Moreover, pathway enrichment analysis was performed with the Kyoto Encyclopedia of Genes and Genomes (KEGG) Pathway Database. Results: We found a total of 207 significant differentional differentially methylated positions (DMPs) of cfDNA between the GCB and non-GCB groups, identified with a p value of 0.001 (Fig. 1A). Of these, 65 presented at least 10% (|Δbeta| &gt; 0.1) difference in the methylation level between GCB and non-GCB. 29 (44.6%) were found hypermethylated in GCB DLBCL, while 36 (55.4%) appeared hypomethylated (Fig. 1B). The distribution of the DMPs identified according to their location relative to CpG islands (CGI) were represented in Fig. 1C. Unsupervised clustering performed on DNA methylation values for the 207 DMPs identified is presented in Fig. 1D. These results highlight the differences between GCB and non-GCB samples. There are 1549 significant DMPs of DNA from FFPE between the GCB and non-GCB groups, identified with a p value of 0.001 (Fig. 1E). Of these, 1512 presented at least 10% (|Δbeta| &gt; 0.1) difference in the methylation level between GCB and non-GCB . 1370 (90.6%) were found hypermethylated in GCB DLBCL, while 142 (9.4%) appeared hypomethylated (Fig. 1F). The distribution of the DMPs identified according to their location relative to CpG islands (CGI) were represented in Fig. 1G. Unsupervised clustering performed on DNA methylation values for the 1549 DMPs identified is presented in Fig. 1H. These results highlight the differences between GCB and non-GCB in FFPE samples which according with that in serum. The KEGG pathway enrichment analysis of DNA from FFPE tissue methylation revealed that the process "PI3K/Akt, Ras, MAPK signaling pathway" and "Human papillomavirus infection" are likely major contributors to Hans pathological type. In addition, the enrichment analysis of cfDNA methylation revealed that the process "MAPK signaling pathway" is likely the most important factor. Furthermore, we also have analyzed the methylation level between refractory or relapsed (R/R) DLBCL patients and individuals with a good prognosis. The differential methylation patterns were also found both in serums and FFPE tissues. Conclusions: The DNA methylation differs in GCB and non-GCB DLBCL patients. MAPK signaling pathway plays an important role in it. The mechanism needs to be further explored. Figure 1 Disclosures No relevant conflicts of interest to declare.

Potential Factors That Impact Lenalidomide/R-CHOP Efficacy in Previously Untreated Diffuse Large B-Cell Lymphoma in the ROBUST and ECOG-ACRIN 1412 Studies

Potential Factors That Impact Lenalidomide/R-CHOP Efficacy in Previously Untreated Diffuse Large B-Cell Lymphoma in the ROBUST and ECOG-ACRIN 1412 Studies

PS1076 PROGNOSTIC IMPACT OF BCL2 AND MYC EXPRESSION AND TRANSLOCATION AMONG NEWLY DIAGNOSED DIFFUSE LARGE B‐CELL LYMPHOMA PATIENTS TREATED WITH R‐CHOP IN THE REAL WORLD

Background:Diffuse large B‐cell lymphoma (DLBCL) patients with dual expression (DE; BCL2 and MYC expression via immunohistochemistry [IHC]) or dual hit (DH; BCL2 and MYC translocation via fluorescence in situ hybridization [FISH]) have poor prognosis.Aims:Given that some heterogeneity exists in IHC and FISH methods across labs in clinical practice, we investigated the real‐world prognostic ability of BCL2/MYC in first‐line (1L) DLBCL patients treated with rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R‐CHOP). If prognostic, this supports exploration of local testing methods as an alternative to centralized testing to expedite time from diagnosis to treatment and limit selection bias for less aggressive patients in investigational studies that require patient selection based on these biomarkers.Methods:The Flatiron Health electronic health record (EHR)‐derived database includes structured and unstructured patient data from &gt;280 cancer clinics. We identified 1L DLBCL patients diagnosed from 2011 to 2018 treated with R‐CHOP. We classified them by BCL2 and MYC expression and translocation based on values extracted by Flatiron Health, for both IHC and FISH, using the most recent report prior to 1L treatment. We also classified patients by cell of origin (COO; germinal b‐cell [GCB], activated b‐cell [ABC]/non‐GCB) based on IHC. Univariate Cox models estimated hazard ratios (HRs) and 95% confidence intervals (CIs) for overall survival (OS) and time to next treatment (TTNT). Kaplan–Meier methods generated survival curves.Results:Median (interquartile range) age at diagnosis was 68 (58–76) years. Most patients were white (69%) and male (55%); 28% were GCB, 19% ABC/non‐GCB, 53% were missing COO. IHC‐assessed BCL2 and MYC were available for 25% and 28% of patients, respectively (60% and 17%, respectively, for FISH). Rates of BCL2/MYC expression and translocation were within the expected range. Characteristics did not differ for patients who were unknown or untested. Inferior OS and TTNT were observed among patients with DH, DE, high BCL2 expression, and MYC translocation, although the association between DE and OS was only marginally significant (Table 1). When stratifying on COO, this association held regardless of COO for DH, BCL2 expression, and MYC translocation, but DE did not significantly predict shorter OS in GCB patients. Inferior TTNT, but not OS, was observed among patients with high MYC expression and BCL2 translocation (Table 1). When stratifying on COO, the association between high MYC expression and shorter TTNT was only marginally significant within each subgroup, likely due to lower power. The association between BCL2 translocation and shorter TTNT was only significant within GCB patients. Multivariate analysis will be presented.Summary/Conclusion:In this real world dataset, we found that BCL2 expression and MYC translocation had strong prognostic ability among DLBCL patients treated with R‐CHOP as 1L treatment, comparable with previous findings. DE, DH, high BCL2 expression, and MYC translocation predicted shorter OS and TTNT, while high MYC expression and BCL2 translocation predicted shorter TTNT only, a proxy for progression. Our findings in this real world dataset, despite key limitations, including significant missing data inherent to EHRs, imply robustness of these biomarkers to identify high‐risk patients in 1L DLBCL. Efforts to improve interpretive accuracy and external quality assurance programs should be considered to strengthen consistency in testing of these biomarkers in routine practice.image

Deubiquitinase OTUD7B Is Potential Prognostic Biomarker in Diffuse Large B-Cell Lymphoma

Background: Diffuse Large B-cell lymphoma (DLBCL) is the most common subtype of lymphoma and most patients can be cured under immune-chemotherapy. One third of patients will relapse and acquired resistance to chemotherapy and turned to uncurable. The identification of biomarkers of clinical behavior may aid in the implementation of novel therapeutic strategies. OTUD7B is a deubiquitinase and is a pivotal regulator of the noncanonical NF-κB pathway. The deficiency of OTUD7B can activate the noncanonical NF-κB pathway and mediate important biological functions, including lymphoid organogenesis, B lymphocyte function, and cell growth and survival. Activation of NF-κB pathway has been an adverse prognostic index in DLBCL. The prognostic value of OTUD7B expression in lymphoma patients remains unclear. We conducted a retrospective study to investigate the impact of OTUD7B expression in DLBCL patients. Methods: Histologically confirmed DLBCL patients (N=100) treated at Fudan University Shanghai Cancer Center with available diagnostic biopsy material were identified. Forty-two patients were treated with CHOP and fifty-eight patients with R-CHOP. OTUD7B expression was determined and quantified by immunohistochemistry (IHC) with the percentage of positive cells. Tumors were considered positive when at least 20% of tumor cells expressed OTUD7B. Kaplan-Meier method was used to calculate progression free survival (PFS) and overall survival (OS) and curves were compared with the long-rank test in both groups. Correlation between the OTUD7B expression and clinical variables were tested by Pearson Chi Square test and a two-sided P value of <0.05 was considered to be statistically significant. Result: the OTUD7B expression was detected in 79/100(79%) DLBCL patients and 21% patients shown OTUD7B deficiency. It seems the OTUD7B positive patients had a better outcome, But there was no significance (p=0.107), The PFS and OS were 81% and 85% in OTUD7B positive group and 67% and 72% in OTUD7B negative group. In patients treated with CHOP regimen, the OTUD7B deficiency was associated with a worse PFS compared with OTUD7B positive patients (20% vs 78%, p=0.004). the OS of OTUD7B deficiency patients also shown a worse trend compared with the positive group (40% vs 79%, p=0.057). In patients treated with R-CHOP regimen, it is surprising, no differences between the OTUD7B positive group and negative groups were observed in terms of PFS or OS. Rituximab may overcome this worse effect caused by OTUD7B deficiency. The OTUD7B deficiency were 15% in GCB patients and 25% in non-GCB patients (p=0.31). The basic characteristics were no difference between OTUD7B positive and negative groups. Conclusion: This is the first report demonstrating that, OTUD7B deficiency is a negative predictor of clinical outcome. OTUD7B is a potential novel target to treat lymphoma and further investigation of OTUD7B need to be done in the near future. (Research supported by National Natural Science Foundation of China (no.81670177, no.81001048). [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Polatuzumab Vedotin (Pola) Plus Bendamustine (B) with Rituximab (R) or Obinutuzumab (G) in Relapsed/Refractory (R/R) Diffuse Large B-Cell Lymphoma (DLBCL): Updated Results of a Phase (Ph) Ib/II Study

Polatuzumab Vedotin (Pola) Plus Bendamustine (B) with Rituximab (R) or Obinutuzumab (G) in Relapsed/Refractory (R/R) Diffuse Large B-Cell Lymphoma (DLBCL): Updated Results of a Phase (Ph) Ib/II Study

Whole-Exome Analysis Reveals Novel Somatic Genomic Alterations Associated with Cell of Origin in Diffuse Large B-Cell Lymphoma

Gene expression profiling has shown that diffuse large B-cell lymphoma (DLBCL) clusters into three major subtypes based on similarity in expression patterns to their cell of origin (COO): germinal center B-cell-like (GCB-DLBCL), activated B-cell-like DLBCL (ABC-DBLCL) and primary mediastinal B-cell lymphoma (PMBCL). These subtypes of DLBCLs are associated with distinctly different overall survival rates after standard immunochemotherapy. However, clinical and prognostic heterogeneity remains within COO subsets and strategies are needed to further stratify patients to identify and target high-risk subsets. A comprehensive genomic analysis of COO on a clinically defined set of DLBCL cases has not been performed and the aim of this study was to use whole-exome sequencing (WES) data from 58 paired tumor-normal DLBCL samples to assess association of known DLBCL genomic alterations with cell COO as well as for identification of novel and relevant genetic biomarkers.To investigate genomic alterations associated with DLBCL subsets, we analyzed WES and genome wide copy number data from 58 paired tumor-normal DLBCL tumors. Gene expression profiling or Hans classification was performed to determine DLBCL COO subtype; 31 patients were classified as GCB and 27 as non-GCB. The WES data were used to 1) assess the association of known DLBCL genomic alterations with COO, and 2) identify novel alternations associated with COO. Statistical analysis was performed and the data were ranked by significance (p≤ 0.05) within each DLBCL subtype. In total, 45 genomic abnormalities were analyzed for their association with either GCB, non-GCB or both. Mutations in CREBBP, EZH2, MEF2B, FOXO1 and REL have also been reported as GCB driver mutations and we observed GCB patients with these mutations, but the mutation clustering was not wholly associated with GCB. MYC double-hits were exclusively found in the GCB-subtype group. For the non-GCB cases we found that mutations in MAP2K3 and MYD88 were significantly associated with this subtype(p< 0.05). In addition to mutational patterns, we identified several copy number alterations (CNA) across both groups. Chromosomal losses in GCB patients were found at chromosomes 10q11.21-10q24.23, 4q12-4q35.2, 3q12.1-3q29, 4p12-4p16.3, 10p11.21-10p15.3, and 14q11.2-14q24.3 whereas gains were localized to 7q11.1-7q36.3, 7p11.2-7p22.3, and 1q21-1q32.1 (p < 0.05). No CNA was observed to directly associate with non-GCB patients, however, a loss at 9p21 and gains at 9q24.1 and 18q21.33 trended with the non-GCB subtype, supporting previous reports. Loss at 10q23.31 or a gain in 2p13-2p12 have been reported as being specific for GCB and our data confirmed the association of 10q23.31 with GCB while a gain at 2p13-2p12 (REL)was found in both subtypes. To further understand genomic differences between DLBCL subtypes, we evaluated the relative percentage of each genomic feature. 18/45 (40%) were only observed in GCB patients whereas 2/45 (5%) were specific to the non-GCB subtype. The majority (25/45, 55%) overlapped between the two subtypes.Throughout our analysis we noted that 7 non-GCB cases lacked any of the driver mutations analyzed in the study. While all cases carried mutations, they consisted of low frequency mutations that were not specifically associated with COO. 2/7 cases had a gain at 9p24.1 that included CD274 and JAK2. Because 9p24.1 gains have not been fully defined in DLBCL, we reviewed all cases and identified 4 (7%) with a 9p24.1 gain, 3 of which were non-GCB and 1 GCB. One non-GCB case was EBV+ and none of the cases showed evidence of PMBCL. Of the 9p24.1 cases, three had RNAseq data available and we found that PDL1 and JAK2 expression was elevated (12 fold p< 0.01 and 7 fold p< 0.01, respectively) when compared to the 9p24.1 normal cases (n=32). While outcome was not the focus of the study, we did note that 6/7 cases that lacked driver mutations achieved event free survival at 24 months (EFS24).Taken together, this analysis has further characterized the genetic profile of each COO subtype and has identified novel GCB CNAs which require independent replication. Additionally, we identify a subgroup of non-GCB DLBCL patients that do not harbor known driver mutations and require further genomic study to better resolve the biology of these tumors. Together, these data provide insight on the genetic heterogeneity of DLBCL and identify genetic variants that may inform subtype specific therapy. DisclosuresNowakowski:Morphosys: Research Funding; Celgene: Research Funding; Bayer: Consultancy, Research Funding. Rimsza:NCI/NIH: Patents & Royalties: L.M. Rimsza is a co-inventor on a provisional patent, owned by the NCI of the NIH, using Nanostring technology for determining cell of origin in DLBCL..