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Venetoclax, a potent B-cell lymphoma-2 (BCL-2) inhibitor, has demonstrated clinical efficacy in chronic lymphocytic leukemia (CLL). VENICE II is an open-label, single-arm, phase 3b study (NCT02980731) evaluating the impact of venetoclax monotherapy (400 mg once daily) for ≤2 years on health-related quality of life (HRQoL) of patients with relapsed/refractory CLL. The primary endpoint was mean change in the global health status (GHS)/quality of life (QoL) subscale of the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire Core 30 (EORTC QLQ-C30) from baseline to Week 48. Overall, 210 patients received ≥1 dose of venetoclax; median treatment duration was 67.4 weeks. The primary endpoint was met with mean improvement of +9.3 points (n = 156, 95% confidence interval 6.1-12.5; p=.004) in GHS/QoL. At Week 48, clinically meaningful improvements were observed for role functioning, fatigue, and insomnia domains of EORTC QLQ-C30, suggesting venetoclax monotherapy has a positive impact on HRQoL. No new safety signals were reported.

Background Neutropenia is a common hematologic Grade (Gr) 3+ adverse event (AE) recorded in patients with chronic lymphocytic leukemia (CLL) receiving venetoclax (VEN). In this analysis, we evaluated fixed duration VEN monotherapy given to patients with relapsed or refractory (R/R) CLL with and without pre-existing Gr3+ neutropenia. Methods This post-hoc analysis pooled data from patients in the ongoing Phase 3b trials VENICE-I and VENICE-II with R/R CLL who had received ≥1 dose of VEN monotherapy (ramp-up to 400 mg QD). Gr4 hematologic AEs and Gr3+ neutropenia ( Results At data cutoff (June 30, 2019), 44/468 (9%) patients had Gr3+ neutropenia at baseline (BL; Gr3+ neutropenia group), 80% of whom received G-CSF during the study vs 38% of those with Conclusion In this large post-hoc analysis, discontinuation due to neutropenia was rare (1%) in the overall population and accounted for 3/11 AE discontinuations in the Gr3+ neutropenia group; 10 patients had a serious infection. Patients with pre-existing neutropenia can be managed on VEN, though concurrent use of G-CSF is likely to be required. Additional data to follow. Abstract was previously published at EHA25.

In the present study, we describe the human and mouse RFP2 gene structure, multiple RFP2 mRNA isoforms in the two species that have different 5' UTRs and a human-specific antisense transcript RFP2OS. Since the human RFP2 5' UTR is not conserved in mouse, these findings might indicate a different regulation of RFP2 in the two species. The predicted human and mouse RFP2 proteins are shown to contain a tripartite RING finger-B-box-coiled-coil domain (RBCC), also known as a TRIM domain, and therefore belong to a subgroup of RING finger proteins that are often involved in developmental and tumorigenic processes. Because homozygous deletions of chromosomal region 13q14.3 are found in a number of malignancies, including chronic lymphocytic leukemia (CLL) and multiple myeloma (MM), we suggest that RFP2 might be involved in tumor development. This study provides necessary information for evaluation of the role of RFP2 in malignant transformation and other biological processes.

We report the primary characterization of a new gene KCNRG mapped at chromosome band 13q14.3. This gene includes three exons and has two alternatively spliced isoforms that are expressed in normal tissues and in some tumor cell lines. Protein KCNRG has high homology to tetramerization domain of voltage-gated K+ channels. Using the patch-clamp technique we determined that KCNRG suppresses K+ channel activity in human prostate cell line LNCaP. It is known that selective blockers of K+ channels suppress lymphocyte and LNCaP cell line proliferation. We suggest that KCNRG is a candidate for a B-cell chronic lymphocytic leukemia and prostate cancer tumor suppressor gene.

Mutational status of immunoglobulin variable region genes (VH-genes) is known as the strongest predictor of long term prognosis in B-CLL. However, applications in the routine clinical practice are time consuming, and therefore some other predictions are required. In this study, we have compared prognostic values of real time PCR quantification of the expression levels of four genes previously shown to be differentially expressed in V(H)-unmutated and mutated B-CLL subtypes: ZAP-70, ZBTB20, DMD and LPL. The study included 134 B-CLL patients. Expression levels of LPL and DMD genes were significantly correlated to mutational status, while expression levels of of ZAP-70 gene correlated only in CD19+ selected cases (N = 40). No correlation was observed for ZBTB20 gene. Expression levels of LPL and DMD predicted overall survival in the entire cohort of patients. Prognostic values of LPL gene expression levels were significant even for CLL patients with stage A. Quantitative RT-PCR assays for measuring LPL gene expression are robust enough to be introduced into routine clinical practice.

Abstract 263 RS represents the development of a diffuse large B-cell lymphoma (DLBCL) in the context of chronic lymphocytic leukemia (CLL). The pathogenesis of RS is still largely unknown. Analysis of RS has been often focused on the study of lesions previously identified in de novo DLBCL. However, we have previously shown that RS lacks many of the typical genetic lesions shown in DLBCL (Rossi et al, Blood 2011). Here, we have applied an unbiased, genome-wide approach, searching for DNA copy number alterations in a large series of RS, comparing them with de novo DLBCL, CLL-phase of RS and untransformed CLL. Methods. The study included 50 RS, 30 CLL-phases preceding RS, 100 DLBCL, 318 CLL. All RS were classified as DLBCL. DNA was extracted from frozen biopsies and was processed using the Affymetrix GeneChip Human Mapping SNP6 arrays. Differences in frequencies between subgroups were evaluated using Fisher's exact test. Clinical data were available in half of the cases and genomic lesions were evaluated for their impact on clinical outcome with the log-rank test. Results. In RS, the most common gains were +12 (40%), +4q12 (14%), +2p25.3-25.2 (13%), +8q23.3-qter (12%), +1q32.1, +2p16.1-13.2 (BCL11A/REL), +15q22.31-26.3, +18q21.33 (BCL2) (10%), and +3q24.1-q26.3 (9%). Amplifications were observed at the BCL2 locus and at 1q32.1. The most common losses were at 17p13.1 (TP53; 29%), 8p21.3 (TNFRSF10A/B; 15%), 8p23.3 (16%), 11q22.3 (ATM) (15%), 14q23.3 (13%), 7q33–35 (11%), 14q32.11 (10%), 14q32.31-32.33 (TRAF3), and 9p21.3 (CDKN2A) (9%). The latter locus was also the target of recurrent homozygous deletions. RS appeared intermediate between CLL-phase and de novo DLBCL in terms of number of recurrent lesions. When compared with de novo DLBCL, RS presented less -6q23 (TNFAIP3; P=2.54E-05), -15q15.1-q21.1 (B2M) (P=5.64E-04), +7/7q (P=5.71E-04), -6q21 (PRDM1; P=6.53E-04), +2p16.1 (BCL11A; P=3.22E-03), -1p36 (TNFRSF14; P 9.57E-03), copy-neutral LOH at 6p25.3-21.32 (P =1.01E-02), +5p (P=1.39E-02), +11q14.1-q25 (P=1.63E-02), +1q22-q41 (P 2.13E-02), +6p21 (P 2.48E-02). Conversely, RS presented more often +4q12 (P=8.96E-03), -7q33-35 (P=2.04E-02), -11q22.3 (ATM; P=2.14E-02), -14q23.3 (P=2.69E-02). No statistical differences were observed for lesions such as +3/+3q, +8q, -8p, - 9p21.3 (CDKN2A), +12q, +15q, -17p (TP53). Compared to CLL-phase, RS had significantly more gains at 1q32.1 (P=2.28E-02), 11q23.3 (P=5.38E-03), 11q24 (P=1.10E-02) and 9p21.3 losses (CDKN2A; P=1.10E-02). Compared to the large series of CLL with no history of transformation, CLL-phase samples presented less frequently losses at 13q14.3 (DLEU2, MIR15A, MIR16-1a; 6.58E-03), more commonly gains at 12 (P=2.66E-02), 4q12 (P=1.103E-02), +2p (P=2.97E-02), and loss at TP53 locus (P=2.23E-03). The presence of losses of CDKN2A (P=4.6E-03), TNFRSF10A/B (P=3.2E-02) and BCL2 gains (P=4.0E-02) determined a poor overall survival in RS. Conclusions. DNA copy number changes in RS are intermediate between de novo DLBCL and CLL-phase. Recurrent lesions were identified that appeared more common in RS than in DLBCL, and that seem to be involved in the progression from CLL to an aggressive lymphoma. Also, the acquisition of mono- and bi-allelic losses of CDKN2A appears as one of the most important event in the transition from CLL-phase to RS. CDKN2A and TNFRSF10A/B losses and BCL2 gains were associated with a poor outcome. AR & EC equally contributed to the work. Disclosures: No relevant conflicts of interest to declare.

Chronic lymphocytic leukemia (CLL) represents 22-30% of all leukemia cases, thus being the most commonly diagnosed form of adult leukemia in the Western world. On a cellular level, the disease progresses due to the prolonged survival of B-cell CLL cells arrested in the G₀ stage of the cell cycle. The current standard treatment for CLL is a combination regimen containing purine analogues and monoclonal antibodies. Although response rates to such regimens in previously untreated patients are high, patients with CLL invariably experience relapse and often acquire high-risk chromosomal abnormalities. Therefore, the search for novel avenues in CLL treatment is warranted. In this manuscript, we will describe theoretical premises and some preliminary data making the case for inhibitors of the potassium currents as possible proapoptotic agents that warrant investigation as a potential pharmacologic target in CLL.

Abstract 4406 We have found only one study aimed to find prognostic significance of abdominal lymph nodes in CLL (E. Montserrat et al, 2007). It was shown that Rai O CLL patients with enlarged abdominal lymph nodes had inferior survival. It merely means that CT scan could early reveal patients with enlarged lymph nodes. This cohort of patients is included in stage 0 only due to the fact of unavailability of CT scan at the times Rai and Binet staging were described (1975, 1977). The aim of the study was to evaluate prognostic significance of abdominal lymph nodes enlargement for the response to FC and FCR Patients and methods 71 untreated CLL patients were included according to the intent to treatment by fludarabine containing regimens (FC and FCR). Staging was performed according to Binet system. Stage B and C patients were included. Lymph nodes were revealed by CT and ultrasound before therapy (Lymph nodes more than 10mm in diameter were considered as abnormal and more than 100 mm in diameter were considered as bulky). CD38 was performed by CytoFlow (cut off 30%). Mutation analysis was done with PCR and sequencing (E.Nikitin, Hematological centre, Moscow), (cut off 98%). CR was defined according to NCI criteria. The patients were treated and followed-up by the single team of physicians, there were no losses of follow-us. SPSS - 17 statistical program was used for analysis. Results Overall response (OR) was 86% (61 pts), the rate of CR was 55% (39 pts), the rate of PR 31% (22 pts). In the whole group, FC: OR 81%, CR 46%, PR 35%. FCR: OR 91%, CR 67%, PR 24%), median of PFS was 26 mons (21 mons for FC and 35 for FCR, p=0.006). CD38 positivity, unmutated type and stage C had adverse prognostic significance in the whole group (p= 0,04; p=0,05; p=0,02), in FCR subgroups, mutation status and CD38 had marginal significance in FCR(p=0.9, p=0.75). Abdominal lymph nodes were enlarged in 41 pts (58%), 16 pts had bulky EAL(23%). PFS was lower in the group of patients with EAL in comparison to patients without EAL(18 and 48 mons, p 0.001). Difference in PFS has been also revealed in FC and FCR subgroups: 16 and 25 mons, p=0,003 for FC; 22 and 56 mons, p=0,01 for FCR (Fig.1) All fludarabine containing regimens resistant patients appeared to have EAL (10 pts). The duration of CR and PR were longer in patients without EAL(resp.p = 0.03 and 0.06, Fig. 2). The differences of PFS were also studied inside the groups defined by standard prognostic markers. CD38-, EAL+ patients had median PFS of 23mons, CD38-, EAL- patients-median not achieved (p= 0.01) CD 38+EAL+patients had median PFS of 12 mons, CD38+EAL- patients – 30 mons (p=0,04) Thus, CD38+ patients without EAL had even better PFS than CD38- patients with EAL. VH-mut+ EAL+ patients had median PFS of 18 mons, VH-mut+ EAL- patients – 58 mons (p=0,06) VH-mut- EAL+ patients had median PFS of 11 mons, VH-mut- EAL- patients – 18 mons (p=0,06) Thus, unmutated patients wit EAL had similar PFS with mutated patients without EAL. Stage B EAL+ patients had median PFS of 20 mons, Stage B EAL- patients – 58 mons (p=0,004) Stage C EAL+ patients had median PFS of 15 mons, Stage C EAL- patients – 21 mons (p=0,05) Thus, patients with stage B without EAL had PFS four times better, than patients with stage C, EAL+. Median PFS is similar in patients stage B EAL+ and stage C EAL-. The response equal to PR and more than PR after three cycles of chemotherapy was more frequent in patients without EAL (65% vs 18%, p < 0,001) Conclusion Enlarged abdominal lymph nodes(EAL) is an adverse prognostic marker for response both to FC and FCR. Differences in PFS according to EAL were also revealed inside standard prognostics subgroups (mutation status and CD38,stage). Even more, PFS of patients with standard adverse prognostic factors without EAL were similar to patients without standard adverse prognostic with EAL. CLL with EAL is a separate entity of CLL. Disclosures: No relevant conflicts of interest to declare.

Abstract 2377 Poster Board II-354 Fludarabine containing regimens has become the gold standard for first-line treatment of CLL. The prognosis for the response is usually done by mutation analysis and cytogenetics. These types of analyses are time consuming and could not be available in general practice. Staging is very easy tool for predicting the response, although it is not a matter of individual prognosis. The aim of the study was to find clinical data predicting the response. As a model, we have evaluated the response after 3 fludarabine containing regimens. Patients and methods 75 patients treated by fludarabin containing regimens (FC and FCR) were included (2002-2008). Staging was performed according to Binet (1977). Stage B and C patients were included. Lymph nodes were evaluated by CT and ultrasound. CD38 was done by CytoFlow (cut off 30%). Mutation analysis was studied by PCR and sequencing with cut off 98%. Early response(ER) was defined as PR and CR after 3 cycles of therapy, others were considered as nER. CR was defined according to NCI criteria. The patients were treated and followed-up by the single team of physicians, there were no losses of follow-us.SPSS - 17 statistical program was used for analysis. Results. In the whole group, median PFS was 25 mons . There were 40 ER pts(33 - PR and 7 - CR).Median PFS in ER patients was 38mons, in other patients (nER) – 17mons (p= 0.0001). After 6 cycles of therapy, the overall response was 100%; CR was 83% and 7% PR I ER pts, in nER- resp- 67%, 23% and 41% ( p < 0,001). Differences of median PFS according to standard prognostic factors is presented in table 1: Thus, ER patients with standard adverse prognostic factors did better than nER with similar prognostic factors. ER patients in comparison to nER had higher median PFS on both FC (28 mons vs 14 mons, p=0.001) and FCR (58 mons vs 18 mons, p=0.018) regimens. Conclusion. The early response (PR and CR) after three fluadarabine containing regimens is a good predictor for PFS. More intensive treatment should be considered for patients without early response. Disclosures: No relevant conflicts of interest to declare.

Richter syndrome (RS) occurs in up to 15% of patients with chronic lymphocytic leukemia (CLL). Although RS, usually represented by the histologic transformation to a diffuse large B-cell lymphoma (DLBCL), is associated with a very poor outcome, especially when clonally related to the preexisting CLL, the mechanisms leading to RS have not been clarified. To better understand the pathogenesis of RS, we analyzed a series of cases including 59 RS, 28 CLL phase of RS, 315 CLL, and 127 de novo DLBCL. RS demonstrated a genomic complexity intermediate between CLL and DLBCL. Cell-cycle deregulation via inactivation of TP53 and of CDKN2A was a main mechanism in the histologic transformation from CLL phase, being present in approximately one half of the cases, and affected the outcome of the RS patients. A second major subgroup was characterized by the presence of trisomy 12 and comprised one third of the cases. Although RS shared some of the lesions seen in de novo DLBCL, its genomic profile was clearly separate. The CLL phase preceding RS had not a generalized increase in genomic complexity compared with untransformed CLL, but it presented clear differences in the frequency of specific genetic lesions.