Increase In Absolute Lymphocyte Count Research Articles

Omission of Anti-CD20 Antibody in Chronic Lymphocytic Leukemia during COVID-19 Pandemic: Twelve Month Course of Venetoclax Is a Safe and Effective Alternative Regimen

Introduction The current treatment for patients with advanced stages of Chronic Lymphocytic Leukemia (CLL) is the combination of the Rituximab - an Anti-CD 20 Antibody and Venetoclax - a BCL-2 inhibitor. However, during COVID-19 Pandemic, we were concerned that the immunodeficiency state induced by anti-CD20 antibody infusions would increase the risk of COVID-19 complications. Also, we were concerned about exposing cancer patients to long term anti-neoplastic treatments when there were so many unknowns about the new Sar-CoV-2 virus. Thus, in order to minimize these deleterious effects, and long exposure to cancer treatment, the CLL treatment was modified through omitting Rituximab, and using a 12-month course of Venetoclax as a single agent, with the goal of preventing COVID complications, but still achieving durable remissions. Methods Between February 2020 and September 2022, 13 patients with advanced stage CLL who met the treatment criteria, were treated with single agent Venetoclax with the mean age of 77 ranging from ages 56-99 and a Male to Female ratio of 10:3. Indications of treatment were as follows: bulky lymphadenopathy (8), markedly elevated lymphocyte count (3), and bone marrow failure (2). Among these 13 patients, there were 7 treatment naive patients and 6 had previously received between 1 to 3 treatments. These treatments included: Chlorambucil-Obinutuzumab, Ibrutinib, Fludarabine-Cyclophosphamide-Rituximab, and Bendamustine-Rituximab. Following are the FISH analysis results of these patients: 13Q14, 12+, TP53- , 11Q22-. Single agent venetoclax was administered to patients as a 5 week ramp up schedule per standard dosing guidelines, with the goal being to achieve 400 mg by week 5 of the treatment. The 400 mg dosage was continued for another 11 months and patients were monitored monthly for treatment response and toxicity. After the conclusion of 12 months of treatment, patients were followed up clinically at regular intervals to monitor durability of treatment response. Patients were informed that if there was progression noted during Venetoclax treatment, a new regimen that included Anti-CD20 Antibody will be started. Results Eleven out of 13 patients achieved full dosage escalation to 400 mg by week 5. Of the 2 patients that did not escalate the full dose, one patient - age 99- only achieved dosage up to 100 mg due to physician discretion, and the other patient achieved dosage up to 200 mg due to extreme fatigue. Common side effects amongst the patients were fatigue and mild cytopenia whilst the toxicity overall was manageable. One patient required treatment interruption for Autoimmune Hemolytic Anemia (AIHA) and only one patient contracted COVID-19. This patient recovered, however, quickly with no complications and treatment of Venetoclax was resumed at full dosage. Temporary increase in absolute lymphocyte count was noted in 8 patients. Mild persistent anemia was shown in 2 patients at the completion of treatment and mild thrombocytopenia was present in 7 patients. Neutropenia was not observed in any patients. Overall, treatment response was noted in 12 out of 13 patients with 11 complete remission (CR) and 1 partial remission (PR). Time to response was 3 months from the start of treatment for all responding patients. All patients who achieved CR and PR remain in remission during follow up with a median remission duration of 15 months. One patient had stable disease while on treatment but progressed immediately after discontinuation of Venetoclax. This patient then received the combination of Obinutuzumab-Acalabrutinib and achieved complete remission. Conclusion Thus, we can conclude that a 12-month course of Venetoclax administered without anti-CD 20 antibody, is an effective treatment regimen in advanced stage CLL. The avoidance of the anti-CD 20 antibody resulted in preservation of the immunity as noted in the one patient who contracted COVID-19 during treatment but recovered quickly from infection without any COVID-19 associated complication. We believe that although Rituximab with Venetoclax remains the standard of care in advanced stage CLL, but in unusual circumstances such as during the COVID-19 pandemic, short, limited course of single agent Venetoclax is a viable option of treatment to achieve durable remission.

Effect of Three Unani Formulations (Tiryaq Wabai, Arq-e-Ajeeb, and Habb-e-Loban) as an Adjuvant to Standard Care Treatment in the Management of Mild-to-Moderate COVID-19 Patients—A Placebo-Controlled, Clinical Trial

IntroductionMore than 768 million cases, including 6.95 million deaths due to COVID-19, have been reported until now. Various Unani formulations were described as effective in the reduction of COVID-19 symptoms. This study aims to investigate the effect and safety of Unani formulations as an adjuvant to standard care for COVID-19. MethodsA randomised placebo-controlled clinical study was conducted with a total of 92 patients with mild-to-moderate COVID-19 disease. Three Unani formulations, (1) Tiryaq wabai 2 g once a day orally, (2) Arq-e-ajeeb two drops intranasal four times a day, and (3) Habb-e-loban one tablet twice a day, are used as an intervention, while the control group received placebo for the Unani formulations. Both arms received standard care treatment. The primary outcomes included a 50% increase in absolute lymphocyte count (ALC), while secondary outcomes involved World Health Organisation (WHO) modified ordinal scale improvement, RT-PCR negativity, SpO2/FiO2 ratio, time to symptom resolution, and hospital stay duration. Relevant statistical tests were used for the analysis. ResultsThe intervention group showed statistically significant improvements in ALC count, SpO2/FiO2 ratio, WHO modified ordinal scale, and RT-PCR negativity. The time taken to be discharged from the hospital showed no significance. No significant adverse drug reactions or events were reported, and liver and kidney function tests were normal in both groups. ConclusionsUnani herbal formulations, when used as an adjuvant to standard care treatment, demonstrated improvements in ALC count, SpO2/FiO2 ratio, earlier reduction of COVID-19 symptoms, and RT-PCR negativity in patients with mild-to-moderate COVID-19.

Determining clinical biomarkers to predict long-term SARS-CoV-2 antibody response among COVID-19 patients in Bangladesh.

Information on antibody responses following SARS-CoV-2 infection, including the magnitude and duration of responses, is limited. In this analysis, we aimed to identify clinical biomarkers that can predict long-term antibody responses following natural SARS-CoV-2 infection. In this prospective study, we enrolled 100 COVID-19 patients between November 2020 and February 2021 and followed them for 6 months. The association of clinical laboratory parameters on enrollment, including lactate dehydrogenase (LDH), neutrophil-lymphocyte ratio (NLR), C-reactive protein (CRP), ferritin, procalcitonin (PCT), and D-dimer, with predicting the geometric mean (GM) concentration of SARS-CoV-2 receptor-binding domain (RBD)-specific IgG antibody at 3 and 6 months post-infection was assessed in multivariable linear regression models. The mean ± SD age of patients in the cohort was 46.8 ± 14 years, and 58.8% were male. Data from 68 patients at 3 months follow-up and 55 patients at 6 months follow-up were analyzed. Over 90% of patients were seropositive against RBD-specific IgG till 6 months post-infection. At 3 months, for any 10% increase in absolute lymphocyte count and NLR, there was a 6.28% (95% CI: 9.68, -2.77) decrease and 4.93% (95% CI: 2.43, 7.50) increase, respectively, in GM of IgG concentration, while any 10% increase for LDH, CRP, ferritin, and procalcitonin was associated with a 10.63, 2.87, 2.54, and 3.11% increase in the GM of IgG concentration, respectively. Any 10% increase in LDH, CRP, and ferritin was similarly associated with an 11.28, 2.48, and 3.0% increase in GM of IgG concentration at 6 months post-infection. Several clinical biomarkers in the acute phase of SARS-CoV-2 infection are associated with enhanced IgG antibody response detected after 6 months of disease onset. The measurement of SARS-CoV-2 specific antibody responses requires improved techniques and is not feasible in all settings. Baseline clinical biomarkers can be a useful alternative as they can predict antibody response during the convalescence period. Individuals with an increased level of NLR, CRP, LDH, ferritin, and procalcitonin may benefit from the boosting effect of vaccines. Further analyses will determine whether biochemical parameters can predict RBD-specific IgG antibody responses at later time points and the association of neutralizing antibody responses.

Influence of exercise on quantity and deformability of immune cells in multiple sclerosis.

The study aimed to investigate the effect of exercise on immune cell count and cell mechanical properties in people with multiple sclerosis (pwMS) on different disease-modifying treatments (DMT) vs. healthy controls (HCs). A cohort of 16 HCs and 45 pwMS, including patients with lymphopenia (alemtuzumab and fingolimod) as well as increased lymphocyte counts (natalizumab), was evaluated for exercise-mediated effects on immune cell counts and lymphocyte deformability. As exercise paradigms, climbing stairs at normal speed or as fast as possible and cycling were used, while blood samples were collected before, immediately, and 20 as well as 60 min post-exercise. Immune cell subtypes and lymphocyte deformability were analyzed using multicolor flow cytometry and real-time deformability cytometry. An increase in lymphocytes and selected subsets was observed following exercise in HCs and all pwMS on different DMTs. Patients with lymphopenia exhibited an increase in absolute lymphocyte counts and immune cell subsets till just below or into the reference range. An increase above the upper limit of the reference range was detected in patients on natalizumab. Exercise-induced alterations were observable even in low and more pronounced in high-intensity physical activities. Lymphocyte deformability was found to be only mildly affected by the investigated exercise regimes. People with multiple sclerosis (PwMS) treated with alemtuzumab, fingolimod, and natalizumab respond to acute exercise with a comparable temporal pattern characterized by the increase of immune cell subsets as HCs. The magnitude of response is influenced by exercise intensity. Exercise-mediated effects should be considered when interpreting laboratory values in patients on immunomodulatory therapy. The impact of exercise on biophysical properties should be further elucidated.

Intravenously administered interleukin-7 to reverse lymphopenia in patients with septic shock: a double-blind, randomized, placebo-controlled trial

BackgroundProfound lymphopenia is an independent predictor of adverse clinical outcomes in sepsis. Interleukin-7 (IL-7) is essential for lymphocyte proliferation and survival. A previous phase II study showed that CYT107, a glycosylated recombinant human IL-7, administered intramuscularly reversed sepsis-induced lymphopenia and improved lymphocyte function. Thepresent study evaluated intravenous administration of CYT107. This prospective, double-blinded, placebo-controlled trial was designed to enroll 40 sepsis patients, randomized 3:1 to CYT107 (10 µg/kg) or placebo, for up to 90 days.ResultsTwenty-one patients were enrolled (fifteen CYT107 group, six placebo group) at eight French and two US sites. The study was halted early because three of fifteen patients receiving intravenous CYT107 developed fever and respiratory distress approximately 5–8 h after drug administration. Intravenous administration of CYT107 resulted in a two–threefold increase in absolute lymphocyte counts (including in both CD4+ and CD8+ T cells (all p < 0.05)) compared to placebo. This increase was similar to that seen with intramuscular administration of CYT107, was maintained throughout follow-up, reversed severe lymphopenia and was associated with increase in organ support free days (OSFD). However, intravenous CYT107 produced an approximately 100-fold increase in CYT107 blood concentration compared with intramuscular CYT107. No cytokine storm and no formation of antibodies to CYT107 were observed.ConclusionIntravenous CYT107 reversed sepsis-induced lymphopenia. However, compared to intramuscular CYT107 administration, it was associated with transient respiratory distress without long-term sequelae. Because of equivalent positive laboratory and clinical responses, more favorable pharmacokinetics, and better patient tolerability, intramuscular administration of CYT107 is preferable.Trial registration: Clinicaltrials.gov, NCT03821038. Registered 29 January 2019, https://clinicaltrials.gov/ct2/show/NCT03821038?term=NCT03821038&draw=2&rank=1.Graphical

CD49d Expression Identifies a Biologically Distinct Subtype of Chronic Lymphocytic Leukemia with Inferior Progression Free Survival on BTK Inhibitor Therapy

Background: Acalabrutinib, a highly selective Bruton tyrosine kinase inhibitor (BTKi), is effective and well tolerated for patients with chronic lymphocytic leukemia (CLL). During treatment with BTKis, transient lymphocytosis is observed due to the efflux of CLL cells from lymphoid organs. CD49d, the alpha chain of the CD49d/CD29 integrin very late antigen-4 (VLA-4), is a key mediator in CLL microenvironmental interactions. High CD49d expression has been associated with inferior response to ibrutinib (Tissino, 2018). Here, we investigated the role of CD49d in response to acalabrutinib in 48 patients with CLL enrolled in a single-center, phase 2 study (NCT02337829). Methods: Patients were divided into CD49d positive (CD49d+, n = 27, 56%) and CD49d negative (CD49dneg, n = 21, 44%) groups based on the established 30% cutoff, as assessed by flow cytometry. CD49d activation was measured as VLA-4 receptor occupancy (RO), as described (Tissino, 2018). Time to progression was estimated using the Kaplan-Meier method. RNA sequencing (RNA-seq) of CD19+ selected CLL cells from CD49d+ (n = 10) and CD49dneg (n = 10) subsets was performed at baseline and 6 months on acalabrutinib. Results: After the first month on acalabrutinib, the median increase in absolute lymphocyte count (ALC) for CD49d+ CLL was 6.4% vs. 50.4% for CD49dneg (P = .02). However, the increase in ALC on day 3 of 42% and 48%, respectively, was comparable (P = .8). A subset of patients showed bimodal CD49d (bimCD49d) expression, with concomitant presence of CD49d positive and negative subpopulations. In these cases, the fraction of CD49d+ CLL cells increased significantly on day 3 (P = .002), but decreased at 6 months (P = .004), suggesting that CD49d+ cells may be better able to remain tissue bound on chronic therapy or reenter lymphoid tissues at higher rates than CD49dneg cells. While constitutive VLA-4 activation was reduced in patients on acalabrutinib, BCR and CXCR4 stimulation still increased VLA-4 RO on CLL cells even after 6 months of treatment. PI3K, in addition to BTK signaling, is implicated in inside-out activation of VLA-4. In vitro, addition of the PI3K inhibitor duvelisib to acalabrutinib-treated CLL cells completely blocked BCR- but not CXCR4- induced VLA-4 activation. The overall response rate of the study was 95.8% (Sun, 2020). Sixteen patients progressed at a median of 42.5 months, 3 with transformation. Time to progression for patients with CD49d+ and CD49dneg CLL (applying 30% cutoff) did not differ (P = .4). However, at 72 months 50% of the combined CD49d+ and bimCD49d patients (n = 37) were estimated progression-free vs 90% of patients with a homogeneous CD49dneg CLL population (n = 11, P = .02). Of 13 patients progressing with CLL, 12 (92%) were CD49d+ or bimodal CD49d and 11 (85%) had BTK and/or PLCG2 mutations. To dissect the impact of CD49d expression on CLL pathobiology and the response to acalabrutinib, we compared transcriptomes of CD49d+ and CD49dneg subsets by RNA-seq. A distinct gene expression profile separated the groups by principal component analysis. Using gene set enrichment analysis, CD49d+ CLL showed constitutively more immunoreceptor, NF-kappa B, and cytokine/JAK-STAT signaling, as well as enhanced cell survival, adhesion and migration capacity compared to CD49dneg CLL (FDR <.25). After 6 months on acalabrutinib, all 20 patients achieved a clinical response and all samples showed downregulation of B-cell receptor and NF-kappa B target genes, as expected for BTKi therapy. Interestingly, on acalabrutinib, CD49d+ samples not only maintained a higher level of activation compared to CD49dneg CLL, but differences in the degree of cytokine (IL-6, IL-10) and STAT3 signaling became more prominent. Conclusion. CD49d+ and CD49dneg CLL cells are rapidly mobilized out of lymph nodes at the start of acalabrutinib therapy, but CD49d+ cells appear able to reenter or persist in lymphoid tissues better than CD49dneg cells. Bimodal CD49dneg and CD49d+ cases share an increased risk of disease progression, with most patients having detectable BTK or PLCG2 mutations at the time of progression. The transcriptional program of CD49d+ CLL reflects upregulation of survival and activation pathways that may increase tolerance to BTKi therapy and set the stage for clonal evolution. Thus, CD49d/VLA-4 emerges as a microenvironmental factor that contributes to BTKi resistance and a possible therapeutic target to improve on BTKi therapy.

Gestational age-specific hematological patterns in preterm infants following necrotizing enterocolitis

Objective To investigate gestational age (GA) specific hematological and transfusion response patterns in preterm infants following necrotizing enterocolitis (NEC) Design A retrospective study comparing hematological/transfusion information in three GA groups’ infants: Group A ≤ 28 weeks. Group B 28–32 weeks, Group C > 32 weeks following necrotizing enterocolitis. Results Group A infants responded with significantly higher WBC count, thrombocytopenia, higher absolute neutrophil, and higher absolute monocyte and lower absolute lymphocyte counts following NEC onset, received more blood transfusions before NEC onset (59.8 versus 30.0%; p = .007), and had higher odds of surgical NEC (OR 3.39 [95% CI 1.19–10.38]; p = .02) than group C. One unit increase in absolute lymphocyte count on the day, and 24 h following NEC was significantly associated with lower surgical NEC odds than groups C. Conclusion The infant’s in-group A had significantly different hematological response patterns following NEC than infants with higher gestational age (groups B and C).

Phase 1b/2 study of GX-I7 plus pembrolizumab in patients with refractory or recurrent (R/R) metastatic triple-negative breast cancer (mTNBC): The KEYNOTE-899 Study.

1081 Background: GX-I7 (efineptakin alfa) is a hybrid Fc-fused long-acting recombinant human IL-7 which plays an essential role in the development and homeostasis of T-cells. GX-I7 can potentially enhance the anti-tumor effect of pembrolizumab via induction of T-cell activity. Here, we report results of phase 1b/2 study of GX-I7 plus pembrolizumab in patients with R/R mTNBC. Methods: Eligible patients had R/R mTNBC that failed up to 3rd lines of chemotherapy in the metastatic setting. Phase 1b patients received GX-I7 in 5 dose levels ranging from 360 µg/kg to 1,440 µg/kg every 9 (Q9W) or 12 (Q12W) weeks plus pembrolizumab 200 mg Q3W (n=51). Phase 2 is an expansion cohort where 33 patients were treated with the recommended phase 2 dose (RP2D). The primary objective was to determine the RP2D for phase 1b and to assess the objective response rate (ORR) for phase 2. Results: The study included 84 patients (phase 1b, n=51; phase 2, n=33) and 53.6% (45/84) of patients have received 2nd to 3rd lines of previous therapy. In phase 1b, one dose-limiting toxicity (DLT; grade 3 skin rash) was reported in the 1,440 µg/kg cohort and GX-I7 1,200 µg/kg Q9W was selected as RP2D. The ORRs were 15.7% [95% confidence interval (CI): 7.0 – 28.6] for phase 1b (n=51) and 21.2% [95% CI: 9.0 – 38.9] for phase 2 (n=33). Median PFS was 2.4 months (95% CI: 2.1 – 2.7) at the median follow-up of 10.4 months for all patients combined (n=84). GX-I7 induced up to 3.6-fold (range 1.2 – 8.1) increase in absolute lymphocyte counts (including CD4+ and CD8+ T cell) in all dose levels. The most common treatment-related adverse events (AEs) of any grade were injection site reaction (50.0%), ALT increased (39.3%), pyrexia (38.1%) and rash (35.7%). The additional correlative study data will be presented. Conclusions: GX-I7 in combination with pembrolizumab demonstrated a manageable safety profile with promising anti-tumor activity in patients with R/R metastatic TNBC. Clinical trial information: NCT03752723. [Table: see text]

A Phase I Study Of The Oral Btk Inhibitor ONO-4059 In Patients With Relapsed/Refractory and High Risk Chronic Lymphocytic Leukaemia (CLL)

IntroductionThe B-cell receptor (BCR) pathway plays a central role in signal transduction pathways that regulate survival, activation, proliferation and differentiation of B-lineage lymphoid cells. Bruton's tyrosine kinase (Btk) is a critical kinase in BCR signal transduction and recent studies support that targeting Btk is effective in the treatment of B-cell malignancies. ONO-4059 is a highly potent and selective oral Btk inhibitor with an IC50 in the sub-nmol/L range that has demonstrated anti-tumour activity in several pre-clinical models (Yasuhiro T et al, AACR 2013). MethodsThis Phase I study was initiated to determine the safety, tolerability, dose-limiting toxicity (DLT), pharmacokinetics and pharmacodynamics of ONO-4059 given as monotherapy to patients with relapsed/refractory, high risk CLL for whom no therapy of higher priority was available. In this safety-driven, dose-escalating 3+3 design, ONO-4059 was administered as an oral, daily dose (flat dose) given continuously initially for up to 6 months, with the option of additional dosing up to 2 years. We present the safety and efficacy data on 16 evaluable patients, median age 67 yrs (range 40-77), with high risk cytogenetics (17p [38% (6/16)] or 11q [19% (3/16)], 12/16 displaying unmutated IgVH status, 8/16 displaying TP53 mutation), median baseline tumour burden 4,534mm2 [461-14,492 mm2]. Patients had received a median of 3 prior therapies [2-7], including fludarabine (16/16) and rituximab-containing therapy (13/16). Baseline median haematology parameters included hemoglobin 11.3 g/dL [8.5 -15.2], WBC 56.02 x 109/L [2.06-394.74], platelets 95.5 x 109/L [23-173], lymphocytes 50.6 x 109/L [0.5-390], neutrophils 2.46 x 109/L [0.5-11.1]. Patients were treated with ONO-4059 at doses ranging from 20mg-320mg (cohorts 1-5) and the study is currently ongoing with additional dose escalation cohorts to be completed. ResultsONO-4059 was found to be well tolerated, with no dose limiting toxicities (DLTs). A total of 15 ONO-4059-related Grade 1 (n=10) and 2 (n=5) adverse events were reported in 8 out of 16 patients, including one event of diarrhea (G2) and two events of rash (G1, G2). ONO-4059-related G3 or G4 haematological toxicities were infrequent and independent of dose; neutropenia (n=3 [G4 x1, G3 x 2]), febrile neutropenia (n=1 [G3]) occurring in 2 patients at the 20mg dose level only. Only one Grade 3 ONO-4059-related non-haematological event (pyrexia) was reported. Infection rate was low with only one event of bronchitis reported (G2, related). Three ONO-4059-related SAEs were reported in two patients (febrile neutropenia [G3] and pyrexia [G3] in one patient [20 mg]; rash [G2] in another [80 mg]). The pharmacokinetics of ONO-4059 reflects rapid absorption and elimination, a half-life of ∼6 hours, a dose dependent increase in exposure with no accumulation of ONO-4059 exposure and low inter- or intra-patient variability; with Btk occupancy in peripheral blood (as measured by phosphorylated Btk) being maintained for at least 24 hours across all dose levels.Responses have occurred across all dose levels. All patients experienced rapid reductions in lymphadenopathy observed early in treatment, between D1-D28 of the first cycle, accompanied by an increase in absolute lymphocyte count in some, but not all patients. Best overall response rate according to IWCLL criteria (including modified PR with lymphocytosis) was 70% [based on CT-scan and P/E for 7/10 evaluable patients]; with 2 PR, 5 PR with lymphocytosis (For 7 responding patients, median reduction of lymph nodes was 67% [Range 61%-79%]), 2 SD, 1 patient PD (possible Richter's). All patients showed an improvement in haematological parameters after ≥3 months on treatment. Patients (ONO-4059 20mg [1pt] and 40mg [1pt] respectively) who had grade 3 thrombocytopenia at study baseline had a significant improvement in platelet count (>100 x 109/L) with ONO-4059 treatment. To date, 15 of 16 patients remain on treatment with a median progression-free survival of 99 days [Range 8-268].In conclusion, ONO-4059 is a highly potent and selective oral Btk inhibitor with a favourable safety profile along with promising efficacy in this heavily pre-treated population, with responses observed in relapsed, refractory and 17p deleted patients. Disclosures:Salles:Janssen: Honoraria; Gilead: Honoraria; Celgene: Honoraria. Karlin:Janssen: Honoraria; Celgene: Honoraria. Morschhauser:ONO Pharma: Honoraria; Roche: Honoraria; Celgene: Honoraria; Mundipharma: Honoraria. Dyer:ONO Pharma: Honoraria, Research Funding. Hutchinson:ONO Pharma: Research Funding. Fegan:ONO Pharma: Honoraria. Cartron:ONO Pharma: Honoraria. Knurowski:ONO Pharma: Consultancy. Saunders:ONO Pharma: Consultancy; Pharmacyclics: Consultancy. Wright:ONO Pharma: Consultancy. Honda:ONO Pharma: Employment. Mazur:ONO Pharma: Consultancy. Yoshizawa:Ono Pharma: Employment. Kawabata:Ono Pharmaceutical Co., Ltd.: Employment. Birkett:Ono Pharma UK: Employment.

IMMUNO-BIOCHEMICAL STATUS OF SWEETS AT CORRECTION OF REPRODUCTION VIOLATIONS WITH THE APPLICATION OF «E-SELENA» AND «OVARIOVITA» IN THE CONDITIONS OF THE INDUSTRIAL COMPLEX

The aim of the authors was to determine the effect of the selenium-containing preparation «E-selenium» and the homeopathic preparation «Ovariovit» on the reproductive indices of sows, as well as the tasks: to study the morpho-biochemical and immunological status of clinically healthy animals; evaluate the performance of the sows under investigation. The experiment on the basis of a large industrial pig complex in the Omsk region using healthy sows selected according to the principle of analogues allowed us to establish that the simultaneous application of the studied complex of drugs allows achieving high productive results. The positive effect of the preparations is indicated by the production parameters of the experimental group with a significant difference from the control group, consisting in a shortening of the farrowing period (P &lt;0.01); an insignificant increase in the number of live-born piglets, the weight of the nest at birth, the average daily gain, and the average live weight of the piglets during weaning. The introduced complex «E-selenium» and «Ovariovit» with the purpose of correction of reproductive functions of sows positively influences blood indices, so on the 30th day after administration of drugs the amount of creatinine in the sows of the experimental group significantly decreased to 320.18 ± 8.69 against 401, 5 ± 13.43 in the control group (P≤0.001); a significant increase in the content of total protein in the blood of the test animals was observed by 9 % (P≤0.05). Immunological studies have confirmed the beneficial effect of selenium-containing and homeopathic preparations, namely on the 30th day after the introduction of the complex it was found that in the sows of the experimental group there was an increase in the number of leukocytes by 50 % and erythrocytes by 25.4 % (P &lt;0.05) with values in the control group of animals. There was also a significant increase in absolute lymphocyte count in the peripheral blood. At the same time, an increase in the number of lymphoid cells was due to all populations. Especially it is necessary to allocate cytotoxic T-lymphocytes, the concentration of which relative to control increased by 2.6 times (P &lt;0.05).

Single-Agent Ibrutinib Versus Chlorambucil-Obinutuzumab As First-Line Treatment in Patients with Chronic Lymphocytic Leukemia or Small Lymphocytic Lymphoma (CLL/SLL): Results of a Cross-Trial Comparison

Single-Agent Ibrutinib Versus Chlorambucil-Obinutuzumab As First-Line Treatment in Patients with Chronic Lymphocytic Leukemia or Small Lymphocytic Lymphoma (CLL/SLL): Results of a Cross-Trial Comparison

Abstract 1596: Phase I trial of recombinant human Interleukin 15 (rhIL-15) administered as continuous intravenous infusion (CIV) for 10 days (240 hours) in patients with refractory metastatic cancers

Abstract Twenty-seven refractory metastatic cancer patients received continuous intravenous recombinant human Interleukin 15 (CIV rhIL-15) for 10 days (240 hours) in a phase 1 dose escalation trial. Six dose levels (0.125, 0.25, 0.5, 1, 2 and 4 micrograms/kilogram/day [mcg/kg/day]) were assessed and 2 mcg/kg was the maximum tolerated dose Common adverse events included fevers, myalgias, decreased albumin, anemia, decreased leukocytes, lymphocytes and platelets and elevations of ALT, AST and bilirubin most often grade 1 or 2 in severity. Serious adverse events were grade 5 (gr5) visceral ischemia, lymphocytic gastritis (gr2), hospitalization for abdominal pain (gr2), papilledema (gr3), duodenal hemorrhage (gr3), bronchial hemorrhage (gr3) and unexplained sudden death (gr5) at home post-treatment in a patient who stopped treatment early for toxicity and symptomatic disease progression. Patients treated at the higher doses developed a striking end of treatment peripheral blood lymphocytosis showing a 24X increase in absolute lymphocyte count, 10X increase in CD8 cells and 75X increase in NK cell numbers. No patients developed anti-IL-15 antibodies. Steady state serum IL-15 level was seen within the first 12 to 24 hours of therapy and was maintained for several days. Dramatically lower IL-15 levels (≈ 8% of early Cmax) were measured during the last 2 days of treatment suggesting a large reservoir of non-circulating IL-15 receptor positive cells. None of the patients achieved a partial response, but 2 patients met the criteria for treatment beyond cycle 2 (≥ 15% decrease in measurable disease). Evidence for tumor directed immune effects included several patients who developed unilateral pleural effusions in their disease involved hemithorax, complete regression of a cutaneous chondrosarcoma metastasis and decreased CA19-9 (125.7 → 47.3 U/mL) in a patient with pancreatic cancer. Histologic analysis of pleural fluid from these patients showed largely lymphocytes and other cells that were consistent with the patients known cancers. Lymphocytic infiltration of the regressing cutaneous sarcoma metastasis was predominantly CD4 cells, with some CD8 cells and a few NK cells. Immunohistochemical analyses of pre and on treatment tumor biopsies obtained from several other patients are currently being performed to better characterize the immunologic effects of this regimen. The results of this trial indicate CIVrhIL-15 treatment generates considerable in vivo immune activation and has important synergistic potential for combination therapy with other Immunotherapeutics. Citation Format: Kevin C. Conlon, Sigrid Dubois, Chyi-chia Richard Lee, Thomas Fleisher, Jennifer I. Hsu, Thomas A. Waldmann. Phase I trial of recombinant human Interleukin 15 (rhIL-15) administered as continuous intravenous infusion (CIV) for 10 days (240 hours) in patients with refractory metastatic cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1596. doi:10.1158/1538-7445.AM2017-1596

High neutrophil to lymphocyte ratio measured before starting ipilimumab treatment is associated with reduced overall survival in patients with melanoma

There is an unmet need to identify markers predictive of response to ipilimumab in patients with melanoma because the number of responders to ipilimumab is low and its cost is very high. An increase in absolute lymphocyte count (ALC) or low neutrophil/lymphocyte ratio (NLR) just before the third infusion has been reported to be associated with better overall survival (OS). Our aim was to determine whether NLR measured before the first infusion was associated with OS. Data were collected on a consecutive series of 58 patients treated with ipilimumab in four hospitals, including 51 at stage M1c and four at stage M1b. The influences of the NLR and other factors such as lactate dehydrogenase (LDH), performance status, ALC, absolute neutrophil count (ANC) and corticosteroids on survival were studied. We also assessed this association with NLR categorized as a binary variable. The cut-off value for the NLR was determined with time-dependent receiver operating characteristic (ROC) analysis. Univariate and multivariate analyses were performed using Cox regression models. High NLR (≥) 4, high ANC LDH levels (>2), performance status ≥2, symptomatic brain metastases, and corticosteroids before week 1 were associated with poorer survival on univariate analysis. Using multivariate analysis, a significant association between high NLR (continuous variable) and poorer survival was demonstrated and remained significant after adjustment on potential confounders [hazard ratio (HR) = 1·21, 95% confidence interval (CI) 1·07-1·36]. NLR ≥4 was an independent prognostic factor (HR = 2·2, 95% CI 1·01-4·78). Intake of corticosteroids before week 1 was not an independent prognostic factor (HR = 1·28, 95% CI 0·54-3·06). High NLR (≥4) before initiating ipilimumab treatment in patients with metastatic melanoma is an independent prognostic indicator of poor survival.

In Vivo Ibrutinib Abrogates BCR-Dependent Adhesion in Tumor Cells of Patients with Chronic Lymphocytic Leukemia

Ibrutinib, now FDA approved for use in both previously treated mantle cell lymphoma (MCL) and chronic lymphocytic leukemia (CLL), induces objective clinical responses and is well tolerated (Advani, JCO 2013; Byrd, NEJM 2013). As with other BCR-directed kinase inhibitors, (such as fostamatinib and idelalisib) ibrutinib leads to a treatment-induced transient lymphocytosis that resolves over the first 2-3 months on therapy in patients with CLL. The initial increase in absolute lymphocyte count (ALC) has been shown to be due to the release of tumor cells from the tissue compartments; specifically the lymph nodes, into the peripheral blood (Herman, Leukemia 2014). In vitro ibrutinib reduces the ability of CLL cells to migrate towards chemokines and prevents adhesion of BCR stimulated CLL cells (Ponader, Blood 2012; de Rooij, Blood 2012). We hypothesized that the initial rise in ALC is due to inhibition of CLL cell adhesion. We tested this ex vivo in patients enrolled on a single agent phase II trial of ibrutinib in CLL (NCT01500733) and investigated the mechanism.In order to evaluate the effect of ibrutinib on CLL cell adhesion in vivo we compared peripheral blood tumor cells collected from patients pre-treatment and on day 28 of treatment. Fluorescently labeled cells were plated on fibronectin-coated plates and allowed to adhere for 1 hour. Adhesion was then evaluated using florescence microscopy and quantified using the ImageJ software. After 28 days on ibrutinib, CLL cells displayed a severely reduced adhesion ability (99% reduction, P<0.001, n=13). Next, we tested the effect of ibrutinib on CLL cell adhesion to a confluent stromal cell layer. Although less dramatic, we found a similar reduction in the ability of cells to adhere to the stromal cell layer in all three patients evaluated. As the initial rise in ALC occurs very rapidly in some patients, we evaluated the ability of cells collected on day 2 of ibrutinib therapy and again found a significant inhibition of cell adhesion to fibronectin (81% reduction, P=0.005, n=10). This suggests that even one dose of ibrutinib is sufficient to disrupt CLL cell adhesion.We next sought to determine if ibrutinib could directly interfere with adhesion by causing cells to detach from fibronectin. We found that after one hour of 1µM ibrutinib treatment in vitro there was a significant reduction in the number of CLL cells that remained attached (33% reduction, P=0.03, n=10). This suggests that ibrutinib can not only prevent CLL cell adhesion but can release adhered cells from the microenvironment, consistent with influx of cells from the lymph node into the peripheral blood within hours of starting treatment.To determine whether the observed defect in cell adhesion is due to changes in the expression of CD29/CD49d (VLA-4 or α4β1 integrin), or CD18/CD11a (LFA-1), we measured the expression of these known regulators of CLL cell adhesion in samples obtained pre-treatment and during treatment. While there was inter-patient variability, overall there was no significant change in expression of CD29 after 28 days on treatment. In contrast albeit minor, we found reductions in both CD49d and CD18 (P<.05). When analyzing cells obtained on day 2, we found no significant changes suggesting that alteration of adhesion molecule expression does not account for the rapid increase in ALC. Integrin α4β1 is activated by PKC downstream of BTK. We thus sought to determine whether stimulation with PMA, an activator of PKC, could restore the ability of CLL cells to adhere to fibronectin. Indeed, PMA stimulation of CLL cells obtained from patients after 28 days on ibrutinib partially restored their ability to adhere to fibronectin, suggesting that reduced α4β1 integrin activity contributes to inhibition of adhesion by ibrutinib.In conclusion, ibrutinib abolished CLL cell adhesion at least in part due to direct inhibition of BTK- and BCR-mediated α4β1 integrin activity. Additionally, ibrutinib not only prevented adhesion but also caused the detachment of adhered cells from the extracellular matrix. Together, this suggests that disruption of cellular adhesion is a major contributor to the observed rise in ALC early on treatment.This work was supported by the Intramural Research Program of NHLBI, NIH. We thank our patients for donating blood samples to make this research possible. DisclosuresWiestner:Pharmacyclics Inc.: Research Funding.

The Bruton’s Tyrosine Kinase (BTK) Inhibitor ONO-4059: Promising Single Agent Activity and Well Tolerated in Patients with High Risk Chronic Lymphocytic Leukaemia (CLL)

IntroductionBruton’s tyrosine kinase (BTK) is a critical kinase involved in B-cell receptor signal transduction. ONO-4059, a highly potent and selective oral BTK inhibitor has demonstrated anti-tumour activity in pre-clinical models (Yasuhiro et al, AACR2013) and in the clinic in both CLL and NHL patients (Morschhauser et al, EHA 2014; Rule et al, EHA 2014). Here, we present data from 25 CLL patients enrolled in the ongoing Phase I study ONO-4059POE001.MethodsTwenty five CLL patients were administered ONO-4059 as monotherapy, given once daily (QD) to determine safety, pharmacokinetics and pharmacodynamics, and preliminary efficacy. Patients can receive ONO-4059 for up to a maximum of 3 years and upon completion of the first 6 months of treatment, intra-patient dose escalation is permitted. Here, we present data for 25 evaluable patients treated with ONO-4059 at doses ranging from 20-600mg (8 cohorts).Patients had a median age 67 yrs [range 40-79], median baseline tumour burden 3,943 mm2 [461-19,750 mm2], median of 4 prior therapies [2-8], including fludarabine (23/25) and rituximab-containing therapy (23/25). Baseline median haematology parameters included lymphocytes 34.6 x 109/L [0.1-391], haemoglobins 11.1 g/dL [8.5-15.2], platelets 97 x 109/L [23-185], neutrophils 2.39 x 109/L [0.5-11.1]. 9/24 patients were 17p deleted [38%] and 6/24 had the 11q deletion [25%], 20/24 were IgVH unmutated status [83%], with 13/24 [54%] displaying TP53 mutation).ResultsTo date, 19 of 25 patients remain on treatment with a median duration of treatment of 363 days [27-659]. ONO-4059 was found to be well tolerated with few adverse reactions over a long duration. A total of sixty two ONO-4059-related Grade 1- 2 adverse events were reported in 19 out of 25 patients. Eight dose independent ONO-4059-related G3 or G4 haematological toxicities were reported in 5 patients; with neutropenia in 3 patients, [G3 x 1, G4 x 2]), febrile neutropenia in 1 patient [G3] and leucopenia in 2 patients [G3 x 4]. Six Grade 3 ONO-4059-related non-haematological events were reported (pyrexia x 1, Hepatitis E reactivation x 1, Lymphocytic infiltration, Purpura, Urinary tract infection and Haematoma in just one patient). Nine ONO-4059-related SAEs were reported in 6 patients (febrile neutropenia [G3] and pyrexia [G3] at 20 mg, rash [G2] at 80 mg, hepatitis E reactivation [G3] at 320mg, neutropenia [G4] at 320 mg, purpura x 2 [G2, G3] and lymphocytic infiltration [G3] at 400 mg, haematoma [G2] at 500mg).All patients experienced rapid reductions in lymphadenopathy observed early in treatment between Cycle1 and Cycle3, accompanied by an increase in absolute lymphocyte count in 72% of patients. Almost all patients have durable response over a long duration. Best overall response rate according to IWCLL criteria (including modified PR with lymphocytosis) was 84% [based on CT-scan and P/E for 21/25 evaluable patients with 17 PR, 4 PR with lymphocytosis (for 21 responding patients, median reduction of lymph nodes was 83% [51-100]), 1 SD and 2 PD], with 89% response rate on 17p deleted. The early tumour shrinkage was associated with a rise in haemoglobin and platelet count. An increase in exposure (as assessed by Cmax and AUC) that was proportional to an increase in dose was seen for all doses of ONO-4059 ranging from 20-600mg.ConclusionONO-4059 is a highly potent and selective oral Btk inhibitor with a favourable safety profile along with promising efficacy over a long duration in this heavily pre-treated population, with responses observed in relapsed, refractory and currently poor prognostic 17p deleted patients. DisclosuresFegan:ONO PHARMA: Honoraria. Salles:celgene: Honoraria, Membership on an entity’s Board of Directors or advisory committees; roche: Honoraria, Membership on an entity’s Board of Directors or advisory committees; genetech: Honoraria, Membership on an entity’s Board of Directors or advisory committees. Karlin:Janssen: Honoraria; celgene: Consultancy, Honoraria; Sandoz: Consultancy. Rule:Pharmacyclics, J&J: Consultancy, Membership on an entity’s Board of Directors or advisory committees. Morschhauser:ONO PHARMA: Honoraria; Roche: Honoraria; Celgene: Honoraria; Mundipharma: Honoraria. Dyer:ONO PHARMA: Research Funding. Cartron:ONO PHARMA: Honoraria. Honda:Ono Pharmaceutical Co., Ltd.: Employment. Yasuhiro:Ono Pharmaceutical Co., Ltd.: Employment. Yoshizawa:Ono Pharmaceutical Co., Ltd.: Employment.

Functional Evidence from Deuterated Water Labeling That the Bruton Tyrosine Kinase Inhibitor Ibrutinib Blocks Leukemia Cell Proliferation and Trafficking and Promotes Leukemia Cell Death in Patients with Chronic Lymphocytic Leukemia and small Lymphocytic Lymphoma

BACKGROUND: In patients with chronic lymphocytic leukemia (CLL), the Bruton tyrosine kinase(BTK) inhibitor ibrutinib induces a rapid reduction in lymphadenopathy, accompanied by a transient lymphocytosis. Continuation of therapy leads to a reduction in peripheral blood disease burden with achievement of objective responses in a majority of patients over time. Inhibition of B cell receptor (BCR) and chemokine receptor signaling are considered the mechanisms for redistribution lymphocytosis and CLL cell death. However, these mechanisms of action have not been formally documented directly in patients. To address this, before initiating therapy with ibrutinib, we labeled the DNA of proliferating CLL cells with deuterium (2H) by asking patients to drink deuterated water (2H2O) and then determined the effects of ibrutinib on leukemia cell kinetics (proliferation and death rates) and mobilization of cells from lymphoid tissues (trafficking) after ibrutinib administration.METHODS: 30 previously untreated CLL/SLL patients were enrolled between December 2012 and June 2013 at MD Anderson Cancer Center. Patients required treatment per iwCLL guidelines, had adequate hematopoietic (platelets > 50,000/μL, ANC > 750/μL) and organ functions. Patients drank 50 mL of 70% 2H2O 3 times a day for 5 days, followed by 60 mL daily for a total of 4 weeks (“labeling phase”). After a 6-12 week “washout phase”, patients started once-daily ibrutinib 420 mg continuously on 28 day cycles. Study objectives were to determine the impact on CLL cell proliferation and death rates, before and after ibrutinib, and on CLL cell trafficking after ibrutinib.PATIENT CHARACTERISTICS: Median age of the 30 patients was 64 years (range 48–78) with 19 males. 16 patients exhibited early stage (Rai stage 0-2), and 14 advanced stage (Rai stage 3-4) disease. 17 patients expressed unmutated IGHVs, 11 patients had mutated IGHVs, and 2 had inconclusive IGHV results. 8 were CD38+ and 15 ZAP-70+ by immunohistochemistry. FISH cytogenetics revealed 11 patients with del13q, 6 with trisomy 12, 3 with del17p or TP53 mutation, 4 with del11q, and 6 without abnormalities. Median β2 microglobulin level was 2.8 mg/L (1.6 - 7.4).RESPONSE TO THERAPY: At a median follow up of 13 months, 28 of 30 patients continued on therapy without disease progression. All patients were evaluable for response assessment per 2008 IWCLL guidelines: 28 (93%) achieved partial remission, 1 (3%) complete remission, and 1 (3%) had stable disease, yielding an ORR of 97%. Two patients came off study at 184 and 480 days, respectively.CLL CELL KINETICS AND TRAFFICKING: The average CLL cell birth rate determined before ibrutinib therapy was 0.42% per day (range 0.32 – 1.42%). After initiating ibrutinib, there was a rapid increase in absolute lymphocyte count in the blood in 26 of 30 patients; this contained previously labeled CLL cells rather than newly-divided unlabeled cells. Over time, the proportion of circulating CLL cells that were labeled did not decrease appreciably despite a concomitant fall in circulating lymphocytes, indicating that newly divided cells were not entering the circulation and strongly suggesting that ibrutinib had a major inhibitory effect on leukemia cell proliferation. In 3 of the 4 patients in whom there was a rapid influx of unlabeled cells with the onset of ibrutinib treatment, maintenance of circulating labeled cells concomitant with a significant fall in circulating lymphocytes suggested proliferation of CLL cells was inhibited with extended treatment. Prior to ibrutinib therapy, the measured proliferation rate of CLL cells was 0.42 % per day. Because after ibrutinib therapy both cell proliferation and exiting from the blood were markedly reduced, the elimination rate should equal the death rate. Notably, with treatment the elimination rate of CLL cells was much faster than before ibrutinib, revealing a “true” death rate of 1.48 % of the clone per day.CONCLUSIONS: Using 2H-labelling of dividing leukemic cells, we have shown for the first time directly in CLL patients that ibrutinib significantly inhibits leukemia cell proliferation, leads to an efflux of CLL cells from tissues into the blood, and promotes a remarkably high death rate of CLL cells in an apparently indirect manner, likely due to the interruption of survival signals from the BCR and other receptors that are engaged in lymphoid tissues. DisclosuresBurger:Pharmacyclics, Inc.: Research Funding; Janssen: Membership on an entity’s Board of Directors or advisory committees. Li:KineMed, Inc.: Employment, Equity Ownership. O’Brien:Pharmacyclics, Inc.: Membership on an entity’s Board of Directors or advisory committees, Research Funding. Hellerstein:KineMed Inc: Consultancy, Employment, Equity Ownership, Membership on an entity’s Board of Directors or advisory committees. Turner:KineMed, Inc.: Employment, Equity Ownership. Emson:KineMed, Inc.: Employment, Equity Ownership. Chiorazzi:Pharmacyclics, Inc.: Research Funding; KineMed: Equity Ownership.

Abstract PL02-03: Immune modulation for cancer therapy: Assessing antagonists and agonists

Abstract PL02-03: Immune modulation for cancer therapy: Assessing antagonists and agonists

Phase 1 Study Of Single Agent CC-292, a Highly Selective Bruton's Tyrosine Kinase (BTK) Inhibitor, In Relapsed/Refractory Chronic Lymphocytic Leukemia (CLL)

IntroductionCC-292, an oral, highly selective, small-molecule irreversible-inhibitor of Btk is under investigation for the treatment of CLL and other B-cell malignancies. This phase 1 trial investigated the safety, dose limiting toxicities (DLT), and clinical activity of CC-292 monotherapy in subjects with relapsed or refractory (R/R) CLL or non-Hodgkin's lymphoma. This interim analysis focused on the safety and clinical activity in subjects with CLL and small cell lymphocytic leukemia (SLL). MethodsEligible subjects with R/R (≥ 1 prior therapy) CLL/SLL were treated with monotherapy CC-292 in a dose-escalation study with doses ranging from 125 mg to 1000 mg QD and BID dose levels of 375 mg and 500 mg. As a maximum tolerated dose was not established, CLL patients have been enrolled in an early dose expansion cohort of 750 mg QD and preliminary recommended phase 2 dose expansion cohort at 500 mg BID. All subjects received continuous dosing in 28-day cycles until progressive disease or intolerable toxicity. Clinical activity was investigator assessed per the 2008 iwCLL criteria. Results83 subjects with R/R CLL/SLL have been enrolled as of June 30, 2013. Baseline characteristics include median age of 67 years (34-89), 52% Rai stage 3/4 disease, median 3 prior therapies (1-12), and 34% refractory to last treatment. Poor-risk factors were present in 67% of subjects, including del11q (22%), del17p (24%), and unmutated IgVH (54%). 67% of subjects remain on study. Subjects have received a median of 6 treatment cycles (0.2-21.6).The most frequent grade 3/4 adverse events (AEs) included neutropenia (21%), thrombocytopenia (15%), pneumonia (10%), and anemia (8%). Rates of febrile neutropenia were low (4%). The most common treatment-emergent AEs (≥ 10% of subjects) were diarrhea (59.7%), fatigue (37.5%), neutropenia (26.4%), thrombocytopenia (26.4%), nausea (26.4%), pyrexia (22.2%), headache (19.4%), cough (19.4%), upper respiratory infection (16.7%), peripheral edema (15.3%), abdominal pain (15.3%), dizziness (13.9%), muscle spasms (13.9%), contusion (13.9%), anemia (12.5%), pneumonia (12.5%), sinusitis (12.5%), and urinary tract infection (11.1%). One CLL patient at the 500 mg BID dose level has experienced a drug-related adverse event of grade 4 thrombocytopenia during Cycle 1 that was assessed as a DLT. The number of patients discontinuing study treatment due to AEs was low (2.4%).Results are summarized for efficacy-evaluable patients treated at the 4 dose levels where at least a partial response (PR) was achieved (750 mg, 1000 mg, 375 mg bid and 500 mg bid) (n = 55). During cycle 1, 89% experienced a ≥ 25% increase in absolute lymphocyte count (ALC), which usually resolved with continued treatment. The table below details the best responses achieved to date. In subjects with del11q, del17p, unmutated IgVH, and no high-risk genomic factors, the PR rate was 42% (5/12), 25% (3/12), 44% (7/16), and 47% (7/15), respectively. Importantly, nodal responses were induced in the majority of subjects receiving BID dosing (375 mg: 67%; 500 mg: 62%) with an overall PR rate of 40%. When achieved, nodal responses were typically observed by cycle 2 and were sustained with continued treatment. Although the sample size is small, subjects treated at 375 mg or 500 mg BID showed continued lymph node size reduction over time from cycle 2 (mean reduction of 42% and 45%, respectively) to cycle 7 (mean reduction of 60% and 71%, respectively). ConclusionCC-292 is well tolerated as an oral daily therapy. Single-agent therapy with CC-292 is sufficient to achieve high nodal and partial response rates in relapsed/refractory CLL subjects, including those with high-risk genomic features. These results support continued development of CC-292 for the treatment of patients with CLL/SLL. Disclosures:Brown:Pharmacyclics: Consultancy; Genentech: Consultancy; Celgene: Consultancy, Research Funding; Emergent: Consultancy; Onyx: Consultancy; Sanofi Aventis: Consultancy; Vertex: Consultancy; Novartis: Consultancy; Genzyme: Research Funding. Off Label Use: The abstract shows scientific information on CC292 which is an investigational product developed by Celgene. This investigational product is not approved by any health authority for any indication. Harb:Celgene: Research Funding. Hill:Celgene: Honoraria, Research Funding. Sharman:Celgene: Consultancy, Research Funding; Genentech: Consultancy, Research Funding; Gilead: Consultancy, Research Funding; Pharmacyclics: Consultancy, Research Funding; Seattle Genetics: Consultancy, Research Funding; Cylene Pharmaceuticals: Consultancy, Research Funding; Avila Pharmaceuticals: Consultancy, Research Funding. Barr:Celgene: Consultancy. Foran:Celgene: Research Funding. Burger:Pharmalytics: Research Funding; Gilead: Research Funding. Mahadevan:Novartis: Speakers Bureau; Millennium: Speakers Bureau. Ma:Genentech: Consultancy; Abbvie: Consultancy. Barnett:Celgene: Employment, Equity Ownership. Marine:Celgene: Employment, Equity Ownership. Nava-Parada:Celgene: Employment, Equity Ownership. Azaryan:Celgene: Employment, Equity Ownership. Mei:Celgene: Employment, Equity Ownership. Kipps:Celgene: Membership on an entity's Board of Directors or advisory committees, Research Funding.

Update On The Safety and Efficacy Of The Pan Class I PI3K Inhibitor SAR245408 (XL147) In Chronic Lymphocytic Leukemia and Non-Hodgkin's Lymphoma Patients

BackgroundConstitutive activation of phosphatidylinositol 3-kinase (PI3K)/ mammalian target of rapamycin (mTOR) pathway by various mechanisms has been implicated in the pathogenesis of chronic lymphocytic leukemia (CLL) and non-Hodgkin's lymphoma (NHL). There is mounting evidence suggesting that along with PI3Kγ, PI3Kα may be involved in CLL/NHL. SAR245408 is a potent and selective inhibitor of all α, γ and δ class I PI3K isoforms. It has been shown to inhibit PI3K signaling and impact tumor growth in preclinical tumor models. The impact of SAR245408 on safety, tolerability, pharmacokinetics, pharmacodynamics and anti-tumor effect was evaluated in patients with relapsed/refractory CLL and NHL from the Sanofi sponsored phase 1 single-agent study (NCT00486135) [Brown et al. ASH Annual Meeting Abstracts 2011. 118 (21): 2683]. MethodsSAR245408 was administered orally, once daily, with continuous dosing in monthly cycles. A total of 25 patients were enrolled at the maximum tolerated dose identified in solid tumor patients as part of the expansion cohort in relapsed/refractory lymphoproliferative malignancies. Plasma cytokines and chemokines were evaluated using the Myriad RBM Human Discovery MAP250+ panel (> 250 analytes) and ELISA assays. ResultsAmong the 25 patients (pts), 40% (n=10) had refractory CLL and 60% (n=15) had various relapsed/refractory lymphomas (R/RL), including follicular lymphoma (FL) (n=5), diffuse large B-cell lymphoma (DLBCL) (n=4), Waldenstrom's macroglobulinemia (WM) (n=3), Hodgkin lymphoma (n=2) and B-cell prolymphocytic leukemia (n=1). The median age was 65 years (range 28–83), and 56% were female. Eighty percent of pts had stage III-IV disease and 48% had bulky disease. Five pts were categorized as having refractory disease and the median number of prior regimens was 1 (range 1-7) for CLL pts and 3 (range 0-9) for R/RL pts. For all 25 patients, the median starting absolute lymphocyte count was 1.15 x 103/μL (range 0.3–37.2), the median starting hemoglobin was 11.4 g/dL (range 9.1–15) and the median platelet count was 162 x 103/μL (range 60–431). The median number of cycles administered in CLL pts was 10 (range 5-24) and 5 (range 1-26) in R/RL pts. Six CLL pts experienced an increase in absolute lymphocyte counts within 2-4 weeks of treatment, as has been seen with other PI3K inhibitors. According to modified International Workshop on Chronic Lymphocytic Leukemia (IWCLL) response criteria, 4 CLL pts had partial response (PR), 1 had nodal PR with increased lymphocytosis, and 5 had stable disease (SD); the progression free survival (PFS) in the responding pts was 22, 21.2, 15.6 and 15.4 months while in the SD pts was 12.5, 9.2, 7.4, 5.6, 4.6 and 3.6 months. According to the modified International Working Group response criteria, 3 PRs were reported in R/RL pts [1 WM, 1 DLBCL (transformed) and one FL with PFS of 23.7, 18.4 and PFS 4.8 months respectively]. One hundred percent of CLL and 80% of R/RL pts reported grade 3 or higher AEs, with the most common (≥ 10% of patients) including neutropenia, diarrhea, anemia and hypotension. SAR245408 induced a reduction in levels of chemokines involved in lymphocyte trafficking in CLL subjects (n=8), including CXCL13, CCL3, CCL22 and CCL19 (64, 58, 52 and 54% reduction, respectively, p<0.05) similar to what was reported with PI3K δ specific inhibitors. In addition, a reduction of tumor necrosis factor receptor 2 (TNFR2) and interleukin-2 receptor alpha (IL-2Rα) levels was observed (63% reduction each, p<0.01). ConclusionsThe recommended phase 2 dose of SAR245408 in solid tumor patients was confirmed as safe and tolerable in patients with CLL and R/RL. Single agent SAR245408 demonstrates clinical activity in patients with relapsed or refractory CLL and promising pharmacodynamic effects on chemokine levels involved in lymphocyte trafficking. Disclosures:Brown:Emergent: Consultancy; Onyx: Consultancy; Sanofi Aventis: Consultancy; Vertex: Consultancy; Novartis: Consultancy; Genzyme: Research Funding; Avila: Consultancy; Celgene: Consultancy, Research Funding; Genentech: Consultancy; Pharmacyclics: Consultancy. Off Label Use: The abstract shows scientific information on SAR245408 which is an investigational product developed by Sanofi. This investigational product is not approved by any health authority for any indication. Egile:Sanofi: Employment. Ruiz-Soto:Sanofi: Employment. Awan:Lymphoma Research Foundation: Research Funding; Spectrum Pharmaceuticals Inc.: Speakers Bureau.

Clinical, Phenotypic, and Genetic Characteristics of Non-CLL Type Monoclonal B Lymphocytosis (MBL)

Abstract 5209Monoclonal B-cell lymphocytosis (MBL) has been established as a diagnostic category to denote the finding of clonal B lymphocytes in otherwise healthy and asymptomatic individuals. MBL likely represents early lesions in lymphoid neoplasms. Immunophenotypically, MBL is divided into CD5+ CLL-type and CD5- non-CLL type MBL, respectively. While most studies have focused on CLL-type MBL and provided evidence of progression of MBL to clinical CLL, the biological nature and clinical relevance of non-CLL type MBL remain unclear. In this study, we have identified and evaluated a series of 18 patients with non-CLL type MBL. This group comprised 10 males and 8 females with a median age of 74 years (range 53–95). All the patients presented initially with a mild to moderate absolute lymphocytosis. Clinical and laboratory evaluations were performed with a follow-up period of 1–20 years (median 8.5). Clonal B lymphocyte populations were confirmed by flow cytometry analysis. All the MBL clones showed negative for CD5, CD10 and CD103, positive for CD19 and CD20, and heterogeneous for CD23, CD25, CD79b, FMC7 and CD38. The phenotypic findings excluded CLL, mantle cell lymphoma and hairy cell leukemia, but may overlap with marginal zone lymphoma or lymphoplasmacytic lymphoma. Cytogenetics were available for 9 patients. Of the cytogenetic abnormalities, 7q deletion was found in two clones, 17 q deletion in two other clones, 11q deletion in one clone and t(2;7) in another clone. An overall tendency was shown toward clonal expansion with an increase in absolute lymphocyte count (ALC) over time (initial mean ALC of 4.9 ± 0.46 × 109/L to a recent mean ALC of 8.3 ± 2.18 × 109/L, p = 0.151). Three patients had an initial ALC >8 × 109/L which remained stable in one patient and decreased to <6 × 109/L in 2 patients. The patient with the longest follow-up of 20 years underwent a completely benign course, while the CD5- clonal B cell population was persistent but non-progressing. Of the progressive clones, three patients had an increase in ALC >10 × 109/L; one patient’s ALC progressed over a period of 9 years to 35 × 109/L that required treatment. Clinically, three patients developed splenomegaly and one patient progressed with lymphadenopathy. Unlike the findings from population screening, this study represents a series of clinical MBL of non-CLL phenotype, and our results indicate the heterogeneity in genetic changes and clonal progression. Disclosures:No relevant conflicts of interest to declare.