ALBERT

Description: Aim/Background CNS relapse in DLBCL is a devastating complication and the optimum strategy for prevention remains unclear. Methods We performed a multi-centre, retrospective analysis of CNS relapse rates in patients (pts) identified at risk of CNS relapse according to the type of CNS-directed prophylaxis administered. Pts receiving initial therapy for DLBCL between 1996 and 2011 (to allow ≥2 years of follow up) were included; DLBCL after histologic transformation of low-grade lymphoma and HIV-associated DLBCL were included, however pts with Burkitt or Burkitt-like lymphoma or CNS involvement at diagnosis were excluded. Selection for CNS prophylaxis strategy was by the primary managing hematologist if they fulfilled ≥2 of the following criteria: 1) multiple extranodal sites 2) raised serum LDH 3) B-symptoms, OR involvement of specific high-risk anatomical sites. We compared 3 prophylaxis strategies: prior to 2003 intrathecal (IT) methotrexate (MTX) in conjunction with CHOP chemotherapy “group 1” was the main strategy; from 2003 onwards, R-CHOP (mostly) with IT MTX was followed by two cycles of high dose intravenous (IV) MTX (1-3g/m2) “group 2”; patients

Description: Abstract 2652 Introduction Despite improvements in cure rates for patients with diffuse large B-cell lymphoma (DLBCL), up to 40% relapse after achieving initial remission, mostly within 18 months from treatment. There is no consensus as to role, or most appropriate form of post-remission surveillance. Our aim was to explore the role of Positron Emission Tomography combined with computer tomography (PET-CT) scanning in the follow up of patients with diffuse large B-cell lymphoma (DLBCL) achieving complete metabolic response (CMR) after primary therapy, identify patterns of relapse and define a risk-adapted strategy. Results We included 116 patients with de novo DLBCL treated at our centre between 2002 and 2009 with a negative post-treatment PET-CT, and at least one surveillance PET-CT scan. International Prognostic Index (IPI) was

Description: Background: Pre-existing thrombocytopenia in MDS/AML is worsened during the initial cycles of azacitidine (AZA) therapy, resulting in bleeding risk and possible platelet transfusion. Eltrombopag (EPG) is an oral TPO-receptor agonist. In vitro, it has anti-proliferative effects on AML blasts. Aim: 
To assess the safety of escalated doses of EPG in patients undergoing AZA for MDS/AML Method
An investigator-initiated phase-II, single arm, study of EPG with AZA. Inclusion: relapsed or de-novo MDS/CMML/AML (blasts 5-30%); or symptomatic cytopenia; or blasts 31-50% if 〉/=65 years or previously-treated disease; and platelets Result
Of 25 patients, 10 had prior therapy, which was chemo in 7; of these 6 had blasts 〉/=10%, 2 with blasts 20% or above. Of the 15 de-novo patients, 7 had blasts 〉/=20% (AML) and a further 2 had blasts between 10 and 19%. The median platelet count was 38x10^9/L (range 8-127). A median 11(2-24) cycles AZA and 6 cycles of EPG were delivered. One patient developed GrII EPG-related LFT abnormalities (resolved). Grade 3 fatigue was attributed to the combination in one patient. Thrombocytosis (〉450 x109/L) resulting in EPG cessation occurred in 6 (at 50, 50, 150 and three at 200mg), without complications. 10 patients experienced reversible skin yellowing. Response/improvement was seen in 18 (72%): 7CR, 3CRm, 5HI-P,1 HI-N,2 with 〉50% blast reduction from 〉20% (8%). 5 patients had progression at first response. There were 19 events in total (17 progressions, 2 with worsening haematology). Median PFS was 12.0 months (95% CI 5.6 to 24.3 months). Median OS was 15.3 months (95% CI 11.9 to 31.7 months). Platelet improvement was seen in 54% (13/24) of patients with baseline platelets

Description: Background Patients (pts) with TrIL have inferior outcomes compared with de novo diffuse large B-cell lymphoma (DLBCL), and their optimum follow up is not well defined. We sought to determine the utility of surveillance PET-CT in pts with TrIL achieving CMR after primary therapy and identify patterns of relapse. Methods We performed a retrospective analysis of pts with TrIL treated at Peter MacCallum Cancer Centre between 2002 and 2012 who achieved CMR after primary therapy who had ³1 subsequent surveillance PET-CT. In the period analysed, departmental protocol recommended 6-monthly scans for pts in CMR for the first 2 years, then annually until 5 years after completion of therapy, if there was intention to intervene on detection of subclinical relapse. A positive scan suggested relapsed lymphoma, with true positive results requiring either biopsy confirmation or unequivocal scan progression. A false positive scan was refuted by biopsy and/or follow up showing resolution of areas of increased FDG uptake. A negative scan was interpreted as negative for relapsed lymphoma: true negatives had no clinical relapse and false negatives manifest relapse within three months from the date of the scan. Indeterminate scans were recorded if determination could not be made. Results The cohort included 55 pts with TrIL: 38 underwent stem cell transplant (autologous, n= 37; allogeneic, n=1) as consolidation; 17 did not. (Table 1). After a median follow-up of 34 (range 3 – 101) months, the actuarial 3-year progression free (PFS) and overall survival (OS) were 77% (95% CI 62 – 86%) and 88% (75 – 94%) respectively. Multiple potential prognostic factors including IPI, stage, serum LDH, timing of transformation (simultaneous diagnosis of transformation versus delayed) and type of indolent histology were not predictive of PFS. Of 180 surveillance PET-CT scans, there were 153 true negatives, 4 false positives, 1 false negative, 7 indeterminate and 15 true positives (Table 2). Considering indeterminate scans as false positives, the specificity of PET-CT for detecting relapse was 93%, sensitivity 93%, positive predictive value 54% and negative predictive value 99%. Of the 15 pts who experienced disease relapse, 7 (47%) were subclinical (i.e. detected by surveillance PET-CT scans) whilst 8 (53%) were suspected on the basis of clinical symptoms. Although 5% of scans in the first 2 years detected a subclinical relapse, all of these were either biopsy or clinically shown to be low-grade lymphoma. All 8 symptomatic relapses (at 2 – 42 months), in contrast were DLBCL. Conclusion In pts with TrIL achieving CMR, PET-CT detects subclinical relapses of low-grade histology with high sensitivity but with a moderate false-positive rate. This is of limited clinical benefit as the initiation of further therapy in these circumstances is rarely based on imaging findings alone. In contrast, all DLBCL relapses in our cohort were accompanied by clinical symptoms. Thus, surveillance imaging of pts with TrIL achieving CMR is not indicated. PET-CT should be reserved for evaluation of suspected relapse only. Disclosures: No relevant conflicts of interest to declare.

Description: Abstract 4180 High risk acute myeloid leukaemia (AML) and myelodysplastic syndrome (MDS) may be amenable to immunotherapy with adoptive transfer of chimeric antigen receptor (CAR) T cells. However, persistence and generation of central memory (CM) are essential for ongoing clinical responses (Morgan et al Science 2006). Our prior work showed that the carbohydrate antigen LewisY (LeY) is expressed in 46% of AML patients (n=33) to varying degrees, making it a suitable and novel target for CAR therapy. In our phase I study, we have assessed the trafficking, persistence and functional capacity of CAR-T cells transduced with an anti-LeY chimeric receptor gene in high-risk patients with LeY positive AML or MDS. METHOD: The CAR comprised extracellular humanized scFv recognizing the LeY Ag, linked to an extracellular CD8 hinge region, a transmembrane and cytoplasmic CD28 and CD3 zeta signalling domain. The humanized anti- LeY scFv-CD28-ζ receptor vector was produced as described previously (Westwood J et al PNAS 2005). T cells, harvested from the patient, were transduced with LeY scFv-CD28-ζ pSAMEN retroviral vector. Bulk transduced T cells (LeY-T) were re-infused into the patient after lymphodepleting fludarabine chemotherapy. AML immunophenotyping and cytogenetics were used to monitor minimal residual disease (MRD). PB and BM samples were collected prior to and post CAR-T adoptive transfer. An optimized PCR assay (sensitivity 1:1e5) for the presence of the LeY transgene (TG) was performed on genomic DNA extracted from each sample. To measure both CAR-T cell functional polarization and persistence of CAR expression, PB or BM cells were co-cultured with a cell line expressing LeY (OVCAR-3), in the presence or absence of MHC class I or II blocking antibodies. Culture supernatant was collected at 24 hours and assessed by Luminex assay for Th1 (IFN-g, IL-2), Th2 (IL-4, IL-10), pro-inflammatory (IL-6, IL-17 and TNF) cytokines and TGF-b. RESULTS: Five AML patients have been enrolled to date. Four patients have had sufficient CAR-T cells generated to provide doses of 0.5 × 109 −1.3 × 109T cells (transduced T cells: 8 – 30% of total T-cells]) with a viability of 96.1– 98.5%. 2 patients (01 and 04) had progressive disease 1 and 2 months after infusion. 2 patients (02 and 05) remain in morphologic remission with stable cytogenetic MRD, 3 and 14 months post infusion. Patients 01, 02 and 04 all had LeY TG in PB and BM samples at all time points measured, up to day 49 in patient 01 (Figure 1) and up to 10 months in patient 02. In patient 04 LeY TG was also detected in a skin biopsy (day +6) of leukemic skin infiltrate. Co-culture supernatant cytokine analysis showed LeY-T cells from patient 01, 02 and 04 secreted high levels of IFN-g (805.5 ± 198.9 pg/ml) and low levels of IL-2 (3.14 ± 0.65 pg/ml) prior to adoptive transfer. In contrast, post adoptive transfer, co-culture with OVCAR-3 cells, and MHC class I and II blocking antibody, induced these PB LeY-T cells to secrete IL-4 (24.13 ± 12.47pg/ml) and IL-10 (9.35 ± 0.44 pg/ml); no IFN-g or IL-2 was detected. This Th2 polarization of CAR-T cells was shown in PB samples taken at days 5 to 28 (patient 01), day 28 to 8 months (patient 02) and day 1 to day 28 (patient 04) post-adoptive transfer (Figure 2). In addition, when Th1/Th2 cytokines were measured in patient 02 PB and BM plasma, Th1 cytokines were detected immediately post-adoptive transfer (IFN-g=39 pg/ml, IL-2=8.6 pg/ml at peak); however these decreased to basal levels by day 3 (IFN-g=1.8 pg/ml) and day 2 (IL-2=2.6 pg/ml). The same trend in serum cytokines was observed for patient 01. Therefore, our studies reveal that, after adoptive transfer, LeY CAR-T cells persist in vivo, maintain their expression of the CAR, but show rapid polarization from a Th1 to a Th2 phenotype. Disclosures: No relevant conflicts of interest to declare.

Description: Aim/Background HDACi induce cancer cell apoptosis and are in use for T-cell lymphoma. Prior studies have indicated that HDACi are immune-suppressive. In this study, we address this issue by examining the effects of two different HDACi on human monocyte derived dendritic cell (hmDC) biology. Methods hmDC were activated by TLR stimuli (LPS or poly:IC). HDACi effect on TLR-induced hmDC activation was assessed by up regulation of co-stimulatory molecules CD80, CD86 and CD83 along with secretion of IL-12p70, IL-10 and IL1β. To address the mechanism for drug-induced altered cytokine secretion mRNA for IL-12p70, IL-10 and IL1β were assessed followed by mir-155 mRNA and SOCS-1 protein. The functional outcome was assessed in an allogeneic MLR. Results At doses sub-optimal to induce myeloma cell death, both drugs significantly inhibited TLR-induced hmDC increase in CD80 and CD83, but not CD86. TLR-induced secretion of IL-10 and IL12p70 was reduced while IL-1β secretion was increased. This occurred regardless of drug sequencing. HDACi did not significantly alter expression of IL-12p70, IL-10 or IL-1β mRNA despite changes in protein levels. To reveal the mechanism for the HDACi-induced hmDC IL-12p70 defect, the upstream molecules miR-155 and SOCS-1 were examined. Interestingly, panobinostat but not romidepsin induced mir-155 down-regulation and SOCS-1 protein increase. Subsequently, allogeneic T-cells co-cultured with HDACi/TLR-ligand-treated hmDC secreted lower levels of cytokines (decreased secretion of IFN-γ, IL-2, TNF, IL-4 and IL-17) compared to those allogeneic T cells co-cultured with control hmDC. Conclusion In this study, we show that whilst HDACi impair DC maturation and modulate subsequent T cell responses. Our findings suggest that panobinostat induces the IL-12p70 effect via the mir-155/SOCS-1 pathway, whereas romidepsin induces the effect via a mir-155 independent pathway. This data suggests that HDACi affect the hmDC maturation response to pathogen, and that these drugs may alter the ability of treated patients to raise an effective immune response to pathogens or, indeed, tumour antigens. Disclosures: No relevant conflicts of interest to declare.

Description: Background: RG7388 is a new, potent, oral, nutlin-class MDM2 antagonist. This trial evaluated its use in AML to determine the recommended phase 2 dose (RP2D), dose-limiting toxicities (DLTs), pharmacokinetics (PK), pharmacodynamics and clinical responses. Methods: This multicenter, open-label phase 1/1b dose escalation (DE) study evaluated RG7388 as monotherapy (daily ´ 5 d q28d) (Part 1 DE) and in combination with cytarabine (ara-C 1 g/m2 IV x 6 d q28d) (Part 2 DE). Extensions (Ex) were initiated at the RP2D. Part 1Ex (RG7388) included pts 〉 70 y or 〉 60 y with comorbidities. Part 2Ex pts (RG7388 ± ara-C) had relapsed/refractory (R/R) AML, ≤ 2 regimens and no antecedent hematologic disorders (AHD) or transplant (ASCT). Blood and marrow analyses included PK, TP53 mutations (mt) by AmpliChip, serum MIC-1 and MDM2gene expression. Results: To date, 86 pts have been treated. DE is complete, Part 1Ex was discontinued after 9 pts and 34 of 40 planned pts are enrolled in Part 2Ex. One DLT of prolonged myelosuppression was reported. RP2D is 1200 mg/d (600 mg bid) x 5 d q28d for both mono- and combination therapy due to diarrhea not formally a DLT but felt to be dose-limiting. The most common adverse events were GI (diarrhea reported by 〉 85% of pts) or infection-related (〉 70% of pts). Twenty pts received monotherapy during Part 1 DE at 400 (n = 2), 800 (n = 6) and 1600 mg (n = 12) daily x 5 d. Median age was 68.5 y (range 28-83 y), median prior therapies = 2 (range 0-4), 1 pt had ASCT and 8 had AHD. Four pts died in the first 30 days. Five pts achieved either a CR (n = 2) or CRi/MLFS (n = 3). In Part 1Ex, 9 pts were treated with RG7388 at the RP2D. Median age was 75 y (range 66-83), 8 pts had AHD (6 prior hypomethylators, 3 pts lower intensity therapy) and 1 had prior solid tumor. Best responses reported were 1 CRi/MLFS, 1 PR, 1HI, 3 PD. Three pts died in the first 30 days. Further enrollment in this arm was discontinued as induction of prolonged myelosuppression increased the risks of infection and early deaths. Combination treatment with lower RG7388 doses may be better tolerated in this fragile elderly population. Twenty-three pts were enrolled in Part 2 DE (RG7388 + ara-C) at daily doses of 400 (n = 10), 800 (n = 7), and 1200 mg (n = 6); median age 64 y (range 32-76); median prior therapies = 1.5 (range 0-5); prior ASCT 2; AHD 4; 5 had prior malignancies. Four pts died in the first 30 days. Six relapsed AML pts achieved CRs (4 received prior ara-C and 2 had prior hypomethylators). To date, 34 of 40 planned pts with R/R AML have been enrolled to Part 2Ex (RG7388 ± ara-C); 23 pts have responses reported, with 4 CRs, 1 CRi, 1 PR, 1 HI, 16 PD. Three pts died in the first 30 days. T1/2 is ≈ 1 d, irrespective of age, concomitant azoles or ara-C. Bone marrow levels are ≈ 70% of plasma drug levels at steady state. CRs were seen in diverse pts, including varied risk groups, prior AHD, therapy-related AML (t-AML), p53 mt, R/R and de novo AML. All pts who achieved a CR during DE were relapse free for 〉 60 d. One pt on monotherapy and 2 on combination therapy remained relapse free for 〉 400 d and 〉 200 d from start of study, respectively. A potential predictive gene expression signature correlated with RG7388 therapy (AUC = 0.73, p = 0.021). Conclusions: We report the Ph1/1b PK, safety and clinical activity of a new, potent MDM2 antagonist in AML. CRs occur rapidly and are durable (〉 60 d) in elderly AML pts with RG7388 monotherapy and in R/R pts with combination therapy. Table Patient characteristics by response to treatment Part 1 DE (N = 20) 16 responses Best response 2 CR; 3CRi/MLFS 3 PR 4 HI 4 PD TP53 status WT WT 1 MT, 3 WT 1 V ELN* AHD(responders) 4 I, 1 A 3 (MDS, MF, CMML) 3 I 2 (MDS, CMML) Part 1Ex (N = 9) 6 responses Best response 1 CRi/MLFS 0 PR 2 HI 3 PD TP53 status WT WT, MT 2 WT, 1 U ELN* AHD(responders) I ET Part 2 DE (N = 23) 20 responses Best response 6 CR 2 PR 2 HI 10 PD TP53 status 1 MT WT WT 1 MT, 1 V ELN* AHD(responders) 1 F, 3 I, 2 A 1 t-AML 1 I, 1 A Part 2Ex (N = 34) 23 responses Best response 4 CR; 1 CRi/MLFS 1 PR 1 HI 16 PD TP53 status 4 WT, 1 U WT WT 1 MT, 5 WT, 10 U ELN*(responders) 1 F, 3 I, 1 A A CR, complete remission; CRi, complete remission with incomplete recovery; HI, hematologic improvement; MLFS, morphologic leukemia-free state; PD, progressive disease; PR, partial response; WT, wild type; V, splice variant; U, unknown/pending; AHD, includes myelodysplastic syndrome (MDS), essential thrombocythemia (ET), chronic myelomonocytic leukemia (CMML), myelofibrosis(MF) *ELN: Risk by European Leukemia Net: favorable (F), Intermediate I and II (I), or adverse (A) Disclosures Yee: Roche: Research Funding. Martinelli:Pfizer: Consultancy, Speakers Bureau; BMS: Consultancy, Speakers Bureau; Novartis: Consultancy, Speakers Bureau; ARIAD: Consultancy, Speakers Bureau. Vey:Roche: Honoraria. Assouline:Roche: Honoraria, Research Funding; Janssen: Honoraria. Drummond:Novartis: Consultancy, Honoraria, Speakers Bureau; Celgene: Consultancy, Honoraria, Speakers Bureau. Blotner:Roche: Employment. Higgins:Roche: Employment. Middleton:Roche: Employment. Nichols:Roche: Employment. Chen:Roche: Employment. Zhong:Roche: Employment. Pierceall:Roche: Employment. Zhi:Roche: Employment. Chen:Roche: Employment.

Description: Geostationary satellites have provided routine, high temporal resolution Earth observations since the 1970s. Despite the long period of record, use of these data in climate studies has been limited for numerous reasons, among them that no central archive of geostationary data for all international satellites exists, full temporal and spatial resolution data are voluminous, and diverse calibration and navigation formats encumber the uniform processing needed for multisatellite climate studies. The International Satellite Cloud Climatology Project (ISCCP) set the stage for overcoming these issues by archiving a subset of the full-resolution geostationary data at ~10-km resolution at 3-hourly intervals since 1983. Recent efforts at NOAA's National Climatic Data Center to provide convenient access to these data include remapping the data to a standard map projection, recalibrating the data to optimize temporal homogeneity, extending the record of observations back to 1980, and reformatting the data for broad public distribution. The Gridded Satellite (GridSat) dataset includes observations from the visible, infrared window, and infrared water vapor channels. Data are stored in Network Common Data Format (netCDF) using standards that permit a wide variety of tools and libraries to process the data quickly and easily. A novel data layering approach, together with appropriate satellite and file metadata, allows users to access GridSat data at varying levels of complexity based on their needs. The result is a climate data record already in use by the meteorological community. Examples include reanalysis of tropical cyclones, studies of global precipitation, and detection and tracking of the intertropical convergence zone.

Description: Geostationary satellites have provided routine, high temporal resolution Earth observations since the 1970s. Despite the long period of record, use of these data in climate studies has been limited for numerous reasons, among them: there is no central archive of geostationary data for all international satellites, full temporal and spatial resolution data are voluminous, and diverse calibration and navigation formats encumber the uniform processing needed for multi-satellite climate studies. The International Satellite Cloud Climatology Project set the stage for overcoming these issues by archiving a subset of the full resolution geostationary data at approx.10 km resolution at 3 hourly intervals since 1983. Recent efforts at NOAA s National Climatic Data Center to provide convenient access to these data include remapping the data to a standard map projection, recalibrating the data to optimize temporal homogeneity, extending the record of observations back to 1980, and reformatting the data for broad public distribution. The Gridded Satellite (GridSat) dataset includes observations from the visible, infrared window, and infrared water vapor channels. Data are stored in the netCDF format using standards that permit a wide variety of tools and libraries to quickly and easily process the data. A novel data layering approach, together with appropriate satellite and file metadata, allows users to access GridSat data at varying levels of complexity based on their needs. The result is a climate data record already in use by the meteorological community. Examples include reanalysis of tropical cyclones, studies of global precipitation, and detection and tracking of the intertropical convergence zone.