ALBERT

Description: Introduction: Hematide™, a novel synthetic PEGylated peptidic compound, binds to and activates the erythropoietin receptor. It is being developed for the treatment of anemia associated with chronic kidney disease and cancer. Objective: To assess the subcutaneous (SC) Hematide™ dose required to increase hemoglobin (Hgb) by ≥1 g/dL in anemic cancer patients on chemotherapy. Safety, PD and PK assessment of Hematide™ are also planned. Method: In an ongoing phase 2, openlabel, multicenter, dosefinding study, Hematide™ is being given SC in 15 patient cohorts every 3 wks for 4 doses. The initial dose is 0.10 mg/kg. Entry criteria include confirmed solid tumor malignancy or lymphoma, ≥9 weeks of chemotherapy, baseline (BL) Hgb ≥8 and

Description: Introduction: Multiple myeloma (MM) is a B-cell neoplasia caused by the proliferation of clonal plasma cells (PCs). MM and benign monoclonal gammopathy of unknown significance (MGUS) are routinely distinguished on the basis of paraprotein concentration, level of PCs infiltration and the presence or absence of other clinical features. Flowcytometric detection of PCs according to the expression of CD38 and CD138 has limitation in discrimination between normal and abnormal PCs, but this is possible in multicolor phenotypic analysis. The aim of this study is to compare the numbers of normal and abnormal PCs in MGUS and MM subjects and to find some parameter useful for evaluation PCs distribution. Materials and methods: 51 newly diagnosed untreated MM patients (63,9±10,0 years old) and 31 non-treated MGUS subjects (64,2±13,8 years old) were analysed. Lysed whole bone marrows were analysed by flowcytometric immunophenotyping and PCs were indentified by expression of CD38, CD138, CD45 and also CD56 and CD19. Results: Discrimination between normal CD19+ PCs and abnormal CD56+ PCs was done on CD38+CD138+ population. Ratio of normal PCs count and abnormal PCs counts (normal/abnormal=N/A) was used to describe a distribution of PCs. Subjects with MGUS had 0,97±1,65% (average±SD) of CD38+CD138+ cells (median 0,45; range 0,03–7,47%) with average N/A ratio 2,91±3,94 (median 1,36; range 0,01–18,44). Newly diagnosed MM patients had 4,96±9,94% of CD38+CD138+ cells (median 0,85; range 0,02–41,80%) with average N/A ratio 1,70±3,03 (median 0,13; range 0–11,5). In 9,7% MGUS subjects evaluation of distibution of PCs showed transformation of MGUS into MM. In some newly diagnosed MM patients (31,4%) CD38+CD138+ plasma cells were positive for CD19 although they were aberrant and these PCs were mostly CD45+. In some patients (9,7% of MGUS; 17,6% of MM) PC did express neither CD19 nor CD56 and this fact may complicate further evaluation of PCs. Conclusions: Results confirmed that in MGUS subjects we can find lower numbers of PCs which are mostly CD45+CD19+. A majority of plasma cells in newly diagnosed MM patients are abnormal CD56+ PCs and these plasma cells are usually CD45−. Further follow-up of patients can confirm the N/A ratio as a predictive factor for transformation of MGUS into MM and its value for evidence of relapse or progression to active myeloma.

Description: Introduction: Hematide™ a novel, synthetic, PEGylated peptidic compound, binds to and activates the erythropoietin receptor. It is in development for treatment of anemia associated with chronic renal failure and cancer. Objective: To assess the subcutaneous (SC) Hematide™ dose required to increase hemoglobin (Hb) by ≥ 1 g/dL in ≥ 50% of anemic cancer patients (pts) on chemotherapy. Safety, pharmacodynamics and pharmacokinetics were also assessed. Method: In a phase 2, open label, multi-center, dose finding study, up to four doses of Hematide™ were given SC every 3 wks (Q3W) to 4 cohorts of fifteen pts at 0.05, 0.10, 0.15 or 0.2 mg/kg each. Entry criteria included confirmed solid tumor malignancy or lymphoma, 〉9 weeks of chemotherapy, baseline (BL) H ≥ 8 and

Description: Introduction: The aim of our study was to analyze the impact of angiotensin-I converting enzyme inhibitors (ACEI) on the outcome of patients with multiple myeloma (MM) treated by autologous peripheral blood stem cell transplantation (APBSCT), as it has been reported that low angiotensin-converting enzyme (ACE) activities are associated with poor prognosis in multiple myeloma (MM). Patients and Methods: Patients with MM who underwent APBSCT (n=168) were studied retrospectively. Patients taking ACEI alone or in combination with other antihypertensive agents during the APBSCT were allocated to the ACEI group (n=25; 15%). The ACEI group was compared both to all other patients (n=143; 85%) and to patients with hypertension who were taking other or no antihypertensive medication (n=44). The induction chemotherapy consisted of 4 cycles of VAD. For stimulation, cyclophosphamide was given, followed by G-CSF. High-dose melphalan was used as conditioning. Results: The ACEI and non-ACEI groups were similar in terms of age, initial levels of albumin, beta2-microglobulin, CRP, LDH, performance status, stage according to IPI and Durie-Salmon criteria, myeloma type, and maintenance treatment. Patients with hypertension (n=69) and patients without hypertension (n=99) had similar overall survival (OS) after APBSCT (65.5 months versus 60.9, respectively; p=0.561), and progression-free survival (PFS) after APBSCT (32.6 months versus 29.6 months; p=0.635). Patients taking ACEI had significantly worse OS after APBSCT (32.7 months) compared to patients not taking ACEI (65.2 months; p=0.033). Among patients with hypertension, OS was significantly shorter in the ACEI group than in the non-ACEI patients (p=0.024). Statistically significant differences were also observed in PFS between hypertensive patients treated by ACEI and other hypertensive patients (19.3 months versus 48.6 months, respectively; p=0.041). Most deaths in both groups were myeloma-related. Renal failure was a main or contributing cause of death in 3 patients from the ACEI group and in 8 patients from the non-ACEI group (p=0.233). Also, there was a trend favoring non-ACEI hypertensive patients in comparison to ACEI hypertensive patients in the number of complete responses after PBSCT (40% versus 17%; p=0.058). Conclusion: In this retrospective study we have found that patients with MM and hypertension treated with ACEI at APBSCT have worse survival in comparison to patients with MM receiving other or no antihypertensive drugs. Further research in this area could lead to important insights into the pathophysiology of MM and to the reevaluation of ACE activity as a prognostic factor in MM.

Description: Tumor suppressor p53 is a powerful transcription factor responsible for cell cycle regulation, which can bind about 300 different promoter elements in the human genome. Disruption of p53 function by mutation or deletion belongs to the most frequent alterations observed in human cancers, in B-CLL ranging from 10 to 15%. In our cohort of 230 B-CLL patients, we detected 10.5% patients with Tp53 mutation, 67% of these exhibited a point mutation (single nucleotide switch) largely combined with deletion of the second allele. The detected mutations are mostly localized in the DNA binding domain of p53 gene and, in accordance with the IARC database, are clustered into two hot-spot regions between amino acids 234–249 (32% of the mutations) and 273–282 (25% of the mutations). B-CLL patients with mutant Tp53 have mostly non-mutated IgVH (95%) and a significantly worse prognosis (17% of patients with Tp53 mutation and only 3% of patients having wt-p53 died within 2 years after diagnosis). In addition, 92% of patients with mutated p53 required therapy; in the non-mutated group this number represented only 54% of the respective patients. Due to these findings detection of p53 status became an essential part of B-CLL diagnostics in our laboratory. However, recent results suggest existence of significant differences in the properties of specific mutants, especially regarding their DNA-binding ability to activate particular promoters (e.g. Pospisilova et al., Mol. Cancer Res. 2: 296–304, 2004). Presence of mutation need not represent a total loss of protein activity but, to the contrary, some amino acid substitutions can evoke a gain of function of the respective p53 mutant. Therefore, it seems to be very important to determine the specific type of amino acid substitution in a particular B-CLL patient and detect ability of the respective p53 mutant to induce cell cycle arrest and/or apoptosis. We used functional protein arrays bearing p53 mutants to determine their DNA-binding activity towards promoter sequences of different p53-response genes such as mdm2, p21/waf1, pig3 or bax. Several analyzed p53 mutants displayed DNA-binding ability similar or even higher than wt-p53 towards all studied promoters (e.g. mutant Arg337Cys) suggesting gain of function. Interestingly, not only the amino acid localization but also type of substitution in the same position can influence protein activity. The most frequent mutational hotspots in B-CLL and majority of other cancers, Arg248 and Arg273, displayed significantly higher DNA-binding activity of mutant variants Arg248Gln and Arg273His in comparison to Arg248Trp and Arg273Cys. These experimental results were validated by protein modeling clearly demonstrating respective structural constraints, such as prevented binding of Trp248 to the promoter region DNA groove for sterical and hydrogen-bonding reasons. In conclusion, our results of B-CLL patient analysis demonstrate importance of the presence of p53 mutation for B-CLL treatment and prognosis. We also document the significance of localization and type of amino acid substitution for p53 mutant activity. These factors represent a potentially valuable marker for grading of p53 mutant functionality in B-CLL patients and tumor prognosis.

Description: Because information on management and outcome of AML relapse after allogeneic hematopoietic stem cell transplantation (HSCT) with reduced intensity conditioning (RIC) is scarce, a retrospective registry study was performed by the Acute Leukemia Working Party of EBMT. Among 2815 RIC transplants performed for AML in complete remission (CR) between 1999 and 2008, cumulative incidence of relapse was 32% ± 1%. Relapsed patients (263) were included into a detailed analysis of risk factors for overall survival (OS) and building of a prognostic score. CR was reinduced in 32%; remission duration after transplantation was the only prognostic factor for response (P = .003). Estimated 2-year OS from relapse was 14%, thereby resembling results of AML relapse after standard conditioning. Among variables available at the time of relapse, remission after HSCT 〉 5 months (hazard ratio [HR] = 0.50, 95% confidence interval [CI], 0.37-0.67, P 〈 .001), bone marrow blasts less than 27% (HR = 0.53, 95% CI, 0.40-0.72, P 〈 .001), and absence of acute GVHD after HSCT (HR = 0.67, 95% CI, 0.49-0.93, P = .017) were associated with better OS. Based on these factors, 3 prognostic groups could be discriminated, showing OS of 32% ± 7%, 19% ± 4%, and 4% ± 2% at 2 years (P 〈 .0001). Long-term survival was achieved almost exclusively after successful induction of CR by cytoreductive therapy, followed either by donor lymphocyte infusion or second HSCT for consolidation.