Granulocyte Growth Research Articles

The Genetic Adjuvants Interleukin-12 and Granulocyte-Macrophage Colony Stimulating Factor Enhance the Immunogenicity of an Ebola Virus Deoxyribonucleic Acid Vaccine in Mice.

In previous studies, we showed that deoxyribonucleic acid (DNA) vaccines expressing codon-optimized filovirus envelope glycoprotein genes protect mice and nonhuman primates from viral challenge when delivered by intramuscular (IM) electroporation (EP). To determine whether we could achieve equivalent immunogenicity and protective efficacy by a simplified delivery method, we generated DNA vaccine plasmids expressing genetic adjuvants to potentiate immune responses. We tested the Th1-inducing cytokine interleukin-12 and the granulocyte growth factor granulocyte-macrophage colony stimulating factor, both of which have demonstrated significant adjuvant effect when included in clinical DNA vaccine formulations. In addition, because interferon (IFN)-αβ is required for DNA vaccine-induced immunity, we tested inclusion of a potent stimulator of the IFN-αβ pathway. Our data suggest that IM vaccination of mice with plasmid DNA encoding genetic adjuvants enhances vaccine immunogenicity, resulting in increased anti-Ebola virus (EBOV) immunoglobulin G and T-cell responses. Codelivery of genetic adjuvants also improved EBOV neutralizing capability compared with vaccine alone. Finally, IM vaccination with plasmid EBOV and genetic adjuvants provided complete protection against EBOV challenge. Overall, our data suggest that codelivery of genetic adjuvants with filovirus DNA vaccines using IM delivery can provide comparable efficacy to the same DNA vaccines when delivered using IM-EP devices.

Monoinstitutional real world experience in management of Vinflunine as second line therapy for transitional cell carcinoma of the urothelium.

Vinflunine is the only cytotoxic agent tested as a second line therapy in transitional cell carcinoma of the urothelium in a phase III trial. It is not largely employed in clinical practice because of the high incidence of grade 3–4 toxicity. We evaluated efficacy and safety of Vinflunine at the dose of 280 mg/m2 every 3 weeks associated with primary prophylaxis with granulocyte growth factors and laxatives for patients progressed after platinum + Gemcitabine. Overall survival was 8.5 months, progression-free survival 4.33 months and response rate 25%, with disease control rate 57.2%. Grade III-IV neutropenia occurred in 10.7% of the patients, grade III-IV anemia and grade III thrombocytopenia in 10.7% and 7.2%, respectively. Among non haematological toxicity, grade I-II constipation was reported in 14.2% of the patients, without grade III-IV adverse events. No discontinuation for toxicity was observed. This study underlines that Vinfluinine at a dose of 280 mg/m2 associated with primary prophylaxis for neutropenia and constipation is effective and with a favorable toxicity profile.

Sequential Brentuximab Vedotin (Bv) before and after Adriamycin, Vinblastine, and Dacarbazine (Bv-AVD) for Older Patients with Untreated Classical Hodgkin Lymphoma (cHL): Final Results from a Multicenter Phase II Study

Background: Standard therapeutic regimens for older HL patients (pts) may confer significantly increased toxicity and inferior survival compared with younger pts. Reported complete remission (CR) rates for older pts are 45-76% with 2-year (yr) progression-free survival (PFS) rates of 50-71% (eg, Evens et al. Blood 2012). With a goal of improving outcomes for older cHL pts, this multicenter phase 2 study tested Bv given sequentially before and after standard AVD chemotherapy. Methods: Older pts (aged ≥60 yrs) with stage IIB-IV, untreated cHL were eligible. Pts received 2 9lead-in9 doses of single-agent Bv 1.8 mg/kg (q 3 weeks) followed by 6 cycles of standard AVD. Supportive antibiotics were encouraged & granulocyte growth factor was allowed. Responding pts proceeded to consolidation (Cx) therapy with 4 Bv cycles. Study design was a Simon 2-stage; the primary endpoint was CR rate after AVD (ie, prior to Bv Cx) using revised Cheson with FDG-PET; pts must have received 2 cycles of AVD to be evaluable for response. The study was considered promising if >70% of evaluable pts demonstrated CR. Univariate & multivariate analyses (MVA) were performed using Cox proportional hazard regression for associations between lab and clinical factors (including co-morbidities assessed by the Cumulative Illness Rating Scale (CIRS)) with survival. Results: 48 pts enrolled to the study (8/2012-8/2016) with 41 being evaluable for response. Characteristics for all pts included: median age 69 yrs (60-88); 30M:18F; median ECOG PS of 1 (21% PS=2); 82% stage III/IV disease (bone marrow involved 23%); and IPS 3-7 in 60%. No pts had loss of activities of daily living (ADL) at baseline, while 12% had loss of instrumental ADLs. Median CIRS co-morbidity score at baseline was 6 (0-20). 6 pts received 1 pt were neutropenia (60%); infection (15%), febrile neutropenia (8%); transaminitis (6%); renal insufficiency (6%); urinary infection (6%); pneumonia (6%); hyponatremia (6%); fatigue (6%); febrile neutropenia (6%); diarrhea (4%); pancreatitis (4%), & peripheral neuropathy (PN) (4%). 33% of all pts experienced grade 2 PN (6% motor/27% sensory); the majority were reversible. Treatment related mortality on study was 2% (n=1 pancreatitis). Reasons for study discontinuation were: completed treatment (52%); toxicity/AEs (33%); withdrew consent/refused additional treatment (9%); & progressive disease or death (6%). Finally, for pt prognostication, increasing age (continuous) (P=0.005), female (P=0.05) & increased CIRS (continuous) (P=0.006) were associated with inferior PFS on univariate analysis; on MVA, only increasing age remained significant (HR 1.19 per yr >60, 95%CI 1.02-1.37, P=0.02). Conclusions: Bv-AVD incorporating Bv sequentially before and after chemotherapy represents among the best-reported outcomes to date for untreated older cHL pts. Efforts to maintain these robust remission and survival rates, but with less toxicity, should be a focus of ongoing investigation. This should include response-adapted design and integration of other novel agents (eg, checkpoint inhibitors), especially for pts with advanced ages and/or multiple co-morbidities. Disclosures Evens: Novartis: Consultancy; Affimed: Consultancy; Merck: Consultancy; Kite Pharma: Consultancy; Amgen: Consultancy; Celgene: Consultancy; Millennium: Consultancy; AbbVie: Consultancy; Pharmacyclics: Consultancy; Seattle Genetics: Consultancy; • Spectrum Pharmaceuticals: Consultancy. Advani: Spectrum: Consultancy; Gilead: Consultancy; Agensys: Research Funding; Bayer Healthcare Pharmaceuticals: Research Funding; Merck: Research Funding; Celgene: Research Funding; Seattle Genetics: Research Funding; Kura: Research Funding; Regeneron: Research Funding; Bristol-Myers Squibb: Consultancy, Research Funding; Nanostring: Consultancy; FortySeven: Research Funding; Janssen: Research Funding; Cell Medica: Research Funding; Millennium: Research Funding; Pharmacyclics: Consultancy; Sutro: Consultancy; Juno Therapeutics: Consultancy; Pharmacyclics: Research Funding; Infinity: Research Funding; Genentech: Research Funding. Fanale: ONYX: Research Funding; CELGENE: Honoraria, Membership on an entity9s Board of Directors or advisory committees, Research Funding; TAKEDA: Honoraria, Research Funding; AMGEN: Membership on an entity9s Board of Directors or advisory committees; BMS: Honoraria, Membership on an entity9s Board of Directors or advisory committees, Research Funding; SEATTLE GENETICS: Honoraria, Membership on an entity9s Board of Directors or advisory committees, Research Funding; GENENTECH: Research Funding; MERCK: Membership on an entity9s Board of Directors or advisory committees, Research Funding; MOLECULAR TEMPLATES: Research Funding; ADC THERAPEUTICS: Research Funding. Winter: Merck: Research Funding; Glaxo-Smith-Kline: Research Funding. Gordon: Janssen: Other: Data Monitoring Committee. Hamlin: Celgene: Consultancy, Honoraria; Portola: Consultancy, Honoraria, Other: research support; Incyte: Other: research support; Gilead: Consultancy, Honoraria; Novartis: Other: research support; Seattle Geneitcs: Other: research support.

Features morphofunctional structure indicators of the red bone marrow in senile age

We studied the histology of bone marrow taken by needle biopsy in 18 elderly people (cadavers). Myelogram analysis shows that residents of Karabalta increased number of lymphocytes, segmented and young cell populations, to a lesser extent reduced the number of erythroblasts, granulocyte growth. These changes are due to the influence of exogenous, endogenous and demographic factors.

Potential Mechanisms of Hematological Adverse Drug Reactions in Patients Receiving Clozapine in Combination With Proton Pump Inhibitors.

Clozapine is a second-generation antipsychotic which has proven efficacy in treating the symptoms of schizophrenia. Although clozapine therapy is associated with a number of adverse drug reactions, it is frequently used. One of the most common adverse drug reactions is gastroesophageal reflux disease which is an indication for treatment with proton pump inhibitors (PPIs). Coadministration of clozapine and PPIs increases the risk of hematological adverse drug reactions, including neutropenia and agranulocytosis. The mechanism in idiosyncratic agranulocytosis is not dose related and involves either a direct toxic or an immune-allergic effect. It is suspected that the clozapine metabolites nitrenium ion and N-desmethylclozapine may cause apoptosis or impair growth of granulocytes. Formation of N-desmethylclozapine is correlated with activity of the cytochrome P450 enzymes 1A2 and 3A4 (CYP1A2 and CYP3A4). Nitrenium ion is produced by the flavin-containing monooxygenase system of leukocytes. A drug interaction between clozapine and a PPI is a consequence of the induction of common metabolic pathways either by the PPI or clozapine. Findings to date suggest that indirect induction of flavin-containing monooxygenase by omeprazole through the aryl hydrocarbon receptor increases the expression of the enzyme mRNA and in the long term may cause the increase in activity. Moreover, induction of CYP1A2, especially by omeprazole and lansoprazole, may increase the serum concentration of N-desmethylclozapine, which can accumulate in lymphocytes and may achieve toxic levels. Another hypothesis that may explain hematological adverse drug reactions is competitive inhibition of CYP2C19, which may contribute to increased serum concentrations of toxic metabolites.

Deficiency of Spicing Factor SF3B1 Impairs Human Erythropoiesis and Leads to Changes in Pre-mRNA Splicing

Pre-mRNA splicing is a fundamental process in eukaryotes and emerges as an important co-transcriptional or post-transcriptional regulatory mechanism. More than 90% of multiple-exon genes undergo alternative splicing, enabling generation of multiple protein products from a single gene. In the context of erythropoiesis, one classic example is the splicing of protein 4.1R alternative exon 16. This exon is predominantly skipped in early erythroblasts but included in late stage erythroblasts. In addition, alternative isoforms of various erythroid transcripts have been reported. More recently, we and others have documented that a dynamic alternative splicing program regulates gene expression during terminal erythropoiesis. These findings strongly suggest the roles of alternative splicing and associated regulatory factors in erythropoiesis. However, the studies on the roles of mRNA splicing in erythropoiesis are very limited.RNA splicing is carried out by mRNA splicing machinery known as spliceosome. Each spliceosome is composed of five small nuclear RNAs (U1, U2, U4, U5, U6) and a range of associated proteins. Of note, recent next-generation sequencing studies have identified several mutations involving multiple components of the mRNA splicing machinery, including SF3B1, SRSF2,U2AF1, ZRSR2, PRPF40B, U2AF65, and SF1 in myelodysplastic syndrome (MDS) patients. Out of these splicing factors, SF3B1 is one of the most frequently mutated genes, and mutations in SF3B1 have been found in up to 90% of patients with refractory anemia with ringed sideroblasts (RARS). The specific high frequency of SF3B1 mutations in RARS makes this gene a very strong candidate responsible for the pathogenesis of this subtype of MDS. Given the fact that RARS is mainly characterized by isolated erythroid dysplasia with mild dysplasia in granulocytic or megakaryocytic lineages, we hypothesize that SF3B1 plays important roles in normal erythropoiesis by regulating the alternative splicing of erythroid transcripts and that dysfunction of SF3B1 in RARS may directly account for the erythroid dysplasia of these patients.To test our hypothesis, we first examined the expression of SF3B1 in erythroid cells. We show that SF3B is abundantly expressed in erythroid cells. We then knocked down SF3B1 in human CD34+ hematopoietic stem cells employing shRNA mediated approach to explore the role of SF3B1 in human erythropoiesis. We show that knockdown of SF3B1 resulted in decreased formation of erythroid colonies BFU-E and CFU-E. We further show that knockdown of SF3B1 led to significantly impaired cell growth of erythroid cells with very little effects on the growth of granulocytes and monocytes. The decreased cell growth is accompanied by increased apoptosis. Knockdown of SF3B1 also led to delayed erythroid differentiation, generation of bi/multinucleated late stage erythroblasts and impaired enucleation. To explore the underlying mechanisms for the phenotypic changes following SF3B1 knockdown, we performed RNA-seq analysis on sorted erythroblasts at each distinct developmental stage. Bioinformatics analysis revealed that more than 40 genes were mis-spliced. Bioinformatics analysis also revealed that consistent with the impaired cell growth and increased apoptosis of CFU-E cells, knockdown of SF3B1 led to changes in expression of genes involved in regulation of cell growth and apoptosis in CFU-E cells. Similarly, consistent with generation of bi/multinucleated late stage erythroblasts and impaired enucleation, the expression of genes involved in mitosis and cytokinesis is downregulated in polychromatic and orthochromatic erythroblasts. Together, our findings demonstrated the critical role of SF3B1 in normal human erythropoiesis and identified potential SF3B1 targets in erythroid cells. Our findings not only provide novel insights into regulation of normal erythropoiesis but also have implications in understanding ineffective erythropoiesis in RARS patients with SF3B1 mutation. DisclosuresNo relevant conflicts of interest to declare.

Integrated and Long-Term Safety Analysis of Ibrutinib in Patients with Chronic Lymphocytic Leukemia (CLL)/Small Lymphocytic Lymphoma (SLL)

Integrated and Long-Term Safety Analysis of Ibrutinib in Patients with Chronic Lymphocytic Leukemia (CLL)/Small Lymphocytic Lymphoma (SLL)

PHP38 - Comparison Of Biosimilar Guideline And Real Consumption Among Italian Regions

The aim of this research is to assess biosimilar adoption by the Italian National Healthcare System in 2014 and 2015, analyzing and comparing regional legislative measures and biosimilar penetration, among four therapeutic areas (TNF inhibitors, erythropoietins, granulocyte colony-stimulating factor G-CSF and growth hormones). A desk-based research examined and compared the different regional legislative deliberations on biosimilar. Furthermore, the study was completed with the analysis of molecule consumption at regional level. Consumption data (source IMS Health data) were collected through all the distribution channels of the selected drugs: hospital in-patients distribution, hospital and local healthcare unit direct distribution. The majority of Italian regions have designed legislative measures to foster the adoption of biosimilars as first choice treatment for naïve patients and, in some cases, to address specific saving objective and consumption target. The analysis of the regional consumption shows important differences in market penetration of biosimilar drugs versus their reference products. Regional penetration ratio ranges from a minimum of 15% (anti TNF drugs, recently introduced into the market) and a maximum of 89% (G-CSF). Finally the study shows a continuous increase of biosimilar adoption, with different growth rate among regions and therapeutic areas. Italy lacks of a binding central guide on biosimilar usage, allowing a fragmented regional regulation and penetration. Although there are significant differences among regions, all of them encourage biosimilar uptake by setting switching guidelines, prescribing incentives and tendering schemes. Regional regulation plays a key role in promoting a wider diffusion of biosimilars, bringing relief to limited healthcare budgets.

The unique myelopoiesis strategy of the amphibian Xenopus laevis

Myeloid progenitors reside within specific hematopoietic organs and commit to progenitor lineages bearing megakaryocyte/erythrocyte (MEP) or granulocyte/macrophage potentials (GMP) within these sites. Unlike other vertebrates, the amphibian Xenopus laevis committed macrophage precursors are absent from the hematopoietic subcapsular liver and instead reside within their bone marrow. Presently, we demonstrate that while these frogs’ liver-derived cells are unresponsive to recombinant forms of principal X. laevis macrophage (colony-stimulating factor-1; CSF-1) and granulocyte (CSF-3) growth factors, bone marrow cells cultured with CSF-1 and CSF-3 exhibit respectively archetypal macrophage and granulocyte morphology, gene expression and functionalities. Moreover, we demonstrate that liver, but not bone marrow cells possess erythropoietic capacities when stimulated with a X. laevis erythropoietin. Together, our findings indicate that X. laevis retain their MEP within the hematopoietic liver while sequestering their GMP to the bone marrow, thus marking a very novel myelopoietic strategy as compared to those seen in other jawed vertebrate species.

Abstract A172: Allogeneic dendritic cells (AlloDCs) transduced with an infection-enhanced adenovirus as adjuvant for cancer immunotherapy

Abstract AlloDC cancer immunotherapy utilizes the allogeneic reaction as an immunostimulatory adjuvant. Intratumoral alloDC vaccination reverses the immune suppressed tumor micro-environment and activates anti-tumor immune responses. Herein, we aim to examine the use of Ad5f35PTD, an infection-enhanced serotype 5 adenovirus containing fibers from serotype 35 and cell-penetrating peptides on the hexons, as a vector to deliver tumor-associated antigens to monocyte-derived immature DCs in combination with an established DC maturation cocktail. Ad5f35PTD-transduced, cocktail-matured DCs expressed co-stimulatory and activation molecules and secreted the Th-1 type cytokine IL-12, in vitro. Natural killer (NK) cells migrated better towards medium from Ad5f35PTD-transduced, cocktail-matured DCs than medium from only cocktail-matured DCs, which was in accordance with a higher CXCL-10 chemokine level. The alloDC effect was examined in vivo using mature murine DCs of BALB/c origin (D2SC/I), transduced with Ad5f35PTD encoding the melanoma-associated antigen gp100, injected subcutaneously into C57BL/6 mice. They induced a higher migration of host DCs in the draining lymph node, in comparison to the D2SC/I cells alone. The migrated host DCs efficiently presented the captured gp100 and activated gp100-specific CD8+ T-cells. Moreover, alloDCs transduced with the gp100-encoding Ad5f35PTD reduced the immunosuppressive environment when injected intratumoraly by reducing the ratio of monocytic to granulocytic myeloid-derived suppressor cell compartment and delay tumor growth in a B16 melanoma model, in comparison with the D2SC/I cells alone. Concluding, the Ad5f35PTD transduced alloDCs expressing gp100 showed potency in eliciting tumor-specific immune responses in comparison to the alloDCs alone. Citation Format: Grammatiki Fotaki, Chuan Jin, Alex Karlsson-Parra, Di Yu, Magnus Essand. Allogeneic dendritic cells (AlloDCs) transduced with an infection-enhanced adenovirus as adjuvant for cancer immunotherapy. [abstract]. In: Proceedings of the CRI-CIMT-EATI-AACR Inaugural International Cancer Immunotherapy Conference: Translating Science into Survival; September 16-19, 2015; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(1 Suppl):Abstract nr A172.

Risk Factors for Early Death after Immunochemotherapy in Older Patients with Diffuse Large B-Cell Lymphoma (DLBCL)

Background: DLBCL is a curable malignancy for a majority of patients >65 years old, but they are at higher risk of toxicities and potential early death after chemotherapy. We evaluated risk factors for death and other severe adverse events during the first month of rituximab-based immunochemotherapy among Medicare beneficiaries with DLBCL, using Medicare claims linked to the Surveillance, Epidemiology and End Results (SEER-Medicare) database.Methods: We selected patients aged ≥65 years, diagnosed with DLBCL between 2003 and 2012, excluding cases diagnosed by autopsy, with incomplete Medicare claims, primary central nervous system lymphoma, or first chemotherapy in the inpatient setting. Eligible patients initiated an outpatient regimen containing rituximab, cyclophosphamide, vincristine, with doxorubicin (R-CHOP-like), or a version without anthracycline. The primary outcome was defined as death within 30 days from the start of chemotherapy. Secondary outcomes included: admission to a hospital, critical care unit or a nursing home, and a cardiac event within this 30-day window. Risk factors were studied in multivariable logistic regression models adjusting for age, sex, race, Medicaid (poverty) status, DLBCL stage, B symptoms, and receipt of anthracycline. We stepwise-selected validated indicators of performance status and comorbidities (based on Medicare claims from the preceding year) which showed strongest associations with the outcomes, using Bonferroni correction. Results are expressed as odds ratios (OR) with 95% confidence intervals (CI).Results: We identified 6,425 patients with median age of 76 years, of whom 49% were women, 88% white non-Hispanic, 44% had stage III/IV lymphoma, and 84% received R-CHOP-like regimen. Cumulative incidence of death was 2.1% at day 30, and 12.3% at day 180 after chemotherapy. Prophylactic granulocyte growth factor was administered to 63% of patients during the first treatment cycle.In a multivariable model, the risk of early death was significantly higher for patients older than 75 years (OR vs. 65-70 years, 2.07, CI 1.04-4.14) or ≥80 years (OR, 3.22, CI, 1.66-6.24), and those with B-symptoms (OR, 1.90, CI, 1.13-3.20), but there was no significant difference by sex, race, stage, poverty status or anthracycline use. The risk was also associated with chronic kidney disease (OR, 3.37, CI, 2.07-5.46), poor performance status (OR, 2.08, CI, 1.22-3.54), prior use of walking aids (OR, 2.26, CI, 1.38-3.71), prior hospitalization (OR, 1.68, CI, 1.14-2.47), or a history of upper endoscopy (OR, 1.73, CI, 1.19-2.51). The risk of early death was only 1.1% for patients with <2 of those factors (79% of cases) while it was 7.2% for those with ≥4 factors (6.8% of cases).The risk of other outcomes within 30 days of chemotherapy was: 24% for hospitalization (8% with a diagnosis of febrile neutropenia), 11% for a cardiac event, 8% for critical care, and 3% for nursing home admission. The median time to hospitalization was 9 days (interquartile range, 7 to 14). The risk of early hospitalization was associated with increasing age, female sex (OR, 1.14, CI, 1.01-1.28), B symptoms (OR, 1.49, CI, 1.27-1.74), stage IV lymphoma (OR, 1.20, CI, 1.02-1.40), prior myocardial infarction (OR, 2.56, CI, 1.57-4.18), renal disease (OR, 1.66, CI, 1.31-2.11), prior hospitalization (OR, 1.50, CI, 1.32-1.69) or a history of upper endoscopy (OR, 1.20, CI, 1.05-1.38).Conclusions: Among older DLBCL patients who receive contemporary rituximab-based chemotherapy, 1 in 50 die during the first month of treatment, and 1 in 4 are hospitalized. Easily identifiable factors can distinguish groups at highest risk of early death, who may benefit from preventive strategies such as the prephase treatment (Pfreundschuh, Blood 2010), or from novel, personalized therapeutic approaches. Despite national guidelines, prophylactic granulocytic growth factors are not administered to over 1/3 of patients, indicating an opportunity to lower the risk of adverse events. Withholding doxorubicin was not associated with a lower risk of early death or hospitalization. DisclosuresOlszewski:Genentech, Inc.: Research Funding; Bristol-Myers Squibb, Inc.: Consultancy.

Progression-Free Survival Following Autologous Transplantation for Multiple Myeloma is Not Compromised by the Use of Dose-Adjusted Melphalan

e154 factor for the mobilization of CD34+ progenitor cells. The mobilization protocols attempt to supplant such a condition associating other treatments to the granulocyte growth factor (G-CSF), such as cyclo. The CD34+ cell target for a single transplant has been 4 to 6 x 106 CD34+/kg, the acceptable minimum in avoiding damage upon harvesting the bone marrow being 2x106 CD34+/Kg. ObjectiveTo evaluate the number of CD34+ cells mobilized in the use of GCSF Vs GCSF+cyclo following induction with the schedule cyclo+thal+dexamethasone (dex)(CTD) in MM patients who are eligible for high-dose chemotherapy. The harvesting > 2.0x 106/kg CD34+ cells was considered adequate. Cases and methods A retrospective cohort of MM patients submitted to TACPH at two Brazilian centers from May 2009 to July 2013. Induction 1⁄4 CTDcenter 1 (cyclo 50mg/day, thal 100-200mg/day, dex 160mg/month and center 21⁄4 cyclo 500mg D1, 8 and 15, thal 100mg/day and dex 200-400mg/month, all drugs being administered orally. Mobilization 1⁄4 GCSF + cyclo (GCSF 10-15mcg/kg for 5 days + cyclo34g/m or GCSF15-20mcg/kg for 5 days isolatedly). Results Ninety-two patients were analyzed. In relation to mobilization, 70 patients received isolated filgrastim, 18 received a combination of filgrastim and cyclophosphamide, 2 patients were not submitted to mobilization and in two cases the schedule was not reported. Median general age was 58 years (51-62), 58 years (53-61) for GCSF and 56.5 years (54-60) for GCSF + cyclo. Fifty-two patients were males . Concerning the DS IIIA/B 1⁄4 80 cases (90.9%) and ISS 2 and 3 1⁄4 38 cases (44.7%) and 20 cases (23.5%). The filgrastim+ cyclo group had the highest average number of collected progenitor cells, 6.4 7.7, VS 3.4 1.3 in the isolated filgrastim group (p 1⁄4 0.008). No correlation was observed between better responses and higher number of mobilized cells. The filgrastim utilized was of a similar type for all the presented cases. Conclusions The number of cells mobilized with GCSF + cyclo is superior to the number mobilized with isolated GCSF. However, the average number of CD34+ was sufficient to perform a single TACPH, with no detriment to harvesting. In addition, the type of filgrastim used was biosimilar, conferring a satisfactory response with said agent.

Identification of Dual Natural Inhibitors for Chronic Myeloid Leukemia by Virtual Screening, Molecular Dynamics Simulation and ADMET Analysis.

Chronic myeloid leukemia (CML) is a disease of bone marrow stem cells caused by excessive growth and accumulation of granulocytes in the blood. Aberrant expression of the BCR-ABL proteins in bone marrow stem cells have found out in 95% cases of CML. Tyrosine Kinase domains (SH2 and SH3) of BCR-ABL proteins are the potent targets to inhibit the process. Initially, imatinib is preferred as an efficient inhibitor to control functional activity of disease. Recently, it has been reported that the advanced stage of CML developed resistance against imatinib. In continuation, dasatinib is the first drug to combat against this disease by targeting multiple receptors and proven better as compared to imatinib. Here, an attempt has been made to identify similar analogs of dasatinib. Virtual screening was performed against various natural compound databases to get some potent natural compounds which are able to inhibit more than one receptor. Binding affinity of screened natural compounds was compared with some of the well-known inhibitors like imatinib, dasatinib, nilotinib etc., by analyzing their docking score and binding efficiency with the receptor. Stability of the best ligand-receptor complex was checked by performing 10 ns molecular dynamics simulation. ADMET properties of the obtained screened compounds were analyzed to check drug like property. Based on the aforementioned analysis, it has been suggested that these screened potent compounds are capable to inhibit multiple receptor proteins like ABL and SRC and consequently combat against the deadly disease CML.

Interleukin‐33 Triggers B1 Cell Expansion and Its Release of Monocyte/Macrophage Chemoattractants and Growth Factors

B1 B lymphocytes are natural IgM-producing cells primarily found in peritoneum and mucosal sites. They perform vital functions during the early defence against viral and bacterial infections. Murine B1 cells express IL-33 receptor complex on activation. IL-33 is a new addition to the IL-1 family with a strong role in Th2 immunity. B1 cells have been recognized to exacerbate contact sensitivity by producing IgM and IL-5 in response to interleukin-33. However, the exact response of IL-33/ST2 signalling in B1 cells is not completely understood. In this study, we report that murine B1 cells respond directly to IL-33 in a ST2-dependent manner. This interaction instigates B1b cell proliferation in a time-dependent manner in vivo. Furthermore, it also mediates monocyte/macrophage and granulocyte recruitment via B1 cell release of chemokines (MCP-1 and MIP-1 alpha). It was noted that upon stimulation, B1b cells additionally release an angiogenic inducer vascular endothelial growth factor and granulocyte-monocyte colony-stimulating factor (GM-CSF). Our findings suggest that these IL-33-mediated B1 cells might be able to play a vital role in the recruitment and growth of monocytes and granulocytes.

Moderne Antirheumatika

The term modern disease-modifying antirheumatic drugs (DMARD) includes not only the constantly growing family of DMARDs for chronic inflammatory rheumatic diseases but also the repositioning of established drugs in updated and novel algorithms of the different entities. The usual precursor for these developments is rheumatoid arthritis for which completely revised and updated guidelines have been published not only in Germany but also on the European level. In addition, label extensions to existing drugs have been granted for connective tissue diseases and vasculitides, e.g. anti-CD20 antibodies for antineutrophil cytoplasmic antibodies (ANCA)-associated vasculitides and belimumab for systemic lupus erythematosus. Moreover, several novel drugs, especially of the biologics class, have been either introduced in clinical rheumatology or are close to being licensed and include ustekinumab for psoriatic arthritis, granulocyte growth inhibitors and janus kinase inhibitors for rheumatoid arthritis and atacicept for systemic lupus erythematosus. With the termination of the patent for several biologics, a new momentum also took place: the approval of the so-called biosimilars which has already initiated intensive discussions not only with respect to "similar" effects and side effects but also with respect to their potential impact on economical aspects.

Granulocyte Colony Stimulating Factor for Treatment of Thin Endometrium in Assisted Reproduction Technology Cycles

ABSTRACTGranulocyte colony stimulating factor (G-CSF), a glycoprotein, belongs to colony stimulating factor family and mainly regulates the growth and differentiation of granulocytes. However, it also plays an important role in endometrial stromal cell decidualization, ovulation, implantation, placental metabolism, trophoblast development and endometrial regeneration. It is due to these effects, it has been used in difficult clinical scenarios, such as unresponsive thin endometrium during assisted reproductive technology treatment, repeated implantation failure and recurrent miscarriages. Most of the studies have investigated its use in thin endometrium. In this review, we have summarized the current updated evidence with regards to use of G-CSF in women with thin endometriumHow to cite this articleKamath MS, Lele P. Granulocyte Colony Stimulating Factor for Treatment of Thin Endometrium in Assisted Reproduction Technology Cycles. Int J Infertil Fetal Med 2015;6(3):97-102.

Multicenter Study with Mbvd Regimen in Elderly or Younger Patients with Cardiac Disorders Affected By Hodgkin’s Lymphoma. a Phase II Study on Behalf of the Fondazione Italiana Linfomi (FIL)

Abstract Backgrounds: ABVD is considered the standard of care for Hodgkin’s Lymphoma (HL) patients; however few prospective studies have evaluated his efficacy and safety in elderly patients. Moreover anthracycline-based chemotherapy is not feasible in patients with moderate/severe concomitant heart disease. In comparison with conventional doxorubicin, non–pegylated liposomal doxorubicin (TLC-D99; Myocet ™) has shown a selective uptake and a reduced clearance by the tumor cells resulting in improved cardiac safety. Purpose: Aim of the study was to investigate efficacy and safety of an ABVD-like regimen with non-pegylated liposomal doxorubicin (TLC-D99; Myocet ™) instead of conventional doxorubicin. Patients and Methods : From March 2010 to January 2013, 41 non-frail elderly (age³ 70) and 6 younger cardiopathic patients with untreated HL were enrolled by 22 FIL centers. “Non-frail” definition was based on less than 3 grade CIRS-G co-morbidities, no grade 4 CIRS-G and no geriatric syndrome at diagnosis. Cardiac disorder was defined according to the presence of at least one of the following: left ventricular ejection fraction (LVEF) &lt; 50%, left ventricular hypertrophy, uncontrolled moderate/severe arterial hypertension, history of ischemic cardiopathy, clinically significant ventricular arrhythmia, atrial fibrillation, pulmonary hypertension, moderate/severe mitral valvular disorder, moderate aortic valvular disorder. For advanced disease (IIB-IV) the treatment plan was 6 courses of MBVD ( Myocet 25 mg/m2; bleomycin 10 mg/m2; vinblastine 6 mg/m2; dacarbazine 375 mg/m2) plus radiotherapy (RT) on bulky disease or residual PET positive area; for early stages (I-IIA) the treatment plan was 3 courses of MBVD plus RT involved field (IF). Preventive use of granulocyte growth factors was recommended in elderly patients; erythropoietin treatment was considered for all patients if the haemoglobin value was &lt; 11 gr/dl. The two primary objectives of the study were to evaluate the complete remission (CR) rate according to international criteria (Cheson 2007) and the rate of cardiac events (CE) defined as a reduction of LVEF ³ 15% from baseline or the occurrence of any significant cardiac disorder. Results. Patients’ median age was 75 (range 46–84); thirteen patients (28%) were in early stage and the remaining 34 (72%) in advanced stage. According to CGA-G scale one or more G2-3co-morbidities were present in 28 patients (60%). All the 13 early stages patients regularly ended the three planned courses of chemotherapy in an outpatients setting without complications; 1 of them refused subsequent RT IF. Among the advanced stage, 13 out 34 patients (38%) interrupted their treatment, mainly from course three to six, for the following reasons: 9 for severe toxicity; 1 for disease progression; 2 for poor compliance or consent retirement; 1 for lung cancer. Fifty-one % of patients experienced at least one episode of grade 3-4 haematological toxicity; severe infections were reported in 3 patients (6%); grade 4 acute cardiac toxicity in only 1 patient. A LVEF reduction ³ 15% from baseline was never documented. The final CR rate was 100% in early stage and 68% in advanced stage patients. At a median follow-up of 28 months all patients with early disease are alive and in CR. In advanced stage patients the 30 months actuarial OS rate is 62%, while the 30 months actuarial PFS rate is only 34% with a median PFS of 20 months. So far, 10 advanced patients have died: 3 of HL; 3 of acute toxicity including 1 sepsis, 1 pneumonia and 1 myocardial infarction, resulting in a 6% treatment-related mortality (TRM); moreover 2 patients died of lung cancer diagnosed within 7 months after the end of their treatment; 1 of heart failure and 1 of pneumonia, thrombocytopenia and gastrointestinal bleeding , both 1 year later the end of treatment. Acute and late CE with fatal exit from the study occurred in 2 patients (4%). Conclusions: In elderly patients with advanced disease MBVD shows an unfavourable toxic profile, not really different from ABVD, even if cardiac toxicity was spared. Prospective studies with baseline definition of comorbidity score and new less aggressive strategies are needed. Disclosures Off Label Use: Lenalidomide in MCL. Spina:Teva: Honoraria.

Feasibility of an Attenuated Maintenance Chemotherapy Regimen Directed at Adolescents and Young Adults with Newly Diagnosed Localized Medulloblastoma and Other Central Nervous System Embryonal Tumors

Purpose: Newly diagnosed adolescents and young adults (AYAs) with medulloblastoma experience substantial toxicities with “standard” maintenance chemotherapy regimens that include lomustine, cisplatin, and vincristine. An attenuated maintenance chemotherapy regimen targeting AYAs was evaluated for toxicity and tolerability. Methods: Eleven patients with central nervous system (CNS) malignant embryonal tumors treated between 2004 and 2010 at five institutions were evaluated. Three patients were <17 years old, seven were aged 17–39, and one was 50 years old. Seven were female. Detailed toxicity data were evaluated. Results: Eighty-seven cycles were administered (87% of all planned cycles). Grade 3/4 neutropenia occurred in 60% of cycles, with the use of granulocyte growth factor support in only 15% of cycles. Grade 3/4 anemia and thrombocytopenia occurred in 33% and 29% of cycles, respectively; however, blood and platelet transfusions were administered in only 32% and 21% of cycles, respectively. Grade 3/4 transaminitis occurred in 8% of cycles. There were five grade 2 reports of anorexia, one grade 3 sensory and four grade 3 motor neuropathies, and one grade 3 hypocalcemia and hypomagnesemia. No grade 1–4 ototoxicities or nephrotoxicities were observed. No grades 3 or 4 weight loss was observed. Two patients had single hospitalizations for chemotherapy-related toxicities (1 and 13 days in duration). After 3.5 years median follow-up, only the single patient with atypical teratoid/rhabdoid tumor had relapsed. Conclusion: This attenuated maintenance chemotherapy regimen was well tolerated by AYAs. A prospective North American clinical trial is planned for AYAs with CNS embryonal tumors to determine the efficacy of this regimen.

Autoimmune neutropenia

Autoimmune neutropenia (AIN) is a rare entity caused by antibodies directed against neutrophil-specific antigens. It includes primary and secondary autoimmune neutropenia. Acute autoimmune neutropenia can be related to drug-induced mechanism or viral infections. Chronic autoimmune neutropenias occur in the context of autoimmune diseases, hematological malignancies, such as large granular lymphocyte leukemia, primary immune deficiency syndromes or solid tumors. The therapeutic management depends on the etiology. Granulocyte growth factor is the main therapeutic option, raising the question of their long-term utilization safety. Corticosteroids or immunosuppressive therapy are indicated in infection-related AIN or in case of symptomatic autoimmune disease or LGL leukemia.

Improving care and reducing costs in oncology.

The costs of healthcare are a cause of concern throughout the world. This is the case for Europe, where most of the service is paid for by a state healthcare system, but also in the USA, where private insurance has a more prominent role. Several articles addressing this issue, i.e. the causes of this sharp increase and how to reduce the costs of medicine, have appeared in medical journals in recent months. Medical oncology is one of the disciplines where expenses are most obvious, even if doctors do not generally notice the price tag on the drugs they prescribe. Prices in the order of 5000€ per month per patient for a single anticancer drug are not uncommon. I will not discuss here the reasons why pharmaceutical companies charge such prices, as this question may be less important than generally considered since the duration of treatment is often limited to a few months (progression-free survival is often very short) and, in the case of targeted agents, only a selected patient population is (or should be!) involved. In a recent paper [1], ASCO underlines five points that are often forgotten when we measure the costs of treatments, updating the recommendations published in 2012 [2]. This list is in some aspects surprising since it includes items that are often overlooked when we think about the price of cancer care: expensive antiemetics, combination chemotherapy, target agents when no target is present. Cautionary use of PET for tumour staging, and follow-up and PSA testing have been reiterated as they were already present in the recommendations issued in 2012. Other items in the 2012 list were to avoid chemotherapy in subjects unlikely to benefit from it and to use granulocyte-stimulating factors only when the risk of febrile neutropenia is not negligible. The most important outcome of reading and reflecting on this paper is that it will hopefully encourage oncologists to analyse their clinical habits and to reduce those that are not only expensive, but also of questionable benefit to the patient. Once more it becomes evident that the most important source of expenses is the pen of the doctor [3]. It will be important for doctors to request fewer radiological tests (including ultrasound scans) which will not only save money but will shorten waiting lists – an issue that, in Italy, is one of the most commonly criticised aspects of the healthcare system. The next step is to look for further ways to reduce costs, but a tentative shortlist may include: the use of CT scans in terminal patients, requesting whole-abdomen ultrasound scans when we are only interested in the liver, measuring bone density in every woman treated with aromatase inhibitors, prescribing echocardiograms for all breast cancer patients receiving adjuvant anthracyclines with a cumulative dose that is well below the toxic range, using expensive anticancer agents in the third or fourth line of treatment of advanced cancer, using pegylated granulocyte growth factors when not required and prolonging treatment with erythropoiesis-stimulating agents when no response can be expected. The price of anticancer drugs, however, cannot be overlooked. Of course the pen of the prescribing oncologist is partly to blame for the overuse of expensive drugs [3] but it has more to do with the regulating authorities. They should require evidence not only of “statistical significance” but also of “clinical relevance” of results obtained with new and super-expensive drugs and even after approval they can still intervene to negotiate favourable prices, as NICE (The National Institute for Health and Care Excellence) have done in the UK [4]. Calculating the real value of a test for the patient before prescribing it would be an excellent exercise for every physician, even if many will object that low cost is by definition low quality medicine. In oncology this concern appears irrelevant since it has been shown that when costs were reduced the quality of assistance improved [5],[6]. Medical oncologists in the USA seem to be more conscious about costs than we are in Europe, but it is time for us here in Europe to start considering this element since paying more attention to our everyday practice will result in paying less for medical care while maintaining the same quality.