Erythropoietic Growth Factors Research Articles

Classification of Absolute Cd4+ T Lymphocytes Count in Immunological, Virological and Erythropoietic Growth Factor among HIV Infected Patients

Background: HIV-infection resulted in CD4+ T-cell depletion which is accompanied by an increase in CD8+ T-cells resulting in an inverted CD4/CD8 ratio. Low CD4/CD8 ratio has been identified as a hallmark of inmmunosenescence and a surrogate of mortality in HIV infected patients especially in ART-naïve patients Aim: Classification of absolute CD4+ T-lymphocytes count in immunological, virological and erythropoietic growth factor among HIV infected patients Methodology: One hundred samples each was collected from HIV positive subjects on ART and HIV positive subjects ART naïve. Six milliliters of whole blood was collected from each consented subject, 3ml was dispensed into 5ml K2EDTA bottle for immediate analysis of absolute CD4 count, CD8 count, total white cell count and HIV screening. The remaining 3ml of blood was dispensed into plain bottle; serum was extracted for the analysis of erythropoietin and viral load Results: Mean values of CD8, CD4/CD8, EPO and TWBC in CD4 <200 were significantly (p<0.05) lower compared to CD4 >500 and CD4 200-499. Mean values of VL in CD4 <200 were significantly (p<0.05) higher compared to CD4 >500 and CD4 200-499 among HIV subjects on ART and ART-naïve Conclusion: Immunological and erythropoetic growth factor assessed in this study were decline while viral load was increasing as HIV infection progresses with depletion in absolute CD4 count, this study shows the efficacy of ART on the treated subjects. However, based on this study, absolute CD8 T cells count, CD4/CD8 ratio and erythropoietin can be used as surrogate makers to ascertain pathogenesis in HIV-infected subjects.

Evaluation of Haematoimmunological, Erythropoietic Growth Factor and Virological Status in HIV-Patients

Background: Human Immunodeficiency Virus reduces immune status and diminishes haematological values causing decreased or ineffective haematopoiesis in HIV-infected individuals. Aim: To evaluate haematoimmunological, erythropoietic growth factor and virological status in HIV-patients. Methodology: This study was carried out at Federal Teaching Hospital, Ido-Ekiti. One hundred samples each were collected from HIV positive subjects on ART and ART-naïve, one hundred samples were collected from apparently healthy individuals as control. Six milliliters of whole blood were collected from each subject; 3mls of whole blood was dispensed into K2EDTA bottle for immediate analysis of haematological parameters using haematology analyzer, CD4 and CD8 count were analyzed using flow cytometer. The remaining 3ml of blood was dispensed into plain bottle, serum was extracted for erythropoietin analysis using enzyme-linked immunosorbent assay technique and viral load using polymerase chain reaction. Results: Mean values of RBC indices CD4, CD4/CD8, EPO, TWBC and LYM among ART-naïve were significantly lower (p<0.05) compared to ART and control subjects. Mean values of CD8, VL, NEU, MONO, EOSIN and BASO in ART-naïve were significantly higher (p<0.05) compared to ART and control subjects. Conclusion: HIV infection suppresses erythropoietic growth factor, haematoimmunological, and virological parameters as observed in the study. However, there is an improvement in HIV subjects on ART compared with ART-naïve, this suggested that there is positive response to antiretroviral therapy in HIV patients.

Plant cell-secreted stem cell factor stimulates expansion and differentiation of hematopoietic stem cells

Ex vivo generation of red blood cells (RBCs) from hematopoietic stem cells (HSCs) used for blood transfusion represents one of the focuses in current regenerative medicine. However, massive production of HSCs-based RBCs requires a significant quantity of erythropoietic growth factors, making manufacturing at large scale cost prohibitive. Plant cell culture is proposed to be a promising bioproduction platform for functional human proteins in a safe and cost-efficient manner. This study exploited a proprietary technology, named HypGP engineering technology, for high-yield production of one of the key erythropoietic growth factors--stem cell factor (SCF)--in plant cell culture. Specifically, a designer hydroxyproline (Hyp)-O-glycosylated peptide (HypGP) comprised of 20 tandem repeats of the “Ser-Pro” motif, or (SP)20, was engineered at either the N-terminus or C-terminus of SCF in tobacco BY-2 cells. The (SP)20 tag dramatically increased the secreted yields of SCF up to 2.5 μg/mL. The (SP)20-tagged SCF showed bioactivity in promoting the proliferation of the TF-1 cell line, although the SCF-(SP)20 was 8.4-fold more potent than the (SP)20-SCF. Both the (SP)20-SCF and SCF-(SP)20 exhibited desired function in stimulating the expansion and differentiation of human umbilical cord blood CD34+ cells towards RBCs.

High‐yield production of hematopoietic growth factors by tobacco BY‐2 cells for ex vivo production of red blood cells

Ex vivo generation of clinically available red blood cells (RBCs) from hematopoietic stem cells (HSCs) represents a promising approach for overcoming the limitations associated with the use of donor’s blood, such as shortage of blood and risk of transfusion‐transmitted infections. Both expansion and differentiation of stem cells are highly reliant on growth and differentiation factors, with the activity and purity of these molecules being critical to successful research and clinical applications. The overarching goal of this project is to develop a novel plant cell‐based bioproduction platform for high‐quality (i.e. animal‐free and endotoxin‐free) erythropoietic growth factors (eGFs) that facilitate mass production of RBCs from HSCs at significantly reduced cost. In addressing the low productivity bottleneck of plant cell culture technology, this proposal aims to leverage a unique, plant‐specific, O‐glycosylation process called “hydroxyproline (Hyp)‐O‐glycosylation” for de novo design and engineering of Hyp‐O‐ glycosylated peptides (HypGPs”) that presumably function as a molecular carrier in boosting secretion of conjoined eGFs from cultured plant cells. This will dramatically increase the secreted yields of plant cell‐derived eGFs and simplify the product purification process. Three key eGFs essential for HSCs expansion and differentiation: erythropoietin (EPO), stem cell factor (SCF) and interleukin 3 (IL‐3), are expressed in tobacco BY‐2 cells as fusion with a strategically designed HypGP tag. The secreted protein yields, molecular structure, and Hyp‐O‐glycosylation of the HypGP‐tagged eGFs are characterized. The bioactivities of individual HypGP‐tagged eGF in stimulating proliferation and differentiation of relevant hematopoietic cell lines, includingTF1 erythroleukemic cell and umbilical cord blood‐derived CD34+ cell, are evaluated to establish a suitable design of HypGP tag (size and orientation) for each eGF. A scalable cell culture system fortified sorely with plant cell‐produced eGFs is developed for production of RBCs from CD34+ HSC cells. This project provides a proof‐of‐concept for ex vivo manufacturing of stem cell‐derived RBCs with plant cell‐produced eGFs.Support or Funding InformationNational Science Foundation Directorate for Engineering [1605564]; National Institutes of Health‐Arkansas INBRE [P20GM103429]

Plant Cell‐Secreted Growth Factors for ex vivo Mass Production of Red Blood Cells

Ex vivo generation of clinically available red blood cells (RBCs) from hematopoietic stem cells (HSCs) represents a promising approach for overcoming the limitations associated with the use of donor's blood, such as shortage of blood and risk of transfusion-transmitted infections. Both expansion and differentiation of stem cells are highly reliant on growth and differentiation factors, with the activity and purity of these molecules being critical to successful research and clinical applications. The overarching goal of this project is to develop a novel plant cell-based bioproduction platform for high-quality (i.e. animal-free and endotoxin-free) erythropoietic growth factors (eGFs) that facilitate mass production of RBCs from HSCs at significantly reduced cost. In addressing the low productivity bottleneck of plant cell culture technology, this proposal aims to leverage a unique, plant-specific, O-glycosylation process called “hydroxyproline (Hyp)-O-glycosylation” for de novo design and engineering of Hyp-O-glycosylated peptides (HypGPs”) that presumably function as a molecular carrier in boosting secretion of conjoined eGFs from cultured plant cells. This will dramatically increase the secreted yields of plant cell-derived eGFs and simplify the product purification process. Three key eGFs essential for HSCs expansion and differentiation: erythropoietin (EPO), stem cell factor (SCF) and interleukin 3 (IL-3), are expressed in tobacco BY-2 cells as fusion with a strategically designed HypGP tag. The secreted protein yields, molecular structure, and Hyp-O-glycosylation of the HypGP-tagged eGFs are characterized. The bioactivities of individual HypGP-tagged eGF in stimulating proliferation and differentiation of relevant hematopoietic cell lines, including TF1 erythroleukemic cell and umbilical cord blood-derived CD34+ cell, are evaluated to establish a suitable design of HypGP tag (size and orientation) for each eGF. A scalable cell culture system fortified sorely with plant cell-produced eGFs will be finally developed for mass production of RBCs from CD34+ HSC cells. This project provides a new cost-effective bioproduction platform for eGFs, facilitating manufacturing of HSCs-derived RBCs at large scale for clinical applications. Support or Funding Information Arkansas INBRE-Research Development Grant (RDG) under Grant No. P20GM103429; National Science Foundation under Grant No. 1605564; Arkansas Biosciences Institute Schematic of the proposed research for mass production of red blood cells (RBCs) ex vivo from hematopoietic stem cells (HSCs) with plant produced eGFs (EPO, SCF, IL-3 and IGF-1) All the eGFs will be engineered in plant cells (tobacco BY-2 cell) with a Hyp-O-glycosylated peptide (HypGP) tag that function as a molecular carrier in boosting the secretion of conjoined erythropoietic growth factors (eGFs). This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Exploring erythropoiesis of common carp (Cyprinus carpio) using an in vitro colony assay in the presence of recombinant carp kit ligand A and erythropoietin

The use of in vitro colony assays in mammals has contributed to identification of erythroid progenitor cells such as burst-forming unit-erythroid (BFU-E) and colony-forming unit-erythroid (CFU-E) progenitors, and serves to examine functions of erythropoietic growth factors like Erythropoietin (Epo) and Kit ligand. Here, we established an in vitro colony-forming assay capable of investigating erythropoiesis in carp (Cyprinus carpio), cloned and functionally characterized recombinant homologous molecules Epo and Kit ligand A (Kitla), and identified three distinct erythroid progenitor cells in carp. Recombinant carp Epo induced the formation of CFU-E-like and BFU-E-like erythroid colonies, expressing erythroid marker genes, β-globin, epor and gata1. Recombinant carp Kitla alone induced limited colony formation, whereas a combination of Kitla and Epo dramatically enhanced erythroid colony formation and colony cell growth, as well as stimulated the formation of thrombocytic/erythroid colonies expressing not only erythroid markers but also thrombocytic markers, cd41 and c-mpl. Utilizing this colony assay to examine the distribution of distinct erythroid progenitor cells in carp, we demonstrated that carp head and trunk kidney play a primary role in erythropoiesis, while the spleen plays a secondary. Furthermore, we showed that presumably bi-potent thrombocytic/erythroid progenitor cells localize principally in the trunk kidney. Our results indicate that teleost fish possess mechanisms of Epo- and Kitla-dependent erythropoiesis similar to those in other vertebrates, and also help to demonstrate the diversity of erythropoietic sites among vertebrates.

Treatment of Low-Risk Myelodysplastic Syndrome: Hematopoietic Growth Factors Erythropoietins and Thrombopoietins

The use of erythropoietic growth factors has become standard of care in many countries for lower risk myelodysplastic syndrome (MDS) patients. Throughout a large number of clinical trials, therapy with erythropoietic agents has consistently shown improvement of anemia and reduction of transfusion dependence. There is currently no evidence of safety issues of erythropoietins in MDS, including thrombosis, polycythemia, and progressive disease. Large retrospective comparative analyses have shown no increase in mortality in erythropoietin (EPO)-treated MDS patients. Doses of up to 80,000 IU/wk have successfully been employed and the addition of granulocyte colony-stimulating factor (G-CSF) can benefit previously unresponsive patients. Although several other combination therapies have been tested, apart from G-CSF, none has gained wide clinical acceptance. Thrombopoietic agents can alleviate thrombocytopenia and bleeding symptoms in lower risk MDS patients. However, concerns regarding a higher rate of transformation to acute myeloid leukemia and the fear of increased bone marrow fibrosis during treatment have hampered their clinical development.

Suboptimal endogenous erythropoietin response in chronic hepatitis C patients during ribavirin and PEG interferon treatment

During the treatment of hepatitis C, anaemia may necessitate pegylated-interferon and ribavirin dose reductions with reduced sustained viral response rates. Although erythropoietic growth factors are frequently used to improve anaemia, it is controversial whether endogenous erythropoietic response is insufficient under these circumstances. We aimed to identify risk factors for more pronounced anaemia and to evaluate endogenous erythropoietic response during antiviral therapy. One hundred and forty-five naive chronic hepatitis C patients on pegylated-interferon-ribavirin treatment were evaluated for haemoglobin, haematocrit, serum ribavirin and erythropoietin levels. About 99% of patients developed anaemia, with maximal decrease in haemoglobin of 2.5 ± 1.0 mmol/l (range 0.3-5.5 mmol/l). Older age, lower baseline creatinine clearance, higher baseline haemoglobin, more pronounced haemoglobin decrease after 2 weeks and higher week 24 serum ribavirin concentrations were independent risk factors for more pronounced anaemia. Serum erythropoietin levels increased from median 12 IU/l (range 4-63 IU/l) at baseline to 41 IU/l (range 12-683 IU/l) after 12 weeks of therapy and to 43 IU/l (range 7-3238 IU/l) at week 24 (P<0.001). Erythropoietin levels at baseline, week 12 and week 24 negatively correlated with haematocrit. The erythropoietic response to anaemia in our study population was significantly different from the normal human response to anaemia. Older age, lower baseline creatinine clearance, higher baseline haemoglobin, more pronounced haemoglobin decrease after 2 weeks and higher week 24 serum ribavirin concentrations were independent risk factors for more pronounced anaemia during antiviral therapy. Endogenous erythropoietin production is suboptimal during antiviral therapy, supporting use of erythropoietic growth factors.

Management of Complications in Multiple Myeloma

Multiple myeloma (MM) is characterized by the presence of osteolytic bone disease, renal impairment, anemia, and immune dysfunction. Adequate supportive care is considered an essential part of anti-myeloma therapy. The administration of bisphosphonates has been shown to reduce skeletal related events and hypercalcemia. Bisphosphonates are well tolerated, but preventive steps should be taken to avoid renal impairment and osteonecrosis of the jaw (ONJ). Adequate pain control is of crucial importance for the quality of life of MM patients. Local radiotherapy may rapidly ameliorate symptoms of painful MM bone lesions, and vertebroplasty and kyphoplasty are able to control symptoms and restore the original height of vertebral fractures. Symptomatic chemotherapy-induced anemia should preferentially be treated with erythropoietic growth factors, but further studies are required to confirm the long-term safety of this approach. Light-chain-induced renal impairment should be treated without delay with a highly effective anti-myeloma regimen consisting of novel drugs. Prophylaxis of infections should be considered particularly in patients with poorly controlled disease and documented infections should be treated aggressively as they contribute significantly to morbidity and mortality. The concerted action of these supportive therapies can significantly improve the quality of life of MM patients during the different phases of their disease.

Erythropoietin and microvascular diabetic complications

When recombinant human erythropoietin (EPO) became available for clinical use, the hormone was considered as a specific erythropoietic growth factor and as a more or less unique molecule, since its endogenous production increases in response to reduced oxygen availability. Subsequently, we have witnessed a revolutionary increase in knowledge about the highly conserved ability of virtually all cells to sense oxygen and to translate reductions in oxygen availability into specific patterns of gene expression. This hypoxia response is to a large extent mediated by hypoxia-inducible transcription factors (HIF). EPO is the prime example for a hypoxia-inducible HIF target gene, but more than 100 additional genes have meanwhile also been identified as HIF targets, including genes involved in new vessel formation and anaerobic metabolism. In general, the activation of the hypoxia pathway stimulates responses that increase either oxygen availability or hypoxia tolerance [1]. Consistent with this broader perspective of oxygen sensing and tissue responsiveness to changes in oxygenation, EPOhasmeanwhilealsobeenfoundtohaveactionsbeyond stimulationofredcellprecursors,includingpro-angiogenic andcellprotectivefunctions.HighdosesofEPOconveytissue protection in experimental models of ischaemic brain and kidney injury, and trials testing the clinical utility of such effects are underway. Moreover, low levels of EPOgene expression have been found in various tissues other than liver and kidneys, the two organs that are responsible for the synthesis of circulating EPO. In combination, these findings suggest that EPO—in addition to its function as an erythropoietichormone—canalsoactasaparacrinegrowth factor. However, the pathophysiological relevance of EPO effectsoutsidethebonemarrowremainspoorlyunderstood.

T1015 Rate and Predictors of Erythropoietic Growth Factors Use in Chronic Hepatitis C Patients with Treatment Induced Anemia

analyses to clarify HCV's role in DM risk. Methods: Two reviewers performed searches in multiple databases and abstracted data. Eligibility criteria included: 1) minimum of 200 adults-100 cases, 100 controls; 2) published in English as peer-reviewed manuscript. Studies with only post-transplant data were excluded. We performed random effects meta-analyses that combined studies with similar risk measures and similar control groups. Egger's test was used to assess potential small study bias. Results: We identified 31 unique studies. Many were performed in U.S. (n=10) or Italy (n=6). Control groups included: non-HCV/ HBV infected in 16 studies, and HBV-infected, with other liver diseases (OLD), and HIV mono-infected in n=8, n=3 and n=4 studies respectively. Among studies comparing DM risk in HCV cases to that in non-infected controls, 13 reported odds ratios (OR) and 3 longitudinal studies reported hazard ratios (HR). Pooled estimators for studies with ORs indicated excess risk (ORcrude=2.11, 95% CI 1.48-2.74, ORadjusted=1.76, 95% CI 1.08-2.44). 5 adjusted estimates included BMI, with 4 finding additional excess DM risk. Although Egger's test was significant, it was explained by 1 study's effect. Excess DM risk was also observed by pooling HRs (HRadjusted=1.67, 95% CI, 1.28-2.06). All adjusted HRs included BMI with no evidence of small study bias. Among studies with HBV+ controls, both pooled ORs suggested excess, though the adjusted effect was no longer significant ORcrude=1.69, 95% CI 1.15-2.22, ORadjusted=1.99, 95% CI 0.92-3.06). Although lack of comparability in effect measures (HIV) or in controls (OLD) prevented meta-analysis, most individual studies reported excess DM risk with HCV that was particularly suggestive for HIV. Conclusions: In this meta-analysis, pooled estimators indicated HCV infection increases DM risk, with suggestive evidence that the excess risk persists even after controlling for other risk factors like BMI. The meta-analysis of prospective studies reporting HRs in particular bolsters argument that HCV conveys excess risk. Additional research will be needed to identify other factors that mediate DM risk in HCV-infected cases in comparison to non-infected or HBVinfected controls.

Immunomodulatory Therapy for Myelodysplastic Syndromes

Thalidomide and lenalidomide belong to the proprietary group of immunomodulatory drugs (IMiDs) that display broad biologic and pharmacologic properties. Encouraging results of clinical studies that evaluated the efficacy of thalidomide in patients with myelodysplastic syndromes (MDSs) led to the investigation of its structural analogue, lenalidomide, in patients with lower-risk MDS. The cumulative results of studies that tested lenalidomide in patients with interstitial deletion of chromosome 5q, ie, del(5q), showed a high frequency of both erythroid and cytogenetic responses (approximately 75% of patients), which led to US Food and Drug Administration approval of this agent for this cytogenetically defined MDS subset. A multicenter phase III study (MDS-002) that investigated the frequency of transfusion response in lower-risk non-del(5q) MDS patients showed that lenalidomide had significant erythropoietic activity, albeit less robust in lower-risk MDS without del(5q). These studies established lenalidomide as an active erythropoietic-remitting agent with novel cytogenetic-remitting activity in lower-risk MDS patients who would not otherwise benefit from therapy with erythropoietic growth factors. The National Comprehensive Cancer Network Clinical Practice Guidelines recently added lenalidomide to the therapeutic algorithm for MDS as front-line therapy for lower-risk MDS patients with del(5q) and transfusion-dependent anemia.

16 INVITED Follow-on erythropoietins – Pros and cons

In myeloma bone disease, bisphosphonates have been shown to delay the progression of osteolytic lesions and prevent fractures. In the case of painful vertebral fractures, vertebroplasty and kyphoplasty have become standard procedures to control symptoms and restore the original height of the vertebrae. Adequate pain control is of crucial importance for the quality of life of myeloma patients, as is maintaining adequate hemoglobin levels with the use of erythropoietic growth factors. Infections should be treated aggressively in myeloma patients, as these contribute significantly to morbidity and mortality. In patients with repeated infectious complications, prophylactic measures such as long-term application of antibiotic or antiviral medication or use of intravenous immunoglobulins is recommended. The concerted action of these supportive therapies can significantly improve the wellbeing of myeloma patients in phases of disease progression as well as during phases of remission. In progressive disease, certain measures such as adequate pain control and radiotherapy can ameliorate symptoms until the therapeutic effect of systemic anti-neoplastic therapy becomes evident.

Limited value of routine iron studies prior to initiation of erythropoietin therapy in patients with ovarian cancer

19677 Background: Anemia is a common consequence of cancer which significantly impacts patient quality of life. The mainstay of treatment for cancer- and chemotherapy-related anemia is erythropoietin therapy. However, approximately 30% to 50% of patients will not respond to these growth factors. The literature attributes this lack of response to functional iron deficiency, when iron stores are normal but the body cannot meet the increased rate of erythropoiesis. We evaluated the iron status of patients with ovarian malignancies receiving chemotherapy and erythropoietin therapy to establish a baseline for implementation of an intervention service. Methods: After obtaining Institutional Review Board approval, we identified 55 ovarian cancer patients receiving erythropoietin therapy from January to December 2005. We then performed a retrospective chart review for patients who had iron studies available. Results: Thirty-four patients had complete iron studies performed, while an additional 10 had only a ferritin level obtained. The mean hemoglobin for all patients was 9.9g/dl (6.9–13.1) with a mean MCV (mean corpuscular volume) of 92.7fl. Four (12%) patients were iron deficient based on ferritin &lt;100ng/ml and iron saturation &lt;20%. However, these patients had normal MCVs, indicating iron deficiency was not the etiology of their anemia. A few patients were assessed for B12 and folate deficiency, but none were identified. Interestingly, we had 22 patients with elevated ferritin levels (greater than 322ng/ml), with the highest being 2178ng/ml. Conclusions: Our results identified a few patients who were iron deficient, but none were diagnosed with iron deficiency anemia. Therefore, the role of routine iron screening in patients with a normal MCV prior to initiation of erythropoietin therapy is in question. We believe that functional iron deficiency may contribute to anemia in our population. Therefore, we suggest that all patients receive iron supplementation at erythropoietin therapy initiation. We plan to prospectively assess the optimal route of iron administration in ovarian cancer patients in order to improve the response rate to erythropoietic growth factors. No significant financial relationships to disclose.

Anemia associated with pegylated interferon-?2a and ?2b therapy in hemodialysis patients

Anemia is a well-known side effect of interferon therapy since interferons are potent inhibitors of erythropoiesis. The aim of this study was to compare the anemia associated with pegylated interferon (PEG-IFN) (alpha2a versus alpha2b therapy in hemodialysis patients (HD) with chronic hepatitis C. In order to study the anemia, doses of erythropoietic growth factors (EGF), hemoglobin (Hb) and erythropoietin resistance index (ERI) were compared at baseline and after PEG-IFN-alpha2a or alpha2b therapy in 16 HD patients with chronic C hepatitis. Pharmacokinetic studies were performed in 4 of those treated with PEG-IFN-alpha2b and 2 patients treated with PEG-IFN-alpha2a. Secondary end-points were viral response and serious adverse events. At 4-6 months after the beginning of therapy, both PEG-IFN-alpha induced a significant increment in the erythropoietin resistance index. This increment was significantly higher in patients treated with PEG-IFN-alpha2a when compared with alpha2b (45 vs 9.9, p = 0.012). The pharmacokinetics of PEG-IFN-alpha2a and alpha2b in HD patients were different, the C(max), C(min) and the area under the serum concentration time curve, were all higher in patients treated with PEG-IFN-alpha2a compared with PEG-INF-alpha2b. Discontinuation of therapy occurred in 2 (28.5%) of the 7 patients in the PEG-IFN-alpha2a group and in 4 (44%) of the 9 patients in the PEG-IFN-alpha2b group. Three (42%) subjects in the alpha2a group and 5 (55%) in the alpha2b group had a response at the end of the 48 weeks of therapy. In 4 (44.4%) of the 9 patients treated with alpha2b the viral response was sustained. In summary, patients treated with PEG-IFN-alpha2a have a major inhibitory effect on erythropoiesis. This could be explained by the different pharmacokinetic properties of PEG-IFN-alpha2a and alpha2b. Further studies are needed to clarify how these findings influence the efficacy, safety and cost-effectiveness of the PEG-IFN-alpha2.

P139 WT1 gene expression decreases during erythropoietic growth factor treatment in MDS patients

P139 WT1 gene expression decreases during erythropoietic growth factor treatment in MDS patients

Phase I/II Trial of Adjuvant Dose-Dense Docetaxel/Epirubicin/Cyclophosphamide (TEC) in Stage II and III Breast Cancer

This phase I/II trial investigates the safety and feasibility of six cycles of concurrent taxane, anthracycline and cyclophosphamide on a dose dense schedule. Patients with stage II/III breast cancer were treated with docetaxel (T) 75 mg/m(2), epirubicin (E) 75 mg/m(2) (cohort 1, n = 3) or 100 mg/m(2) (cohort 2, n = 12), and cyclophosphamide (C) 500 mg/m(2) IV on day 1, with pegfilgrastim 6 mg subcutaneously on day 2, every 2 weeks for six cycles. Patients were assessed for toxicity every 2 weeks; cardiac function and response (if neoadjuvant) were assessed after six cycles. All patients in cohort 1 received 100% planned dose intensity; in cohort 2, five of twelve patients received 100% and 11/12 received >80%. There were no dose reductions or delays for day 1 myelotoxicity. Dose reductions as a result of febrile neutropenia (FN) occurred in cohort 2, with six of twelve patients experiencing FN in seven of sixty-nine cycles. Six patients had anemia > or =grade 3; five received RBC transfusion and seven received an erythropoietic growth factor. Four patients required dose reductions for nonhematologic toxicity (two mucositis; one neurotoxicity; one diarrhea + cellulitis). Four patients developed thrombophlebitis, which was associated with FN in one of four. Two of fourteen evaluable patients had asymptomatic decreases in LVEF >10%; all remained within normal range. All four patients receiving neoadjuvant TEC had significant clinical responses (one CR, three PR). No pathologic CRs were seen. Dose dense TEC chemotherapy is feasible, has acceptable toxicity at doses equivalent to TAC (docetaxel 75 mg/m(2), epirubicin 75 mg/m(2), cyclophosphamide 600 mg/m(2)), and has moderate but manageable toxicity using a higher epirubicin dose of 100 mg/m(2), with FN occurring in six of twelve patients at the higher dose.

Hemoglobin variation and blood transfusion rates in patients affected by locally advanced cervical cancer undergoing neo-adjuvant chemotherapy followed by radical surgery: the role of erythropoietic growth factors

The objectives of the present study were to evaluate hemoglobin levels and consequent clinical behaviors related to anemia developed in patients affected by locally advanced cervical cancer treated with neo-adjuvant chemotherapy in the last decade and to evaluate the impact that the introduction of erythropoietic growth factors had in the clinical practice. Blood chemistries, prospectively recorded from 98 cervical cancer patients, treated with neo-adjuvant chemotherapy and, if necessary, erythropoietic growth factors, were compared with matched historical controls before the introduction of growth factors in clinical practice. Hemoglobin level in the study group did not differ significantly during chemotherapy. At the third cycle of chemotherapy and at the end of chemotherapy, hemoglobin level was significantly higher in the study group compared with the control group. Transfusion rates in the study group were significantly lower. The analysis within the study group revealed that hemoglobin level in patients who suffer at diagnosis from anemia tends to increase whereas hemoglobin level in nonanemic patients tends to decrease. Erythropoietic growth factors increase hemoglobin level and reduce blood transfusions in cervical cancer patients undergoing neo-adjuvant chemotherapy followed by radical surgery. An appropriate autologous blood donation program can noticeably reduce homologous blood transfusions.

Outcomes of an Erythropoietic Growth Factor Interchange Program in Hospitalized Chronic Hemodialysis Patients

Background The substantial cost of erythropoietic growth factors (EGFs), darbepoetin alfa and epoetin alfa, has led many institutions to evaluate EGF usage and costs in the hospital setting. The primary objectives of this study were to determine the clinical impact on hemoglobin (Hb) concentration, postdischarge epoetin alfa dose requirements, and the economic impact of a darbepoetin alfa for epoetin alfa therapeutic interchange program in hospitalized hemodialysis patients. Methods A retrospective review was conducted in 79 patients (43 during a control period and 36 during the interchange period). The control group consisted of chronic hemodialysis patients admitted to the hospital before the therapeutic interchange program was implemented (September 2002 through September 2003); the interchange group included patients admitted after implementation of the interchange program (October 2003 through October 2004). Clinical parameters including Hb concentration, concomitant iron therapy, packed red blood cell (PRBC) transfusions, EGF dose and cost were collected beginning 2 months prior to hospitalization, during hospitalization, through 2 months post hospital discharge. Results No differences were observed between groups in baseline demographics. There was a greater decrease in Hb concentration in the control group compared to the interchange group, but this did not achieve statistical significance (0.5 g/dL vs 0.3 g/dL, respectively; P = 0.67). EGF dose requirements did not differ between the two groups, nor did the total EGF cost per patient admission before and after hospitalization. Total cost per patient during hospitalization was $164 ± 144 in the control group and $165 ± 156 in the interchange EGF group ( P= 0.97). Conclusion: In the treatment of anemia in hospitalized chronic hemodialysis patients, there were no differences seen in Hb concentration, EGF dose requirements, or overall EGF cost per patient admission. Implementation of the interchange program allowed for market share gains to permit an overall economic benefit across all patients in the amount of $300,000 annually.

Consumption of C.E.R.A. and Epoetin Beta in a Cellular Assay: UT-7 Consumption Model.

Continuous Erythropoietin Receptor Activator (C.E.R.A.), an innovative erythropoietic agent with unique receptor activity, is currently in development to provide correction of anemia and stable maintenance of hemoglobin (Hb) levels at extended administration intervals up to once monthly in patients with all stages of chronic kidney disease, and is also in development for the treatment of chemotherapy-induced anemia. C.E.R.A. has an equivalent half-life in humans of approximately 130 h following subcutaneous or intravenous administration. Clearance of erythropoietic agents may occur by binding, internalization and degradation in receptor bearing cells in the bone marrow. A functional assay was developed to evaluate the consumption of C.E.R.A. and epoetin beta in a UT-7 cellular system (a human acute myeloid leukemia cell line whose growth is dependent on the presence erythropoietic and other growth factors). UT-7 cells were incubated in the presence of C.E.R.A. (1000 pM) or epoetin beta (100 pM) for 72, 96 or 120 h. The cell supernatants were then harvested and the amounts of remaining growth factor were determined by a validated erythropoietin ELISA. As controls, the growth factors were incubated for the same times in medium only. Five independent identical experiments were conducted for each compound. Mean percentages of C.E.R.A. and epoetin beta were calculated relative to the control values (set at 100%). C.E.R.A. 1000 pM and epoetin beta 100 pM were shown to be equipotent, stimulating the proliferation of UT-7 cells to the same degree. There was a highly significant difference (P=1.9×10−9) between C.E.R.A. and epoetin beta for the time course of concentrations relative to controls. For epoetin beta 100 pM, the relative concentration decreased significantly over time, while for C.E.R.A. 1000 pM no statistically significant changes were observed. Thus, C.E.R.A. was still present in the culture supernatant at close to 100% after 4–5 days' incubation, while the epoetin beta concentration had declined to approximately 20%. In conclusion, at the end of the incubation time the C.E.R.A. concentration was still sufficient to maintain the initial level of cell stimulation. However, at the end of the 96 h incubation the epoetin beta concentration dropped to a level that was insufficient to produce further stimulation. The UT-7 consumption model thus emulates the in vivo situation in humans with regard to receptor mediated elimination of the compounds and stimulation of erythroid progenitors.This study provides further evidence that C.E.R.A. interacts differently with the erythropoietin receptor relative to epoetin beta. Preclinical studies have shown that these properties translate into more continuous stimulation of erythropoiesis in vivo compared with epoetin beta and Phase III data indicate that C.E.R.A. allows predictable Hb maintenance in direct conversion from more frequently dosed agents.