BIOEQUIVALENCE of a BIOSIMILAR FILGRASTIM and REFERENCE FILGRASTIM: IMPACT ON the MOBILIZATION of CD34+ PERIPHERAL BLOOD PROGENITOR CELLS in a RANDOMIZED, PHASE I STUDY.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 4564-4564
Author(s):  
Miguel H Bronchud ◽  
Cornelius F Waller ◽  
Stuart J Mair ◽  
Rodeina Challand
Keyword(s):  
Phase I ◽  
Cell Count ◽  
Progenitor Cells ◽  
Clinical Development ◽  
Post Dose ◽  
Double Blind ◽  
Hematopoietic Stem ◽  
Cell Counts ◽  
Peripheral Blood Progenitor Cells ◽  
Biosimilar Filgrastim

Abstract Abstract 4564 Background Granulocyte colony-stimulating factor (G-CSF) stimulates the proliferation and differentiation of hematopoietic stem and progenitor cells. Recombinant human G-CSF (filgrastim) was developed by Amgen to enhance mobilization of peripheral blood progenitor cells (PBPCs) prior to autologous or allogeneic hematopoietic stem/progenitor cell transplantation. After the patent expiry of Amgen filgrastim (Neupogen®), Hospira developed a biosimilar filgrastim. Preclinical and phase I studies supported the bioequivalence of Hospira filgrastim and Amgen filgrastim in terms of their physicochemical, pharmacokinetic and pharmacodynamic characteristics (Skrlin A, et al. EHA 2009, Abstract 0568; Waller CF, et al. EHA 2009, Abstract 0562). Here we compare the impact of Hospira filgrastim and Amgen filgrastim on CD34+ PBPC mobilization in a randomized, double-blind phase I study. Aim To demonstrate the bioequivalence of Hospira filgrastim and Amgen filgrastim with respect to the mobilization of CD34+ PBPCs. Methods Healthy, male or female volunteers aged 18–50 years were enrolled at Charles River Clinical Services, Edinburgh, UK, between 2 November 2006 and 24 January 2007. Using a computer-generated randomization list, volunteers were first randomized to 5μg/kg or 10μg/kg dose groups, before further randomization to order of agent administration. Subcutaneous injections of Hospira filgrastim or Amgen filgrastim were administered under double-blind conditions on five consecutive days (days 1–5), with crossover to the alternative agent after a washout period of ≥13 days. Blood samples were taken at day 1 (pre dose), day 3 (6 hours [h] post dose), day 5 (6 h post dose), day 7 (48 h post dose) and day 10 (120 h post dose). Mean CD34+ cell counts were evaluated by flow cytometry and bioequivalence was assessed using a mixed effects analysis of variance model. Bioequivalence was concluded if the 90% confidence intervals (CI) for the ratio of ‘test’ (Hospira filgrastim) to ‘reference’ (Amgen filgrastim) mean CD34+ cell counts at day 5 were completely within the conventional bioequivalence limits of 0.80–1.25. Results Twenty-four volunteers were randomized to the 5μg/kg group and 26 to the 10μg/kg group. At both doses, CD34+ cell counts were similar with Hospira filgrastim and Amgen filgrastim across all time points. Regardless of agent or dose, mean CD34+ cell count at day 1 was 2.2–2.8 cells/μl, increasing to a maximum count at day 5. In the 5μg/kg group, mean CD34+ cell count at day 5 (n=24) was 47.2 cells/μl (95% CI: 36.1, 61.7) with Hospira filgrastim and 46.0 cells/μl (95% CI: 33.6, 63.0) with Amgen filgrastim. In the 10μg/kg group, mean CD34+ cell count at day 5 (n=23) was 81.9 cells/μl (95% CI: 64.5, 104.0) with Hospira filgrastim and 77.5 cells/μl (95% CI: 59.4, 101.3) with Amgen filgrastim (Figure 1). At both doses, 90% CIs for the ratios of test to reference mean CD34+ cell counts at day 5 were within the predefined range required to demonstrate bioequivalence. The incidence of adverse events (AEs) was slightly lower with Hospira filgrastim than with Amgen filgrastim at both doses (5μg/kg, 79 vs 83%; 10μg/kg, 77 vs 92%). The most frequently reported AEs of any severity (mild/moderate/severe) with each agent at each dose were back pain (38–62%) and headache (44–58%), which could be treated with standard analgesics. Conclusions Hospira filgrastim and Amgen filgrastim are similar in their ability to stimulate mobilization of CD34+ PBPCs. These data add to a growing body of evidence in support of the bioequivalence of these agents. Hospira filgrastim may provide a useful alternative to Amgen filgrastim as a growth factor to support PBPC mobilization and transplantation. The bioequivalence of these agents may enable Hospira filgrastim to be used without the need to adapt PBPC harvesting protocols. The continued clinical development of Hospira filgrastim is warranted. Acknowledgments Medical writing support provided by Hannah FitzGibbon (GeoMed) with financial support from Hospira. Disclosures: Bronchud: Hospira UK Ltd: Consultancy. Waller:Hospira UK Ltd: Consultancy. Off Label Use: Hospira filgrastim is a biosimilar filgrastim that is in clinical development for the treatment of neutropenia associated with cytotoxic chemotherapy.. Mair:Charles River Clinical Services: Employment. Challand:Hospira UK Ltd: Employment.

In Vitro Manipulation of Peripheral Blood Progenitor Cell Collections

1998 ◽  
Vol 21 (6_suppl) ◽  
pp. 1-10
Author(s):  
C. Carlo-Stella ◽  
V. Rizzoli
Keyword(s):  
Progenitor Cells ◽  
Peripheral Blood ◽  
Progenitor Cell ◽  
High Dose ◽  
Hematopoietic Stem ◽  
Peripheral Blood Progenitor Cells ◽  
Stem And Progenitor Cells ◽  
Therapy Strategies ◽  
Mobilized Peripheral Blood

Mobilized peripheral blood progenitor cells (PBPC) are increasingly used to reconstitute hematopoiesis in patients undergoing high-dose chemoradiotherapy. PBPC collections comprise a heterogeneous population containing both committed progenitors and pluripotent stem cells and can be harvested (i) in steady state, (ii) after chemotherapeutic conditioning, (iii) growth factor priming, or (iv) both. The use of PBPC has opened new therapeutic perspectives mainly related to the availability of large amounts of mobilized hematopoietic stem and progenitor cells. Extensive manipulation of the grafts, including the possibility of exploiting these cells as vehicles for gene therapy strategies, are now possible and will be reviewed.

scholarly journals Phase I Evaluation of ΔvirG Shigella sonnei Live, Attenuated, Oral Vaccine Strain WRSS1 in Healthy Adults

2002 ◽  
Vol 70 (4) ◽  
pp. 2016-2021 ◽  
Author(s):  
Karen L. Kotloff ◽  
David N. Taylor ◽  
Marcelo B. Sztein ◽  
Steven S. Wasserman ◽  
Genevieve A. Losonsky ◽  
...  
Keyword(s):  
Phase I ◽  
Oral Vaccine ◽  
Geometric Mean ◽  
Healthy Adults ◽  
Shigella Sonnei ◽  
Low Grade ◽  
Double Blind ◽  
Antibody Secreting Cell ◽  
Cell Counts ◽  
Group 2

ABSTRACT We conducted a phase I trial with healthy adults to evaluate WRSS1, a live, oral ΔvirG Shigella sonnei vaccine candidate. In a double-blind, randomized, dose-escalating fashion, inpatient volunteers received a single dose of either placebo (n = 7) or vaccine (n = 27) at 3 × 103 CFU (group 1), 3 × 104 CFU (group 2), 3 × 105 CFU (group 3), or 3 × 106 CFU (group 4). The vaccine was generally well tolerated, although a low-grade fever or mild diarrhea occurred in six (22%) of the vaccine recipients. WRSS1 was recovered from the stools of 50 to 100% of the vaccinees in each group. The geometric mean peak anti-lipopolysaccharide responses in groups 1 to 4, respectively, were 99, 39, 278, and 233 for immunoglobulin (IgA) antibody-secreting cell counts; 401, 201, 533, and 284 for serum reciprocal IgG titers; and 25, 3, 489, and 1,092 for fecal IgA reciprocal titers. Postvaccination increases in gamma interferon production in response to Shigella antigens occurred in some volunteers. We conclude that WRSS1 vaccine is remarkably immunogenic in doses ranging from 103 to 106 CFU but elicits clinical reactions that must be assessed in further volunteer trials.

Efficient Gene Transfer into Human CD34+ Peripheral Blood Progenitor Cells Using Pseudotyped Recombinant Adeno-Associated Viral Vectors

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 4627-4627
Author(s):  
Leopold Sellner ◽  
Marlon Veldwijk ◽  
Marius Stiefelhagen ◽  
Jurgen A. Kleinschmidt ◽  
Stephanie Laufs ◽  
...  
Keyword(s):  
Gene Transfer ◽  
Progenitor Cells ◽  
Safety Profile ◽  
Peripheral Blood ◽  
Vector System ◽  
Hematopoietic Stem ◽  
Human Cd34 ◽  
Peripheral Blood Progenitor Cells ◽  
The Difference ◽  
Relevant Gene

Abstract Due to their pluripotency, human CD34+ hematopoietic stem cells are targets of interest for the treatment of many acquired and inherited disorders of the hematopoietic system using gene therapeutic approaches. Human CD34+ peripheral blood progenitor cells (PBPC) can be readily mobilized into the blood and harvested by leukapheresis, providing an easily accessible source of hematopoietic progenitor cells. Unfortunately, for gene transfer into CD34+ PBPC, most current vector systems either lack sufficient transduction efficiency or an acceptable safety profile. Standard adeno-associated virus-based vectors have an advantageous safety profile, yet lack the required efficiency. Therefore a panel of pseudotyped recombinant adeno-associated viral (rAAV2/1 - rAAV2/6) vectors expressing the eGFP gene was screened on human G-CSF-mobilized CD34+ PBPC to determine their efficacy. In addition, the difference in transgene expression between conventional single-stranded rAAV and self complementary rAAV (scAAV) vectors was determined. For each vector n≥6 was performed and data are shown as mean ± SD. Of all screened conventional rAAV vectors, rAAV2/6 proved to be the most efficient (13.5% ± 9.8% GFP+ and CD34+ PBPC; p<0.001 vs other vectors) on human CD34+ PBPC, followed by rAAV2/2 (2.6% ± 2.0% GFP+ cells) and rAAV2/1 (1.4% ± 1.2% GFP+ cells). For rAAV2/3, rAAV2/4 and rAAV2/5 no relevant gene transfer efficiency (<1% GFP+ cells) was observed. Furthermore, the relevance of the single-to-double-strand conversion block in transduction of human PBPC could be shown using scAAV vectors. scAAV2/6 and scAAV2/2 (both p<0.001) showed significantly higher gene expression (38.4% ± 12.2% and 11.8% ± 5.7% GFP+ cells, respectively) compared to their conventional counterparts in this cell entity. Similar results were observed for scAAV2/1 vectors (2.8% ± 1.9% GFP+ cells), though the difference was not significant. Of note, as previously observed using AAV peptide library-derived rAAV vectors (Sellner et al., 2008, Exp Hematol. 36), also here inter-patient variances in CD34+ PBPC susceptibility were found. For the first time we were able to obtain clinically relevant gene transfer and expression levels (>10%) with expression rates up to 60% in human CD34+ PBPC using an AAV-based vector system, thereby providing an efficient alternative vector system for gene transfer into this clinically important target cell.

scholarly journals Altered kinetics of circulating progenitor cells in cardiopulmonary bypass (CPB) associated vasoplegic patients: A pilot study

2020 ◽  
Author(s):  
Sanhita Nandi ◽  
Uma Rani Potunuru ◽  
Chandrani Kumari ◽  
Abel Arul Nathan ◽  
Jayashree Gopal ◽  
...  
Keyword(s):  
Stem Cells ◽  
Logistic Regression ◽  
Cardiopulmonary Bypass ◽  
Hematopoietic Stem Cells ◽  
Progenitor Cells ◽  
Gradient Boosting ◽  
Hematopoietic Stem ◽  
Elective Cardiac Surgery ◽  
Cell Counts ◽  
Clinically Significant

AbstractVasoplegia observed post cardiopulmonary bypass (CPB) is associated with substantial morbidity, multiple organ failure and mortality. Circulating counts of hematopoietic stem cells (HSCs) and endothelial progenitor cells (EPC) are potential markers of neo-vascularization and vascular repair. However, the significance of changes in the circulating levels of these progenitors in perioperative CPB, and their association with post-CPB vasoplegia, are currently unexplored. We enumerated HSC and EPC counts, via flow cytometry, at different time-points during CPB in 19 individuals who underwent elective cardiac surgery. These 19 individuals were categorized into two groups based on severity of post-operative vasoplegia, a clinically insignificant vasoplegic Group 1 (G1) and a clinically significant vasoplegic Group 2 (G2). Differential changes in progenitor cell counts during different stages of surgery were compared across these two groups. Machine-learning classifiers (logistic regression and gradient boosting) were employed to determine if differential changes in progenitor counts could aid the classification of individuals into these groups. Enumerating progenitor cells revealed an early and significant increase in the circulating counts of CD34+ and CD34+CD133+ hematopoietic stem cells (HSC) in G1 individuals, while these counts were attenuated in G2 individuals. Additionally, EPCs (CD34+VEGFR2+) were lower in G2 individuals compared to G1. Gradient boosting outperformed logistic regression in assessing the vasoplegia grouping based on the fold change in circulating CD 34+ levels. Our findings indicate that a lack of early response of CD34+ cells and CD34+CD133+ HSCs might serve as an early marker for development of clinically significant vasoplegia after CPB.

Cell Therapy in a Mouse Model of Immune-Mediated Bone Marrow Failure.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 1689-1689
Author(s):  
Jichun Chen ◽  
Neal S. Young
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
Progenitor Cells ◽  
Stromal Cells ◽  
Stromal Cell ◽  
Adherent Cells ◽  
Hematopoietic Stem ◽  
Cell Counts ◽  
Immune Mediated ◽  
Stem And Progenitor Cells

Abstract Immune-mediated bone marrow (BM) failure has been modeled in the mouse by infusion of lymph node cells from allogeneic C57BL/6 (B6) donors into major or minor histocompatibility antigen-mismatched recipients (Chen et al., Blood 2004; Bloom et al., Exp Hematol 2004, Chen et al., J Immunol 2007). Co-infusion of limited numbers of CD4+CD25+ regulatory T lymphocytes (Tregs) can alleviate clinical manifestations by suppressing the expansion of pathogenic T cells (Chen et al., J Immunol 2007). In the current study, we investigated the effectiveness of Tregs and suppressor cells contained in BM stroma in this fatal disease. Infusion of fewer than 3 × 103 Tregs to each recipient mouse had only a minor effect in preserving BM cells and did not prevent pancytopenia. Fifteen-50 × 103 thymic Tregs was moderately protective: blood WBC, RBC, platelet and BM cell counts at three weeks after cell infusion were 197%, 116%, 155% and 158% of those of control animals that did not receive Treg infusion; 5–10 × 103 B6 splenic Tregs produced the largest effect as WBC, RBC, platelet and BM cell counts were 275%, 143%, 276%, and 198% of controls. Overall, Treg therapy was helpful but its effectiveness was limited and variable among individual recipients as no antigen-specific Tregs can be identified for the treatment of BM failure. Learned about the immunosuppressive effects of mesenchymal stem cells (MSCs), we went on to test the effectiveness of stromal cells as another therapeutic modality for BM failure, since stromal cells contain MSCs. These cells were derived from B6 BM by culture in α-modified Eagle medium at 33°C with 5% CO2 for two weeks. After separating the non-adherent cells, we detached the adherent stromal cells and infused them into TBI + B6 LN-infused C.B10 mice. Injection of 106 stromal cells at the time of LN cell infusion effectively preserved WBCs (3.09 ± 0.51 vs 0.61 ± 0.18), RBCs (8.72 ± 0.14 vs 3.52 ± 0.46), platelets (924 ± 93 vs 147 ± 25) and BM cells (186.6 ± 8.7 vs 52.7 ± 7.8) when compared to LN-cell-infused mice without stromal cell addition. Delayed stromal cell injection at day 9 after LN cell infusion had only a mild effect on the preservation of RBCs (147%), platelets (276%) and BM cells (223%) and no effect on WBCs (64%), and infusion of non-adherent cells from the same stromal cell culture had no therapeutic effect. Stromal cell-infused mice had higher proportion of FoxP3+CD4+ cells in the peripheral blood (59.7 ± 10.7% vs 29.8 ± 5.4%) and more Lin−CD117+CD34− hematopoietic stem and progenitor cells in the BM (591 ± 95 vs 60 ± 43, thousand) in comparison to LN cell infused mice without stromal cell treatment. Mitigation of pathogenic T cells, including both CD4 and CD8 T lymphocytes, is the potential mechanism for the effectiveness of Treg and stromal cell therapies that helped to protect hematopoietic stem and progenitor cells in the BM of affected animals. Figure Figure

Hypoxia Potentiates Notch-Induced Expansion of Human Hematopoietic Stem/Progenitor Cells

Blood ◽  
2016 ◽  
Vol 128 (22) ◽  
pp. 2640-2640
Author(s):  
Jianfei Fu ◽  
Heather D. Huntsman ◽  
Ayla Cash ◽  
Patali S. Cheruku ◽  
Richard H. Smith ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Research Funding ◽  
Ex Vivo ◽  
Post Transplantation ◽  
Hematopoietic Stem ◽  
Nsg Mice ◽  
Cell Counts ◽  
Cd34 Cells ◽  
Number Of Cells

Abstract Activation of Notch signaling in human hematopoietic stem/progenitor cells (HSPCs) by treatment with Notch ligand Delta1 has enabled a clinically relevant ex vivo expansion of short-term HSPCs. In vitro studies have also revealed a role of low O2 tension in HSPC regulation. A molecular link has been demonstrated in several stem/progenitor cell populations between Notch and hypoxia pathways but their interaction has not been investigated in human HSPCs. G-CSF mobilized human CD34+ cells from 4 healthy subjects were cultured in the presence of cytokines (SCF, FLT3L and TPO) in hypoxia (1.5-2% O2) or normoxia (21% O2) in vessels coated with fibronectin alone or combined with increasing concentrations of the immobilized ligand Delta1 (2.5, 5, 10 and 20 µg/mL). After 21 days in culture, cells were counted and characterized using CFU assays, flow cytometry for lineage (Glycophorin A+, CD13+, CD20+, CD3+ and CD41+ cells) and HSC (CD34+ CD38- CD45RA- CD90+ CD49f+ Rholow) phenotypes, and transplantation in immunodeficient (NSG) mice. In normoxia, the total number of cells increased 118-fold compared to baseline in the absence of Delta1 with limited residual CD34+ cells (1.5 ± 0.7%), extensive differentiation toward the myeloid lineage (96.3 ± 0.3% CD13+ cells) and minimal engraftment potential in NSG mice (0.2 ± 0.2% human CD45+ cells). With increasing concentrations of Delta1 in normoxia, consistent with the hypothesis that Delta1 delays differentiation, the total number of cells increased less (41-, 25-, 11- and 7-fold relative to baseline, respectively) CD34+ cells expanded more (4-, 4-, 3- and 2-fold relative to baseline, respectively), and CFU numbers increased more (8-, 7-, 4- and 3-fold relative to baseline, respectively) than without Delta1. However, phenotypically defined HSCs were undetectable or markedly decreased at the lowest Delta1 concentrations used (2.5 and 5 µg/mL) and their numbers were maintained or only minimally increased at the highest Delta-1 concentrations tested (10 and 20 µg/mL) relative to uncultured CD34+ cells. Accordingly, only cells cultured with 10 and 20 µg/mL Delta1 resulted in levels of engraftment in NSG mice (5.5 ± 5.4% and 5.4 ± 0.9% human CD45+ cells, respectively) comparable to uncultured cells (7.0 ± 0.1% human CD45+ cells). In hypoxia, total cell counts increased less than in normoxia both without (8-fold relative to baseline) and with increasing concentrations of Delta1 (11-, 11-, 9-, 9-fold relative to baseline, respectively) due to diminished myeloid differentiation. Total CD34+ cells decreased 1.7-fold in hypoxia in the absence of Delta1, but expanded modestly in the presence of Delta1 (3-, 3-, 2- and 2-fold, respectively). CFU numbers followed a similar trend. However, in hypoxic cultures with 2.5, 5 and 10 µg/mL Delta1, phenotypically defined HSCs increased 2.5-, 6.6- and 1.3-fold, respectively, compared to uncultured cells. Importantly, hypoxia combined with 2.5, 5 and 10 µg/mL Delta1 concentrations resulted in increased human cell engraftment in NSG mice (21.2 ± 4.4%, 29.3 ± 11% and 11.8 ± 5.4% human CD45+ cells, respectively) compared to uncultured cells (7.0 ± 0.1% human CD45+ cells). When 20 µg/mL Delta1 was used in hypoxia, engraftment potential in NSG mice was decreased (1.1 ± 0.6% human CD45+ cells). We next performed limiting dilution analysis to measure the frequencies of long-term repopulating HSCs (LT-HSCs) within the CD34+ cell compartment at baseline and after 21 days in hypoxic or normoxic cultures supplemented with the optimized concentrations of Delta1 (10 µg/mL in normoxia and 5 µg/mL in hypoxia). LT-HSCs in uncultured CD34+ cells were measured at the expected frequency (1 in 7,706; 95% CI of 3,446 to 17,232). When analyzed at 3 months post-transplantation, a limited (1.5-fold) increase in LT-HSC frequency (1 in 5,090; 95% CI 2.456 to 10,550) was obtained from Delta1 normoxic cultures compared to uncultured cells. In contrast, the frequency of LT-HSCs (1 in 1,586; 95% CI 680 to 3,701) was 4.9-fold higher in hypoxic Delta1 cultures compared to uncultured cells, and 4.2-fold higher than in normoxic Delta1 cultures. Similarly, absolute numbers of LT-HSCs per 100,000 Day 0 equivalent CD34+ cells increased from 13 (baseline) to 216 (normoxia) and 694 (hypoxia). Our data indicate that hypoxia potentiates Notch-induced expansion of human HSPCs and may be of benefit in stem cell transplantation and gene therapy applications. Disclosures Cheruku: Novartis: Research Funding. Larochelle:Novartis: Research Funding.

scholarly journals A phase I study of paclitaxel for mobilization of peripheral blood progenitor cells

1999 ◽  
Vol 23 (4) ◽  
pp. 311-315 ◽  
Author(s):  
BA Burtness ◽  
A Psyrri ◽  
M Rose ◽  
E D’Andrea ◽  
C Staugaard-Hahn ◽  
...  
Keyword(s):  
Phase I ◽  
Progenitor Cells ◽  
Peripheral Blood ◽  
Phase I Study ◽  
Peripheral Blood Progenitor Cells
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