Recombinant Granulocyte Colony-stimulating Factor Research Articles

The efficacy and safety of the prophylactic application of PEG-rhG-CSF in radiotherapy with weekly concurrent chemotherapy for cervical cancer

ObjectiveTo evaluate the efficacy and safety of pegylated recombinant human granulocyte colony stimulating factor (PEG-rhG-CSF) in preventing neutropenia during concurrent radiotherapy combined with weekly chemotherapy in patients with cervical cancer.MethodsFrom September 2019 to November 2020, 180 patients with cervical cancer who required concurrent chemoradiotherapy (CCRT) were enrolled in this study. The patients were randomly divided into the following three groups at a ratio of 1:1:1: Group A (PEG-rhG-CSF), Group B (PEG-rhG-CSF + rhG-CSF) and the control group (rhG-CSF). Cisplatin or carboplatin was administered weekly. The primary endpoint was the incidence of grade 3/4 neutropenia. The secondary endpoints were the incidence of febrile neutropenia (FN), delay of radiotherapy, chemotherapy dose reduction, rate of PFS at 2 years and 3 years, and incidence of adverse reactions.ResultsSixty patients were randomly assigned to each group. Two patients in Group B withdrew from the trial for personal reasons. The incidence of grade 1/2 neutropenia in the Control group was significantly lower than that in Group A and Group B (P < 0.001). However, the incidence of grade 3/4 neutropenia in the Control group was significantly greater than that in Group A and Group B (P < 0.001). There was no significant difference in the incidence of FN among the three groups (P = 0.771). The rate of platinum dose reduction in Group A (6.7%) and Group B (5.2%) was significantly lower than that in the Control group (30.0%) (P < 0.001). The delay rate of radiotherapy in Group A (10%) and Group B (13.8%) was obviously inferior to that in the Control group (31.7%) (P = 0.006). The 2-year PFS rates of Group A, Group B, and the Control group were 93.3%, 98.3%, and 91.7%, respectively, with no statistically significant difference (P = 0.270). The 3-year PFS rates were 89.8%, 94.1%, and 84.9%, respectively, with no statistically significant difference (P = 0.178). No treatment-related serious adverse events occurred in any of the patients.ConclusionIn Group A and Group B, The prophylactic application of PEG-rhG-CSF during conventional chemoradiotherapy combined with weekly chemotherapy for patients with cervical cancer can reduce the incidence of grade 3/4 neutropenia and does not increase the incidence of adverse reactions when compared to Control group.Trial registrationchictr.org.cn, ChiCTR1900026309, registered 29 September 2019-prospectively registered.

Differential impact of pegfilgrastim, a recombinant human granulocyte colony stimulating factor, on the neutrophil count of male and female deer mice (Peromyscus maniculatus bairdii)

BackgroundAn increasing body of research implicates inflammatory processes, including alterations in the neutrophil-lymphocyte ratio (NLR), in the pathophysiology of psychiatric illness. The deer mouse (Peromyscus maniculatus bairdii) is commonly studied for its naturalistic expression of compulsive-like behaviour. Towards future efforts to gain an understanding of how innate and adaptive immune processes might be involved in this model, we aimed to study the effects of pegfilgrastim, a pegylated recombinant human granulocyte colony-stimulating factor (g-CSF) analogue, on the NLR of both male and female deer mice.MethodsBriefly, 54 deer mice (equally distributed between sexes) were exposed to a single injection with either control or pegfilgrastim (0.1 or 1 mg/kg) (n = 18 per group). Six mice of each group (three per sex) were euthanized on days two, four and seven post-administration, their blood collected and the NLR calculated. Data were analysed by means of ordinary three-way ANOVA, followed by Bonferroni post-hoc testing.ResultsIrrespective of dose, pegfilgrastim resulted in higher NLR values in mice of both sexes at days four and seven of testing. However, female mice exposed to the higher dose, presented with significantly higher NLR values irrespective of time, compared to male mice exposed to the same.ConclusionThe data generated from this work highlight important dose- and sex-specific aspects of pegfilgrastim with female mice showing heighted elevation of the NLR in response to high-dose pegfilgrastim administration only. Since the innate immune components of male and female deer mice is differentially sensitive to g-CSF stimulation, our results provide a useful basis for further study of sex-specific immunological processes in deer mice.

Demonstration of Physicochemical and Functional Similarity of Biosimilar Pegfilgrastim-cbqv to Pegfilgrastim.

Pegfilgrastim-cbqv/CHS-1701 (UDENYCA®) (hereafter referred to as pegfilgrastim-cbqv) was approved in 2018 by the US Food and Drug Administration as a biosimilar for pegfilgrastim (Neulasta®) (hereafter referred to as pegfilgrastim). Both pegfilgrastim-cbqv and pegfilgrastim are conjugates of recombinant human granulocyte colony stimulating factor (r-metHuG-CSF) with a 20kDa polyethylene glycol (PEG) indicated to decrease the incidence of infection, as manifested by febrile neutropenia, in patients receiving myelosuppressive anticancer drugs. The demonstration of analytical similarity for PEG-protein conjugates presents unique challenges since both the protein and PEG attributes must be characterized. The current study demonstrates the analytical similarity of pegfilgrastim-cbqv and the reference product, pegfilgrastim. In addition to the physicochemical and functional characterization of the protein, the study assessed attributes specific to PEGylation including PEG size and polydispersity, site of attachment, linker composition, and PEGylation process-related variants. The structural, functional, and stability attributes of pegfilgrastim-cbqv and pegfilgrastim were compared using state-of-the-art analytical methods. For the protein, the primary structure, disulfide structure, and secondary and tertiary structures were assessed using traditional protein characterization techniques such as mass spectrometry (MS), circular dichroism (CD), intrinsic fluorescence, and differential scanning calorimetry (DSC), as well as more advanced techniques such as two-dimensional (2D) nuclear magnetic resonance (NMR) and hydrogen deuterium exchange (HDX). For the PEG moiety, the site of attachment, occupancy, linker composition, size and polydispersity were compared using mass spectrometry (both intact and after endoprotease digestion), multiangle light scattering detection (MALS), and Edman degradation. Purity assessments included the assessment of both protein variants and PEGylation variants using chromatographic and electrophoretic analytical separation techniques. The functional similarity between pegfilgrastim-cbqv and pegfilgrastim was compared using both a cell-based bioassay and surface plasmon resonance (SPR). The degradation rates and stability profiles were compared under accelerated and stressed conditions. Biosimilarity was demonstrated by a thorough assessment of physiochemical and functional attributes, as well as comparative stability, of pegfilgrastim-cbqv relative to pegfilgrastim. These studies demonstrated identical primary structure and disulfide structure, highly similar secondary and tertiary structure, as well as functional similarity. The impurity profile of pegfilgrastim-cbqv was comparable to that of pegfilgrastim with only minor differences in PEGylation variants and a slight offset in the PEG molar mass. These differences were not clinically relevant. The degradation profiles were qualitatively and quantitatively similar under accelerated and stress conditions. The structural, functional, and stability data demonstrate that pegfilgrastim-cbqv is highly similar to the reference product, pegfilgrastim.

Guard 01: A randomized, multicentre, controlled trail on the prophylactic use of efbemalenograstim alfa for concurrent chemo-radiotherapy induced neutropenia.

TPS12155 Background: Concurrent chemo-radiotherapy plays a crucial role in the treatment of various advanced-stage malignant tumors. However, it is often associated with significant hematologic toxicity, especially leukopenia and neutropenia, which may lead to treatment interruptions, reduced efficacy, and even severe infections. Efbemalenograstim alfa is a novel long-acting recombinant human granulocyte colony stimulating factor (rhG-CSF) combined of an immunoglobulin (Ig) G2-Fc fragment and two rhG-CSF molecules. Although efbemalenograstim alfa has been confirmed to be effective in the prevention of chemotherapy induced neutropenia by a number of clinical studies, its prophylactic efficacy during concurrent chemo-radiotherapy was seldom investigated. Guard-01 is aimed to evaluate the effectiveness and safety of primary preventive use of efbemalenograstim alfa in concurrent chemo-radiotherapy. Methods: This study is a multicenter, open-label, randomized, controlled clinical trial involving 120 patients who are planned to receive definitive concurrent chemo-radiotherapy with non-small cell lung cancer, small cell lung cancer, esophageal cancer, nasopharyngeal carcinoma, head and neck squamous cell carcinoma, or cervical cancer. Patients will be randomized into efbemalenograstim alfa arm or control arm in a 1:1 ratio on the first day of the first concurrent chemo-radiotherapy cycle. The stratification factor is chemotherapy regimen of the patients. Approximately 48 (±4) hours after the completion of each chemotherapy cycle, patients in the efbemalenograstim alfa arm will receive efbemalenograstim alfa (20 mg per dose, subcutaneous injection) for prevention. Patients in the control arm will receive rhG-CSF (5 μg/kg/day) when the absolute neutrophil count (ANC) is less than 1.0×109/L, and treatment will be discontinued when ANC reaches ≥5×109/L. The primary endpoint is the incidence of grade ≥3 neutropenia (ANC&lt;1.0×109/L) during concurrent chemo-radiotherapy. The trial is opened in November 2023 and will be conducted in 8 hospitals with the aim of recruiting 120 evaluable patients (4 patients are recruited untilled 29-Jan-2024). Clinical trial information: ChiCTR2300077504.

Cytokine polarized, alternatively activated bone marrow neutrophils drive axon regeneration.

The adult central nervous system (CNS) possesses a limited capacity for self-repair. Severed CNS axons typically fail to regrow. There is an unmet need for treatments designed to enhance neuronal viability, facilitate axon regeneration and ultimately restore lost neurological functions to individuals affected by traumatic CNS injury, multiple sclerosis, stroke and other neurological disorders. Here we demonstrate that both mouse and human bone marrow neutrophils, when polarized with a combination of recombinant interleukin-4 (IL-4) and granulocyte colony-stimulating factor (G-CSF), upregulate alternative activation markers and produce an array of growth factors, thereby gaining the capacity to promote neurite outgrowth. Moreover, adoptive transfer of IL-4/G-CSF-polarized bone marrow neutrophils into experimental models of CNS injury triggered substantial axon regeneration within the optic nerve and spinal cord. These findings have far-reaching implications for the future development of autologous myeloid cell-based therapies that may bring us closer to effective solutions for reversing CNS damage.

Clinical Analysis of PEG-rhG-CSF in Mobilization of Autologous Hematopoietic Stem Cells in Hematological Tumors

To investigate the efficiency and optimal time of stem cell apheresis mobilized by pegylated recombinant human granulocyte colony stimulating factor (PEG-rhG-CSF) in autologous stem cell transplantation (ASCT) for hematological malignancies without monitoring pre-collection CD34+ cells. Forty-six patients underwent stem cell mobilization were retrospectively analyzed between August 2017 and January 2022 at the First Affiliated Hospital of Fujian Medical University. 27 patients using high dose chemotherapy combined with PEG-rhG-CSF mobilization were enrolled in the PEG-rhG-CSF group, and other 19 patients mobilized with recombinant human granulocyte colony stimulating factor (G-CSF) were enrolled in G-CSF group. The mobilization and collection effects of the patients in two groups were compared. A total of 46 patients underwent 86 apheresis procedures, the median amount of mononuclear cell (MNC) in the PEG-rhG-CSF group and G-CSF group was 6.54（3.85-12.61）×108/kg and 6.15（1.13-11.58）×108/kg, respectively (P >0.05), the total CD34+ cells of the grafts were 11.44(1.33-65.02)×106/kg and 4.95(0.30-24.02)×106/kg (P < 0.05), with harvest timing of 14(10-20) days and 14(4-22) days, respectively (P >0.05). In the PEG-rhG-CSF group, there was a significant difference between the number of CD34+ cells collected when white blood cells (WBC) ≥10×109/L and WBC<10×109 /L, 19.04(2.85-65.02)×106/kg and 6.22(0.81-34.86)×106/kg, respectively (P < 0.05). Stem cells mobilization with PEG-rhG-CSF was highly efficient with a median mobilization time of 14 days. In the absence of peripheral blood CD34 monitoring, peripheral blood WBC≥10×109/L can be considered as a threshold for a single stem cell apheresis to collect sufficient stem cells.

Impact of Recombinant Granulocyte Colony-Stimulating Factor During Neoadjuvant Therapy on Outcomes of Resected Pancreatic Cancer.

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive disease characterized by chronic inflammation and a tolerogenic immune response. The granulocyte colony-stimulating factor (G-CSF)-neutrophil axis promotes oncogenesis and progression of PDAC. Despite frequent use of recombinant G-CSF in the management and prevention of chemotherapy-induced neutropenia, its impact on oncologic outcomes of patients with resected PDAC is unclear. This cohort study assessing the impact of G-CSF administration was conducted on 351 patients with PDAC treated with neoadjuvant therapy (NAT) and pancreatic resection at a high-volume tertiary care academic center from 2014 to 2019. Participants were identified from a prospectively maintained database and had a median follow-up of 45.8 months. Patients receiving G-CSF (n=138; 39.3%) were younger (64.0 vs 66.7 years; P=.008), had lower body mass index (26.5 vs 27.9; P=.021), and were more likely to receive 5-FU-based chemotherapy (42.0% vs 28.2%; P<.0001). No differences were observed in baseline or clinical tumor staging. Patients receiving G-CSF were more likely to have an elevated (>5.53) post-NAT neutrophil-to-lymphocyte ratio (45.0% vs 29.6%; P=.004). G-CSF recipients also demonstrated higher circulating levels of neutrophil extracellular traps (+709 vs -619 pg/mL; P=.006). On multivariate analysis, G-CSF treatment was associated with perineural invasion (hazard ratio [HR], 2.65; 95% CI, 1.16-6.03; P=.021) and margin-positive resection (HR, 1.67; 95% CI, 1.01-2.77; P=.046). Patients receiving G-CSF had decreased overall survival (OS) compared with nonrecipients (median OS, 29.2 vs 38.7 months; P=.001). G-CSF administration was a negative independent predictor of OS (HR, 2.02; 95% CI, 1.45-2.79; P<.0001). In the inverse probability weighted analysis of 301 matched patients, neoadjuvant G-CSF administration was associated with reduced OS. In patients with localized PDAC receiving NAT prior to surgical extirpation, G-CSF administration may be associated with worse oncologic outcomes and should be further evaluated.

Optimization of human recombinant granulocyte-colony stimulating factor supplementation during in vitro production of porcine embryos to improve the efficiency of resource utilization of poor-quality cumulus-oocyte complexes

Granulocyte colony-stimulating factor (G-CSF), a pleiotropic cytokine, is secreted by the reproductive tract. Furthermore, our previous study indicated that human recombinant G-CSF (hrG-CSF) supplementation during porcine oocyte in vitro maturation (IVM) or during embryo in vitro culture (IVC) improved their quality and development potential when using cumulus-oocyte complexes (COCs) with more than three cumulus cell layers (CCL >3). Thus, in this study, we investigate the optimal conditions of hrG-CSF supplementation throughout the in vitro production (IVP: IVM + IVC) system to improve the embryo production efficiency of “poor-quality (CCL ≤3)” oocytes. COCs were classified into two groups according to the number of CCL (>3 and ≤3) and embryonic viability was analyzed after treatment with hrG-CSF during IVC. The mRNA transcription levels of G-CSF in COCs were compared based on their type and the period of IVM. Finally, developmental capacity and quality were evaluated after treatment with hrG-CSF for different periods of IVP. No marked effects on the developmental potential of embryos when using CCL ≤3 type COCs were observed after supplementing hrG-CSF only during IVC. Moreover, the mRNA transcription level of G-CSF increased gradually with IVM culture time and was higher in CCL ≤3 COCs than in >3. Supplementing hrG-CSF only during the IVM period resulted in the best embryo developmental potential, while supplementing hrG-CSF during the IVP period resulted in the best quality embryos, reflected in the increased total cell number and decreased apoptotic nuclei index of blastocysts. These findings indicate that “poor-quality” COCs may have a greater demand for G-CSF than “good-quality”, meanwhile hrG-CSF supplementation throughout IVP improves resource utilization efficiency in poor-quality COCs.

Pomalidomide, Rituximab, Orelabrutinib, and Minichop-like (PRO-miniCHOP) in Elderly Patients with Newly Diagnosed Diffuse Large-B Cell Lymphoma: Preliminary Results from a Phase II Study

Introduction: Owing to comorbidities and poor tolerance to standard-dose chemotherapy, elderly patients with newly diagnosed diffuse large-B cell lymphoma (DLBCL) have to reduce their chemotherapy dose, ultimately leading to a poor prognosis (Di M, et al. Oncologist, 2021). Therefore, new therapeutic strategies with superior efficacy and less toxicity are urgently needed. The Smart Start study demonstrated that induction therapy with rituximab (R), lenalidomide, and ibrutinib resulted in a high overall response rate (ORR) in patients with newly diagnosed DLBCL (Westin J, et al. J Clin Oncol, 2023). Orelabrutinib (O), as a novel covalent Bruton's kinase inhibitor (BTKi) with high target selection, was reported to preserve the NK-cell-mediated antibody-dependent cellular cytotoxicity induced by R and thus boosted the antitumor effect of R-based regimen (Yu H, et al. Mol Ther-Oncolytics, 2021). The responders to OR induction therapy have been reported to attain synergistic antitumor effect and high complete remission (CR) rate (CRR) when receiving subsequent treatment (Qu C, et al. Hematol Oncol, 2023). Here, a prospective study was conducted to investigate the efficacy and safety of pomalidomide (P; a third-generation immunomodulatory drug), R, O, and miniCHOP-like (PRO-miniCHOP) in elderly patients with newly diagnosed DLBCL. Methods: Patients aged ≥70 years with newly diagnosed DLBCLwere enrolled in this open-label, single-arm, phase II study (NCT05809180). All eligible patients received one 21-day cycle of induction therapy with PRO (P, 4 mg, d1-7; R, 375 mg/m 2, d1; O, 150 mg, QD). Subsequently, patients who achieved at least mini response (miniR, a reduction in tumor lesions by 25%-50%) were administered additional 6 cycles of PRO-miniCHOP regimen (PRO with reduced-dose CHOP regimen [cyclophosphamide, 400 mg/m 2, d2; doxorubicin/liposomal doxorubicin, 25 mg/m 2/15 mg/m 2, d2; vindesine, 2 mg, d2; and dexamethasone, 7.5 mg/m 2, d2-6]). The subsequent treatment was administrated according to the tumor response after 6 cycles of the PRO-miniCHOP regimen, including the end of treatment for CR, 2 years of pomalidomide maintenance therapy for partial remission (PR), and discharge from the study for stable disease or progression disease. The primary endpoints were ORR and CRR after 6 cycles of the PRO-miniCHOP regimen. Secondary endpoints were ORR and CRR at the end of the induction therapy, as well as 2-year progression-free survival, 2-year overall survival and safety. Results: From January 01, 2023 to July 30, 2023, 10 patients were enrolled in this study. The median age was 76.0 (range, 70.0-84.0) years (Table 1). The majority of patients had an Ann Arbor stage of III or IV (7/10, 70.0%), International Prognostic Index score of ≥3 (6/10, 60.0%), and non-germinal center B-cell-like subtype (6/10, 60.0%). Five (50.0%) patients had hypertension, 4 (40.0%) patients presented with extranodal involvement, and MYC/BCL-2 double expression lymphoma (40.0%). After one cycle of induction therapy with PRO, all (10/10, 100.0%) patients achieved at least miniR, including 3 CR, 6 PR, and 1 miniR (Figure 1). Five patients completed ≥3 cycles of the PRO-miniCHOP regimen, among whom 4 (80.0%) achieved a CR and 1 (20.0%) had a PR, with the best ORR of 100%; of these, one sustained CR at the end of cycle 6. During the induction therapy with PRO, 50.0% (5/10) of patients reported adverse events (AEs) of any grade, the majority of which were grade 1-2, with only 2 patients experiencing grade 3-4 neutropenia. Grade ≥3 AEs during the whole treatment period were neutropenia (6/10, 60.0%), thrombocytopenia (1/10, 10.0%), and lymphopenia (1/10, 10.0%), of which one (1/10, 10.0%) patient developed febrile neutropenia. Notably, the use of prophylactic pegylated recombinant human granulocyte colony stimulating factor significantly reduced the recurrence rate of severe neutropenia in the subsequent treatment, and no treatment-associated death was observed. Moreover, despite half of the patients having the comorbidity of hypertension, no off-target related cardiac events such as atrial fibrillation or atrial flutter were observed. At the time of data cutoff, 9 patients were still under treatment. Conclusions: The present study provided preliminary evidence supporting the use of the PRO-miniCHOP regimen in elderly patients with newly diagnosed DLBCL. More clinical data will be updated from this ongoing study.

Aggregation of human plasma and of human blood induced in vitro by pegfilgrastim originator formulation buffer and pegfilgrastim products

PEGylated recombinant human granulocyte colony stimulating factor (pegfilgrastim) is used clinically to reduce the incidence and duration of severe neutropenia in patients who have received chemotherapy treatment. Pegfilgrastim products are administered by subcutaneous injection. We herein report that solutions of pegfilgrastim originator product Neulasta®, of a biosimilar product candidate, and also of the pegfilgrastim originator formulation buffer, induced aggregate formation when mixed in vitro with human plasma, and formation of large membranous aggregated structures when mixed with human blood. Human donor variability in the plasma aggregation induced by pegfilgrastim products was observed. In all donors less aggregation occurred in plasma mixtures with the biosimilar pegfilgrastim product candidate compared to the originator products. Instantaneous aggregation of erythrocytes and formation of large membranous aggregated structures of erythrocytes occurred in mixtures of human blood with pegfilgrastim buffer or pegfilgrastim products. The formation of the large membranous aggregated structures likely involved fusion of erythrocyte membranes; erythrocyte membrane fusion events were observed. Pegfilgrastim proteins in the products accelerated the formation of irreversible erythrocyte aggregated structures. Pegfilgrastim originator formulation buffer (10 mM Na-acetate pH 4.0, 274 mM sorbitol, 0.004 % polysorbate 20) was identified as the main driver of the plasma and erythrocyte aggregation. Lipoprotein aggregation at low pH in the presence of sorbitol and erythrocyte membrane fusion induced by the lipoprotein aggregates, are proposed as the main mechanisms for the formation of plasma and blood aggregates. Such aggregation phenomena may also occur during pegfilgrastim clinical use and may be related to known side effects and individual variability in the efficacy of pegfilgrastim therapy.

Intravenous immunoglobulins and recombinant granulocyte-colony stimulating factor modulate expression of NK cytotoxic receptors

Natural killer cells (NK cells) are a population of innate immune lymphocytes capable of cytolysis of infected or transformed cells without prior sensitization. Natural killers are detected in various organs and tissues and may differ in phenotypic and functional characteristics depending on localization. For example, NK cells are the dominant population (up to 70%) of decidual lymphocytes in early pregnancy. NK cells are able to contact with trophoblast cells, exert cytotoxicity towards them, as well as regulate their invasion, contributing to spiral arteries remodeling and establishment of physiological blood flow between mother and fetus. The contribution of impaired NK cell functional activity to immune mechanisms of the reproductive disorders is widely discussed. Various drugs are used to treat infertility, including intravenous immunoglobulins (IVIG) and recombinant granulocyte colony stimulating factor (G-CSF). Increased rates of embryo implantation and higher frequency of successful gestation have been shown after treatment with these drugs. The effect of these drugs on NK cells phenotype and functional activity is assumed, thus requiring further studies on the effects of IVIG and G-CSF on the receptor profile of NK cells. The aim of this work was to evaluate expression of cytotoxic receptors on the NK-92 cells in presence of IVIG and recombinant G-CSF preparations. NK-92 cells were used as effectors, and trophoblast-derived JEG-3 line served as target population. The cells were co-cultured in presence of drugs, as well as without them. Expression of CD45, CD56, CD215, KIR2DL3, KIR2DS4, NKG2D, NKp44, NKp30 receptors by NK-92 cells was evaluated by flow cytometry. The number of NK-92 cells expressing NKG2D, NKp30, KIR2DL3 receptors and the expression intensity of NKG2D and NKp30 receptors were reduced in presence of IVIG preparations. The numbers of KIR2DL3+ and NKp44+ NK cells were reduced when supplied with G-CSF and trophoblast cells. The obtained results may be associated with both direct and indirect effects of the studied drugs on the NK cell phenotype.

A novel PEGylated form of granulocyte colony-stimulating factor, mecapegfilgrastim, for peripheral blood stem cell mobilization in patients with hematologic malignancies.

The Pegylated recombinant human granulocyte colony stimulating factor (PEG-rhG-CSF) has longer half-life and is given once only, which is more comfortable for patients. We aimed to evaluate the efficacy of mecapegfilgrastim for hematopoietic stem cell (HSC) mobilization in patients with hematologic malignancies and to explore the potential factors related to HSC mobilization. A retrospective analysis was performed on patients who underwent HSC mobilization in the hematology department of Mianyang Central Hospital from April 2016 to November 2022. The number of CD34 + cells collected was compared between the patients receiving mecapegfilgrastim (PEG group) and those receiving recombinant human granulocyte colony-stimulating factor (rhG-CSF group), and the possible factors for mobilization failure were analyzed. The success rates of collecting CD34 + cells in the PEG group and rhG-CSF group were 80.6% and 67.7%, respectively (χ = 1.444, P = 0.229). The median CD34 + cell counts were 3.62 × 10^6/kg and 2.92 × 10^6/kg (P = 0.178), respectively. After combination with plerixafor for mobilization, the median number of CD34 + cells collected in the PEG group and rhG-CSF group were 3.64 × 10^6/kg and 3.92 × 10^6/kg, respectively, with no significant difference (P = 0.754). There was no significant difference in hematopoietic cell recovery or infection between the groups (P > 0.05). Multivariate analysis showed that more than 5 cycles of chemotherapy (OR = 15.897, 95% CI: 1.766-143.127, P = 0.014), a precollection WBC count < 32 × 10^9/L (OR = 14.441, 95% CI: 2.180-95.657, P = 0.006) and a precollection to premobilization lymphocyte ratio < 1.7 (OR = 11.388, 95% CI: 2.129-60.915, P = 0.004) were independent risk factors for HSC mobilization failure. The HSC mobilization efficacy of mecapegfilgrastim in patients with hematologic malignancies was comparable to that of rhG-CSF, and combination with plerixafor for mobilization was feasible and effective. Patients with more than 5 cycles of chemotherapy before HSC mobilization, a precollection WBC count lower than 32 × 10^9/L, and a precollection lymphocyte count less than 1.7 times the premobilization lymphocyte count have a high probability of HSC mobilization failure.

Chemotactic NO/H2S Nanomotors Realizing Cardiac Targeting of G-CSF against Myocardial Ischemia-Reperfusion Injury.

Recombinant granulocyte colony-stimulating factor (G-CSF), with a direct repair effect on injured cardiomyocytes against myocardial infarction ischemia-reperfusion-injury (IRI), displays a poor effect owing to the limited cardiac targeting efficacy. There are almost no reports of nanomaterials that deliver G-CSF to the IRI site. Herein, we propose a way to protect G-CSF by constructing one layer of nitric oxide (NO)/hydrogen sulfide (H2S) nanomotors on its outside. NO/H2S nanomotors with specific chemotactic ability to high expression of reactive oxygen species (ROS)/induced nitric oxide synthase (iNOS) at the IRI site can deliver G-CSF to the IRI site efficiently. Meanwhile, superoxide dismutase is covalently bound to the outermost part, reducing ROS at the IRI site through a cascade effect with NO/H2S nanomotors. The synergistic effect between NO and H2S on the effective regulation of the IRI microenvironment can not only avoid toxicity caused by excessive concentration of a single gas but also reduce inflammation level and relieve calcium overload, so as to promote G-CSF to play a cardioprotective role.

G-CSF rescue of FOLFIRINOX-induced neutropenia leads to systemic immune suppression in mice and humans

BackgroundRecombinant granulocyte colony-stimulating factor (G-CSF) is routinely administered for prophylaxis or treatment of chemotherapy-induced neutropenia. Chronic myelopoiesis and granulopoiesis in patients with cancer has been shown to induce immature monocytes...

Efficacy and safety of dexcitabine combined with HAAG regimen in the treatment of recurrent acute myeloid leukemia

Objective: To investigate the efficacy and safety of dexithabine (DAC) combined with HAAG regimen [harringtonine (HHT), cytarabine (Ara-C), aclarubicin (Acla) and recombinant human granulocyte colony stimulating factor (G-CSF)] in the treatment of acute myeloid leukemia (AML). Methods: The clinical data of 89 AML patients in People's Hospital Affiliated to Shandong First Medical University from January 2019 to January 2021 were retrospectively analyzed. The patients were divided into observation group (n=48) and control group (n=41) according to the treatment plan. The observation group included 25 males and 23 females, aged (44.4±9.3) years old, and was treated with DAC combined with HAAG. The control group included 24 males and 17 females, aged (42.2±10.1) years old, and was treated with DAC regimen. After 3 cycles of treatment, the treatment efficacy of the two groups was judged, including complete remission, partial remission and no remission. The level of serum P-glycoprotein (P-gp) in the two groups was detected by direct immunofluorescence-labeled monoclonal antibody flow cytometry. The enzyme-linked immunosorbent assay was used to detect the level of soluble urokinase-type plasminogen activator receptor (suPAR). Meanwhile, the incidence of adverse reactions such as digestive tract reaction, liver and kidney dysfunction, hemorrhage and infection during treatment were recorded. Results: After 3 cycles of treatment, the observation group had complete remission, partial remission and no remission in 10 cases, 21 cases and 17 cases, respectively, and the control group had 3 cases, 11 cases and 27 cases, respectively. The overall efficacy of the observation group was better than that of the control group (Z=-2.919, P=0.004). The levels of serum P-gp and suPAR in the observation group were (5.2±1.8) % and (464.4±103.4) ng/L, respectively, which were significantly lower than those in the control group [(8.8±1.9) % and (660.6±110.4) ng/L, respectively] (both P<0.05). During the treatment, the incidence of digestive tract reaction, liver and kidney dysfunction, hemorrhage and infection in the observation group was 29.2% (14/48), 22.9% (11/48), 16.7% (8/48) and 33.3% (16/48), respectively, while in the control group was 26.8% (11/41), 21.9% (9/41), 14.6% (6/41) and 24.4% (10/41), respectively, with no statistically significant difference (all P>0.05). Conclusions: The overall efficacy of DAC combined with HAAG in the treatment of AML is better than that of DAC alone. Moreover, the incidence of adverse reactions in DAC combined with HAAG is similar to that of DAC alone, with a high safety profile.

Pegbovigrastim use in periparturient embryo-recipient cows: Effects on health and reproduction

The objective of this randomized clinical study was to evaluate the effects of periparturient administration of pegbovigrastim (recombinant bovine granulocyte colony stimulating factor; PEG) on postpartum health and reproductive outcomes in dairy cows carrying transferred embryos. A total of 60 multiparous Jersey cows (second parity n = 21, >second parity n = 39) that were in vitro-fertilized embryo recipients were enrolled. Animals were selected from a commercial herd and assigned to 1 of 2 treatments: (1) PEG (n = 31), which received a subcutaneous injection of 15 mg of PEG 7 d before expected calving and a second dose within 1 h of parturition; or (2) CON, untreated control (n = 29). Parturition was induced pharmacologically in all cows at 280 d of pregnancy with 500 μg of cloprostenol and 25 mg of dexamethasone intramuscularly. Cow health was monitored daily during the periparturient period and included assessment for retained fetal membranes within 24 h after calving and metritis until 20 d in milk. Descriptive statistics were calculated, to determine the incidence of postpartum diseases, causes of culling, and early-lactation reproductive outcomes, including number of inseminations, days to first heat, days to first service, and interval from calving to conception. The Wilcoxon rank sum test was used to assess treatment effects on early-lactation reproductive performance outcomes. The overall incidence of metritis in this cohort of cows was 17.14%. Animals treated with PEG tended to have more cases of metritis (25%) compared with CON (8.8%). Mastitis incidence in PEG was 19.35% compared with CON at 27.59%. These data suggest that in this dairy, with embryo-recipient cows induced to calve at 280 d of pregnancy, the administration of 2 doses of PEG tended to increase postpartum metritis but had no effect on mastitis incidence, culling rate, or early-lactation reproductive outcomes.

Filgrastim, a Recombinant Form of Granulocyte Colony-stimulating Factor, Ameliorates 3-nitropropionic Acid and Haloperidol-induced Striatal Neurotoxicity in Rats.

Striatal neurotoxicity is the pathological hallmark for a heterogeneous group of movement disorders like Tardive dyskinesia (TD) and Huntington's disease (HD). Both diseases are characterized by progressive impairment in motor function. TD and HD share common features at both cellular and subcellular levels. Filgrastim, a recombinant methionyl granulocyte colony-stimulating factor (GCSF), shows neuroprotective properties in in-vivo models of movement disorders. This study seeks to evaluate the neuroprotective effect of filgrastim in haloperidol and 3-NP-induced neurotoxicity in rats. The study was divided into two: in study one, rats were administered with haloperidol for 21days, filgrastim at the dose of (20, 40, 60µg/kg,s.c.) was administered once a day before haloperidol treatment and the following parameters (orofacial movements, rotarod, actophotometer) were performed to assess TD. Similarly, in the second study, rats were administered with 3-NP for 21days, filgrastim at a dose of (20 and 40µg/kg, s.c.) was administered, and the following parameters (rotarod, narrow beam walk, and open field test) were assessed for HD. On the 22nd day, animals were sacrificed and cortex and striatum isolated for oxidative stress (LPO, GSH, SOD, catalase, and nitrate) marker assessment. Results revealed that haloperidol and 3-NP treatment significantly impaired motor coordination, and oxidative defense inducing TD and HD-like symptoms. Treatment with filgrastim significantly averted haloperidol and 3-NP-induced behavioral and biochemical alterations. Conclusively, the neuroprotective effect of filgrastim is credited to its antioxidant properties. Hence, filgrastim might be a novel therapeutic candidate for the management of TD and HD.

G-CSF Mobilized Apheresis As an Alternative Source of CAR T-Cells

Background Cellular therapies are traditionally manufactured from an apheresis product collected at 'steady state' i.e. without any cytokine administered prior. We sought to evaluate the feasibility of utilising the CD34-negative fraction of recombinant human granulocyte colony stimulating factor (rhG-CSF) mobilised apheresis products for the manufacture of CAR T-cells. The effect of rhG-CSF on CAR T-cell function is largely unknown. We conducted an extensive comparison of phenotypic and functional attributes of CAR T-cells manufactured after rhG-CSF exposure (mobCAR) to those manufactured from a steady state apheresis (ssCAR), in order to test the hypothesis that rhG-CSF mobilised CAR T-cells are functionally equivalent to steady state CAR T-cells. Methods Untransduced T-cells (UTD) and CAR T-cells targeting CD33 (CART33) were manufactured using paired steady state (ss) cells and cells collected after rhG-CSF treatment (mob). On a protocol approved by the institutional review board, 4 healthy volunteers had ss cells collected from peripheral blood and cryopreserved, and later underwent an apheresis collection after 4 days of G-CSF (10ug/kg/daily) to collect and cryopreserve mob cells. CART33 were then manufactured from thawed ss and mob cells in paired expansions. Cells were activated using CD3/CD28 beads, transduced with lentiviral vector and expanded using Ex-Vivo based cell culture media (fortified with IL-7/IL-15 and human serum), and cryopreserved when mean cell volume was less than 300fL. Extensive comparisons between ssCART33 and mobCART33 were completed, including analysis by flow cytometry, CyTOF, single cell RNA sequencing (scRNAseq) and metabolomics by mass spectrometry. In vitro assays of degranulation, cytokine production and killing were performed, and in vivo function was assessed using an immunodeficient murine model after engraftment with a CD33 expressing cell line. Results Paired effector T-cells were manufactured with lower fold expansion of mobCAR [4 vs. 7 population doublings, p<0.01] though a sufficient number of mob-CAR were able to be manufactured. Mean CAR expression was higher in ssCAR though this was not statistically significant [ssCAR 78.6% (55-79%) vs. 64.7% (59-71%), p>0.05], and no significant differences in CD4/CD8 ratio were observed. In vitro degranulation, cytokine production and tumour cytolysis were comparable between ssCAR and mobCAR. Functional equivalence was also seen between ssCAR and mobCAR in mouse xenografts of an acute myeloid leukemia (AML) model, with no differences in body weight, toxicity, or mouse survival observed (Figure 1). While significant inter-donor differences in T-cell subsets by CyTOF were observed, minimal differences were seen between the ss and mob products for a given donor, and integrated analysis of scRNASeq data showed a similar transcriptomic profile for ssCAR and mobCAR cells (Figure 2). Metabolomics analysis is ongoing. Conclusions and future directions We were able to successfully manufacture CAR T-cell products from G-CSF mobilized cryopreserved apheresis products, and found comparable in vivo anti-tumor efficacy and toxicity, with no significant differences in the G-CSF exposed product by multi-omic analyses. Successful completion of this work may support the use of cryopreserved G-CSF stimulated apheresis units for cell therapy applications. This may allow for an adjunct cell therapy to hematopoietic stem-cell transplant to be manufactured from the same mobilized collection, or provide a means by which previously cryopreserved mobilized units may be utilised, noting frequently lower cumulative chemotherapy exposure in historically cryopreserved T-cells. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Effect and safety of pegylated recombinant human G-CSF on hematopoietic reconstitution after autologous hematopoietic stem cell transplantation in lymphoma patients

Objective: Efficacy and safety analysis of pegylated recombinant human granulocyte colony stimulating factor (PEG-rhG-CSF) in promoting hematopoietic recovery after autologous hematopoietic stem cell transplantation (auto-HSCT) in patients with lymphoma. Methods: A total of 149 patients after auto-HSCT in Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine were enrolled in this study from April 2016 to December 2021. There were 75 cases in the PEG-rhG-CSF group who were given a single subcutaneous dose of 100 µg/kg on the first day and +8 d, while 74 cases in the rhG-CSF group were given a dose of 5-10 µg·kg(-1)·d(-1) by subcutaneous injection from +1d continuing to an absolute value of neutrophil (ANC) of more than 1.5×10(9)/L. Results: ①The time of grade 3/4 agranulocytosis and neutrophil implantation in the PEG-rhG-CSF group were significantly different from that in rhG-CSF group (P=0.010, 0.030, 0.007) . There were no significant differences in the platelet implantation time, anemia incidence and duration, and platelet and red blood cell infusion within 1 month after transplantation between groups. ②The agranulocytosis with fever incidence in PEG-rhG-CSF group was similar to that in rhG-CSF group (84.0% vs 82.4% , P=0.798) , but the duration was shorter in the PEG-rhG-CSF group (4.0 d vs 5.5 d, P=0.005) . ③The incidence of infection in the PEG-rhG-CSF and the rhG-CSF groups were 22.7% (17/75) and 31.1% (23/74) , respectively (P=0.247) , and the bloodstream infection incidence were 5.3% (4/75) and 9.5% (7/74) , respectively (P=0.336) . ④The PEG-rhG-CSF group and rhG-CSF group's mean length of hospital stay were 31.5 (23-43) days and 37 (25-75) days, respectively (P<0.001) . ⑤The PEG-rhG-CSF group and rhG-CSF group's disease-free survival rates were (96.4±2.5) % and (94.7±2.6) % (P=0.638) , respectively, and the OS rates were 100.0% and (98.6±1.3) % (P=0.312) , respectively. Conclusion: PEG-rhG-CSF application after auto-HSCT in patients with lymphoma can promote hematopoietic granulocyte reconstruction and shorten hospital stay, but has no significant effect on the incidence of infection, disease-free survival, and overall survival after transplantation.

A phase II, multicenter, open-label, randomized trial of pegfilgrastim for patients with alcohol-associated hepatitis.

SummaryBackgroundIn trials conducted in India, recombinant granulocyte colony stimulating factor (GCSF) improved survival in alcohol-associated hepatitis (AH). The aim of this trial was to determine the safety and efficacy of pegfilgrastim, a long-acting recombinant GCSF, in patients with AH in the United States.MethodsThis prospective, randomized, open label trial conducted between March 2017 and March 2020 randomized patients with a clinical diagnosis of AH and a Maddrey discriminant function score ≥32 to standard of care (SOC) or SOC+pegfilgrastim (0.6 mg subcutaneously) on Day 1 and Day 8 (clinicaltrials.gov NCT02776059). SOC was 28 days of either pentoxifylline or prednisolone, as determined by the patient's primary physician. The second injection of pegfilgrastim was not administered if the white blood cell count exceeded 30,000/mm3 on Day 8. Primary outcome was survival at Day 90. Secondary outcomes included the incidence of acute kidney injury (AKI), hepatorenal syndrome (HRS), hepatic encephalopathy, or infections.FindingsThe study was terminated early due to COVID19 pandemic. Eighteen patients were randomized to SOC and 16 to SOC+pegfilgrastim. All patients received prednisolone as SOC. Nine patients failed to receive a second dose of pegfilgrastin due to WBC > 30,000/mm3 on Day 8. Survival at 90 days was similar in both groups (SOC: 0.83 [95% confidence interval [CI]: 0.57–0.94] vs. pegfilgrastim: 0.73 [95% CI: 0.44–0.89]; p > 0.05; CI for difference: -0.18–0.38). The incidences of AKI, HRS, hepatic encephalopathy, and infections were similar in both treatment arms and there were no serious adverse events attributed to pegfilgrastim.InterpretationThis phase II trial found no survival benefit at 90 days among subjects with AH who received pegfilgrastim+prednisolone compared with subjects receiving prednisolone alone.Fundingwas provided by the United States National Institutes of Health and National Institute on Alcohol Abuse and Alcoholism U01-AA021886 and U01-AA021884.