Recombinant Human Granulocyte Colony-stimulating Factor Research Articles

A Universal CRISPR/Cas9n-Mediated Genome Editing Approach for ELANE-Related Severe Congenital Neutropenia

Severe congenital neutropenia (CN) is an inherited bone marrow failure syndrome characterized by impaired maturation of neutrophil granulocytes. Due to the lack or very low levels of neutrophils in the peripheral blood, patients experience severe, life-threatening bacterial infections as early as birth. Autosomal-dominant ELANE mutations are the most common cause of CN. Although most patients respond to daily treatment with recombinant human granulocyte colony-stimulating factor, about 15% of patients do not respond to this cytokine, and approximately 20% of patients develop myelodysplasia or acute myeloid leukemia. The only curative treatment available thus far is allogenic hematopoietic stem cell transplantation, which is associated with serious side effects. Autologous transplantation of gene-edited patients' HSPCs offers a novel curative therapy. Considering the wide range of mutations distributed throughout the ELANE gene, A universal, high-safety-profile approach could be advantageous for all ELANE-CN patients. Our group has already published a universal CRISPR/Cas9-mediated knockout approach for the ELANE gene; however, targeting the coding sequence region (CDS) of the ELANE gene could lead to the introduction of new disease-causing variants, so it may be harmful for patients. In order to address this concern, we reformatted our previously published knockout concept into a clinical-grade gene therapy strategy. We have screened cis-regulatory regions in the ELANE core promoter and, using CRISPR/Cas9n, introduced two nicks on opposing strands of the Goldberg-Hognes box region (TATA-box) to make the transcription starting process inefficient, as transcription factors and RNApol II cannot recognize the modified sequence. Theproposed strategy targets the non-coding region of the ELANE gene and thus does not generate new unwanted variants. Also, by replacing Cas9 with Cas9-ncikase, the off-target activity should decrease by up to 1000-fold. We have observed a markedly elevated neutrophil differentiation in gene-edited ELANE-CN patients HSPCs (n = 2), as assessed by the percentage of CD45 +CD11b +CD15 +, CD45 +CD15 +CD16 +, and CD45 +CD16 +CD66b + myeloid/granulocytic cells, compared to the mock electroporated ELANE-CN group, while no disruption of granulopoiesis in healthy donor HSPCs (n = 2) was detected. Results validated by the CFU assay. To have nucleotide-level resolution of on-target events, we performed targeted next-generation sequencing of edited loci on gene-edited healthy donors and ELANE-CN HSPCs. Next-generation sequencing results showed >90% on-target efficiency. CRISPR/Cas9n-mediated targeting of the TATA box rescued defective granulopoiesis in vivo, as gene-edited ELANE-CN HSPCs (n = 3) were transplanted in NSG mice and assessed by the percentage of neutrophils (hCD19 -hCD3 -hCD66b Int/lowhCD33 +hCD16 high) after 16 weeks. We have also observed efficient engraftment and preserved the multilineage potential of gene edited HSPCs in immunodeficient NSG mice. The strategy also depicted a safe profile upon applying gene editing to healthy donor CD34 + HSPCs and performing GUIDE-seq and CAST-seq. GUIDE-seq highlighted two potential off-target sites after >=6 mismatches, while no chromosomal translocations were detected by CAST-seq. RNA-sequencing of in vitro-generated neutrophils from gene-edited HSPCs of healthy donors (n = 2) confirmed a >9-fold reduction of ELANE gene expression level. Important to note, none of the other serine proteases or neighboring genes were down-regulated. Taken together, ex vivo CRISPR-Cas9n-based gene editing of ELANE's gene promoter in the setting of autologous stem cell transplantation could be a safe therapeutic approach for all ELANE-CN patients.

Using recombinant human G-CSF to treat chemotherapy-induced neutropenia over 3 decades: What is next?

Chemotherapy-Induced Neutropenia (CIN) is a potentially fatal side effect of cancer treatment, affecting > 50% of cancer patients treated with chemotherapy. Clinical use of recombinant human granulocyte colony-stimulating factor (rhG-CSF) has allowed for primary and secondary prophylaxis of CIN and its sequela (i.e., febrile neutropenia, fatal infection) during myelosuppressive chemotherapy. Here, we review the translation and properties of first, second, and third-generation rhG-CSF molecules, including filgrastim (Neupogen, FDA approved in 1991) and biosimilars, pegfilgrastim (Neulasta, FDA approved in 2002) and biosimilars, and F-627 (Ryzneuta, NMPA approved in 2023), a novel long-acting rhG-CSF agent developed this past decade. Even with the development of increasingly personalized and targeted cancer therapy, chemotherapy, and stem cell transplantation remains a backbone for the majority of patients with advanced cancers, especially in the hematopoietic system. As such, more than 20 million cancer patients have been treated with rhG-CSF drugs since the first approval of filgrastim. In the next decade, we envision third-generation rhG-CSF products such as Ryzneuta lowering costs to patients and healthcare providers, expanding access to this essential medication for cancer patients worldwide, particularly for patients who require more aggressive chemotherapy treatment.

Efficacy and safety of PEG-rhG-CSF versus rhG-CSF in preventing chemotherapy-induced-neutropenia in early-stage breast cancer patients

BackgroundTo compare the clinical value of recombinant human granulocyte colony-stimulating factor (rhG-CSF) and pegylated rhG-CSF(PEG-rhG-CSF) in early-stage breast cancer (EBC) patients receiving adjuvant chemotherapy, compare the efficacy of PEG-rhG-CSF with different dose and explore the timing of rhG-CSF rescue treatment.MethodsPatients in two PEG-rhG-CSF subgroups were given 3 mg or 6 mg PEG-rhG-CSF within 24 ~ 48 h after chemotherapy for preventing myelosuppression, while patients in the rhG-CSF group were given rhG-CSF. Observation indicators include the incidence of febrile neutropenia (FN) and grade 3/4 chemotherapy-induced-neutropenia (CIN), the overall levels and nadir values of white blood cells (WBC) and absolute neutrophil count (ANC), comparison of WBC and ANC curves over time, the incidence of CIN-related complications, the incidence of adverse events in each group and the timing of rescue treatment for rhG-CSF.ResultsThere was no significant difference in the incidence of FN in the first cycle among the groups (P = 0.203). But the incidence of ≥ 3 grade CIN in two PEG-rhG-CSF subgroups was significantly lower than that in the rhG-CSF group (P < 0.001). The overall WBC and ANC levels in the PEG-rhG-CSF group were significantly higher than those in the rhG-CSF group (P < 0.001). In terms of CIN-related complications, less chemotherapy delay rate (1.1 vs. 7.5%, P = 0.092), less dose reduction rate (6.9 vs. 7.5%, P = 1.000), less antibiotic use rate (3.4 vs. 17.5%, P = 0.011) and less proportion of rhG-CSF rescue therapy (24.1 vs. 85.0%, P < 0.001) in the PEG-rhG-CSF group, and there were no significant differences between PEG-rhG-CSF subgroups. In the incidence of adverse events among the groups, there were no statistical differences. All patients undergoing rhG-CSF rescue treatment were mainly 4 grade (63.6%) and 3 grade (25.5%) CIN, and 10.9% of patients with 1 ~ 2 grade CIN who had high infection risk or had been infected.ConclusionPEG-rhG-CSF has better efficacy and equal tolerance compared with rhG-CSF in preventing CIN in EBC patients receiving EC regimen. Moreover, a half-dose 3 mg PEG-rhG-CSF also had good efficacy. Last, patients with ≥ 3 grade CIN and others who have been assessed to be at high risk of infection or have co-infection should consider rhG-CSF or even antibiotic rescue treatment.

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.

Efficacy and safety of anlotinib combined with anthracycline and ifosfamide followed by anlotinib maintenance in advanced soft tissue sarcoma: A single-arm, phase 2 trial.

11553 Background: Anthracycline plus ifosfamide is the standard first line treatment regime for advanced soft tissue sarcoma (STS). Anlotinib, a multikinase angiogenesis inhibitor, has evolved as a standard second line treatment option in Chinese advanced STS patients. This study was performed to evaluate the efficacy and safety of anlotinib combined with anthracycline and ifosfamide followed by anlotinib maintenance in the first line treatment of patients with advanced STS. Methods: This prospective, open-label, single-arm, phase 2 trial was conducted in Shandong Cancer Hospital and Institute (ChiCTR2100054711). The key inclusion criteria included: (a) age was 18-70 years; (b) ECOG performance state of 0-1; (c) histologically confirmed high-grade STS; (d) unresectable or metastatic STS; (e) previously untreated, and sensitive to chemotherapy; (f) having measurable lesions according to RECIST 1.1. All patients received up to 6 cycles of anlotinib (12 mg QD on day1-14, 21 days per cycle) combined with anthracycline (epirubicin 40mg/m2/d on day 1-2, 21 days per cycle; or liposomal doxorubicin 30mg/m2/d on day 1, 21 days per cycle) and ifosfamide (2g/m2/d on day 1-3, 21 days per cycle) followed by anlotinib maintenance until disease progression, unacceptable toxicity or death. Pegylated recombinant human granulocyte colony-stimulating factor was administered following chemotherapy. The primary endpoint was objective response rate (ORR). The secondary endpoints included progression-free survival (PFS), overall survival (OS), and adverse effect (AE). Results: From December 2021 to December 2022, 31 patients (median [range] age, 53 [18-69] years; 20 [64.5%] male) were enrolled. 29 were evaluable for objective response, and the ORR and DCR were 31.03% (95%CI, 17.28-49.23%) and 82.76% (95%CI, 65.45-92.4%), respectively. With a median follow-up duration of 6.3 months (range, 0.8–14.0), the median PFS for all 31 patients was 6.9 months (95% CI, 2.62-11.17). The median OS was not reached. Treatment-related AEs of any grade occurred in all patients. The most common AEs included nausea/vomiting (88.46%), anemia (23.08%), leukopenia (19.23%), venous thrombosis (15.38%), hand-foot syndrome (11.54%) and elevated alanine aminotransferase or aspartate aminotransferase (11.54%). The most frequent ≥ grade 3 AEs were leukopenia (11.54%) and thrombocytopenia (7.69%). 2 serious AEs (1 left heart failure and 1 intestinal obstruction) were recorded. No treatment-related death occurred. Conclusions: This study suggested that anlotinib combined with anthracycline and ifosfamide followed by anlotinib maintenance demonstrated encouraging efficacy with a manageable safety profile in the first line treatment of patients with advanced STS. Clinical trial information: ChiCTR2100054711 .

A multicenter phase II trial of primary prophylactic PEG-rhG-CSF in pediatric patients with solid tumors and non-Hodgkin lymphoma after chemotherapy: An interim analysis.

Pegylated recombinant human granulocyte colony-stimulating factor (PEG-rhG-CSF) can be used in pediatric patients. This study assessed the safety and efficacy of PEG-rhG-CSF as a primary prophylactic drug against neutropenia after chemotherapy in pediatric patients with solid tumors or non-Hodgkin lymphoma (NHL). This phase II study (between October 2020 and March 2022) enrolled pediatric patients with solid tumors or NHL treated with high-intensity chemotherapy and with grade ≥3 myelosuppression for at least 14 days during chemotherapy. Prophylactic PEG-rhG-CSF was given at 100 μg/kg body weight (maximum total dosage of 6 mg) once 24-48 h following chemotherapy for two cycles. The primary endpoint was the incidence of PEG-rhG-CSF-related adverse events (AEs). The key secondary endpoints were the rates of grade 3/4 neutropenia and febrile neutropenia (FN). This study included 160 pediatric patients with a median age of 6.22 (0.29, 18.00) years. Fifty-eight patients (36.25%) were diagnosed with sarcoma. AEs potentially related to PEG-rhG-CSF included bone pain (n = 32), fatigue (n = 21), pain at the injection site (n = 21), and myalgia (n = 20). The rates of grade 3/4 neutropenia and FN during treatment were 57.28% and 29.45%, respectively. PEG-rhG-CSF is well tolerated and effective in pediatric patients with solid tumors or NHL. These findings should be substantiated with further trials. ClinicalTrials.gov identifier: NCT04547829.

Application of Mecapegfilgrastim for Peripheral Blood Hematopoietic Stem Cell Mobilization in Patients With Hematologic Neoplasms and Analysis of Predictors for Poor Mobilization

To evaluate the efficacy of applying mecapegfilgrastim for peripheral blood hematopoietic stem cell (PBSC) mobilization in patients with hematologic neoplasms, and to investigate the influencing factors of PBSC collection. Patients who underwent PBSC mobilization in the Department of Hematology, Mianyang Central Hospital between April 2016 and May 2022 were retrospectively analyzed. The CD34 + cell collection results of two groups, the mecapegfilgrastim group ( n=28), or the PEG group, and the recombinant human granulocyte colony-stimulating factor (rhG-CSF) group ( n=30), were compared, and the influencing factors of collection failure were analyzed. The success rates of CD34 + cells collection in the PEG group and the rhG-CSF group were 75.0% and 63.3%, respectively ( P>0.05). The median CD34 + cell counts were 3.37×10 6/kg and 2.68×10 6/kg, respectively, showing no significant difference. After combined mobilization with plerixafor, the median counts of CD34 + cells collected in the PEG group and rhG-CSF group were 4.23×10 6/kg and 3.26×10 6/kg, respectively, showing no significant difference ( P>0.05). There was no significant difference in hematopoietic system reconstruction and infections between the two groups ( P>0.05). Multivariate analysis found non-plasma cell disease (odds ratio [ OR]=19.697, 95% confidence interval [ CI] : 1.501-258.537, P=0.023), anemia before collection ( OR=18.571, 95% CI: 1.354-254.775, P=0.029) and white blood cell count before collection under 32×10 9 L -1 ( OR=85.903, 95% CI: 4.947-1491.807, P=0.002) to be independent risk factors for PBSC collection failure. The effect of PBSC mobilization with mecapegfilgrastim was comparable to that of rhG-CSF in patients with hematologic neoplasms. Furthermore, combined mobilization with plerixafor was feasible and effective. Patients with leukemia or lymphoma, anemia, and WBC<32×10 9 L -1 before stem cell collection have a high probability of PBSC collection failure.

Effects of pegylated recombinant human granulocyte colony-stimulating factor on lymphocytes and white blood cells of patients with malignant tumor.

We investigated the effect of pegylated recombinant human granulocyte colony-stimulating factor (PEG-rhG-CSF) on lymphocytes and white blood cells of patients with malignant tumors. After PEG-rhG-CSF treatment, the count of lymphocytes increased in 66 cases, remained unchanged in 2 cases, and decreased in 20 cases. The difference in lymphocyte count before and after treatment was statistically significant (P < 0.001). White blood cell changes were positively correlated with lymphocyte changes (r = 0.36, P = 0.001). In the subgroup with increased white blood cells (n = 80), there were 62 cases with increased lymphocytes, 1 case with unchanged lymphocytes, and 17 cases with decreased lymphocytes after PEG-rhG-CSF treatment. There was significant difference in the count of lymphocytes and white blood cells (P < 0.001). In the subgroup with 6 mg of PEG-rhG-CSF (n = 66) and the subgroup with 3 mg of PEG-rhG-CSF (n = 22), the changes of white blood cell and lymphocyte counts before and after treatment were statistically significant (P < 0.001). The two were positively correlated in the 6 mg PEG-rhG-CSF subgroup, with correlation coefficient r = 0.34 (P = 0.002). PEG-rhG-CSF can increase the count of lymphocytes and white blood cells in patients with malignant tumors, and the increase of lymphocytes is positively correlated with the increase of white blood cells.

Efficacy and safety of recombinant human granulocyte colony-stimulating factor in patients with unexplained recurrent spontaneous abortion: A systematic review and meta-analysis

IntroductionPublished data regarding efficacy of intrauterine perfusion of recombinant human granulocyte colony-stimulating factor for patients with unexplained recurrent spontaneous abortion (URSA) is inconclusive. This study aims at evaluating the efficacy and safety of G-CSF in URSA. Materials and methodsElectronic databases were searched including Cochrane Library, PubMed, Embase, China Biology Medicine disc, China Science and Technology Journal Database, Wanfang Database and China National Knowledge Infrastructure Database (last search was performed on Sep 10th, 2022). A systematic review and meta-analysis was conducted with R-language software. Combined relative risk (RRs), and 95% confidence intervals (CIs) were calculated to estimate efficacy and safety. ResultsCompared with placebo, the efficacy of G-CSF in the treatment of URSA patients was significant in conception rate (RR=1.34, 95%CI: 1.03–1.74, P = 0.028), and was none of significance in live birth rate (RR=1.35, 95%CI: 0.99–1.84, P = 0.06). Subgroup analysis showed that the ovulation-period-medication was the protective factor for conception rate, while "Ethnicity Asian" and "ovulation-period medication" were the protective factors for live birth rate. When it comes to the safety of rhG-CSF on URSA, meta-analysis showed that rhG-CSF had no significant effect on the incidence of adverse events (AEs) (RR=1.13, 95% CI: 0.89–1.43, P = 0.322), and subgroup analysis showed that the incidence of AEs in each subgroup did not increase significantly (P > 0.05). ConclusionBased on our meta-analysis, intrauterine perfusion of rhG-CSF in ovulation period is an effective and safe way to improve conception rate in URSA.

Clinical Applications of Recombinant Human Granulocyte Colony-Stimulating Factor in Reproductive Medicine: Expert Consensus

The primary clinical indication for recombinant human granulocyte-stimulating factor (rhG-CSF) is neutropenia of multiple etiologies. In recent years, rhG-CSF has been proved to be effective and safe for mothers and fetus in the clinical application in some reproductive disorders. Until recently, there has been no reliable evidence of authoritative expert consensus or standards. In order to regulate the research and rational application of rhG-CSF in the fields of reproductive medicine, the Committee on Reproductive Immunity of the China Society for the Promotion of Human Health Science and Technology (the Committee) organized experts from relevant disciplines to formulate the Expert Consensus, which is based on the latest international basic study and clinical research evidence. According to evidence-based data, recommendations were established on the applicable population, starting method, and duration of rhG-CSF administration in reproductive medicine. It is recommended that rhG-CSF could be applicated in the patients with thin endometrium which was ineffective to standard treatments, in unexplained recurrent spontaneous abortion (RSA), in unexplained recurrent implantation failure (RIF), and could attempt to be applicated in unexplained RSA with slow rise of human chorionic gonadotropin and/or embryo growth retardation. Meanwhile, the patients are demand of ineffective to the standard therapy and need to sign informed consent for the off-label use of rhG-CSF before treatment. Comprehensive communication between OB/GYN and patients before therapy is essential according to that the rhG-CSF treatment would be inefficient when genetic abnormality, gamete damage, heterotopic pregnancy and other unknown disorder exist.

Impact of rhG-CSF on Clinical Efficacy and Immune Cell Subsets after Initial Induction Chemotherapy in AML.

The impact of recombinant human granulocyte colony-stimulating factor (rhG-CSF) in acute myeloid leukemia (AML) is still controversial. The purpose of this study is to explore the impact of rhG-CSF administration on clinical efficacy and immune cell subsets after initial induction chemotherapy in AML. The clinical efficacy and immune cell subsets were compared in the newly diagnosed patients with AML according to whether rhG-CSF was used after initial induction chemotherapy. Next, rhG-CSF stimulation experi-ments on leukemia cell lines and primary leukemia blasts were performed in vitro. There was no statistical difference between control group and rhG-CSF therapy group in complete remission rate and relapse free survival. The duration of agranulocytosis was significantly shortened in rhG-CSF therapy group compared with control group. The percentage of circulating monocytic myeloid-derived suppressor cells (M-MDSCs) and regulatory T cells (Tregs) were significantly increased after the administration of rhG-CSF. Furthermore, it was found that rhG-CSF did not promote the proliferation of leukemia cell lines and primary leukemia blasts, but increased the proportion of M-MDSCs and Tregs in vitro. Administration of rhG-CSF after initial induction therapy of AML does not affect the clinical remission and relapse rate, but reduces the duration of agranulocytosis and increases the proportion of M-MDSCs and Tregs.

MATHEMATICAL SOLUTION OF A PHARMACOKINETIC MODEL WITH SIMULTANEOUS FIRST-ORDER AND HILL-TYPE ELIMINATION

Mathematical studies on pharmacokinetic models are essentially important for drug development and optimal dose design. Considering the interaction between drug molecules and their receptors, the elimination of drug molecules can exhibit Hill-type kinetics. In this paper, motivated by the recombinant human granulocyte colony-stimulating factor (G-CSF) and the transcendent Lambert <i>W</i> function, we have studied the mathematical solutions of a one-compartment nonlinear pharmacokinetic model with simultaneous first-order and Hill-type (<i>n</i> = 2) elimination for the case of intravenous bolus administration. By introducing three well-defined transcendental functions depending on three different scenarios, we have established the closedform precise solutions of time course of drug concentration, which is a method to calculate drug concentrations at any time point. As a result, we also have derived the explicit expressions of some key pharmacokinetic surrogates such as the elimination half-life <i>t</i>_1/2 and total drug exposure (i.e. area under the concentration curve (AUC)), which are found as dose-dependent. Finally, a case study of a G-CSF drug is quantitatively illustrated to delineate our theoretical results, including the elimination half-life and AUC for different dosages. Our findings can provide an effective guidance for drugs with simultaneous first-order and Hill-type (<i>n</i> = 2) elimination in clinical pharmacology

Refolding of Proteins Expressed as Inclusion Bodies in E. coli.

Microbial-based biotherapeutics that are produced in Escherichia coli (E. coli) can be generated intracellularly in the form of inclusion bodies (IBs) or in soluble active form in periplasmic space or extracellularly. Overexpression of these biotherapeutics in E. coli leads to formation of insoluble aggregates called inclusion bodies. These IBs contain misfolded and inactive form of proteins which need to be refolded to obtain a functionally active form of proteins. Here, we discuss refolding of E. coli-based recombinant human granulocyte colony-stimulating factor (GCSF), expressed as IBs, and highlight some of the key features associated with the refolding kinetic reaction.

Experimental study of intra-peritoneal instilation of recombinant human granulocyte colony-stimulating factor for the treatment of acute peritonitis

Aim. In the experiment, the possibility of intraperitoneal application of recombinant human granulocyte colony-stimulating factor (G-CSF) for the treatment of acute peritonitis (AP) was investigated.Materials and methods. 60 non-linear white rats. AP was modeled by intra-abdominal puncture of 20% autofecal mixture. After 12 hours, a laparotomy and sanitation of the peritoneal cavity were performed. In 30 animals (control), a solution of decamethoxine was used. In the experiment, after sanitation, a solution of G-CSF on NaCl was injected into the peritoneal cavity at a dose of 0.1 million units per 100 g of mass. After 6, 12, 24 and 48 hours, a relaparotomy was performed and the parietal peritoneum was taken for examination. The number of cells was counted on digital copies of histological preparations.The results. After 6 hours: in the control, the number of polymorphonuclear leukocytes (PLL) - 2.7±0.39, in the experiment - 3.9±0.38 (р&lt;0.05); in the control, the number of lymphocytes (LC) - 0.2±0.13, in the experiment - 0.7±0.33 (р&lt;0.05). After 12 hours: in the control, the number of PLL - 3.1±0.62, in the experiment - 4.6±0.45 (р&lt;0.05); in the control, the number of LC - 0-1 in the field of vision, in the experiment - 1.8±0.33; in the experiment, the number of fibroblasts (FB) was 0.9±0.27. After 24 hours: in the control, the number of PLL - 3.3±0.39, in the experiment - 1.3±0.33; in the control, the number of LC - 1.8±0.41, in the experiment - 2.3±0.33; in the control, the number of FB - 0.4±0.16, in the experiment - 1.6±0.31 (р&lt;0.01); in the control, the number of macrophages (MF) - 0.4±0.16, in the experiment - 0.7±0.21; in the experiment, the number of plasma cells (PC) was 1.1±0.28. After 48 hours: in the control, the number of PLL - 2.1±0.27, in the experiment - 1.4±0.31 (р&lt;0.05); in the control, the number of LC - 2.2±0.29, in the experiment - 3.3±0.37 (р&lt;0.05); in the control, the number of FB - 1.9±0.34, in the experiment - 2.9±0.23 (р&lt;0.05); in the control, the number of macrophages (MF) – 2.0±0.36, in the experiment – 3.4±0.22 (р&lt;0.01); in the control, the number of PCs was 0.9±0.28, in the experiment - 3.1±0.27 (р&lt;0.01). Conclusions. In animals with AP models, signs of a delay in the local response of immunocompetent cells for 12-24 hours, regeneration processes for 24-48 hours, and slowing down of the regression of the inflammatory process in the peritoneum are observed after the peritoneal cavity is cleaned with a decamethoxine solution. In animals that received intraperitoneal instillation of G-CSF after rehabilitation, signs of early activation of the local reaction of immune cells and regeneration processes are observed, along with the acceleration of the regression of inflammation. The results of the experiments indicate that intraperitoneal instillation of G-CSF can be used under clinical conditions for the treatment of patients with AP.

Comparable Incidence Rates of Hematologic and Non-Hematologic Malignancies, Thrombotic Events, and Autoimmune Disorders in Unrelated Adult Donors Undergoing Bone Marrow Collection Versus Peripheral Blood Stem Cell Mobilization with Recombinant Human Granulocyte Colony-Stimulating Factor (rHuG-CSF): Results of the Donor Long-Term Follow-up Study By the National Marrow Donor Program (NMDP)

Comparable Incidence Rates of Hematologic and Non-Hematologic Malignancies, Thrombotic Events, and Autoimmune Disorders in Unrelated Adult Donors Undergoing Bone Marrow Collection Versus Peripheral Blood Stem Cell Mobilization with Recombinant Human Granulocyte Colony-Stimulating Factor (rHuG-CSF): Results of the Donor Long-Term Follow-up Study By the National Marrow Donor Program (NMDP)

Comparison of Hematotoxicity of Pegylated Recombinant Human Granulocyte Colony-Stimulating Factor (PEG-rhG-CSF) Combined with Dual-Agent Concurrent Chemoradiotherapy and Cisplatin Concurrent Chemoradiotherapy for Locally Advanced Cervical Cancer

<h3>Purpose/Objective(s)</h3> Concurrent chemoradiotherapy (CCRT) with cisplatin is the standard treatment for locally advanced cervical cancer (LACC), but there are still some patients who have poor efficacy. Though dual-agent concurrent chemoradiotherapy (dCCRT) can improve the efficacy, the higher hematotoxicity makes it difficult to be popularized in clinical. PEG-rhG-CSF has been recommended by relevant NCCN guidelines for CCRT of malignant tumors in recent years, however there are few reports in LACC. The aim of this study was to evaluate the effect of PEG-rhG-CSF on reducing hematotoxicity in LACC with dCCRT. <h3>Materials/Methods</h3> This study retrospectively analyzed the clinical data of patients with LACC treated with CCRT in the Affiliated Cancer Hospital of Guizhou Medical University. According to patients' clinical protocol, all patients were divided into three groups, single-agent concurrent chemoradiotherapy group(sCCRT), dCCRT group and PEG-rhG-CSF combined with dCCRT group (PEG+dCCRT). Patients in sCCRT received CCRT with cisplatin 40 mg/m² for four to six cycles and patients in dCCRT were treated with paclitaxel 135-175mg/m²+cisplatin 60-80mg/m² or lobaplatin 30mg/m² for two cycles. Recombinant human granulocyte colony-stimulating factor (rhG-CSF) 5ug/kg/d was given to patients in these two groups according to the instructions. Patients in PEG+dCCRT were received dCCRT, and PEG-rhG-CSF 6mg was given in 24 to 72 hours after chemotherapy each cycle. The incidence and duration of grade 3-4 leukocytopenia and neutropenia and febrile neutropenia (FN) (according to CTCAE, V5.0) were the primary endpoints. The second endpoints were completion rate of chemotherapy and radiotherapy, time to complete radiotherapy. <h3>Results</h3> A total of 192 patients were enrolled between July 2019 and December 2021. 51 patients in sCCRT, 68 patients in dCCRT and 73 patients in PEG+dCCRT. The three groups were respectively compared for primary and second endpoints, and the results were shown below. The incidences of grade 3-4 leukocytopenia and neutropenia of the three groups (sCCRT, dCCRT and PEG+dCCRT) were 41.2%, 67.2%, 27.4% and 27.5%, 48.6%, 13.7%, respectively. The incidences of PEG+dCCRT were lowest in the three groups(P<0.05). In the three groups, the duration of grade 3-4 leukocytopenia and FN of PEG+dCCRT were observably reduced(P<0.05). There was no statistical difference of other primary and second endpoints in the three groups. <h3>Conclusion</h3> Prophylactic use of PEG-rhG-CSF during dCCRT makes hematotoxicity manageable, which may make dCCRT have better promotion and improves efficacy of LACC. However, further prospective studies are needed to confirm the results.

Effective Amino Acid Sequencing of Intact Filgrastim by Multimodal Mass Spectrometry with Topdownr.

Therapeutic proteins, known as biologicals, are an important and growing class of drugs for treatment of a series of human ailments. Amino acid sequence variants of therapeutic proteins can affect their safety and efficacy. Top-down mass spectrometry is well suited for the sequence analysis of intact therapeutic proteins. Fine-tuning of tandem mass spectrometry (MS/MS) fragmentation conditions is essential for maximizing the amino acid sequence coverage but is often time-consuming. We used topdownr, an automated and integrated multimodal approach to systematically assess high mass accuracy MS/MS fragmentation parameters to characterize filgrastim, a 19 kDa recombinant human granulocyte colony-stimulating factor used in treating neutropenia. A total of 276 different MS/MS conditions were systematically tested, including the following parameters: protein charge state, HCD and CID collision energy, ETD reaction time, ETD supplemental activation, and UVPD activation time. Stringent and accurate evaluation and annotation of the MS/MS data was achieved by requiring a fragment ion mass error of 5 ppm, considering reproducible N- and C-terminal fragment ions only, and excluding internal fragment ion assignments. We report the first EThcD and UVPD MS/MS analysis of intact filgrastim, and these two techniques combined resulted in 98% amino acid sequence coverage. By combining all tested fragmentation modes, we obtained near-complete amino acid sequence coverage (99.4%) of intact filgrastim.

An open-label pilot study of recombinant granulocyte-colony stimulating factor in Friedreich’s ataxia

Friedreich’s ataxia (FA) is an inherited progressive neurodegenerative disease for which there is no proven disease-modifying treatment. Here we perform an open‐label, pilot study of recombinant human granulocyte-colony stimulating factor (G-CSF) administration in seven people with FA (EudraCT: 2017-003084-34); each participant receiving a single course of G-CSF (Lenograstim; 1.28 million units per kg per day for 5 days). The primary outcome is peripheral blood mononuclear cell frataxin levels over a 19-day period. The secondary outcomes include safety, haematopoietic stem cell (HSC) mobilisation, antioxidant levels and mitochondrial enzyme activity. The trial meets pre-specified endpoints. We show that administration of G-CSF to people with FA is safe. Mobilisation of HSCs in response to G-CSF is comparable to that of healthy individuals. Notably, sustained increases in cellular frataxin concentrations and raised PGC-1α and Nrf2 expression are detected. Our findings show potential for G-CSF therapy to have a clinical impact in people with FA.

Anaphylaxis caused by pegylated recombinant human granulocyte colony-stimulating factor.

Pegylated recombinant human granulocyte colony-stimulating factor (PEG-rhG-CSF), is commonly used to prevent and/or treat neutropenia caused by tumor chemotherapy and radiotherapy with reduced clearance, increased plasma half-life, sustained biologic activity, and good safety. PEG-rhG-CSF anaphylaxis was rare. A 55-year-old male Non-Hodgkin lymphoma patient appeared with chest tightness, increased heart rate, and decreased blood pressure after taking PEG-rhG-CSF used as a primary treatment for neutropenia caused by chemoradiotherapy. The patient's anaphylaxis is likely due to PEG-rhG-CSF. 0.9% sodium chloride 250 mL and dexamethasone sodium phosphate injection 5 mg were carried out. After close observation, the symptoms were improved. Two months later, the patient was readministered with PEG-rhG-CSF and anaphylaxis reoccurred. After the last contact with PEG-rhG-CSF, the patient was prescribed rhG-CSF for many times and did not experience any adverse reactions. PEGylated drugs are not free of risk and immune-mediated adverse events are of concern. PEG is a substance capable of triggering an immune response. The mechanism of PEG involved IgE-mediated anaphylaxis and non-IgE-mediated anaphylaxis, and requires further research. PEGylated drugs are not free of risk and immune-mediated adverse events are of concern.

Clinical implications, safety, efficacy of Recombinant Human Granulocyte Colony-Stimulating Factors and Pegylated Equivalent

&#x0D; A wide use of recombinant human granulocyte colony-stimulating factors (G-CSFs) and their pegylated equivalent is a significant step forward in the treatment of both solid tumors and hematological malignancies. Evidence-based use of these molecules resulted in more intensive treatments, safely extended to frail and elderly patients, and development of response- and comorbidity-tailored approaches. The available G-CSFs are filgrastim, and the long-acting PegFilgrastim, which are produced in E. Coli cells, and are chemically different from native human G-CSF, and lenograstim, a molecule produced in mammalian cells, with a chemical structure identical to native human G-CSF. These chemical differences produce a diverse interaction with receptors and stimulated neutrophils. For instance, lenograstim binds to receptors in the same way of endogenous ligand, and neutrophils obtained from stimulation with this G-CSF have a physiological activity profile similar to neutrophils normally generated in humans. Conversely, the different interaction between filgrastim and G-CSF receptor is more frequently associated with morphological abnormalities, reduced motility and chemotaxis and a reduced response to bacterial stimuli in induced neutrophils.&#x0D; On this background, we reviewed available evidence in order to analyze the impact of these chemical and pharmacodynamic differences among G-CSF molecules on safety, particularly in healthy peripheral-blood stem-cells donors, functional qualities of inducted neutrophils, and mobilization of hematopoietic stem cells. &#x0D;