Granulocyte Colony-stimulating Factor Priming Research Articles

O-076 Novel method to enhance preantral follicle growth by granulocyte colony-stimulating factor administration improves embryos and pregnancy rate in poor ovarian reserve: a randomized controlled trial

Abstract Study question Whether priming with granulocyte colony-stimulating factor (G-CSF) before ART improves embryo and pregnancy rate while increasing anti-Müllerian hormone (AMH) in patients with poor ovarian reserve. Summary answer G-CSF priming improved embryonic development and pregnancy rate in ART with poor ovarian reserve, simultaneously increasing serum AMH. Enhanced preantral follicle growth likely was responsible. What is known already Eleven years ago, we treated repeated implantation failure in 10 women with diminished ovarian reserve by administering G-CSF with embryo transfers. No implantation succeeded directly, but serendipitously 3 of the 10 spontaneously became pregnant 2 months later. One of these pregnancies involved twins in a 45-year-old woman without ovulation induction. We hypothesized that G-CSF administration might have stimulated preantral follicle growth, with improved ovulation after 2 months. In animal models with diminished ovarian reserve, G-CSF administration consistently increased ovarian preantral follicles and AMH. Therefore, we designed this clinical trial. Study design, size, duration Patients were prospectively randomized to priming or control groups. All patients initially underwent conventional infertility treatment for 2 consecutive cycles in which the priming group but not controls received a subcutaneous G-CSF priming injection during the early luteal phase. Each group then underwent 1 cycle of IVF/ICSI and fresh embryo transfer (IVF/ICSI-fresh ET), followed by cryopreserved ET if needed until live birth or embryo depletion. AMH was measured before and after priming. Participants/materials, setting, methods Hundred ART patients 20 to 42 years old with AMH below 2 ng/mL were randomized to priming or control groups (50 patients each). None had over 1 ART failure, day-3 follicle-stimulating hormone (FSH) above 30 IU/L, uterine anomalies, or a partner with azoospermia. Main results and the role of chance Fertilization rate, embryonic development, and implantation rate by fresh ET were significantly improved by G-CSF priming. Clinical and ongoing pregnancy rates by IVF/ICSI-fresh ET were significantly higher with priming (30% and 26% in 47 ART patients; 3 delivered with conventional treatment) than in controls (12% and 10% in 49 ART patients; 1 dropped out). With priming, significantly more patients achieved cryopreservation of redundant blastocysts (53% with priming vs. 24% in controls). The cumulative live birth rate was 32% in 50 patients with priming, significantly higher than 14% in 49 controls (relative risk, 2.8; 95% confidence interval, 1.04-7.7). Infants derived from priming had no congenital anomalies, while infant weights, birth weeks, and Apgar scores were similar between groups. Among 4 variables (age, day-3 FSH, AMH, and priming), logistic regression significantly associated age and priming with cumulative live birth. Priming significantly increased serum AMH. The efficacy of G-CSF priming was not associated with genotypes for killer-cell immunoglobulin-like receptors. No adverse effects of priming were observed. Limitations, reasons for caution Enhancement of preantral follicle growth by G-CSF administration needs to be confirmed histologically as this clinical trial could not allow ovarian biopsies for its purpose of infertility treatments. Wider implications of the findings This study proposes a novel, simple, and safe treatment for poor ovarian reserve. In addition, enhancement of preantral follicle growth by G-CSF priming may also improve ovulatory dysfunctions in polycystic ovary syndrome with elevated AMH, reflecting pooling of retarded/arrested growing preantral follicles. Trial registration number UMIN000013956

Effectiveness and safety of granulocyte colony-stimulating factor priming regimen for acute myeloid leukemia: A systematic review and meta-analysis of the Clinical Practice Guideline for the use of G-CSF 2022 from the Japan Society of Clinical Oncology.

The outcomes of relapsed or refractory acute myeloid leukemia (AML) remain poor. Although the concomitant use of granulocyte colony-stimulating factor (G-CSF) and anti-chemotherapeutic agents has been investigated to improve the antileukemic effect on AML, its usefulness remains controversial. This study aimed to investigate the effects of G-CSF priming as a remission induction therapy or salvage chemotherapy. We performed a thorough literature search for studies related to the priming effect of G-CSF using PubMed, Ichushi-Web, and the Cochrane Library. A qualitative analysis of the pooled data was performed, and risk ratios (RRs) with confidence intervals (CIs) were calculated and summarized. Two reviewers independently extracted and accessed the 278 records identified during the initial screening, and 62 full-text articles were assessed for eligibility in second screening. Eleven studies were included in the qualitative analysis and 10 in the meta-analysis. A systematic review revealed that priming with G-CSF did not correlate with an improvement in response rate and overall survival (OS). The result of the meta-analysis revealed the tendency for lower relapse rate in the G-CSF priming groups without inter-study heterogeneity [RR, 0.91 (95% CI 0.82-1.01), p=0.08; I2=4%, p=0.35]. In specific populations, including patients with intermediate cytogenetic risk and those receiving high-dose cytarabine, the G-CSF priming regimen prolonged OS. G-CSF priming in combination with intensive remission induction treatment is not universally effective in patients with AML. Further studies are required to identify the patient cohort for which G-CSF priming is recommended.

Granulocyte colony-stimulating factor priming improves embryos and pregnancy rate in patients with poor ovarian reserve: a randomized controlled trial

BackgroundGranulocyte colony-stimulating factor (G-CSF) administration increased ovarian preantral follicles and anti-Müllerian hormone (AMH) in animal models with diminished ovarian reserve. We investigated whether G-CSF priming before treatment with assisted reproductive technology (ART) improved embryo development and pregnancy rate while increasing serum AMH in patients with poor ovarian reserve.MethodsIn this prospective randomized open-label controlled trial, 100 patients 20 to 42 years old with AMH below 2 ng/mL were randomized to priming or control groups (50 patients each). None had over 1 ART failure, day-3 follicle-stimulating hormone (FSH) above 30 IU/L, uterine anomalies, or a partner with azoospermia. All patients initially underwent conventional infertility treatment for 2 consecutive cycles in which the priming group but not controls received a subcutaneous G-CSF priming injection during the early luteal phase. Each group then underwent 1 cycle of in vitro fertilization/intracytoplasmic sperm injection and fresh embryo transfer (IVF/ICSI-fresh ET), followed by cryopreserved ET if needed until live birth or embryo depletion. AMH was measured before and after priming.ResultsFertilization rate, embryonic development, and implantation rate by fresh ET were significantly improved by priming. Clinical and ongoing pregnancy rates by IVF/ICSI-fresh ET were significantly higher with priming (30% and 26% in 47 ART patients; 3 delivered with conventional treatment) than in controls (12% and 10% in 49 ART patients; 1 dropped out). With priming, significantly more patients achieved cryopreservation of redundant blastocysts. The cumulative live birth rate was 32% in 50 patients with priming, significantly higher than 14% in 49 controls (relative risk, 2.8; 95% confidence interval, 1.04–7.7). Infants derived from priming had no congenital anomalies, while infant weights, birth weeks, and Apgar scores were similar between groups. Among 4 variables (age, day-3 FSH, AMH, and priming), logistic regression significantly associated age and priming with cumulative live birth. Priming significantly increased serum AMH. No adverse effects of priming were observed.ConclusionG-CSF priming improved embryonic development and pregnancy rate during ART treatment and increased AMH in patients with poor ovarian reserve. Enhanced preantral follicle growth likely was responsible.Trial registrationUMIN registration in Japan (UMIN000013956) on May 14, 2014. https://www.umin.ac.jp/ctr/index.htm.

Randomised controlled trial of conditioning regimen for cord blood transplantation for adult myeloid malignancies comparing high-dose cytarabine/cyclophosphamide/total body irradiation with versus without G-CSF priming: G-CONCORD study protocol

IntroductionA better long-term quality of life after umbilical cord blood transplantation (CBT) is observed compared with transplants from other alternative donors, whereas graft failure and relapses after CBT are still...

Bone Marrow Harvest Volume and CD34+ Cell Count in Pediatric Sibling Donors: A Single Centre Retrospective Analysis

INTRODUCTION: Allogeneic hematopoietic stem cell transplantation (HSCT) is the only treatment modality offering cure or long-term survival for many hematologic malignancies, and non-malignant diseases in children. HLA-matched siblings are considered the best donors because of reduced risks of transplant-related complications and better clinical outcome. According to the National Marrow Donor Program Guidelines, the maximum amount of bone marrow harvest is limited to 20ml/kg donor's body weight. The aim of this retrospective study was to assess the optimal bone marrow harvest volume in pediatric donors needed to obtain the desired CD34+ cell count. METHODS: We reviewed medical charts of 553 pediatric (age at harvest <18 years) sibling donors who were harvested for bone marrow from Jan 2007 to Dec 2017 at our institution for pediatric (age at infusion < 14 years) transplant naïve recipients in order to examine the volume harvested per donor's weight, the percentage of harvests that reached the minimum desired CD34+ cell count of ≥3.0X10^6 per Kg of recipient weight, harvest related hospitalization days and side effects related to the procedure and the impact of granulocyte-colony stimulating factor (GCSF) priming on CD34 count of harvested bone marrow. RESULTS: 288 out of 553 donors were male. 155 (28%) were below 5 years of age at harvest, 189 (34.2%) were between 5-10 and remaining 209 (37.8%) were 10 years and above, with a median of 8.4 years (range: 0.2-17.9). Primary indication for transplant among 131 (23.7%) of our pediatric recipients were Malignant Disorders, Non-Malignant Disorders in 214 (38.7%) and Primary Immunodeficiency and Histiocytic Disorders in 208 (37.6%). GCSF priming was carried out in 219 (39.6%) donors. The minimum desired CD34+ cell count of ≥3.0X10^6 per Kg of recipient weight was reached in 517 (93.5%) harvests. Post infusion Absolute Neutrophil Counts (ANC) recovery within Day+28 was recorded among 472 (85.4%) of the transplant naïve recipients, while in 72 (13%) cases ANC never recovered and in remaining 9 (1.6%) time to recovery was beyond Day+28. ANC recovery within Day+28 was significantly associated with CD34+ cell dose of ≥3.0X10^6 per Kg of recipient weight (n=441, 93.4% vs. 31, 6.6%; P-Value<0.001). Median CD34+ cells (X10^6) collected per Kg of donor weight were significantly higher among donors younger than 5 years of age when compared to those 10 and beyond (P-Value <0.001) with a median harvested volume of 13.7, 12.0 and 8.3 mL/Kg (P-Value<0.001, Table 1). On the same note, median CD34+ cells collected per donor weight were significantly higher among donors primed with GCSF in contrast to those who did not (6.02 X10^6 vs. 3.1 X10^6, P-value <0.001). 54 (9.8%) of our donors required PRBC transfusion; among whom 34 (63%) were below 5 years of age at harvest, 15 (27.8%) 5-10 years and remaining 5 (9.3%) were 10 and above (P-Value<0.001). 2 (0.5%) donors were hospitalized for four days, 12 (3.2%) for three, 201 (54.3%) for two and 155 (42%) for one day only. No significant side effects were noted among our donor population. CONCLUSION: Our study confirmed that CD34 cell count were significantly higher among younger donors. The use of Higher CD34 cell dose is significantly associated with engraftment. Priming with G-CSF had significant impact on CD34+ cell count. These large data confirm the suggestion that the volume of bone marrow harvested can be decreased among younger donors without significantly changing the overall CD34 cell count. Disclosures No relevant conflicts of interest to declare.

Granulocyte colony stimulating factor priming chemotherapy is more effective than standard chemotherapy as salvage therapy in relapsed acute myeloid leukemia

Introduction and objectiveTo improve the complete remission (CR) rate of newly diagnosed acute myeloid leukemia (AML) patients and alleviate the severe side effects of double induction chemotherapy, we combined a standard regimen with granulocyte colony-stimulating factor (G-CSF) priming chemotherapy to compose a new double induction regimen for AML patients who failed to achieve CR after the first course. Patients and methodsNinety-seven patients with AML who did not achieve CR after the first course of standard chemotherapy were enrolled. Among them, 45 patients received G-CSF priming combined with low-dose chemotherapy during days 20–22 of the first course of chemotherapy, serving as priming group, 52 patients were administered standard chemotherapy again, serving as control group. ResultsBetween the two groups there were no differences in the French–American–British (FAB) classification, risk status, the first course of chemotherapy, blood cell count or blasts percentage of bone marrow before the second course. But the CR rate was significantly higher and the adverse effect was much lower in the priming group than the control group. Cox multivariate regression analysis showed that WBC level before the second course and the selection of the second chemotherapy regimen were two independent factors for long survival of patients. DiscussionThese results elucidate that standard chemotherapy followed by G-CSF priming new double induction chemotherapy is an effective method for AML patients to improve CR rate and reduce adverse effects.

Upregulated microRNA-146a expression induced by granulocyte colony-stimulating factor enhanced low-dosage chemotherapy response in aged acute myeloid leukemia patients

The selection of chemotherapy regimen for elderly patients with acute myeloid leukemia (AML) remains challenging. Here, we report that granulocyte colony-stimulating factor (G-CSF) upregulates the expression of microRNA (miR)-146a in a nuclear factor kappaB-dependent manner, leading to direct decreases in the expression of the target proteins CXCR4 and Smad4 in AML cells in vitro. The reduction in CXCR4 expression suppressed the migration abilities of leukemia cells. Downregulation of Smad4 promoted cell cycle entry in leukemia cells. Furthermore, an increase in apoptosis was observed when leukemia cells were treated sequentially with G-CSF and cytosine arabinoside in vitro. These findings suggest that G-CSF treatment may disrupt the protection of bone marrow niches from leukemia cells. In a review of data from 78 cases of primary AML, we found that a high miR-146a expression and/or upregulation of this miRNA during G-CSF priming chemotherapy was predictive of better clinical outcomes. Our findings suggest that miR-146a may be a novel biomarker for evaluating the clinical prognosis and treatment effects of a G-CSF priming protocol in elderly patients with AML.

Granulocyte colony stimulating factor priming chemotherapy is more effective than standard chemotherapy as salvage therapy in relapsed acute myeloid leukemia

Introduction and objectiveTo improve the complete remission (CR) rate of newly diagnosed acute myeloid leukemia (AML) patients and alleviate the severe side effects of double induction chemotherapy, we combined a standard regimen with granulocyte colony-stimulating factor (G-CSF) priming chemotherapy to compose a new double induction regimen for AML patients who failed to achieve CR after the first course. Patients and methodsNinety-seven patients with AML who did not achieve CR after the first course of standard chemotherapy were enrolled. Among them, 45 patients received G-CSF priming combined with low-dose chemotherapy during days 20–22 of the first course of chemotherapy, serving as priming group, 52 patients were administered standard chemotherapy again, serving as control group. ResultsBetween the two groups there were no differences in the French–American–British (FAB) classification, risk status, the first course of chemotherapy, blood cell count or blasts percentage of bone marrow before the second course. But the CR rate was significantly higher and the adverse effect was much lower in the priming group than the control group. Cox multivariate regression analysis showed that WBC level before the second course and the selection of the second chemotherapy regimen were two independent factors for long survival of patients. DiscussionThese results elucidate that standard chemotherapy followed by G-CSF priming new double induction chemotherapy is an effective method for AML patients to improve CR rate and reduce adverse effects.

Two versus three day upfront use of granulocyte-colony stimulating factor in healthy bone marrow donors for pediatric bone marrow transplantation

In order to decrease donors’ exposure to granulocyte-colony stimulating factor (G-CSF), we compared the effect of two versus three days of G-CSF priming on CD34+ yield in bone marrow (BM) harvest. Although the number of BM-CD34+ cells was higher in 3day G-CSF priming, we achieved the same number of CD34+ cells per recipient’s weight in 2day G-CSF priming group, too. In addition, the number of total nucleated cells (TNC) harvested from BM were similar with two or three day regimen. But mononuclear cells (MNC) of the BM graft was higher in the 3day G-CSF priming group. Similar to CD34+ cell numbers, BM harvest yielded similar TNC, and MNC numbers per kilogram of the recipient. We also found that, young donors (≤10year) had more peripheral blood MNC, bone marrow MNC and CD34+ cell numbers. Another interesting finding of this study was obtaining adequate number of peripheral blood stem cells for leukapheresis with three day G-CSF administration. Since engrafment times were also similar in two groups, we concluded that 2-days G-CSF priming was resulted in sufficient mobilization of BM stem cells.

Bone Marrow Harvest in Pediatric Sibling Donors: Role of Granulocyte Colony-Stimulating Factor Priming and CD34+ Cell Dose

To ensure optimal clinical outcomes for patients while retaining adequate protection for donors, the National Marrow Donor Program developed guidelines specifying that up to 20 mL/kg of bone marrow can be harvested from donors. These guidelines, originally developed for unrelated adult donors, are followed in children as well. We studied the impact of granulocyte colony-stimulating factor (G-CSF) priming on the cellular composition of harvested bone marrow, sought to develop an algorithm to optimize bone marrow harvest volume from pediatric matched sibling donors, and studied the impact of CD34+ cell dose on clinical outcomes. We analyzed data from 92 bone marrow harvests and clinical outcomes for 69 sibling recipient-donor duos, The mean age of recipients was 9.85 ± 5.90 years, and that of donors was 11.85 ± 6.36 years. G-CSF priming was not associated with higher yield of CD34+ cells/µL. The median CD34+ cell count obtained from donors was 700 cells/µL (range, 400-1700 cells/µL) in donors age <6 years, 360 cells/µL (range, 100-1100 cells/µL) in donors age 6 to 12 years, and 300 cells/µL (range, 80-800 cells/µL) in donors age >12 years (P < .001). The number of CD34+ cells infused had no impact on traditional clinical outcomes; however, it was significantly related to graft-versus-host disease/relapse/rejection-free survival. Our investigation revealed that ultimately, a CD34+ cell count of approximately 3 to 5 × 106/kg was a threshold beyond which increasing CD34+ cell dose did not impact outcome. In this study, we addressed the broad question of whether harvesting up to 20 mL/kg of bone marrow from a child donor is truly necessary for optimal outcomes in every pediatric case.

Homeostatic γδ T Cell Contents Are Preserved by Granulocyte Colony-Stimulating Factor Priming and Correlate with the Early Recovery of γδ T Cell Subsets after Haploidentical Hematopoietic Stem Cell Transplantation

Emerging evidence from graft manipulations and immunotherapeutic treatments has highlighted a favorable effect of γδ T cells in the setting of allogeneic hematopoietic stem cell transplantation (alloHSCT). However, γδ T cell subsets and their distinct features in the allograft have not been characterized. Additionally, whether homeostatic γδ T cell fractions are influenced by treatment with granulocyte colony-stimulating factor (G-CSF) remains elusive. We initially compared the phenotypes of γδ T cell subsets, including CD27+, CD27-, Vδ1+, Vδ2+, Vδ1+CD27+, Vδ1+CD27-, Vδ2+CD27+, and Vδ2+CD27- cells, in the peripheral blood of 20 healthy donors before and after G-CSF mobilization. The effects of G-CSF on the cytokine production capacities of γδ T cell subsets were also detected. Moreover, the correlation between donor homeostatic γδ T cell content and the early recoveries of γδ T cell subgroups after haploidentical HSCT was investigated in 40 pairs of donors and recipients. We found that both the proportions and IFN-γ secretion capacities of peripheral γδ T cell subsets were preserved in G-CSF–primed grafts. Homeostatic Vδ1 and Vδ2 proportions of donors significantly correlated with the early recoveries of Vδ1 and Vδ2 cells after haploidentical HSCT. Interestingly, a higher day 30 Vδ1 concentration was associated with a lower incidence of cytomegalovirus reactivation in recipients. These results not only clarify the preservation of γδ T cell phenotypes and functional features by G-CSF mobilization but also suggest the importance of homeostatic γδ T cell content for immune recovery after alloHSCT.

Immunological effects of vaccines combined with granulocyte colony-stimulating factor on a murine WEHI-3 leukemia model.

Granulocyte colony-stimulating factor (G-CSF) mobilizes regulatory T cells (Tregs) from bone marrow into the peripheral blood, by reducing the expression of stromal cell-derived factor-1α (SDF-1α). However, G-CSF has rarely been studied in acute myeloid leukemia (AML) immunotherapy. The present study performed a Transwell migration assay in vitro to determine the contribution of SDF-1α to the migration of leukemia cells, and the effects of G-CSF were evaluated. The effects of G-CSF on SDF-1α and Tregs in the AML microenvironment were examined, by employing a WEHI-3-grafted BALB/c mouse AML model (AML-M4). It is evident that G-CSF reversed immunosuppression of the AML microenvironment by reducing SDF-1α in bone marrow and elevating Tregs in the peripheral blood in in vivo studies. Furthermore, AML mice treated with vaccines combined with G-CSF achieved a longer survival time than those treated with vaccines without G-CSF, showing the efficiency of the regimen. The present study demonstrates the effects of G-CSF on the mobilization of leukemia cells and Tregs into the peripheral blood. In addition, immunotherapy with G-CSF priming represents a promising therapeutic strategy of targeting the immunosuppression.

Decitabine before Low-Dose Cytarabine-Based Chemotherapy Combined with Human Leukocyte Antigen–Mismatched Stem Cell Microtransplantation Improved Outcomes in Elderly Patients with Newly Diagnosed Acute Myeloid Leukemia

The optimal treatment for elderly patients with acute myeloid leukemia (AML) remains a great challenge. Establishing a more feasible, acceptable, accessible and safe treatment strategy for elderly patients is urgently needed. We conducted a prospective study of 23 elderly patients (median age, 68 years; range, 60 to 87 years) with newly diagnosed AML to evaluate the efficacy and toxicity of decitabine plus granulocyte colony–stimulating factor priming, low-dose aclarubicin, and cytarabine (DCAG) chemotherapy combined with HLA-mismatched stem cell microtransplantation (SC-MST) without graft-versus-host disease (GVHD) prophylaxis. After the first cycle, the overall response and the complete remission (CR) rates were 86.4% and 81.8%, respectively. CR was achieved in 90.9% of the normal karyotype group and in 80.0% of patients with unfavorable karyotypes at baseline. The median overall survival (OS) and disease-free survival rates were 17 and 13 months, respectively, with a 2-year OS of 34.8%. The median OS of the patients who received ≥3 cycles of SC-MST was significantly longer than those who received only 1 or 2 cycles of treatment. The regimen was well tolerated with a 4-week mortality of 4.3%, and no GVHD was observed. The most common adverse events were hematologic toxicities. Our data suggest that the innovative combination of DCAG with SC-MST may optimize the clinical strategy for elderly patients with newly diagnosed AML.

Up-Regulated Mir-146a Expression in Leukemia Cells Induced By G-CSF Correlates with Enhanced Sensitivity to Cytarabine Treatment: A Putative Novel Predictive Biomarker for Chemotherapy

BackgroundsLeukemic stem cells (LSCs) have emerged as one of reasons for relapse to chemotherapy. To eradicate these quiescent LSCs is of great importance. Administration of granulocyte colony stimulating factor (G-CSF) with low-dose chemotherapy, as a priming strategy, has shown satisfactory effect in acute myelocytic leukemia treatment, especially in elder patients. It is important to investigate the specific signaling pathways and explore useful biomarker to monitor clinical response to therapy. MiR-146a and its downstream putative targets CXCR4 and Smad4 may sustain quiescent LSCs in bone marrow niches. The objective of this study is to further explore the possibility of miR-146a as a marker for sensitivity detection to G-CSF priming chemotherapy.Patients and MethodsMiR-146a over-expression (OE), knock-down (KD) and empty virus (EV) HL-60 cells were constructedin vitro. Leukemia cells were pre-treated with G-CSF for 48h or not. MiR-146a, p50, p65, CXCR4 and Smad4 expression levels were detected. Human stromal cell line HS-5 and HL-60 cells were co-cultured to simulate interaction between bone marrow microenvironment and leukemia cells. Chemotaxis, cell viability, cell cycle and apoptosis were estimated individually. Relationship between miR-146a expression and prognostic factors were analyzed in 46 new diagnosis AML patients, including overall survival (OS) rates, relapsed-free survival (RFS) rates and complete remission (CR) rates.ResultMiR-146a-OE cells showed lower CXCR4 and Smad4 levels than miR-146a-EV cells (p=0.011 and 0.01), while miR-146a-KD cells showed increased CXCR4 and Smad4 expression (p=0.0002 and 0.001), both in mRNA and total protein levels. Higher miR-146a expression was observed in G-CSF pre-treated group. p50 and p65 mRNA and total protein levels were both elevated in response to G-CSF treatment (p<0.01). Meanwhile, the amount of cells migrating to medium containing SDF-1a or HS-5 cells decreased significantly. More cells turned into G1 phase with accelerated proliferative rates, and more apoptotic cells were detected in response to cytarabine treatment.Taking median miR-146a value (3.085) as boundaries, patients with high miR-146a expression at diagnosis (n=23) had longer 2-year OS rates (56.5% vs 26.1%, p=0.042), and longer 2-year RFS rates (52.2% vs 13%, p=0.044) than low-expression group (n=23) (Fig 1). After first course of CAG regimen treatment, sensitive respondents with significantly increased miR-146a (n=24) showed longer OS (83.3% vs 40.9%, p<0.01), longer RFS (62.5% vs 22.7%, p=0.032), and higher CR rates (70.8% vs 40.9%, p=0.042) than those with no molecular response (n=22) (Fig 2). According to their response to G-CSF treatment, we divided patients with low miR-146a expression at diagnosis into 2 subgroups. It was proved that significant miR-146a up-regulation after G-CSF treatment promoted more patients into CR courses with longer OS (80% vs 23.1%, p<0.01) and RFS (50% vs 15.4%, p=0.023).Conclusions1. CXCR4 and Smad4 are both downstream targets of miR-146a.2. G-CSF up-regulates miR-146a expression via activating upstream NF-KB signaling pathway, thus altering a series of biological processes including migration, proliferation and cell cycle of leukemia cells, and eventually sensitized them to chemotherapy.3. High miR-146a levels at diagnosis might predict better outcomes.4. Patients with significantly increased miR-146a levels after G-CSF treatment often have longer OS and RFS rates.5. MiR-146a might be a novel biomarker for evaluating clinical prognosis and treatment effect. [Display omitted] [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Effect of granulocyte colony-stimulating factor priming combined with low-dose cytarabine and homoharringtonine in higher risk myelodysplastic syndrome patients

As sensitization of leukemia cells with granulocyte colony-stimulating factor (G-CSF) can enhance the cytotoxicity of chemotherapy in myeloid malignancies, a pilot study was conducted in order to evaluate the effect of G-CSF priming combined with low-dose chemotherapy in patients with higher risk myelodysplastic syndrome (MDS). The regimen, G-HA, consisted of cytarabine (Ara-C) 7.5mg/m2/12h by subcutaneous injection, days 1–14, homoharringtonine (HHT) 1.5mg/m2/day by intravenous continuous infusion, days 1–14, and G-CSF 150mg/m2/day by subcutaneous injection, days 0–14. 56 patients were enrolled, 34 patients (61%, 95% confidence interval: 51.44–70.56%) achieved complete remission (CR). Median duration of neutropenia was 7days (ranging from 2 to 16days). Grade 1–2 nonhematologic toxicities were documented, including nausea and vomiting (5%), liver function abnormality (5%), and heart function abnormality (2%). No central nervous system toxicity was found. Mortality within the first 4 weeks was 4%. The G-HA regimen is effective in remission induction for higher risk MDS patients and well tolerated due to the acceptable toxicity in maintenance therapy in the patients who cannot undergo Hematopoietic cell transplantation (HCT).

Hematopoietic, Mesenchymal, and Immune Cells Are More Enhanced in Bone Marrow than in Peripheral Blood from Granulocyte Colony-Stimulating Factor Primed Healthy Donors

The use of granulocyte colony-stimulating factor (G-CSF) primed bone marrow (G-BM) has been recently considered as an alternative to mobilized hematopoietic stem cells from peripheral blood (G-PB), especially in the haploidentical transplant setting. The purpose of this study was to compare the effect of in vivo G-CSF priming on BM and PB hematopoietic, mesenchymal (MSC), and immune cells. Forty healthy donors undergoing BM harvest for haploidentical transplant were given subcutaneous recombinant human G-CSF for 7 days. BM and PB samples were harvested on days −7 and 0. The hematopoietic stem/progenitor cells increased significantly after G-CSF priming in both BM and PB with a selective rise of BM CD34+CD38− cell subset. A striking enhancement of the mesenchymal progenitors was detected in G-BM. CD3+, CD4+, CD8+, and CD19+ cell fractions; the naive CD4+ and CD8+ subpopulations; and natural killer and regulatory T cells increased in G-BM, whereas only slight changes were detected in PB. Myeloid dendritic cells (DC1) were significantly up-regulated in both G-BM and G-PB, whereas DC2 increased only in G-BM. In conclusion, our results show substantial differences in the biologic effects exerted by G-CSF at BM and PB levels on hematopoietic cells and immune cell fractions. Furthermore, the impressive rise of MSC progenitors in G-BM might also be relevant to provide MSCs for several clinical use.

Infusion of a non-HLA-matched ex-vivo expanded cord blood progenitor cell product after intensive acute myeloid leukaemia chemotherapy: a phase 1 trial

The intensive chemotherapy regimens used to treat acute myeloid leukaemia routinely result in serious infections, largely due to prolonged neutropenia. We investigated the use of non-HLA-matched ex-vivo expanded cord blood progenitor cells to accelerate haemopoietic recovery and reduce infections after chemotherapy. We enrolled patients with a diagnosis of acute myeloid leukaemia by WHO criteria and aged 18-70 years inclusive at our institution (Fred Hutchinson Cancer Research Center) into this phase 1 trial. The primary endpoint of the study was safety of infusion of non-HLA-matched expanded cord blood progenitor cells after administration of clofarabine, cytarabine, and granulocyte-colony stimulating factor priming. The protocol is closed to accrual and analysis was performed per protocol. The trial is registered with ClinicalTrials.gov, NCT01031368. Between June 29, 2010, and June 26, 2012, 29 patients with acute myeloid leukaemia (19 newly diagnosed, ten relapsed or refractory) were enrolled. The most common adverse events were fever (27 [93%] of 29 patients) and infections (25 [86%] of 29 patients). We observed one case of acute infusional toxicity (attributed to an allergic reaction to dimethyl sulfoxide) in the 29 patients enrolled, who received 42 infusions of expanded progenitor cells. The following additional serious but expected adverse events were observed (each in one patient): grade 4 atrial fibrillation, grade 4 febrile neutropenia, lung infection with grade 4 absolute neutrophil count, colon infection with grade 4 absolute neutrophil count, grade 4 changed mental status, and one death from liver failure. No unexpected toxicity or graft-versus-host disease was observed. There was no evidence of in-vivo persistence of the expanded progenitor cell product in any patient beyond 14 days or induced alloimmunisation. Infusion of the expanded progenitor cell product seemed safe and might provide a promising treatment method for patients with acute myeloid leukaemia. Biomedical Advanced Research and Development Authority in the US Department of Health and Human Services and Genzyme (Sanofi).

A low-dose cytarabine, aclarubicin and granulocyte colony-stimulating factor priming regimen versus a daunorubicin plus cytarabine regimen as induction therapy for older patients with acute myeloid leukemia: A propensity score analysis

This retrospective analysis compared the efficacy of intensive induction therapy consisting of daunorubicin and cytarabine (DNR-AraC) to that of less-intensive therapy including low-dose cytarabine, aclarubicin and granulocyte colony-stimulating factor priming (CAG). Patients aged 60 years or older who were newly diagnosed as acute myeloid leukemia (AML) were analyzed. Sixty-four and 48 patients were treated with DNR-AraC and CAG, respectively. The complete remission rates, 3-year overall survival and event-free survival in the DNR-AraC group were significantly superior to those in the CAG group (65.6% vs. 29.2%, p<0.001, 38.4% vs. 12.3%, p=0.0033, and 20.3% vs. 7.8%, p=0.0030, respectively), although these differences were not statistically significant in multivariate analyses. Next, we calculated a propensity score for selecting the CAG regimen from six factors. The DNR-AraC regimen was associated with better survival than the CAG regimen in a low propensity score group, but there was no difference in survival between regimens in a high propensity score group. Intensive therapy should be performed for patients with sufficient general and comorbid conditions, but less-intensive therapy may be sufficient for patients with higher age, myelodysplasia-related changes, and lower white blood cell counts, which were relevant factors in the propensity score calculation.

The priming induction regimen of HAG as a low dose chemotherapy strategy in AML clonal evolution.

Chemotherapy employs chemical substances to interfere with the growth of cancer cells, and is a major treatment strategy in human cancer including acute myeloid leukemia (AML). Although they often effectively kill fast-dividing tumor cells, chemotherapeutic drugs also profoundly affect mutation spectrum of the tumor genome. Each chemotherapeutic drug has a working concentration within an appropriate range. Concentrations beyond this range will increase drug toxicity without adding clinical benefit, while concentrations below the range will result in an insufficient drug effect. The conventional strategy in oncology is to give a patient drugs at the maximum tolerated dose (MTD) in order to kill as many tumor cells as possible. However, this inevitably causes a high rate of toxicity to normal tissue, as well as resulting in new mutations and genome instability which can lead to drug resistance and relapse. The priming induction regimen of HAG (low dose of cytarabine (Ara-C) and Homoharringtonine (HHT) with granulocyte colony-stimulating factor (G-CSF) priming), has proven to be effective in subgroup of AML patients in small scale clinical studies. It is of great medical significance to explore whether low dose chemotherapy can reduce the side effect of chemotherapeutic drugs while maintaining the therapeutic benefits, and whether such alternative therapeutic strategy is advantageous compared to conventional intensive treatment regarding the development of drug resistance by affecting leukemic genome mutation load and clonal evolution in subgroup of AMLs.

MiRNA Expression Profiling in Mesenchymal Stromal/Stem Cells Derived from G-CSF Primed Bone Marrows

Regulatory microRNAs (miRNAs) are involved in the molecular control of hematopoiesis and diseases that result from aberrations during this process. Bone marrow (BM) transplantation is a commonly used therapy for patients with certain hematopoietic diseases. Prior to BM transplantation, donors are occasionally treated with granulocyte colony-stimulating factor (G-CSF) to increase the number of hematopoietic stem or progenitor cells in the collected stem cell product. Here we aimed to assess the effects of G-CSF priming on miRNA expression of BM derived mesenchymal stromal/stem cells (BM-MSCs) from healthy donors.Previously isolated and characterized BM-MSCs from 3 days G-CSF (10 μg/kg/d) treated (n=5) and untreated (n=8) healthy donors were used for miRNA expression profiling with Affymetrix GeneChip 2.0 Array. Following Robust Multi-array Average (RMA) normalization, data analysis was carried out in Partek Genomics Suite software package (Partek Incorporated). Differentially expressed miRNAs between G-CSF primed and unprimed groups were determined (p<0.05, fold change ±1.5).A total of 2156 miRNAs were upregulated whereas 2436 miRNAs were downregulated after in-vivo G-CSF priming. The expression of hp_hsa-mir-1238 and has-miR-3180-3p non-coding RNAs was significantly increased in G-CSF primed BM-MSCs (p<0.001). Modular enrichment analysis of hsa-miR-3180-3p predicted target genes by GeneCodis 3 software revealed that some may play a role in different types of cancer such as small cell lung cancer (Adjusted p-value: 8.47338e-7, 0.000382029). Our data indicate that G-CSF treatment changes miRNA expression of bone marrow stromal cells (MSCs) and this change might be important to evaluate for long-term biosafety. DisclosuresNo relevant conflicts of interest to declare.