Influence Of Race and Ethnicity On The Collection Of G-CSF Mobilized Peripheral Blood Stem Cells From Unrelated Donors, a Center For International Blood and Marrow Transplant Research Analysis

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 3273-3273
Author(s):  
Jack W. Hsu ◽  
John R. Wingard ◽  
Brent R. Logan ◽  
Pintip Chitphakdithai ◽  
Bronwen E. Shaw ◽  
...  
Keyword(s):  
Stem Cell ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Conflicts Of Interest ◽  
Collection Efficiency ◽  
Univariate Analysis ◽  
Categorical Variables ◽  
Continuous Variables ◽  
Cd34 Cells ◽  
Unrelated Donors

Abstract Peripheral blood stem cell (PBSC) collection is increasingly used in allogeneic stem cell transplantation. However, a small percentage of healthy donors have a poor mobilization response to G-CSF. Very little information exists on the effect of donor race or ethnicity on PBSC mobilization. We analyzed 10776 unrelated donors from the National Marrow Donor Program (NMDP) who underwent G-CSF mobilized PBSC collection from 2006–2012. We investigated the effect of self-reported donor race/ethnicity on collection efficiency, defined as number of CD34+ cells/L (of donor blood processed), number of mononuclear cells (MNC)/L and CD34+ cells/MNC collected on the first day of apheresis. Categorical variables were analyzed by the Chi-square test and the Kruskal-Wallis test was used for continuous variables. A linear regression model was used to compare the various race/ethnic groups while controlling for potential confounding factors (such as age, BMI, gender, and year of apheresis). The result of our analysis is shown in Table 1. Univariate analysis revealed statistically significant differences in CD34+ cells/L, MNC/L and CD34+/MNC in all races analyzed. In general, African Americans (AA) had the highest collection efficiency while Caucasians had the lowest. Other races/ethnicities had collection efficiencies between the two groups. On multivariate analysis, statistically significant differences in CD34+ cell/L were seen in Hispanics, AA and Asian/Pacific Islanders (API), primarily in the obese (Hispanic, AA, API) and overweight (AA, API) donors. In the API group the differences in collection efficiency were predominately seen in males. No differences were seen between Caucasians and Native Americans. This study reveals significant racial/ethnic differences in the efficiency of collection of CD34+ cells in unrelated donors. Although these differences do not appear to interfere with the ability to collect adequate numbers of PBSC, it is currently unknown why they exist. This is an area for continued research. Disclosures: No relevant conflicts of interest to declare.

Is There a Role for Autologous Stem Cell Transplantation (Auto SCT) for Patients with Acute Myelogenous Leukaemia? A Retrospective Analysis.

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 1689-1689
Author(s):  
Nicolas Novitzky ◽  
Valda Thomas ◽  
Cecile du Toit ◽  
Andrew McDonald
Keyword(s):  
Stem Cell ◽  
Stem Cell Transplantation ◽  
Cell Transplantation ◽  
Mononuclear Cells ◽  
Conflicts Of Interest ◽  
Univariate Analysis ◽  
Entire Group ◽  
Myeloablative Conditioning ◽  
Post Transplant ◽  
Significant Difference

Abstract Abstract 1689 Introduction: To better counsel our patients on the role of auto SCT for patients with AML we studied consecutive individuals in CR 1 who had tissue compatible siblings and underwent allogeneic (allo) SCT with subjects who had no HLA donor and actually underwent auto SCT. Methods: Patients were in CR 1 following induction combinations containing 7 days of cyatarabine and etoposide with 3 days daunorubicin followed by similar consolidation therapy. The choice for the type of graft was based on availability of HLA identical siblings. Allogeneic donors underwent PBPC mobilisation with filgrastim and for GvHD prophylaxis grafts were exposed ex vivo to alemtuzumab 1mg/1010 mononuclear cells. Patients were prescribed cyclosporin until day 90 post transplant. Individuals lacking a donor underwent PBPC mobilization with etoposide 2 gr/m2 and harvested PBPC were cryopreserved. Patients received similar myeloablative conditioning followed by infusion of the grafts. Patients were stratified by clinical and laboratory factors as well as cytogenetic risk. The end points were TRM, DFS and OS. Results: The median presentation age for both transplant groups was 35 (14-60) years. Of the 112 consecutive patients achieving remission 37 had HLA identical siblings, but 3 relapsed and donors became unavailable in 2. Thus, autologous or allogeneic grafts were actually transplanted to 43 and 32 patients respectively. There was no significant difference in the presentation clinical features, laboratory parameters, marrow morphology or proportion of low and intermediate cytogenetic risk for both transplant options. Treatment mortality as well as relapse rate was similar (14 and 15%; 39 and 27%, respectively). At a median of 1609 and 1819 post transplant days, 56% and 63% in each group survive. In univariate analysis performance status, cytogenetic risk, morphological features of dysplasia, blast count and LDH were significant factors for survival. While for the entire group there was no difference in survival between both modalities, all patients with unfavourable cytogenetics receiving an autologous graft died of disease recurrence (3 year survival 35% vs 0%; p= 0.05). Conclusions: We conclude that patients with AML who have low or intermediate cytogenetic risk undergoing myeloablative conditioning followed by autologous or allogeneic T-cell depleted stem cell transplantation appeared to have similar outcome. However, those with unfavourable karyotype are unlikely to be cured with autologous grafts and are candidates for experimental modalities. Disclosures: No relevant conflicts of interest to declare.

Carfilzomib and Bortezomib Containing Regimens Yield High Rates Of MRD Negative Autologous Hematopoietic Progenitor Cell (HPC) Grafts In Multiple Myeloma (MM) and High Risk Smoldering Myeloma (SMM)

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 2030-2030
Author(s):  
Nishant Tageja ◽  
Neha Korde ◽  
Constance Yuan ◽  
Kristen Cole ◽  
Jennifer Hsu ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Stem Cell ◽  
High Risk ◽  
Tumor Cell ◽  
Peripheral Blood ◽  
Conflicts Of Interest ◽  
Hematopoietic Progenitor Cell ◽  
Myeloma Cells ◽  
Cd34 Cells ◽  
The Mean

Abstract Background Regimens incorporating modern anti-myeloma drugs, such as carfilzomib (CFZ) and bortezomib (BOR), produce rapid, deep and durable responses in newly diagnosed myeloma patients but their effect on collection of autologous HPC is not well known, including minimal residual disease (MRD) testing of stem cell grafts. Employing older induction regimens (such as VAD), less sensitive flow cytometry techniques detected circulating myeloma cells in 38-46% of autologous HPC grafts (Stewart, et al. JCO. 2001 and Bourhis, et al. Haematologica. 2007). We hypothesized that the use of modern CRd combination therapy including Carfilzomib (CFZ)-Lenalidomide (LEN)-Dexamethasone (DEX) would significantly lower the rates of HPC product contamination. Methods Thirty-six patients, including 29 with MM and 7 with high-risk SMM, underwent HPC mobilization and collection following induction with CRd (n=30), LEN-BOR-DEX (RVd, n=4), Cyclophosphamide-BOR-DEX (CyBorD, n=1) and Cyclophosphamide-BOR-Prednisone (CyBorP, n=1). For HPC mobilization, all patients received 5 days of filgrastim at 10-16 mcg/kg/dose. A combination of the patient’s weight and a peripheral blood CD34 count after 4 doses was used to determine the likelihood of collecting > 4 x106 CD34+ cells/ kg in a single apheresis procedure after a fifth filgrastim dose, according to a previously published algorithm from our institution. Only subjects predicted to require > 1 apheresis by the algorithm received Plerixafor (PLX) at 240 mcg/kg/dose on the fifth day along with the fifth filgrastim dose. HPC collection occurred on day 6, 8 hours after the last mobilizing agent(s) administration. Product contamination with myeloma cells (i.e. MRD status) was evaluated using multi-parameter flow cytometry with a minimum of 3 x 106 events obtained (sensitivity detection rate 1 x 10-5) to examine expression of 9 antigens by the plasma cells. Results The median age at mobilization was 56.2 years (range 40-73) and 19 (53%) were male. At the time of HPC collection, 20 (55%) patients were in sCR/CR/nCR, 11 (30%) had VGPR with 4 PR (11%) and 1 SD (3%). The mean CD34+ cells in the peripheral blood were 33/uL on day 5 and 55/uL on day 6 for the whole cohort. Thirteen (36%) patients did not need PLX. Interestingly, the mean CD34+ count dropped by a mean of 2% from D5 to D6 in patients not receiving PLX while, as expected, it increased by 304% in those who did. The median number of CD34+ cells collected was 6.86 million/kg (range 2.6-12.5) for the whole cohort, (6.6 million/kg without PLX and 7.52 million with PLX p=0.46). Thirty-three of 36 patients (92%) achieved a collection of > 4 million cells /kg in a single apheresis procedure. The 30 patients treated with CRd had a median of 5 (range = 3-7) prior cycles containing LEN with a median of 12 days (range 1-34) between mobilization and last LEN dose. Only 2 of 36 (5%) products were found to have evidence of tumor cell contamination (i.e. MRD positive) using sensitive multiparameter flow cytometry, one patient in PR after 6 cycles of CRd and a second patient in CR after 5 cycles of RVd. Conclusions Modern anti-myeloma therapies, such as CRd and RVd, allow adequate HPC collection in a single apheresis procedure in most cases and improve the quality of the HPC product with greatly reduced tumor cell contamination compared to historical controls. Indeed, 34/36 (94%) patients treated with modern anti-myeloma therapy collected an MRD negative HPC product. Future prospective studies are needed to assess whether autologous stem cell transplants (ASCT) using tumor-free HPC products collected in the era of modern induction therapies have better outcomes. Disclosures: No relevant conflicts of interest to declare.

Donor-to-Recipient Weight Ratio Is Independently Associated with CD34+ Yield in Healthy Donors Undergoing Peripheral Blood Stem Cell Collection for Allogeneic Transplantation

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 2456-2456 ◽  
Author(s):  
Mark A Fiala ◽  
Soo Park ◽  
Camille N. Abboud ◽  
Amanda F. Cashen ◽  
Meagan Jacoby ◽  
...  
Keyword(s):  
Diabetes Mellitus ◽  
Stem Cell ◽  
Peripheral Blood ◽  
Univariate Analysis ◽  
Weight Ratio ◽  
Allogeneic Transplantation ◽  
Healthy Donors ◽  
Cd34 Cells ◽  
Stem Cell Donation ◽  
Reduced Intensity

Abstract Background: Over the past two decades, peripheral blood stem cells (PBSC) have surpassed bone marrow as the preferred graft source for adult allogeneic transplantation due to its more rapid engraftment and potentially better graft-vs-tumor effects, and because PBSC collection is much less invasive for the donor. The optimal CD34+ PBSC dose is ≥ 4.0x106cells/kg, but doses ≥ 8.0x106cells/kg are suggested by some for reduced-intensity conditioning and haploidentical transplants. There is no established minimum CD34+ PBSC dose, but doses below 2.0x106cells/kg have been associated with a higher risk of engraftment delay and failure. There is significant inter-donor variability in the ability to mobilize PBSCs. Several factors have been identified as predictors of PBSC mobilization in healthy donors including: gender, age, weight, body mass index (BMI), and blood counts before and after mobilization. The impact of the donor’s comorbidities on mobilization is currently unknown. Patients/Methods: We performed retrospective chart review of 488 consecutive adult patients who underwent apheresis for allogeneic stem cell donation at Washington University School of Medicine from 2006 through 2013. Patients who received any collection regimen other than 10mcg/kg of G-CSF daily with 20 liters (+/-10%) apheresis on Day 5 were excluded. Patients who had undergone a previous apheresis for stem cell donation were excluded. Univariate analysis was performed to identify predictors of CD34+ PBSC collection in a single apheresis. Variables analyzed were: gender; age; weight; BMI; donor-to-recipient weight ratio; pre and post-mobilization blood counts (white blood count [WBC], hematocrit, platelets, neutrophils, lymphocytes, and monocytes); pre-mobilization glucose and triglyceride levels; post-mobilization peripheral blood (PB) CD34+count; and medical history significant for hypertension, hyperlipidemia, or diabetes mellitus. Subsequently, a linear regression multivariate analysis was performed with all variables found to be significant in the univariate analysis. 2-tailed tests for significance were used throughout the analysis. Results: 304 patients met the eligibility criteria for analysis. The median age was 53 years (range 18-76), 90% were Caucasian, and 50% were male. The median number of CD34+ cells collected was 7.4 x106/kg (range 0.8-27.1). 97% (295) collected ≥ 2.0x106 CD34+cells/kg, 81% (247) collected ≥ 4.0x106 CD34+cells/kg, and 44% (134) collected ≥ 8.0x106 CD34+ cells/kg. Post-mobilization PB CD34+ count (r= 0.841, p <0.001) and donor-to-recipient weight ratio (r= 0.439, p <0.001) were the strongest correlates with CD34+ collection. Weak correlations were seen with post-mobilization neutrophils (r= 0.360, p <0.001), WBC (r= 0.353, p <0.001), and hematocrit (r= 0.207, p <0.001); weight (r=0.280, p <0.001); BMI (r= 0.229, p <0.001); and age (r= -0.207, p <0.001). Male donors collected 9.1x106 CD34+ cells/kg compared to 7.5 x106 CD34+ cells/kg for females, on average (p = 0.003). Hypertension, hyperlipidemia, and diabetes mellitus were not associated with Day 1 CD34+ collection. We performed a multivariate model with the variables: post-mobilization PB CD34+ count; donor-to-recipient weight ratio; gender; post mobilization neutrophil, WBC, and hematocrit; weight; BMI; and age. In this analysis, post-mobilization PB CD34+count, donor-to-recipient weight ratio, and age were all independently significant. Conclusion: After one apheresis, the majority (81%) of donors collected ≥ 4.0x106 CD34+ cells/kg, the optimal number of CD34+ cells for standard allogeneic transplant, but only 44% were able to collect ≥ 8.0x106, the suggested number of CD34+ cells for reduced-intensity or haploidentical transplants. As these reduced-intensity and haploidentical transplants increase in frequency, the need for more accurate predictors of CD34+cell collection will also increase. Donor-to-weight ratio could be a useful tool to stratify potential donors for reduced-intensity and haploidentical transplants. In this study, 63% of donors with a donor-to-recipient weight ratio > 1.0 collected ≥ 8.0x106 CD34+cells/kg, while only 20% of donors with a donor-to-recipient weight ratio ≤ 1.0 did. While gender, weight, and BMI have previously been reported as predictors for CD34+ collection, they potentially were just surrogates for donor-to-recipient weight ratio. Disclosures No relevant conflicts of interest to declare.

The Tetraspanin CD9 Regulates Engraftment and Mobilization of Human CD34+ Hematopoietic Stem/Progenitor Cells and Modulates VLA-4 Activity

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 1169-1169
Author(s):  
Kam Tong Leung ◽  
Karen Li ◽  
Yorky Tsin Sik Wong ◽  
Kathy Yuen Yee Chan ◽  
Xiao-Bing Zhang ◽  
...  
Keyword(s):  
Stem Cell ◽  
Progenitor Cells ◽  
Peripheral Blood ◽  
Scid Mouse ◽  
Conflicts Of Interest ◽  
Chimeric Mice ◽  
Hematopoietic Stem ◽  
Cell Engraftment ◽  
Cd34 Cells

Abstract Migration, homing and engraftment of hematopoietic stem/progenitor cells depend critically on the SDF-1/CXCR4 axis. We previously identified the tetraspanin CD9 as a downstream signal of this axis, and it regulates short-term homing of cord blood (CB) CD34+ cells (Leung et al, Blood, 2011). However, its roles in stem cell engraftment, mobilization and the underlying mechanisms have not been described. Here, we provided evidence that CD9 blockade profoundly reduced long-term bone marrow (BM; 70.9% inhibition; P = .0089) and splenic engraftment (87.8% inhibition; P = .0179) of CB CD34+ cells (n = 6) in the NOD/SCID mouse xenotransplantation model, without biasing specific lineage commitment. Interestingly, significant increase in the CD34+CD9+ subsets were observed in the BM (9.6-fold; P < .0001) and spleens (9.8-fold; P = .0014) of engrafted animals (n = 3-4), indicating that CD9 expression on CD34+ cells is up-regulated during engraftment in the SDF-1-rich hematopoietic niches. Analysis of paired BM and peripheral blood (PB) samples from healthy donors revealed higher CD9 expressions in BM-resident CD34+ cells (46.0% CD9+ cells in BM vs 26.5% in PB; n = 13, P = .0035). Consistently, CD34+ cells in granulocyte colony-stimulating factor (G-CSF)-mobilized peripheral blood (MPB) expressed lower levels of CD9 (32.3% CD9+ cells; n = 25), when compared with those in BM (47.7% CD9+ cells; n = 16, P = .0030). In vitro exposure of MPB CD34+ cells to SDF-1 significantly enhanced CD9 expression (1.5-fold increase; n = 4, P = .0060). Treatment of NOD/SCID chimeric mice with G-CSF decreased the CD34+CD9+ subsets in the BM from 79.2% to 62.4% (n = 8, P = .0179). These data indicate that CD9 expression is down-regulated during egress or mobilization of CD34+ cells. To investigate the possible mechanisms, we performed a VCAM-1 (counter receptor of the VLA-4 integrin) binding assay on BM CD34+ cells. Our results demonstrated that CD34+CD9+ cells preferentially bound to soluble VCAM-1 (17.2%-51.4% VCAM-1-bound cells in CD9+ cells vs 12.8%-25.9% in CD9- cells; n = 10, P ≤ .0003), suggesting that CD9+ cells possess higher VLA-4 activity. Concomitant with decreased CD9 expression, MPB CD34+ cells exhibited lower VCAM-1 binding ability (2.8%-4.0% VCAM-1-bound cells; n = 3), when compared to BM CD34+ cells (15.5%-37.7%; n = 10, P < .0130). In vivo treatment of NOD/SCID chimeric mice with G-CSF reduced VCAM-1 binding of CD34+ cells in the BM by 49.0% (n = 5, P = .0010). Importantly, overexpression of CD9 in CB CD34+ cells promoted VCAM-1 binding by 39.5% (n = 3, P = .0391), thus providing evidence that CD9 regulates VLA-4 activity. Preliminary results also indicated that enforcing CD9 expression in CB CD34+ cells could enhance their homing and engraftment in the NOD/SCID mouse model. Our findings collectively established that CD9 expression and associated integrin VLA-4 activity are dynamically regulated in the BM microenvironment, which may represent important events in governing stem cell engraftment and mobilization. Strategies to modify CD9 expression could be developed to enhance engraftment or mobilization of CD34+ cells. Disclosures No relevant conflicts of interest to declare.

Peripheral-blood stem-cell collections after paclitaxel, cyclophosphamide, and recombinant human granulocyte colony-stimulating factor in patients with breast and ovarian cancer.

1995 ◽  
Vol 13 (7) ◽  
pp. 1714-1719 ◽  
Author(s):  
T Demirer ◽  
S Rowley ◽  
C D Buckner ◽  
F R Appelbaum ◽  
K Lilleby ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Stem Cell ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Granulocyte Colony Stimulating Factor ◽  
Colony Stimulating Factor ◽  
Breast And Ovarian Cancer ◽  
Peripheral Blood Stem Cells ◽  
Cd34 Cells ◽  
Stimulating Factor

PURPOSE Here we evaluate Taxol (paclitaxel; Bristol-Myers Squibb, Princeton, NJ) and cyclophosphamide (CY) with recombinant human granulocyte colony-stimulating factor (rhG-CSF) for mobilization of peripheral-blood stem cells (PBSCs) for autologous stem-cell transplantation in patients with breast and ovarian cancer. PATIENTS AND METHODS PBSCs were collected from 17 patients with breast (n = 11), ovarian (n = 5), and gastric (n = 1) cancer after administration of Taxol (170 mg/m2 x 1) and CY (4 g/m2 x 1) followed by rhG-CSF (10 micrograms/kg/d). PBSC collections after Taxol and CY were compared with PBSC collections from nine patients with stage IV breast (n = 8) or stage III ovarian (n = 1) cancer who had received CY (4 g/m2 x 1) followed by rhG-CSF (16 micrograms/kg/d) for mobilization. RESULTS Mean WBC and platelet counts on the first day of apheresis were 6.3 x 10(9)/L (range, 1.9 to 22.1) and 35 x 10(9)/L (range, 19 to 77), respectively. The median numbers of CD34+ cells, peripheral-blood mononuclear cells (PBMNC), and peripheral-blood total nucleated cells (PBTNC) collected were 13.02 x 10(6)/kg (range, 5.4 to 57.8; mean, 16.02), 6.86 x 10(8)/kg (range, 1.9 to 51.2), and 17.41 x 10(8)/kg (range, 2.4 to 106.6), respectively. In the comparison group, the median yield of CD34+ cells was 6.39 x 10(6)/kg (range, 0.2 to 28; mean, 10.01; P = .01). The mean daily yield of CD34+ cells/kg/collection was 3.5 (range, 0.8 to 28.9) after Taxol and CY, as compared with 1.3 (range, 0.1 to 7.0) for patients who received CY alone (P = .01). All patients who received CY and Taxol reached a target level of 5 x 10(6) CD34+ cells/kg, as compared with five of nine patients (55.5%) who received CY alone (P = .03). CONCLUSION These data suggest that Taxol and CY followed by rhG-CSF mobilizes PBSCs in patients with advanced breast and ovarian cancer more effectively than this regimen without Taxol.

Impact of CD34+ Cell Dose in Peripheral Blood Stem Cell Transplantation from Unrelated Donors for Acute Leukemia Patients.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 2065-2065
Author(s):  
R. Marks ◽  
A. Spyridonidis ◽  
G. Ihorst ◽  
H. Bertz ◽  
J. Finke
Keyword(s):  
Stem Cell ◽  
Stem Cell Transplantation ◽  
Cell Transplantation ◽  
Peripheral Blood ◽  
Peripheral Blood Stem Cell ◽  
Hazard Ratio ◽  
Blood Stem Cell ◽  
Cd34 Cells ◽  
Unrelated Donors ◽  
Blood Stem Cell Transplantation

Abstract In a retrospective study we analysed the impact of the graft composition and several clinical criteria on the outcome of adult AML (n=104) and ALL (n=29) patients undergoing first allogeneic peripheral blood stem cell transplantation from unrelated donors at the University Hospital in Freiburg. The median age of the patients was 52 years (range 18–74) and of the donors 35 years (range 20–58). All patients above 55 years of age received a preparative regimen consisting of fludarabine, melphalan and carmustin (FBM) whereas patients <55y received standard busulphan and cyclophosphamide. Graft-versus-host disease (GvHD) prophylaxis consisted of rabbit ATG (40–60 mg/kg; Fresenius) and cyclosporine combined with MTX in BuCy treated patients or mycophenolate mofetil in patients treated with FBM. There was a trend in the younger donors to mobilize more CD34+ cells than older ones (r=−0.147, p=0.11). A median of 6.1x106 CD34+ cells/kg (range 1.5–17.0) and 3.1 x108CD3+ cells/kg (range 1.0–20.0) were infused. There was no association between the numbers of CD34+ and CD3+ cells in the transplanted peripheral blood stem cell graft (r=0.097, p=0.29). The dose of CD3+ cells infused correlated with the occurence of acute GvHD (aGvHD) II–IV (p=0.02) but not with the development of chronic GvHD (cGvHD). The risk of chronic or acute GvHD was not different between patients receiving more than the 75. percentile (p75) of CD34+ cells (>8x106cells/kg) compared to those receiving < p75 CD34+ cells. In multivariate analysis of all leukemia patients, acute GvHD, chronic GvHD and age were independent prognostic factors in OS. But in older patients (>50y), higher (>p75) CD34+ cell dose in the PB graft was associated with a trend to better OS at one year both in univariate (hazard ratio 0.508, 95% CI, 0.231 to 1.114, p=0.0908) and multivariate analysis (hazard ratio 0.521, 95% CI, 0.235 to 1.155, p=0.1084), while CD34+ cell dose did not have any impact in the OS of younger (<50y) patients. Univariate analysis of 104 patients with myeloid diseases revealed that patients sex, disease status at HCT, HLA match, or the preparative regimen (BuCy versus FBM) did not have any recognizable effect on OS, resulting in a 1-year survival of 65–70% in all analysed subgroups. Multivariate analysis showed that occurence of aGvHD was associated with significantly increased mortality in younger patients (BuCy treated, hazard ratio 3.287, 95% CI 1.126 to 9.596, p=0.03) as compared to FBM patients (hazard ratio 1.52, 95% CI 0.552 to 4.188, p=0.41). Relapse rates were lower in patients who experienced cGvHD as compared to those without cGvHD resulting in a significantly better OS (hazard ratio 0.276, 95%CI, 0.102 to 0.750, p=0.011). In conclusion the data show that in leukemia patients treated with peripheral blood stem cell transplantation from unrelated donors, older patients receiving grafts with higher CD34+ cell counts show a trend to better survival, while in the same group FBM conditioning does not increase the treatment related mortality.

Characterisation of Side Population (SP) Cells Obtained from Different Hematopoietic Progenitor/Stem Cell Compartments in Human Bone Marrow and Peripheral Blood.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4170-4170
Author(s):  
Dag Josefsen ◽  
Lise Forfang ◽  
Marianne Dyrhaug ◽  
Gunnar Kvalheim
Keyword(s):  
Bone Marrow ◽  
Stem Cell ◽  
Cell Population ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Hematopoietic Stem ◽  
Cell Compartments ◽  
Cd34 Cells

Abstract Side population (SP) cells are characterised by their ability to exclude Hoechst 33342 dye from the cells. Using this method, it has been demonstrated that cells within the SP+ fraction of mononuclear cells from both murine and human hematopoietic systems are enriched for primitive hematopoietic stem- and progenitor cells. Moreover, most of the SP+ cells did not express CD34, indicating the presence of a CD34 negative hematopoietic stem cell population. To explore this further, we have examined SP+ cells obtained from different cell compartments in human bone marrow and peripheral blood. Human bone marrow (BM) was obtained from healthy volunteer donors by iliac crest aspiration after informed consent. Mononuclear cells (MNC) were obtained by Ficoll grade centrifugation. CD34+ cells were then isolated from MNC. Highly enriched CD34+ cells were isolated from PBPC obtained from patients with Hodgkin lymphoma. To identify the SP+ cells, the cells were stained with Hoechst 33342 dye. Using flowcytometric techniques (FACStar+, FACSDiva, Becton Dickinson, San Jose, CA) we were able to visualize the dye efflux in SP+ cells. SP+ cells were functionally confirmed using Verapamil. Phenotypical characterisation of the different cell populations using flow cytometric methods was performed. The level of SP+ cells in BM-MNC was 1,3% (mean, n=3) In line with previous findings, we observed that SP+ cells obtained from BM-MNC lack expression of several lineage committed markers, including CD15 and CD19. Most of the cells were CD34− (mean=2,2%), which was lower than in the main population (MP; mean=5%). The level of CD133 expression was low and similar in both populations. Furthermore we found a higher fraction of CD3+ T-cells in the SP fraction than in the MP fraction (mean: 69% vs 51%). To further investigate the SP+CD34+ cell fraction, we examined CD34+ cells isolated from both human bone marrow and peripheral blood. The percentage of SP+CD34+ cells varied from 0,4 up to 18% of the total CD34+ cell population obtained from PBPC (n= 16), whereas the level of SP+CD34+ cells obtained from bone marrow was 5% of the total CD34+ cell population (n=3). Expression of lineage committed markers, including CD10, CD15 and CD19 was less then 10% of the whole CD34+ cell population obtained from PBPC, whereas we found a higher level of expression of these markers in CD34+ cells isolated from bone marrow. However, when we examined the SP+CD34+ cells from either PBPC or bone marrow, we observed that the phenotypic profile of these cells were similar with almost no expression of lineage markers. The frequency of LTC-IC was markedly increased in SP+MNC, in line with previous findings. In addition we also observed a marked increase in LTC-IC in SP+CD34+ cells compared to SP-CD34+ cells in both BM and PB (BM: 7-fold increase; PB: 3–4 fold). In conclusion, SP cells are present in different hematopoietic progenitor cell populations, including BM-MNC, BM-CD34+ cells and PB-CD34+ cells. In SP+CD34+ cell fractions from both BM and PB we observed an increased expression of stem cell markers like CD90 and CD133, whereas in SP+MNC we found low levels of CD34, CD90 and CD133 expression. However, the LTC-IC frequency was markedly higher in all SP+fractions compared to MP fractions, suggesting that sorting of SP+ cells from different hematopoietic stem- and progenitor cell compartments identify immature hematopoietic cells.

Screen for Small Molecules Capable of Expanding Human Hematopoietic Stem Cell Ex Vivo

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 1919-1919
Author(s):  
Iman Hatem Fares ◽  
Jalila Chagraoui ◽  
Jana Krosl ◽  
Denis-Claude Roy ◽  
Sandra Cohen ◽  
...  
Keyword(s):  
Stem Cell ◽  
Hematopoietic Stem Cell ◽  
Mononuclear Cells ◽  
Conflicts Of Interest ◽  
Ex Vivo ◽  
Fold Increase ◽  
Hematopoietic Stem ◽  
Cd34 Cells

Abstract Abstract 1919 Hematopoietic stem cell (HSC) transplantation is a life saving procedure whose applicability is restricted by the lack of suitable donors, by poor responsiveness to mobilization regimens in preparation of autologous transplantations, by insufficient HSC numbers in individual cord blood units, and by the inability to sufficiently amplify HSCs ex vivo. Characterization of Stemregenin (SR1), an aryl hydrocarbon receptor (AHR) antagonist that promotes HSC expansion, provided a proof of principle that low molecular weight (LMW) compounds have the ability to promote HSC expansion. To identify novel putative agonists of HSC self-renewal, we initiated a high throughput screen (HTS) of a library comprising more than 5,000 LMW molecules using the in vitro maintenance of the CD34+CD45RA- phenotype as a model system. Our study was based on the fact that mobilized peripheral blood-derived CD34+CD45RA- cells cultured in media supplemented with: stem cell factor, thrombopoietin, FLT3 ligand and interleukin 6, would promote the expansion of mononuclear cells (MNC) concomitant with a decrease in CD34+CD45RA- population and HSC depletion. LMW compounds preventing this loss could therefore act as agonists of HSC expansion. In a 384-well plate, 2000 CD34+cells were initially cultured/well in 50μl medium comprising 1μM test compounds or 0.1% DMSO (vehicle). The proportions of CD34+CD45RA− cells were determined at the initiation of experiment and after a 7-day incubation. Six of 5,280 LMW compounds (0.11%) promoted CD34+CD45RA− cell expansion, and seventeen (0.32%) enhanced differentiation as determined by the increase in proportions of CD34−CD45RA+ cells compared to control (DMSO). The 6 LMW compounds promoting expansion of the CD34+CD45RA− cell population were re-analyzed in a secondary screen. Four out of these 6 molecules suppressed the transcriptional activity of AHR, suggesting that these compounds share the same molecular pathway as SR1 in stimulating HSC expansion, thus they were not further characterized. The remaining 2 compounds promoted, similar to SR1 or better, a 10-fold and 35-fold expansion of MNC during 7 and 12-day incubations, respectively. The expanded cell populations comprised 65–75% of CD34+ cells compared to 12–30% determined for DMSO controls. During 12-day incubation with these compounds, the numbers of CD34+ cells increased ∼25-fold over their input values, or ∼ 6-fold above the values determined for controls. This expansion of CD34+ cells was associated with a ∼5-fold increase in the numbers of multilineage CFC (granulocyte, erythroid, monocyte, and megakaryocyte, or CFU-GEMM) compared to that found in DMSO control cultures. The ability of the 2 newly identified compounds to expand functional HSCs is currently being evaluated in vivo usingimmunocompromised mice. In conclusion, results of our initial screen suggest that other mechanism, besides inhibition of AhR, are at play for expansion of human HSC. Disclosures: No relevant conflicts of interest to declare.

First Reported Use of a Biosimilar G-CSF in Allogeneic Stem Cell Mobilisation

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 4571-4571
Author(s):  
Nabih Azar ◽  
Sylvain Choquet ◽  
Alice Garnier ◽  
Damien Roos-Weil ◽  
Véronique Leblond
Keyword(s):  
Body Weight ◽  
Stem Cell ◽  
Peripheral Blood ◽  
Conflicts Of Interest ◽  
Median Number ◽  
Large Hospital ◽  
Originator Product ◽  
Healthy Donors ◽  
Stem Cell Mobilisation ◽  
Cd34 Cells

Abstract Abstract 4571 OBJECTIVES: Biosimilar granulocyte colony-stimulating factor (G-CSF) has been approved on the basis of comparable quality, safety and efficacy as the originator product. Approval of biosimilar G-CSF is in the same indications as the originator, including autologous and allogeneic peripheral blood stem cell (PBSC) mobilisation, for which it is being used throughout our large hospital group in Paris, France. Concerns have been raised by professional bodies over use of biosimilar G-CSF in allogeneic transplants. To our knowledge, this is the first reported use of biosimilar G-CSF in healthy donors for allogeneic transplantation. METHODS: Healthy related donors received biosimilar G-CSF (EP-2006, Sandoz Biopharmaceuticals) 10 μg/kg/day for PBSC mobilisation. G-CSF was administered on days 1–4 with the first leukapheresis performed on day 5. If the required number of CD34+ cells per recipient body weight (4 × 106 CD34+ cells/kg) was not collected, G-CSF administration was continued and a second PBSC collection was performed on day 6. Further G-CSF administration and a third leukapheresis was done on day 7 if required. RESULTS: Twelve healthy donors (7 male, 5 female; mean ± SD age 61.0 ± 7.8 years, range 51–73) received biosimilar G-CSF for PBSC mobilisation. Median donor body weight was 82 kg (range 55–115 kg). Median CD34+ cell count on day 5 was 75/ml (range 23–140). A single leukapheresis after 4 injections of biosimilar G-CSF was sufficient to collect the CD34+ cell dose required in six donors with peripheral blood (PB) CD34+ counts ranging from 80–140 ml. Six donors underwent a second leukapheresis after 5 days of biosimilar G-CSF, one of whom (a 73-year old female) underwent a third leukaphereses on day 7 (PB CD34+ at first apheresis 23 ml). Median number of CD34+ cells per recipient bodyweight was 5.5 × 106 cells/kg (range 2.6–8.9). Five donors reported bone pain (4 mild, 1 moderate severity) which was effectively treated with paracetamol. No other adverse events were reported. Blood measurements were normal in all donors when assessed one week after the end of mobilisation. These findings are consistent with our previous use of originator G-CSF (Neupogen®) in allogeneic stem cell mobilisation. CONCLUSION: These preliminary findings suggest biosimilar G-CSF is effective and well-tolerated in allogeneic stem cell mobilisation. Further studies are required to assess longer-term safety outcomes. The use of biosimilar G-CSF may provide important cost-savings when compared with the originator product. Disclosures: No relevant conflicts of interest to declare.

Allo-HSCT From Unrelated Donors Improves The Outcome Of Elderly AML Patients In CR1

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 2166-2166
Author(s):  
Yosuke Nagahata ◽  
Yuichiro Ono ◽  
Yotaro Ochi ◽  
Yusuke Koba ◽  
Yasuhiro Kazuma ◽  
...  
Keyword(s):  
Multivariate Analysis ◽  
Stem Cell ◽  
Complete Remission ◽  
Conflicts Of Interest ◽  
Intensive Therapy ◽  
Univariate Analysis ◽  
Unrelated Donor ◽  
Hematopoietic Stem ◽  
Elderly Aml ◽  
Unrelated Donors

Abstract Background Recent reports have suggested that allogeneic hematopoietic stem cell transplantation (allo-HSCT) from HLA-matched related donors improves the outcome of elderly acute myeloid leukemia (AML) patients in first complete remission (CR1) (Kurosawa et al BBMT 2011). However, the efficacy of allo-HSCT from alternative donors for the management of elderly AML patients in CR1 remains to be clarified. In this study, we examined the efficacy of allo-HCST for the management of elderly AML patients. Methods We retrospectively collected the data of consecutive AML patients who were aged 50-70 years and diagnosed in our hospital between January 1, 2000 and May 31, 2013. Patients with acute promyelocytic leukemia (APL) and patients who could not receive intensive therapy or died within 60 days after diagnosis were excluded. The primary endpoint of this study was overall survival (OS), which was defined as the interval between diagnosis and the last follow up. The unadjusted probabilities of OS were estimated using the Kaplan-Meier method. To compare OS between the treatment groups, the log-rank test was used. Multivariate analysis were performed with Cox proportional hazard regression models and 95% confidence intervals (CIs) were calculated. Results A total of 131 elderly AML (non-APL) patients were identified. Except for 14 patents, all received chemotherapy. 12 patients who died within 60 days were excluded from the study. Among the remaining 105 patients, 41 could not be induced into complete remission (CR) and showed a dismal outcome; the probabilities of 2 years (2y)-OS were 0% and 4.4% for 5 patients who received allo-HSCT and 36 patients who did not, respectively. In contrast, the probability of 2y-OS of 64 patients who achieved CR1 was 48.5%. Of these 64 patients, 20 proceeded to allo-HSCT. 13 patients were transplanted in CR1, 3 in second CR (CR2), and 4 in relapse. The probabilities of 2y-OS were 56.1% and 45.0% for patients who received and who did not receive allo-HSCT, respectively (p=0.27). When the disease status at allo-HSCT was considered, the 2y-OS of patients who received allo-HSCT in CR1 reached 83.3%, whereas that of patients transplanted in CR2 or relapse reached only 14.3%. Next, we performed univariate and multivariate analysis of OS of patients who achieved CR1. In addition to whether allo-HSCT was performed in CR1 or not, the following factors were also considered as covariates: age and leukocyte counts at diagnosis, cytogenetic classification according to National Comprehensive Cancer Network guidelines, presence or absence of preceding hematological diseases, and the number of induction regimens required for achieving CR1. Univariate analysis showed that allo-HSCT in CR1 was associated with improved OS (2y-OS: 83.3% vs 40.6%, p=0.013) (figure1). Multivariate analysis also showed that allo-HSCT in CR1 was the only favorable prognostic factor for OS (Hazard ratio=0.25, 95%CI: 0.077-0.82, p=0.022). Stem cell sources of allo-HSCT done in CR1 were 1 HLA-matched related donor and 12 unrelated donors including umbilical cord blood from 1 donor. Conclusion Allo-HSCT, even from an unrelated donor, improves the outcome of elderly AML patients in CR1. However, allo-HSCT for those with relapsed or refractory disease could hardly extend their survival. Disclosures: No relevant conflicts of interest to declare.

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