bluuebunny/crossref_metadata_2025_split huggingface dataset:
Phosphorylation Levels of BCR-ABL, Stat5, and CrkL Proteins in Imatinib-Resistant CML Patients as Compared to Previously Untreated CML Patients: Implication of Poor Drug Binding and Activation of Alternate Pathways.
Authors: Gorre M. , Kantarjian H. , Jilani I. , Cortes J. , Salvado A. , Rae P. , Alland L. , Dugan M. , Ottmann O. , Bhalla K. , Giles F. , Albitar M. 
Published: October 2019

Abstract
Disease progression in patients with chronic myeloid leukemia (CML) while on imatinib therapy has been reported, and ex-vivo studies have suggested that this resistance can be seen despite adequate suppression and deactivation of the BCR-ABL fusion protein (Donato et al. Blood. 2003;15:690). Whether the levels of BCR-ABL phosphorylation differ between imatinib-resistant and imatinib-sensitive patients is not known. We compared the relative phosphorylation of BCR-ABL protein (ratios of phospho-tyr245-BCR-Abl:total BCR-ABL protein and phospho-thr735-BCR-ABL:total BCR-ABL protein) in 54 previously untreated CML patients and 62 CML patients with progressive disease who were resistant to imatinib. Resistant patients had significantly lower levels of phospho-thr735-BCR-ABL protein than previously untreated patients, with a median ratio of 0.64 (range, 0.03–1.63) vs. 0.80 (range, 0.40–1.67) (Wilcoxon test, P <0.001). Ratios of phospho-tyr245-BCR-ABL were also lower in patients with resistant progressive disease, with a median of 0.71 (range, 0.02–2.18) vs. 0.80 (range, 0.30–2.15) in previously untreated patients (P=0.03). Furthermore, when quantitative flow cytometry (QuantiBRITE) was used to measure CrkL phosphorylation, resistant patients had significantly lower levels of phosphorylated CrkL/cell than did previously untreated patients (median of 4204 mol/cell Vs. 5982 mol/cell; P=0.01), despite an increase in the overall percentage of p-CrkL-positive cells due to an increase in blasts. Similarly, the levels of AKT phosphorylation as measured by quantitative flow cytometry showed significantly lower intensity p-AKT in resistant patients than in previously untreated patients (median of 4131 mol/cell vs. 5949 mol/cell; P=0.008), again despite an increased percentage of p-AKT-positive cells due to an increase in blasts. There was no significant difference in Stat5 phosphorylation as measured by quantitative intensity, but the percentage of p-Stat5-positive cells was significantly higher in resistant patients than in previously untreated patients (median 22% vs. 8%; P=0.02). This data confirms that in imatinib-resistant CML patients, the disease resistance is not necessarily dependent on higher activity in the BCR-ABL and AKT pathways, but is more likely due to inability of imatinib to bind and suppress BCR-ABL (possibly due to mutations) or to the activation of other pathways that render the disease resistant. Further studies are needed to focus on specific mutations and the activation of other pathways.
Heat Shock Protein 70 over Expression Is Associated to Imatinib Resistance in Chronic Myelogenous Leukemia.
Authors: Pocaly Marion , Lagarde Valérie , Etienne Gabriel , Ribeil Jean-Antoine , Bonneu Marc , Claverol Stéphane , Moreau-Gaudry François , Hermine Olivier , Turcq Béatrice , Mahon François-Xavier , Pasquet Jean-Max 
Published: October 2019

Abstract
Imatinib is an effective therapy for chronic myeloid leukemia (CML), a myeloproliferative syndrome characterised by the expression of the recombinant oncoprotein Bcr-Abl. Imatinib inhibits Bcr-Abl tyrosine kinase activity leading to apoptosis of leukemic cells sparing normal hematopoiesis. Several mechanisms of resistance to imatinib have been identified both in vitro and in vivo: Bcr-Abl mutations, an over-expression of the Bcr-Abl kinase itself or other tyrosine kinase bypass. To identify unknown mechanism, we used an imatinib resistant cell line (K562-R) generated from the erythroblastic cell line K562 (K562-S) (Blood, 2000; 93: 1070–1079) for which all described mechanisms of resistance have been previously invalidated. Previous results from a proteomic study identified some chaperon proteins such as heat shock proteins with an increased expression level in K562-R. One of them, the heat shock protein 70, Hsp70, has a 3 fold increase expression level in K562-R cells, results which have been confirmed by western-blot analysis. To characterise the role of Hsp 70 in imatinib resistance, we inhibit Hsp 70 expression by RNA silencing (siRNA) in K562-R cells and over-express it in K562-S cells. Inhibition of Hsp70 protein expression by siRNA decrease Hsp70 expression rapidly over 90% at day 4 which is associated with a significant reduction of viability (66 ± 6%, n = 5, p < 0.03). Over expression of Hsp 70 in K562-S cells induced a significant increase of resistance to imatinib since the addition of imatinib only increases mortality by 27 ± 5 % in comparison to 52 ± 4 % for K562-S cells (n = 4, p < 0.001). Detection of HSF-1 phosphorylation, the major transcription factor involved in Hsp 70 expression, did not show significant differences between K562-S and K562-R cells although over a 3 fold increase is detected in the mRNA level of Hsp 70 in K562-R cells by quantitative PCR. Furthermore, the comparison of Hsp70 expression in mononuclear cells of 7 CML patients before imatinib treatment and at the relapse time shows that Hsp 70 is increased in imatinib resistant patients suggesting it could also play a role in resistance in vivo. Present study confirmed that over expression of Hsp 70 in the cell line K562-R is involved in the mechanism of imatinib resistance in vitro. Moreover, the correlation between the increase of Hsp 70 in CML patient cells and resistance suggests it could be an interesting marker and potentially a therapeutic target.
Adoptive Immunotherapy with Donor Lymphocyte Infusions after T-Cell Depleted Allogeneic Stem Cell Transplantation for Patients with Acute Myeloid Leukemia.
Authors: Barge Renee M.Y. , Steel Eva , Starrenburg Ingrid , Beaumont Floor , Smit Willem M. , Schaafsma Martyn Ronald , Posthuma Ward , Fibbe Willem E. , Falkenburg Frederik , Willemze Roelof 
Published: October 2019

Abstract
In this study we analyzed 40 patients with AML who received donor lymphocyte infusions (DLI) after allogeneic stem cell transplantation (alloSCT). Patients with a median age of 42 (range, 11–66) were transplanted with an in vitro T-cell depleted (Campath in the bag) graft from an HLA matched sibling following a myeloablative (n=35) or a nonmyeloablative conditioning regimen (n=5). Patients received DLI for relapsed AML (n=28) or for mixed chimerism (n=12). All patients with relapsed AML were treated with a cytarabine-containing chemotherapy regimen or with gemtuzumab ozogamycin immediately prior to DLI treatment and received daily 3 x 106 U α-IFN sc. The DLI dose administered was depending on the protocol used. Fourteen relapsed patients received high dose DLI with a median of 50 x 106 CD3+ cells/kg and 14 low dose DLI with a median of 5 x 106 CD3+ cells/kg. Ten of 28 patients with relapsed AML obtained a complete remission after the combination of chemotherapy and DLI. One patient obtained a partial remission and fourteen patients showed no response. Three patients were not evaluable. No effect of cell dose was observed on the clinical response after DLI. The median percentage of donor chimerism in patients with relapsed AML before DLI was 74% (range, 5–96%) and increased after DLI to 97% (range, 5–100%). Six patients with relapsed AML became complete chimera after DLI. DLI caused in 12 of 28 relapsed patients grade 3–4 GVHD. cGVHD was observed in 3 patients of whom 2 developed extensive GVHD. With a median follow up of 11 months only 1 pt in the high dose and 4 pts in the low dose group are alive in CR and 23 have died due to progressive disease (16) or GVHD (7).
Due to this low rate of long-term survival of relapsed AML patients treated with DLI, we recently developed a new DLI protocol for AML patients. Twelve patients with mixed chimerism at 6 months after T-cell depleted alloSCT were pre-emptively treated with 3 x 106 CD3+ cells/kg. Complete chimeric response was observed in 6 of these 12 patients. Two patients showed increasing mixed chimerism and three patients showed no response. Two of 12 patients treated for mixed chimerism developed aGVHD gr 1–2. No severe aGVHD was observed in this group. Two patients developed chronic GVHD, both after nonmyeloablative conditioning. Incidence of GVHD was significantly lower in this group compared to the relapsed patients. Currently, 7 patients are alive in CR. In conclusion: 1) Long term survival of patients with overt relapse AML after alloSCT treated with DLI remains poor 2) High dose DLI appeared to be less effective than low dose DLI due to increased toxicity 3) DLI for AML may be more successful when it is administered pre-emptively for mixed chimerism after T-cell depleted alloSCT.
Serum Free Light Chain [FLC] Assay in Multiple Myeloma Patients Who Achieved Negative Immunofixation after Allogeneic Stem Cell Transplantation.
Authors: Moesbauer Ulrike , Schieder Heike , Renges Helmut , Ayuk Francis , Zander Axel , Kröger Nicolaus 
Published: October 2019

Abstract
The quantitative assay for free light chains [FLC] has been reported to be sensitive and specific for detecting and monitoring free light chain diseases such as multiple myeloma. To evaluate the sensitivity of FLC for monitoring patients in complete remission for early detection of relapse, the measurement of more than 250 serum free light chains were performed with the commercial available Freelite TM kit [Binding Site] in 26 patients who achieved complete remission with negative immunofixation after dose reduced allogeneic stem cell transplantation. The patient groups were divided in those who remained immunofixation negative [n=12, group 1] during follow-up of at least 1 year and those who had been immunofixation negative but became positive during follow-up [n=9, group 2] and those who had achieved near complete remission with positive immunofixation but then became immunofixation negative during follow-up [n=5, group 3]. In group 1 the measuring of 105 FLC concentration and kappa/lambda ratio was performed in 12 patients. In 10 patients [83 %] free light concentration of kappa or lambda remained within the normal range during follow-up of more than 1 year. In 2 patients [17 %] kappa or lambda FLC concentration was above the normal range, but remained stable without any signs of increasing amount. Group 2 consisted of 9 patients who had been immunofixation negative but became positive during follow-up. In all patients an increase of the corresponding free light chain could be observed in serum. In 4 patients a very close monitoring of immunofixation and free light assay was performed and an at least 25 % increase of the free light concentration in serum was observed at a median of 97 days before immunfixation became positive. In group 3 five patients who had been immunofixation positive became negative during follow-up. In all of the patients the free light concentration was within the range at time of negative immunofixation. The corresponding free light concentration dropped down and reached normal level at a median of 38 days before the patients had achieved negativity of immunofixation. These results suggest that serum free light chain assay allows monitoring of patients with complete remission and might detect early relapse before immunofixation becomes positive. Thus, an early increase of free light chain assay in immunofixation negative patients after allogeneic transplantation might be an useful guide for adoptive immunotherapy strategies to prevent clinical relapse.
Outcome of Allogeneic Stem Cell Transplantation for Patients with Severe Aplastic Anemia after Conditioning with Procarbazine, ATG and Cyclophosphamide: A Single Center Experience.
Authors: Seong Ki , Lee Jong Wook , Kim Yoo Jin , Kim Hee Je , Ki Chang , Lee Seok , Yong Sung , Sung Woo , Choo Kim Chun 
Published: October 2019

Abstract
Background: Cyclophosphamide (CY, 200 mg/kg) plus ATG seems to be accepted as a standard conditioning regimen in HLA-matched sibling stem cell transplantation (SCT) for severe aplastic anemia (SAA). We report herein the outcome of allogeneic SCT for the patients with SAA conditioned with procarbazine (PCB), ATG and CY.
Methods: Between January 1995 and March 2004, consecutive one hundred and sixty two patients with SAA received matched sibling SCT after conditioning with PCB, ATG and CY. The median age of patients was 28.5 (16~50) and median interval between diagnosis and SCT was 10.5 months (1~216). Fifty-five patients (34%) had a history of immunosuppressive therapy before SCT. The conditioning regimen consisted of PCB (6.25 mg/kg/day, 6 days), CY (50 mg/kg/day, 4 days) and ATG (1.25 mg/kg/day, 3 days). No graft manipulation was performed in 92 patients (52%), and donor marrow expansion and megadose transplantation (bone marrow plus CD34+ isolated PB as a stem cell source) were done in 35 patients (21.6%). All patients received of cyclosporine and methotrexate as GVHD prophylaxis.
Results: The median dose of CD34+ and CD3+ cells infused were 4.8 x 106/kg (0.8~49.6) and 5.1 x 107/kg (0 ~ 38), respectively. Median time to neutrophil recovery more than 500/μL and platelet recovery more than 20,000/μL were 13 days (0~49) and 19 days (0~150), respectively. All patients achieved successful primary engraftment, but delayed engraftment failure developed in 18 patients (11%). Ten among 18 patients who developed graft failure received second SCT, resulting survival of 9 patients. The incidence of acute (more than grade II) and chronic GVHD were 9.3% (n=15) and 13.0 % (n=21), respectively. DFS and overall survival (OS) at 6 years was also 90%. Factors influencing on DFS on univariate analysis were patient and donor’s age, development of acute and chronic GVHD, and times of pregnancy (over 2 times). Development of acute and chronic GVHD, and occurrence of rejection were factors that influence DFS on multivariate analysis. Causes of death (n=15) were pneumonia (n=4), rejection (n=4), GVHD±infection (n=2), chronic GVHD (n=1), MOF (n=1), CVA (n=1), autoimmune hemolytic anemia (n=1), and unknown (n=1).
Conclusions: These data suggest that the conditioning regimen used in this study is feasible for patients with SAA who received matched sibling SCT. GVHD and rejection were the most important factors that influence DFS.
Fludarabine Based Conditioning for Allogeneic Transplantation in Severe Aplastic Anemia.
Authors: Chandy Mammen , George Biju , Viswabandya Auro , Mathews Vikram , Bajel Ashish , Srivastava Alok 
Published: October 2019

Abstract
Patients with severe aplastic anemia (SAA) who are multiply transfused or septic have a poor outcome after allogeneic stem cell transplantation. Forty three patients (31 males and 12 females) with SAA underwent allogeneic BMT using a fludarabine based conditioning regimen between 1998 and 2005. The median age was 20 years (range: 4–38) with 11 children and 32 adults. All donors were 6 antigen matched HLA identical sibling or family donors. Co morbidities seen included bacterial sepsis in 15 patients, fungal pneumonia in 4 and a recent intracranial bleed in 5 patients. Seven patients had failed Antithymocyte or antilymphocyte globulin (ATG/ALG) and two patients had failed their first transplant. The median time from diagnosis to transplant was 12 months (range: 2 – 96) and the median transfusions prior to BMT was 28 (range: 2 – 380). Conditioning therapy consisted of: Fludarabine (Flu) 180 mg/m2 over 6d + Busulfan (Bu) 8 mg/kg over 2d + ATG 40 mg/kg/day over 4 d in 17 patients, Flu 180 mg/m2 over 6d + Cyclophosphamide (Cy) 120 mg/kg over 2d ± ATG 40 mg/kg/day over 4d in 17 patients, Flu/TBI/OKT3 in 4, and Cy 120 + Flu 150mg/m2 in 5 patients. Graft versus host disease (GVHD) prophylaxis consisted of Cyclosporine and mini methotrexate. Graft source was peripheral blood stem cells in 39 patients and G-CSF stimulated bone marrow in 4. The median cell dose was 5.2 x 108 MNC/kg (range: 2.1 – 13.6) for PBSC and 5.2 x 108 TNC/kg (range: 3.7 – 6.8) for bone marrow. Five patients expired within the first 10 days due to sepsis. Thirty seven patients engrafted with a mean time to ANC > 500 of 11.6 days (range: 8 – 18) and median platelet engraftment time of 13 days (range: 8 – 32). One patient had primary graft failure and expired on day 64 due to fungal pneumonia despite a second transplant. Acute GVHD was seen in 14 patients (38%) with Grade III–IV GVHD in 4 (10.5%). Chronic GVHD was seen in 10 patients with 6 having limited and 4 with extensive GVHD. Two patients had secondary graft rejection on day + 24 and +60 respectively and expired due to fungal pneumonia. At a median follow up of 17 months (range: 5 – 78); 29 patients (67.7%) are alive and well. Among patients treated with Flu/Bu/ATG, 12/17 (70.5%) are alive and well while the DFS is 82% (14/17) in patients treated with Flu/Cy ± ATG. Comparison with patients conditioned with Cy/ATG during 1990–2004 is given in the table. This comparison suggests that a fludarabine based conditioning regimen may be better, with less rejection, than Cy/ATG for allogeneic BMT in sick patients with SAA who are infected and multiply transfused at the time of BMT.
Comparative data Fludarabine Cy/ATG Number 43 26 Rejection 3 (7%) 7 (27%) DFS 29(67.7%) 11 (46%)
Birth Order and Outcome in HLA-Identical Sibling Donor Hematopoietic Stem Cell Transplants. Impact of a Sequential Fetomaternal - Maternofetal Cell Transfer?.
Authors: Bucher Christoph M. , Heim Dominik , Buser Andreas , Passweg Jakob R. , Gratwohl Alois 
Published: October 2019

Abstract
Fetomaternal and maternofetal cell transfer have been described. Their clinical relevance is unknown. We hypothesized that firstborn siblings could tolerize their siblings born thereafter through sequential fetomaternal-maternofetal cell transfer. Hence, stem cell transplants within a family from a firstborn sibling (group A) should result more graft-versus-host disease (GvHD) and worse overall survival than transplants to a firstborn donor (group B). Results of a retrospective single center cohort analysis of 321 HLA-identical sibling donor hemopoietic stem cell transplants showed a survival of 48.3% (+/−10.9%) at 10 years in the group with firstborn donors (group A, 110 patients) as compared to 63.2 (+/−10.6%) in the group with firstborn recipients (group B, 105 patients; p<0.02) and a RR of death after adjustment for other risk factors of 2.6 (CI 1.45–4.66; p<0.001) for the group with firstborn donors. These results support the concept of a clinically relevant tolerizing effect of birth order, possibly mediated by fetomaternal cell transfer in man.
Patients characteristics and outcomes Birth Order Donor First Sibling Recipient First Sibling n 110 105 Donor Age median (range) 30.1 (4.9–68) 25.4 (0.4–59.1) p=0.005 Recipient Age median (range) 25.4(3.2–62.1) 30.8 (2.2–63.0) Number of Siblings 2 67 69 n.s. 3 26 22 >3 17 14 Diagnosis n.s. Acute myeloid leukemia n 33 26 Acute lymphoblastic leukemia n 24 24 Chronic myeloid leukemia n 23 23 Lymphoproliferative disorders n 14 13 Severe aplastic anemia n 11 12 Myelodysplastic syndromes n 5 7 Stem Cell Source n.s. Bone Marrow n 71 73 PBSCT n 39 32 Conditioning n.s. With totoal body irradiation n 18 22 Without total body irradiation n 92 83 Outcomes Survival at 10 yrs % 48.3 63.2 p<0.02 Relapse at 3 yrs. % 26 20 n.s. Acute GvHD p=0.017 < Grade II n 46 61 >= Grade II n 64 44 Chronic GvHD n.s. none n 28 30 n.a. n 29 17 Limited n 32 30 Extensive n 21 23 Figure 1: Kaplan Meyer estimate of cumulative survival of groups A(firstborn donor: gray line) and B (firstborn recipient: black line). + indicates censored patient. Figure 1:. Kaplan Meyer estimate of cumulative survival of groups A(firstborn donor: gray line) and B (firstborn recipient: black line). + indicates censored patient.
Double Unit Cord Blood Transplantation in Children with Acute Leukemia.
Authors: Yoo Keon Hee , Kang Hyoung Jin , Lee Soo Hyun , Jung Hye Lim , Sung Ki Woong , Koo Hong Hoe , Shin Hee Young , Ahn Hyo Seop 
Published: October 2019

Abstract
Cell dose is one of the most important factors predicting successful outcome after cord blood transplantation (CBT), and the University of Minnesota group recently reported results of transplantation using 2 partially HLA-matched cord blood units for adult patients. Although a significant proportion of pediatric recipients would benefit by cell dose augmentation, data concerning multiple unit CBT in children are lacking by far. We investigated the feasibility of double unit CBT for pediatric recipients. If the cryopreserved nucleated cell dose of the largest available unit was 3.5x107 or less per kg recipient body weight, double unit CBT were scheduled. Twelve patients with acute leukemia (7 myeloid, 3 lymphoblastic, 2 mixed) aged 6.2–16.9 y (median 13.3 y) received 2 cord blood units at our two cooperative institutions. All patients except one were in complete remission before transplant (4 CR1, 6 CR2, 1 CR3, 1 in 3rd relapse). Patients were conditioned with intravenous busulfan- or TBI-based myeloablative regimens. All transplanted units were 4–6/6 and 3–6/6 HLA-matched to the recipient and to the other co-transplanted unit, respectively. Cyclosporine was used for GVHD prophylaxis with combined use of short-course methotrexate (n=3), methyl-prednisolone (n=4), or mycophenolate mofetil (n=5). Median infused nucleated cell dose was 3.13x107/kg (range, 0.72–6.2x107/kg). All patients engrafted at a median of 23 days (range, 15–34 days). All of the 9 patients who were followed-up longer than 90 days achieved platelet engraftment at a median of 47 days (range, 24–81 days). The initial chimerism analysis performed at 1 mo revealed donor-derived hematopoiesis in all patients including 5 patients with 2 donor mixed chimerism, and one patient showed persistent 2 donor mixed chimerism beyond 1 year following transplant. Grade 2–4 and grade 3–4 acute GVHD were developed in 8 (66.7%) and 2 (16.7%) patients, respectively. Three out of the 9 evaluable patients developed limited chronic GVHD. There were 3 events (2 transplant-related deaths and 1 relapse) and estimated disease-free survival was 70.7% at 18 mo. We conclude that double unit CBT is also safe and feasible in pediatric recipients without a single unit containing adequate cells.