bluuebunny/crossref_metadata_2025 huggingface dataset:
Phase I Study of AVE9633, an AntiCD33-Maytansinoid Immunoconjugate, Administered as an Intravenous Infusion in Patients with Refractory/Relapsed CD33-Positive Acute Myeloid Leukemia (AML).
Authors: Giles Francis , Morariu-Zamfir Rodica , Lambert John , Verstovsek Srdan , Thomas Deborah , Ravandi Farhad , Deangelo Dan
Published: October 2019

AVE9633 is an immunoconjugate created by conjugation of the cytotoxic maytansinoid, DM4, to the monoclonal IgG1 antibody, huMy9-6 (average of 3.5 molecules of DM4 per antibody). The huMy9-6 antibody is a humanized version of a murine monoclonal antibody, My9-6, which is specific for the CD33 antigen expressed on the surface of myeloid cells, including the majority of cases of AML. Because CD33 has little expression outside the hematopoietic system, it represents an attractive target for antibody-based therapy in patients with AML. The humanized antibody, huMy9-6, binds to the CD33 antigen with an apparent KD in the range of 10−10 M. Maytansinoids are anti-mitotics that inhibit tubulin polymerization and microtubule assembly, inhibiting cells during the G2/M phase of the mitotic cycle. In order to link maytansinoids to antibodies via disulfide bonds, a new thiol-containing maytansinoid (DM4) was synthesized. Attachment of potent maytansinoids to an antibody via disulfide bonds provides a satisfactory stability in the bloodstream. After the conjugate is bound at the specific tumor site it is internalized and the cytotoxic agent is released within the target cell. A phase I study of AVE9633 is being conducted in patients with refractory/relapsed CD33+ AML. The study regimen consists of AVE9633 IV infusion on Day 1 of a 3 weeks cycle. To date dose levels of 15 (N=3), 30 (N=5), 50 (N=4), 75 (N=4), 105 (N=2), 200 (N=3) and 260 (N=1) mg/m2 have been investigated. Hypersensitivity reactions during perfusion were noted, requiring prophylaxis with steroids. No other AVE9633- attributable extramedullary Grade 3 AE has been observed to date. Free DM4, measured by LC/MS/MS was detectable from the 75 mg/m2 dose level; its Cmax (at the end of infusion) increased from 10 ng/mL at the 75 mg/m2 dose level to 70 ng/mL at 200 mg/m2. Neither AVE9633-associated myelosuppression nor responses have been noted. Using Flow Cytometry Assay on peripheral blasts and monocytes, total saturation and down regulation of CD33 were observed following administration of doses ≥ 30 mg/m2. AVE9633 exposure (measuring, by ELISA method, all antibodies containing at lease one molecule of DM4) increased proportionally with the administered dose in the dose range 15 to 200 mg/m2. Updated PK results and potential explanations for the lack of efficacy using this treatment schedule will be presented.
The Effect of Rituximab in Combination with MEVP Chemotherapy and rhG-CSF in Old Patients of Refractory and Terminal Non-Hodgkin’s Lymphoma.
Authors: Qian Meihua , Wang Wensong , Lan Jianping , Wang Lianming , Zhan Shumei , Zhao Zhonsen , Xu Wenjian
Published: October 2019

It is very important and difficult to study and research the safety and effective methods of treatment in the Terminal or refractory Non-Hodgkin’s Lymphoma ( NHL)patients, especially old patients. To evaluate the safety and efficacy of Rituximab in Combination with MEVP Chemotherapy and rhG-CSF in old patients of Refractory and Terminal Non-Hodgkin’s Lymphoma, 4 old patients of Refractory and Terminal Non-Hodgkin’s Lymphoma( disease history about 6~8 years),3 females and 1 male, between 66~75 years of age (mean age 70.5 years), 2cases IVB, 1 cases IVA, 1case IIIA., Superficial and deep multiple lymph nodes enlarged obviously in all patients and 3 cases of them showed multiple lymph node enlargement and marked lumps in abdominal cavity, 2 cases of them with hydrothorax and ascites, 2 cases of them with bone marrow infiltration and heart disease, 1 case of them with multiple lumps in liver and another of them with autoimmune hemolytic anemia (AIHA), All patients had been treated with chemotherapy included COP (or COPP), CHOP, BCAOP (or BECAOP), MIV (or ESHAP) etc. and 2 patients of them had been treated by radiotherapy before they were treated by Rituximab Combination with MEVP Chemotherapy and rhG-CSF. In this experiment, they were treated by Rituximab plus MEVP Chemotherapy and rhG-CSF,( Rituximab 375 mg/m2/d1, Mitoxantrone 8~10mg/m2/d3, Wumon 0.1/d3~d5 (or VP16 O.1/d3~d7), Vinorelbinum 20mg/m2/d3, Prednison 60mg/d3~d9), rhG-CSF 100~300/d were injected when the number of WBC was <2.0×109/L after chemotherapy. Each cycle was 28~35days. The result is, the minimum of WBC after chemotherapy were 0.6~1.4×109/L, A patient of NHL with AIHA after a treating cycle achieved complete remission (CR), all of enlarged lymph nodes disappeared and Hemoglobin return to normal. Two of patients obtained part remission (PR), most of enlarged lymph nodes disappeared or reduced obviously and hydrothorax and ascites in patients disappeared. The treatment of one patient was inefficacious.

Our study result indicates: The therapy method of Rituximab in Combination with MEVP Chemotherapy and rhG-CSF for the most of patients of Refractory and Terminal Non-Hodgkin’s Lymphoma is safe and effective, no significant toxicity was found during treatment, this treatment method are suitable for most of patients of refractory and Terminal Non-Hodgkin’s Lymphoma and old patients of refractory and Terminal Non-Hodgkin’s Lymphoma.
Primitive Hematopoietic Cells Resist HIV-1 Infection Via P21Waf1/Cip1/Sdi1.
Authors: Zhang Jie Lin , Crumpacker Clyde S. , Scadden David T.
Published: October 2019

Hematopoietic stem cells are resistant to HIV-1 infection. We have identified a novel mechanism by which the cyclin-dependent kinase inhibitor, p21Waf1/Cip1/Sdi1 (p21), known for its regulation of stem cell pool size (1,2), restricts HIV-1 infection of primitive hematopoietic cells in a non-cell cycle dependent manner. Knocking down p21 by siRNA increased HIV-1 infection and induction of p21 expression by phorbol ester (TPA) blocked HIV-1 replication. P21 did not affect the overall levels of cDNA synthesis, but significantly blocked viral integration and resulted in marked increase in 2-LTR circles, a surrogate marker of abortive integration. Consistent with these observations, p21 coimmunoprecipitated with viral integrase and both were detected in the preintegration complex (PIC). Furthermore, silencing p27Kip1 and p18INK4C, cyclin dependent kinase inhibitors related to p21 that affect cell cycle, revealed no impact on viral DNA integration. A closely related dual-tropic lentivirus with a distinct integrase, SIVmac-251 and the other cell-intrinsic inhibitors of HIV-1, Trim5a, PML, Murr1, and IFN-a were unaffected by p21. These results indicate a new function for p21, participating in prevention of HIV integration into the cellular genome. Therefore p21 is an endogenous cellular component in stem cells that provides a unique molecular barrier to HIV-1 infection and may explain the basis for these cells being an uninfected ‘sanctuary’ in HIV disease.
Prognostic Value of Seric Immunoglobulins in CLL: Analysis of 331 Patients with a Median Follow-Up Time of 7.5 Years.
Authors: Garidi Réda , Guilhaume Marie-Noëlle , Vaida Ioana , Brevet Marie , Mazière Jean-Claude , Desablens Bernard
Published: October 2019

Prognostic value of seric immunoglobulins in CLL has been rarely studied. So we look at all our CLL and we find 331 pts with an initial dosage of IgG, IgA and IgM without M component at diagnosis or during the first year of survey. Ages at diagnosis range from 38 to 93 yrs and median age is 66 yrs. Sex ratio M/F is 1.55 and staging according to Binet’s system notes 264 stages A (188 stages A0), 43 stages B and 24 stages C. IgG levels range from 0.9 to 22.2 g per l with a mean value of 9.38±3.18 g. In logistic regression, IgG level is linked with sex, age, number of lymphoid area according to Binet’s system, blood lymphocytosis and hemoglobin levels used as continuous values. IgA range from 0.1 to 7.8 g (mean value=1.66±1.03 g) and is linked with the same parameters and platelet level too. At least, IgM range from 0 to 3.4 g (mean value=0.78±0.63 g) and is linked with age, number of lymphoid area, blood lymphocytosis and platelet levels. Immunoglobulins levels are also linked with LDH, β2-microglobulin, sCD23 and sCD25 values.

Looking at “corrected” survival after exclusion of not CLL-related deaths including those due to a solid tumor because this complication is independent of initial prognostic factors of CLL, we find that the best cut-off values are 7 g for IgG and 2 g for IgA. For IgG, median survival time is 14.6 yrs for the 268 pts with initial IgG ≥ 7 g versus 6.5 yrs for the 63 other pts (p=0.00004). For IgA, the best cut-off value is 2 g: median survival time is >13.3 yrs for the 100 pts with IgA ≥ 2 g versus 10 yrs for the 231 other pts (p=0.04). These 2 cut-off values are independent and so we can build a prognostic system: 98 pts with IgG ≥ 7 g and IgA ≥ 2 g - 172 pts with IgG < 7 g or IgA < 2 g - 61 pts with low IgG and IgA levels. Median “corrected” survival times of these 3 groups are statistically different: > 13.3, 10.6 and 6.8 yrs (p=0.0009) and when we look at global survival, rates are more different: >13.3, 8.6 and 5.7 yrs (p=0.00009). Same data are noted among stage A patients only and among stage A0 too. So when we opposite this system and with Binet’s and Rai’s classifications, only Rai’s system is ruled out by Cox model looking at “corrected” or global survival. Curiously, this new system has no impact of initiation on treatment among stage A pts while it is linked to evolution to stage C (p=0.05). Finally, we find no correlation with occurrence of an M component or an immunological complication, and also with occurrence of a zoster infection or a Richter syndrome while it is a trend for occurrence of a second neoplasia (p=0.06).
Fludarabine Monophosphate as First Line Therapy for Chronic Lymphocytic Leukemia (CLL) - 11 Years Clinical Experience.
Authors: Novoa J.E. , Rojo A.L. , Beñaran B. , Draper R. , Calvo H. , Iriondo N. , Diaz L. , Rivero M. , Cabrera A. , Pebet M. , Brignoni S. , Luongo A. , De Bellis R.
Published: October 2019

Background: fludarabine (F) has become the standard first line therapy for chronic lymphocytic leukemia (CLL) in younger patients. Treatment of early stage patients with chlorambucil without risk stratification has not been shown to prolong survival. In recent years effective and potentially curative approaches such as nucleosides analogues, stem cell transplantation or monoclonal antibodies have been developed. The attraction of monoclonal antibodies is based on selective targeting of tumor - relevant surface markers and a distinct mechanism of action (antibody-dependent cellular cytotoxicity).

Aims: to assess the efficacy, safety and quality of life of F in previously untreated B-cell CLL patients in a group of medical institutions in Uruguay during 11 years (1995–2006).

Methods: 168 patients between the period 1995 – 2006 were evaluated.120 of them received F intravenous formulation (1995–2006) and 48 the oral one (2002–2006). Age: 48 – 85 years old, media 67 years old. Gender: male 90, female 78. Inclusion criteria for B-cell CLL was Binet stages B, C and A progressive (Ap), 18 to 85 years old, non multiorganic failure, performance status 0 – 2 (WHO), written informed consent. First condition was non previous treatment. Staging: Binet A 12/168, B 116/168 & C 40/168. Treatment: as first line therapy all the patients received (minimum): 6 cycles of i.v. Fludarabine (Fludara®, Schering) 25 mg/m2/daily (5 days) e/30 days or Oral Fludarabine, 40 mg/m2/daily (5 days), 6 cycles.

Results: on this B-cell CLL cohort the overall response rate (ORR) was 78% (CR+PR), 80% of them have immunophenotypic response. Safety: on the 1100 cycles in 168 patients, the toxicity was: 1 AIHA, 2 pancytopenia, 3 plaquetopenia. Grade 3–4 infection rate was 1,3%. No alopecia was observed in any patient. Kaposi sarcoma (0,7%). Mortality rate: 1,7% (3/168 patients). Other adverse factors to overall survival were, age over 65 (p=0,0001) and hepatic impairment (p=0,0001). Toxicity: (WHO>2): granulocytopenia 28%, thrombocytopenia 8%, infection 2%. Although fludarabine-treated patients experienced more significant myelosuppression, no difference in the treatment group was demonstrated. Causes of death: Richter 12%, sepsis 5%, associated disease 34%, second malignancy 17% and others 30%. Comparing oral with intravenous formulation in overall survival the results were: CLL 34% vs 36% (p= NS).

Conclusions: fludarabine monofosfate (Fludara®) looks like an effective and safe treatment for B-cell CLL. The oral and intravenous formulations have a similar response rate in elderly and young patients. The challenge remains to integrate new information to apply novel therapies in a disease-specific and risk-adapted maner. A longer follow up and a larger trial, might be needed to confirm these results.
Metabolism, Disposition and Pharmacokinetics of PR-171, a Novel Inhibitor of the 20S Proteasome.
Authors: Yang Jinfu , Fonseca Fabiana , Ho Mark , Kirk Christopher J. , Sehl Louis S. , Lewis Evan R.
Published: October 2019

Clinical studies using the boronic acid-based proteasome inhibitor bortezomib have validated the proteasome as a therapeutic intervention point for the treatment of multiple myeloma and non-Hodgkin’s Lymphoma. PR-171 is a novel peptide epoxyketone inhibitor of the 20S proteasome that is currently in phase I clinical trials. Herein, the metabolism, disposition and pharmacokinetics of PR-171 are described. Incubation of 3H-PR-171, labeled in the phenylalanine ring, with freshly prepared rat, monkey and human hepatocyte preparations showed similar metabolite profiles with six predominant radiolabeled metabolites observed in all three species; no metabolites unique to human hepatocytes were detected. The diol of PR-171, formed via epoxide ring hydrolysis, was the most abundant observed metabolite in hepatocyte preparations of each of the three species. Single dose IV bolus PK studies using unlabeled PR-171 in Sprague Dawley rats (2 to 9 mg/kg) and cynomolgus monkeys (1 to 4 mg/kg) showed approximately dose linear PR-171 exposure for both species as measured by plasma AUCs. In both species, PR-171 plasma clearance values (554 and 227 mL/min/kg in rat and monkey, respectively) exceeded hepatic blood flow implying that extrahepatic clearance mechanism(s) must be involved. While the in vitro hepatocyte results suggest that hepatic clearance mechanism(s) are also likely to be involved, in vivo hepatic clearance cannot be conclusively inferred from available data. As predicted by the in vitro hepatocyte results, PR-171 diol was detected at abundant levels in plasma from both rat and monkey. Human pharmacokinetics are similar to those of rat and monkey. Mass balance and tissue distribution were also investigated in rats using 3H-PR-171. Radioactivity was widely distributed following a single IV bolus dosing and reached maximum concentrations at times ranging from 0.5 to 24 hours in different tissues. At 168 hours post-dose, more than 44% of the administrated radioactivity remained in tissues. Excretion via urine and feces accounted for 14% and 18%, respectively, of administered radiation. The distribution of radioactivity may not be indicative of the true distribution of PR-171 as much of the radioactivity may reflect metabolic incorporation of the radiolabel and/or 3H exchange with water.
Effectiveness of GVHD and GVL of Donor-Derived NK Cell of MHC Haplotype-Mismatched BMT in Mice.
Authors: Wang Chunyan , Tan Huo , Kunyuan Guo
Published: October 2019

Objective: To study Effectiveness of GVHD and GVL of donor-derived NK cell of MHC haplotype-mismatched BMT in mice.

Methods: CB6F1 H-2b/d mice model of EL9611 (H-2d) erythroleukemia was developed by injection of EL9611 (H-2d) cells in tail vein, CB6F1 H-2b/d mice as the recipient, and C57BL/6H-2b mice as the donor. 70 CB6F1 H-2b/d mice were randomly separated into 7 groups (10 mice per group). Control groups (4 groups): the first group was the one without treatment, the second one was simple-irradiation group, the third one was the Ara-c-treated group which Ara-c were injected into mice at 50mg/(kg·one)×6d followed by infusion of EL9611(H-2d) 5 days, the fourth one was haplotype-mismatched GVHD-control group which bone marrow cells and spleen cells of C57BL/6H-2b mice were injected into the mice after 4 hours irradiation. Experimental groups(2 groups): Irradiated 9Gy, mice were injected C57BL/6H-2b NK cells(1×106) and then BM cells after 4 hours in the first experimental group. After irradiation of 9Gy, mice were injected C57BL/6H-2b NK cells(1×106) and then BM cells and spleen cells after 4 hours secondly. The effect was assessed by blood routine test, survival time, body weight, and histopathology in the recipients.

Results: [circ1]Life span: the survival time was (10.10±0.88) days, (9.80±0.92)days, (22.70±3.23) days and (20.10±1.73) days in the first, second, third and fifth control groups respectively, (30.10±15.95) days in the fourth control group in which the survival time of 2 mice was more than 30 days. The survival time was (39.10±18.11) days and (49.30±17.24) days in the first and second experimental groups respectively. The survival time in 4 mice of the first experimental group was more than 30 days and 7 mice of the second experimental group. The survival time of the first of experimental group was much longer than that in the first, second, third and fifth control groups (P<0.01). The survival time of the second experimental group was much longer than that in other groups (P<0.05). [circ2]Histopathology change: It was observed that the liver and spleen were enlarged and destroyed in the mice died from leukemia by leukemia cells. [circ3]It was noted that chimerism of Y chromosome appeared in mice of experimental groups of long survival time.

Conclusion: The findings provide that donor-derived NK cell has the ability of antileukemia and can reduce GVHD in C57BL/6H-2b→CB6F1 H-2b/d of erythroleukemia mice (EL9611, H-2d).
Allogeneic SCT with Matched Related and Unrelated Donors for Patients (pts) with Refractory Anemia (RA): Improved Survival after Introduction of Reduced Intensity Conditioning (RIC) Regimens.
Authors: de Witte T.M. , Brand Ronald , van Biezen Anja , Mufti Ghulam , Ruutu Tapani , Finke Jurgen , von dem Borne Peter , Vitek Antonin , Delforge Michel , Alessandrino Paolo E. , Harhalakis Nicholas , Russell Nigel H. , Carreras Enric , Verdonck Leo F. , Niederwieser Dietger W.
Published: October 2019

Early alloSCT of pts with MDS before transformation usually results in a 50% EFS, but the nonrelapse mortality (NRM) has precluded a general application of alloSCT for patients with RA. Introduction of RIC has allowed the application of alloSCT to more advanced ages. This study evaluated the impact of recipient age, transplant year, interval between diagnosis and SCT (Dx-SCT), cytogenetic characteristics, T-cell depletion (TCD), type of donor, and the intensity of the conditioning regimen on the outcome of alloSCT. The study population consisted of 374 pts, 244 of whom have received a transplant from an HLA-id. sibling. None of the pts had progressed to advanced stages of MDS. 83 Pts received TCD grafts and 102 pts received SCT after RIC. 58 Pts were <20 yrs, 52 between 20–30, 66 between 30–40, 95 from 40–50 and 103 pts >50 yrs. 86 Pts received SCT before 1996, 125 in 1997–2001, and 163 pts more recently. 184 Pts have been transplanted with a Dx-SCT of <12 months, and 190 >12 months. Cytogenetic data were available for 197 pts, 85 of whom had cytogenetic abnormalities. The overall 5-year survival was 50% (SE: 3%). Relapse occurred in 44% pts resulting in a 5-year relapse risk of 21% (SE: 3%). NRM was the cause of failure in 120 pts resulting in a 5-year NRM of 41% (SE: 3%). The survival after SCT with matched related and unrelated donors was 54% and 48% resp. Young age was associated with better survival in the univariate model (p=0.04). An association exists between various factors such as between age and regimen type, transplantation year, or donor type. Therefore the outcome of the multivariable analyses was based on Cox models with age, transplant year, type of donor, Dx-SCT, stem cell source, and regimen type as main factors. Variables with a significant impact on survival were transplant year with an improved survival in more recently performed SCT (HR=0.95/year; p=0.05), TCD (HR=1.5; p=0.03) and Dx-SCT >12 months (HR=1.4; p=0.05) while higher age (HR=1.1/10 years; p=0.08) and RIC (HR=1.0; p=1.0) did not influence outcome significantly. The HR of RIC for relapse-free survival was 1.2; p=0.5. The fully adjusted estimate for relapse after RIC was HR=4.1 (p=0.002). The NRM was lower after RIC: HR 0.7 (P=0.22), but this difference was not significant. The donor type and the source of stem cells did not influence significantly any of the outcome variables. This data show that alloSCT results in excellent outcome nowadays, even at older age and using matched unrelated donors. The relatively high relapse rate observed after RIC warrants prospective studies.
Marrow Stromal Cell (MSC) Growth from Long Term Cryopreserved Bone Marrow.
Authors: Lane Thomas A. , Garls Davina , Mackintosh Ellen , Cramer Steven C.
Published: October 2019

Autologous MSC are under active investigation for a variety of potential clinical indications in regenerative medicine and gene therapy. The potential to derive autologous MSC from long-term cryopreserved bone marrow has not been fully elucidated. We report here the growth characteristics of MSC from bone marrow that was cryopreserved for more than 10 years.

Methods: We identified 3 deceased patients whose cryopreserved autologous bone marrow that was originally intended for autologous transplant, was to be discarded after fulfilling all accepted criteria for discard. The pts were age 46, 47, and 60 at the time of harvest, one had high risk and 2 had metastatic breast cancer (both to bone) and all had had multiple chemotherapy regimens prior to harvest. The marrow was cryopreserved between 1994–1995 in 10% DMSO using a controlled-rate freezer and stored in the vapor phase of liquid nitrogen until June, 2006 (11–12 years). The marrow was thawed in a 37 C waterbath, diluted with an equal volume of PBS, washed × 3 in PBS, resuspended in culture medium with 20% fetal bovine serum and plated at 5 × 10E6 mononuclear cells (mnc)/cm2 in 2 – 4 T175 flasks. Culture medium was changed in 48 hours and the plates were observed during subsequent culture medium changes every 3–4 days.

Results: At 2 – 4 days only scattered round cells were observed, but all samples grew scattered spindle-shaped cells were between days 4 – 12. Six of 8 primary cultures became > 70% confluent between days 11 and 21; 2 remained at < 40% confluence until replated (then achieving > 70% confluence at day 31). First passage cultures (n=12) became > 70% confluent in 4 – 10 days (median = 7) and 2nd passage cultures were > 70% confluent in 7 – 10 days (median = 9). Second or 3rd passage cultures yielded 2.1 – 4.2 × 10E6 MSC/T175 (n= 14; median 3.15) and were 89 – 96 % viable. Flow cytometry (n = 3) confirmed that 3rd passage cells stained positive for CD105 and CD73 and negative for CD14 and CD45. Additional characterization of the differentiation potential of these presumptive MSC are in progress.

Conclusion: These studies indicate that cells typical for MSC can be readily grown from marrow that has been cryopreserved for > 10 years and suggest that even long-term stored bone marrow may serve as a source of cells for regenerative medicine.
Hematopoietic Stem Cell Transplantation (HSCT) Using Plasma Depleted Umbilical Cord Blood (UCB) That Were Not Red Cell Depleted.
Authors: Chow Robert , Tan Patrick , Jaing Tang-Her , Rosenthal Joseph , Nademanee Auayporn , Karanes Chatchada , Graham Michael , Eames Gretchen , Chan Lee Lee , Wang Brian , Chow Tiffany , Wu Tina , Tan Ah Moy , Tan Poh-Lin , Lin Hai-Peng , Gjertson David , Petz Lawrence
Published: October 2019

UCB is an attractive source for HSCT; however, limiting cell dose has hampered its widespread use. The red cell depletion (RCD) techniques that are widely used incur significant nucleated cell loss after processing; therefore, to minimize cell loss and still reduce volume during processing, a technique was devised to deplete plasma (PD) but not red blood cells. A large racially diverse inventory of over 25,000 PD UCB is available on stem cell registries. Of the 265 PD UCB transplanted into 240 patients up to May 2006, an updated retrospective audited analysis of all 205 patients with engraftment and/or survival data was performed. The characteristics for the patients were: median age 8.8 yo, range 0.3–59, 76 ≥16 yo (32%); median weight 29 kg, range 5–137, 84 ≥50kg (35%); male 59%; median # HLA ABDR matches 4.0; median pre-freeze TNC dose 5.31 × 107/kg (n=229); transplant center reported median post-thaw TNC dose 4.92 × 107/kg (n=112); median pre-freeze CD34 dose 1.7 × 105/kg; malignant indications 71%; transplants outside U.S. 39%; double transplant 23%; and non-myeloablative 15%. For all engrafted patients, the median time to engraftment for ANC 500 (n=192), platelet 20K (n=174) and 50K (n=169) were 22.0 days (range 7–64), 49.5 days (range 12–181), and 63.5 days (range 21–374) respectively. Kaplan-Meier estimates of engraftment of ANC500, platelet 20K and 50K engraftment were 88±3%, 82±4% and 76±4% respectively. The incidence of grade III–IV acute GVHD and extensive chronic GVHD were 13% and 14% respectively. One-year relapse rate was 23±4% for all patients (n=190), and TRM was 29±3% (n=203) at 1 year. With a median follow-up of 178 days (range 4–1402 days) for the surviving patients, the Kaplan-Meier estimates of 1-year OS (n=203) and DFS (n=190) are 59±4% and 54±4% respectively for all patients. These results demonstrate that HSCT using PD UCB can be performed safely and effectively.
Quantitative Monitoring of Lineage-Specific Chimerism after Dual Hematopoietic Stem Cell Transplantation (Umbilical Cord Blood with Co-Infusion of Third Party Donor CD34+ Cells).
Authors: Sanchez-Hernandez Noemi , Manzano Cintia , Barroso Diana , Iglesias Gustavo , Balsalobre Pascual , Serrano David , Carrion Rafael , Gomez-Pineda Alfonso , Diez-Martin Jose L. , Buño Ismael
Published: October 2019

Background: Allogeneic umbilical cord blood (UCB) stem cell transplantation (SCT) shows some advantages (HLA matching requirements or availability) respect to SCT using other sources of matched unrelated donor (MUD) stem cells. However, it is correlated with slower engraftment, increasing risk of infections and early mortality. It has been recently shown that co-infusion of third party donor (TPD) CD34+ cells (dual SCT) is useful to speed up engraftment.

Objective: To evaluate the usefulness of lineage-specific chimerism quantification in the management of this transplant setting.

Patients and methods: 8 dual SCT (Tables 1, 2) in 7 patients (1 CML-BC, 2 AML-M2, 1 AML-M4, 1 ALL-Ph+, 1 biphen. ALL, 1 NHL). Chimerism was analyzed by STR-PCR (AmpFlSTR SGM Plus, Applied Biosystems; sensitivity 1%) and quantitative real-time PCR (qrt-PCR) of null alleles and insertion/deletion polymorphisms (Light Cycler, Roche; sensitivity 0,01%). Peripheral blood (PB) and leukocyte lineages (T cells, CD3+, and myeloid cells, CD15+), isolated by positive selection using automated immunomagnetic technology (AutoMACS, Miltenyi Biotec), were analyzed weekly. Bone marrow (BM) was analyzed at days +30, +100, +180 and +365).

Results: 7/8 cases showed initially a high proportion of TPD cells in PB which were progressively replaced by UCB cells. UCB complete chimerism (UCB-CC, absence of recipient or TPD cells even in qrt-PCR assays) was acquired in a median of 22.5 days (range 18–39). In one patient, fully HLA-mismatched with the TPD, no TPD cells were observed after dual SCT. 4/8 cases showed recipient cells in PB after dual SCT during a median period of 12 days (range 4–18 days). In 3/8 cases, recipient cells were found after CC had been acquired, which allowed early diagnosis of 1 graft rejection and 2 relapses. T cells (CD3+) are mainly of UCB origin early after dual SCT and reach UCB-CC a median of 7 days (range 0–21) before PB. However, myeloid cells (CD15+) derive primarily from the TPD and reach CC together with PB. TPD cellularity favoured early engraftment (before UCB-CC took place) in 4 cases. In this context, only one important infectious complication (hepatosplenic tuberculosis) was observed, which resolved with the appropriate treatment.

Conclusions: Lineage-specific chimerism quantification allowed a close monitoring of the dynamics of engraftment of cells of both donors which is of key importance in this SCT setting. Moreover, lineage-specific chimerism analysis was useful to diagnose one graft rejection and two relapses (the patient with NHL showed a ganglionar relapse in CC).

Table 1. Transplantation characteristics. Table 2. Transplantation results. Median (range).
Allogeneic Stem Cell Transplantation for Adult Acute Lymphoblastic Leukaemia: Local Results of Modern Transplant Practice.
Authors: Butler Jason P. , Kennedy Glen A. , Hill Geoff R. , Morton James , Western Robyn , Hutchins Cheryl , Durrant Simon
Published: October 2019

Aims: To report the single institution outcomes for recent allogeneic transplantation for adult acute lymphoblastic leukaemia (ALL), and explore variables predictive of successful outcome.

Methods: All patients with ALL transplanted at the Royal Brisbane Hospital between 1st January, 2000 and 31st December, 2005 were included in the study sample. Survival analysis was performed using the Kaplan-Meier product-limit method. Univariate and multivariate analyses were performed using Cox proportional hazards modelling. Follow-up was censored at 30th June, 2006.

Results: A total of 41 patients were included in the study cohort, including 28 males and 13 females. Median age at transplantation was 27.6 years (range 15.2 to 64.5). 34 patients had precursor-B phenotypes; the remaining 7 had T-lineage ALL. Based on cytogenetics, patients were high-risk (n=19), intermediate risk (n=12), good risk (n=2), or indeterminate due to inadequate diagnostic sampling (n=8). Patients were transplanted in CR1 (n=23, 2 patients demonstrating delayed CR), 1st relapse (n=5, 2 with early relapse, 1 with untested frank relapse, and 2 with refractory disease), CR2 (n=9, 2 with treated isolated CNS relapse), CR3 (n=1) and primary refractory disease (n=3). Conditioning regimen was CyTBI, with the exception of 1 patient who received fludarabine (125mg/m2) and cyclophosphamide (120mg/kg). All patients received cyclosporine and full-course methotrexate for GVHD prophylaxis. Donor source was matched sibling (n=17) and matched unrelated donor (n=24). 5 patients received unprimed bone marrow grafts; the remaining patients received peripheral blood stem cells. At a median follow-up for survivors of 1.8 years (range 0.5 to 6.4), overall and progression-free survival at 2 years is 61% (95% CI 43 to 75%) and 74% (95% CI 55 to 85%) respectively. 32 patients developed acute GVHD, of whom 27 (66%) developed Grades II–IV. 21/40 evaluable patients developed chronic GVHD; all were extensive stage. 9 patients relapsed during follow-up at a median of 0.47 years post transplant (range 0.02 to 1.46); 1 remains alive in remission following reinduction therapy. 6 patients died from therapy-related causes - 3 from pulmonary toxicity, and 1 each from fungal sepsis, leukoencephalopathy, and hepatic GVHD. On both univariate and multivariate analyses of overall survival, younger age, transplantation in CR1, and development of chronic GVHD were predictors for an improved outcome.

Conclusions: Allogeneic stem cell transplantation for ALL is associated with excellent long-term outcome with respect to both disease control and treatment related toxicity. From these data, transplantation in CR1, younger age, and the development of chronic GVHD are covariates associated with prolongation of overall survival.
Febrile Neutropenia in Lymphoma Patients Is Associated with Substantial Resource Use and Healthcare Costs in Clinical Practice in Spain.
Authors: López A. , Alonso J.D. , Gómez-Codina J. , Novo A. , Herrera C. , Lara N. , Gasquet J.A. , Sánchez J. ,   on Behalf of the ENIA Study Group
Published: October 2019

Background

Despite the significant impact of chemotherapy-induced febrile neutropenia (FN) on patients (pts) with cancer and its consequences for health care costs, there have been no studies in common clinical practice in Spain assessing the burden and economic impact of this complication.

Methods

This is a sub-analysis of lymphoma pts included in a multicentre, retrospective, chart review of adult pts from 16 Spanish hospitals who suffered from at least one FN episode related to cytotoxic chemotherapy (CT). Resource use and subsequent costs including days of hospitalization, number of transfusions, number and type of complementary tests, use of colony-stimulating factors (CSFs), and use of antibiotics and other drugs to manage FN were assessed for each episode. The impact of FN on planned CT was also analysed in terms of dose delays (DD) and/or reductions (DR).

Results

Medical charts from 194 pts were reviewed, 67 (34.5%) of whom had lymphoma, which accounted for 87 documented FN episodes included in this analysis. The median (range) age of patients was 62 (19–85) years, 31.7% had aggressive NHL, and 58.2% were treated with CHOP-like CT. FN appeared during first CT cycle in 61.2% of the pts. Hospitalization was required in 100% of the pts and the median length of hospital stay due to FN was 8 days (p25:6–p75:11). During an FN episode, 42% of pts required ≥1 transfusion, 100% needed a blood test and 98.9% a blood culture. Microbiologically documented infection appeared in 33% of FN episodes. All pts were treated with antibiotics (69.3% with cephalosporins) and CSFs were used in 64.8% of pts. In 40.9% of episodes, FN impacted on planned CT dose and/or schedule: DR was observed in 16.7% of pts, DD in 24.0% and CT withdrawal in 15.2%.

Conclusions

FN has a substantial impact on resource use and associated costs in pts with lymphoma. Hospitalization and antibiotic treatment were the main drivers of the cost associated with the management of FN in current clinical practice. Furthermore, FN has a meaningful effect on planned CT dose and/or schedule, with potential consequences for treatment outcome.

Mean (SD) healthcare costs per FN episode (All cost data expressed as €) Hospitalization Transfusions Complementary Tests CSFs Antibiotics and Other Drugs Total 3,557.17 (3,050.44) 43.24 (58.59) 162.77 (135.36) 223.39(231.40) 527.67 (448.56) 4,514.24 (3,392.20)
In Vivo Studies of Autologous Platelets Stored at Room Temperature (22°C), 4°C, and 4°C with Galactosylation.
Authors: Slichter Sherrill J. , Wandall Hans H. , Hoffmeister Karin M.
Published: October 2019

Background: Previous studies in a mouse model with “chilled” mouse platelets stored at 4°C for two hours with and without galactosylation showed protection against the adverse effects of chilling with galactosylation (Science2003;301:1531). However, prior studies with human platelets demonstrated that the adverse effects of 4°C storage became irreversible only after 18 to 24 hours of 4°C storage (J Lab Med1978;6:971). Therefore, we performed viability studies using human platelets stored for two days to exceed the period of potential reversibility of any cold-induced platelet injury.

Methods: Apheresis platelets were collected using Haemonetics MCS+ equipment from four normal volunteers. After collection, the platelets were pooled into one bag, and then the platelets were divided equally between two storage bags. Half of the platelets from each of the four collections were stored at 4°C with galactosylation, and the other half of the platelets were stored at 4°C without galactosylation (two collections) or at 22°C using standard storage conditions (two collections). After storage, samples from each of the two types of platelets stored from a volunteer were radiolabeled with either 51Cr or 111In, re-injected, and serial blood samples were drawn to determine post-transfusion recoveries and survivals of the radiolabeled autologous platelets.

Results: Results of the radiolabeled platelet studies are shown in the table.

In Vivo Radiolabeled Autologous Platelet Recoveries And Survivals 4°C GALACTOSYLATED PLATELETS 4°C STORED PLATELETS 22°C STORED PLATELETS Normal Volunteer Recovery (%) Survival (Days) Recovery (%) Survival (Days) Recovery (%) Survival (Days) 1 46 2.8 48 2.9 2 64 2.8 58 2.8 3 46 2.2 59 6.8 4 26 1.0 34 6.9 Ave. ± 1 S.D. 46 ± 16 2.2 ± 0.9 53 ± 5 2.9 ± 0.1 47 ± 13 6.9 ± 0.1

No adverse events were reported with the transfusion of the galactosylated platelets, there was no evidence of the development of anti-platelet antibodies against galactosylated or non-galactosylated platelets, and samples of the galactosylated platelets obtained after storage verified galactosylation of the platelets.

Conclusions: The data show that, following two days of 4°C storage, the recoveries and survivals of the galactosylated platelets are no different than the non-galactosylated 4°C stored platelets from the same volunteer. Although the recoveries of the 4°C stored platelets with and without galactosylation are well-maintained compared to the 22°C stored platelets, the survivals are markedly reduced as has been previously shown for 4°C stored platelets (Br J Haematol1976;34:403).
A Randomized, Double-Blind, Placebo-Controlled Trial of Thalidomide Plus Dexamethasone Versus Dexamethasone Alone as Primary Therapy for Newly Diagnosed Multiple Myeloma.
Authors: Rajkumar S. Vincent , Hussein Mohamad , Catalano John , Jedrzejcak Wieslaw , Sirkovich Svetlana , Olesnyckyj Marta , Yu Zhinuan , Knight Robert , Zeldis Jerry , Blade Joan
Published: October 2019

Purpose: Updated results, including response rate data, from a randomized, double-blind, placebo-controlled study comparing thalidomide plus dexamethasone versus dexamethasone alone as primary therapy for newly diagnosed multiple myeloma (MM).

Methods: Patients (pts) with previously untreated, symptomatic MM were eligible. Pts were randomized to Thal/Dex (Arm A) or placebo plus Dex (Arm B). Pts in Arm A received Thal 50 mg PO daily, escalated to 100 mg on day 15, and to 200 mg from day 1 of cycle 2; Dex 40 mg PO was given on days 1–4, 9–12, and 17–20. Pts in Arm B received placebo instead of Thal, and Dex as in Arm A. Each cycles was 28 days long. Treatment was continued until progression or undue toxicity. The primary endpoint was time to progression (TTP) defined using EBMT criteria. All analyses were conducted on an intent to treat basis. An independent data monitoring committee recommended release of results.

Results: 470 pts were enrolled: 235 were randomized to Thal/Dex and 235 to placebo/Dex. Median follow up is 18 months. Median age was 65 yrs. The response rate (CR+PR) was significantly higher with Thal/Dex compared to placebo/Dex, 59% vs 42%, respectively, P<0.001. Eighty-two percent of patients have gone off-study. TTP was significantly superior with Thal/Dex vs placebo/Dex, median TTP not reached with Thal/Dex versus 8.1 months (95% CI: 6.5–12.8 months) with placebo/Dex, P<0.001. Major grade 3–4 events are summarized in the Table below. Deep vein thrombosis (DVT) was higher with Thal/Dex vs placebo/Dex, 15.4% vs 4.3%, respectively.

Conclusions: Thal/Dex yields superior response rates and TTP compared to Dex alone when used as first-line therapy for multiple myeloma.

Major Grade 3–4 adverse events Adverse Event Thal/Dex (n=234) Placebo/Dex (n=232) Venous thromboembolism (Grade >=3) 17.9% 4.3% Cerebral ischemia (Grade >=3) 3.0% 1.3% Myocardial ischemia (Grade >=3) 3.8% 2.2% Neuropathy (Grade >=3) 3.0% 0% Any toxicity (Grade >=4) 30.3% 22.8%
Tendency towards Minor Bleeding in Women Is Associated with a Reduced Risk for Major Bleeding, without Apparent Differences in Level of Anticoagulation during Treatment with Vitamin K Antagonist.
Authors: Veeger Nic J.G.M. , Piersma-Wichers Margriet , van der Meer Jan
Published: October 2019

Easy bruising is frequently found in healthy women. This mild bleeding tendency may be enhanced by treatment with vitamin K antagonists. This may result in more intense monitoring of the level of anticoagulation (INR) to improve the individual time within target range (ITTR) and consequently to reduce the risk of major bleeding. We performed a retrospective study in 6758 consecutive patients receiving vitamin K antagonists for primary or secondary prophylaxis of venous or arterial thrombosis (mean age 67, 51% male and 189762 INRs in 6681 person-years) to evaluate differences in gender on the occurrence of minor and major bleeding, as well as the ITTR.

The incidence rates (IR, per 100 person-years) of minor and major bleeding were assessed and survival analysis performed to obtain adjusted hazard ratios (HR, 95%CI) for major bleeding in men versus women with or without prior minor bleeding, controlling for significant patient characteristics. Minor bleeding occurred more frequently in women than men (IR=33.2 versus 21.1, respectively; HR=1.5, 95%CI 1.4–1.7, p<0.001), whereas major bleeding showed a tendency towards a higher incidence in men (IR=1.6 versus 2.0; HR=1.3, 95%CI 0.9–1.8, p=0.20).

Figure Figure

However, in women with prior minor bleeding, the incidence of major bleeding was low (IR=1.3, 95% CI 0.7–2.1), when compared to men with minor bleeding (3.3, 95%C 2.2–4.7) (HR =2.7, 95% CI 1.4–5.0, p=0.002). Men and women without prior minor bleeding had comparable incidences of major bleeding (IR 1.8, 95% CI 1.3–2.5, and 1.5, 95% CI 1.1–2.1, respectively), but both were higher than in women with prior minor bleeding (HR 1.4 (0.8–2.5, p=0.30 and 1.2 (0.7–2.2, p=0.55)). These differences were even more pronounced in patients at high risk of major bleeding due to a strongly reduced ITTR (ITTR below 30%, lowest quartile).

Considering the actually achieved level of anticoagulation, there were no differences between men and women with or without prior minor bleeding that could explain the reduced risk of major bleeding in women with prior minor bleeding. With 42% to 43%, the ITTR was comparable, as was the time spend above INR 5.0 (6% to 7%).

In conclusion, more frequent minor bleeding in women was associated with a reduced incidence of major bleeding, whereas in men, such a reversed association was not present. Differences in intensity of INR monitoring in patients with or without minor bleeding and as a result in actual level of anticoagulation, could not be demonstrated.
Liquid Heparin Assay: A Straightforward Approach for Monitoring the Clinical Use of Heparins.
Authors: Kung Chun , Gabbeta Jagadeesh , Krougliak Valeri , Quiazon Emmanuel , Triscott Mark
Published: October 2019

There are number of commercial heparins available in the market. The choice of heparin may depend on the clinical condition of the patient. For example, patients with deep-vein thrombosis (DVT), low molecular weight heparins (LMWH) (such as enoxaparin) may be used; whereas for pulmonary embolism (PE) cases, unfractionated heparins (UFH) may be the heparin of choice. In clinical studies, the synthetic heparin pentasaccharide, Fondaparinux (commercially known as Arixtra) has been shown to be efficacious in the treatment of both types of patients. Monitoring heparin activity in patients under treatment is critical in reducing the patient risk of bleeding or thromboembolism. The majority of current available tests allow measurement of heparins in the range of 0.0 – 1.0 U/mL. In many clinical laboratories, the analysis of each type of heparin requires a separate calibration curve, which makes the assay more complex and time consuming. In most cases these calibrators are “home made” and are not related to a standard, which may contribute to variability in testing. The reagents are usually provided in lyophilized form and require prior reconstitution which can introduce additional variation and increase handling time. An assay developed here, based on ready to use liquid reagents and designed for use on ACL TOP™, ACL Futura®/, ACL Advance™ ACL Elite™/ACL Elite Pro™ and ACL™ 9000/10000, addresses these issues and minimizes testing time. The Liquid Heparin Assay is built on the one stage principle of heparin detection, utilizing endogenous antithrombin III. The assay kit consists of two reagents, a liquid Factor Xa and a chromogenic substrate for Factor Xa, S2732. The composition of the kit and assay parameters are optimized to achieve overlay of the calibration curves for UFH and LMWH in the range from 0.0 to 2.0 U/mL of heparin activity. Furthermore, the assay is evaluated with respect to the various commercial heparins including Enoxaparin, Fragmin and Arixtra. The calibration curves for WHO UFH standard, WHO LMWH standard, Enoxaparin, and Fragmin are observed to overlap, with a maximum difference of 10% at each calibration point. The recoveries of heparin activities in samples spiked with various heparins are also within 10% of the expected values. The weight concentration (μg/mL) of Arixtra in plasma samples and its corresponding heparin activity (U/mL) can also be measured using this assay. The potential utility of the Universal Calibrator, composed of the mixture of UFH and LMWH is also evaluated and shown to be promising. Thus, the Liquid Heparin Assay permits simultaneous determination of various commonly used heparins in patient samples using a single calibration curve, and ready to use reagents. This can simplify analysis, reduce time and improve assay reliability. Since one assay is sufficient to measure different heparins, this assay could also reduce the overall costs of heparin analysis in the clinical laboratory.
Successful Overcoming of Imatinib Resistance by Synergistic Effect of Imatinib and Simvastatin in Imatinib Resistant Chronic Myelogenous Leukemia Cell Line.
Authors: Oh Bora , Kim Tae Young , Min Hyun Jung , Yoon Sung-Soo , Lee Dong Soon
Published: October 2019

Statins (HMG-CoA reductase inhibitors) are known to show antiproliferative effects and are anticipated as a potential drug in the treatment of malignancies including acute myelogenous leukemia (AML) and multiple myeloma (MM). Using cell proliferation assays, we determined the effects of simvastatin in CML cells. In the current study we show that simvastatin inhibits the proliferation of CML cell lines (The IC50 to simvastatin of K562, Kcl22, and LAMA84 were 13 uM, 19 uM, and 31 uM, respectively). Moreover, synergistic interactions between simvastatin and imatinib were observed in imatinib-resistant CML cell lines (K562-r, KCL22-r, and LAMA84-r). The cytotoxic effect of simvastatin occurred via the induction of apoptosis and inhibited Bcr-Abl tyrosine kinase (TK) activity, as evidenced by annexin V assay and Western blot analyses. Co-treatment with imatinib and simvastatin decreased the amount of Bcr-Abl protein and stimulated the import of Abl protein in the nuclei in K562 cells. Simvastatin may be a potential candidate for the treatment of imatinib-resistant CML patients and the effective dose of imatinib could be reduced in a combined treatment with simvastatin. Mean combination index (CI) values at 50%, 75%, and 90% growth inhibition (IC50, IC80) derived from combined treatment experiments

Cell line Drug combination Drug ratio CI: IC50 CI: IC75 CI: IC90 Gradual levels of synergy are marked as recommended by the manufacture of the Calcusyn software: +++, strong synergism; ++, moderate synergism; + slight synergism; ±, nearly additive; −, slight antagonism: and − −, moderate antagonism K562-s Imatinib + Simvastatin 1:40 0.61+++ 0.49+++ 0.40+++ K562-r Imatinib + Simvastatin 1:10 0.60+++ 0.46+++ 0.41+++ Kcl22-s Imatinib + Simvastatin 1:100 0.96± 1.32− 1.83− − Kcl22-r Imatinib + Simvastatin 1:1 0.42+++ 0.35+++ 0.30+++ LAMA84-s Imatinib + Simvastatin 1:200 1.20− 1.0± 1.0± LAMA84-r Imatinib + Simvastatin 1:20 0.99± 0.8+ 0.84+ Figure Figure