bluuebunny/crossref_metadata_2025 huggingface dataset:
The Renin-Angiotensin Axis Regulates the Development of a Yolk Sac-Like Hemangioblastic Progenitor of Primitive and Definitive Hematopoiesis from Human Pluripotent Stem Cells.
Authors: Zambidis Elias T. , Park Tea Soon , Tam Ada , Levine Michal , Peault Bruno
Published: October 2019

We have defined serum-free (SF) culture conditions for efficiently generating clonogenic hemangioblasts from pluripotent human embryonic stem cells (hESC). We also report that angiotensin-converting enzyme (ACE), a critical physiologic regulator of blood pressure, angiogenesis, and inflammation, is a novel marker for identifying and purifying hemangioblastic stem-progenitors from differentiating hESC. Using in vitro clonal assays of primitive and definitive hematopoietic potential, we demonstrated that ACE+ human embryoid body (hEB) cell populations contain a common yolk sac (YS)-like progenitor for not only endothelium, but also both primitive and definitive hematopoieses. Thrombopoietin (TPO) and basic fibroblast growth factor (FGF2) were identified as critical growth factors for expanding ACE+ hemangioblasts. ACE+CD34-CD45- human embryoid body (hEB) cells contained the highest number of these progenitors. Further maturation of ACE+ hemangioblasts in a stromal environment produced definitive-type lympho-hematopoietic progenitors, albeit with a late YS, and not AGM phenotype. The developmental kinetics of YS-like hematopoiesis generated from hEB cells was remarkably in parallel to the actual timeline of human YS development, which occurs during weeks 2–6 of human gestation. We also showed that ACE and the renin-angiotensin system (RAS) axis plays a direct role in hESC-derived hemangioblastic differentiation, and is directly regulated via signaling through the two major angiotensin peptide receptors (AGTR1 and AGTR2). Moreover, directed differentiation of hemangioblasts toward either endothelium, or multipotent primitive hematopoietic progenitors was efficiently enhanced via modulation of either AGTR1 or AGTR2 signaling. RAS axis manipulation may therefore be exploited for directing the differentiation of transplantable hESC-derived hemangioblastic stem-progenitors, thus providing novel opportunities for human tissue engineering. The key molecular and cellular events of human hemato-endothelial genesis can now be delineated in a manner that was previously impossible due to inaccessibility of human embryonic tissue.

Figure Figure
Biologic Features and Treatment Outcome of Secondary Acute Lymphoblastic Leukemia - Review of 101 Cases.
Authors: Shivakumar Ranga , Tan Wei , Wilding Gregory , Wang Eunice S. , Wetzler Meir
Published: October 2019

Secondary acute lymphoblastic leukemia (sALL) is a rare disease and its biologic features are not well described. Data suggested that sALL occurs more frequently at older age. Since leukemia at older age in general is associated with worse outcome, we wanted to assess the biology and outcome of patients with sALL by age at time of primary diagnosis. We describe a cohort of 7 patients and found additional 94 cases in the literature on whom biological parameters were described. Patients were stratified (at least 5 patients per strata) according to their age at initial diagnosis (<18, 18–59, and ≥60 years of age), initial diagnosis [acute myeloid leukemia (AML), Hodgkin’s disease (HD), neuroblastoma, breast and prostate cancers], cytogenetic groups [diploid, t(9;22), 11q23 aberrations, complex karyotype], immunophenotypes (B vs. T), and Burkitt’s defined by either morphology and/or cytogenetic analysis demonstrating c-myc rearrangement. A total of 101 patients were evaluated; 29 were <18, 54 were 18–59 and 18 were ≥60 years old. The distribution of primary diagnoses was as expected: neuroblastoma was seen only in the <18 age group (P=0.003), HD was more common in the 18–59 age group (75% of all HD cases; P=0.084) while breast (P=0.003) and prostate (P=0.005) cancers were prevalent only in the >18 year old patients. The time interval to develop sALL was similar among the three age groups (3, 2.2 and 1.8 years, P=0.561). However, the time interval to develop sALL was longer for HD (5.5 years) and neuroblastoma (3.7 years) as compared to AML (1 year), breast (1.6 years) and prostate (1.98 years) cancers (overall P=0.0003). Further, the time interval to develop sALL was significantly longer for patients with complex karyotype (5.3 years) as compared to all other aberrations [11q23 − 1.78; t(9;22) − 1.9; diploid − 1.98 years; overall P=0.0497]. Disease characteristics at diagnosis were as follows: T cell immunophenotype was more common in the <18 age group (P=0.016) and the presence of 11q23, t(9;22), complex and normal karyotypes was equally distributed among the three age groups (P=0.2, 0.073, 0.635 and 0.271 individually). Complete remission was infrequent in the ≥60 age group (22.22%) compared to the other groups (73.9% for <18 and 67.7% for 18–59; P=0.025). Even though only patients <60 years old were transplanted (33.3% for <18 and 19.4% for 18–59; P=0.102), the overall survival was poor in all age groups [probability of survival at 1 year for <18=0.222, 18–59=0.226 and ≥60=0.3 (P=0.7941)]. Primary diagnoses, cytogenetic subgroups and immunophenotype did not affect outcome. In summary, the time interval to develop sALL is significantly longer for HD, neuroblastoma and complex karyotype. However, sALL is associated with very poor outcome regardless of age and any of the biologic features. Therefore, identification of prognostic factors to prevent the occurrence of sALL is needed.
Longitudinal Assessment of Nutritional Status in Adults with Acute Leukemia.
Authors: Serralde Aurora , Crespo-Solis Erick , Lopez-Karpovitch Xavier , Damasco Ericka , Rosas-Lopez Adriana , Aguayo Alvaro
Published: October 2019

RATIONALE: The nutritional status of patients with acute leukemia (AL) is frequently impaired during intensive induction chemotherapy. Previous studies have suggested that malnutrition may have an adverse effect on prognosis. The aim of the study was to determine the prevalence of malnutrition during induction chemotherapy treatment and its impact on the outcome.

METHODS: The study was performed over a period of 18 consecutive months. Acute leukemia patients were prospectively studied during first remission induction chemotherapy. The Subjective Global Assessment (SGA), anthropometric and laboratory data were determined at diagnosis and on day 30 of the treatment in order to evaluate nutritional status. Patients were prospectively followed until 60 days after diagnosis. An analysis was made including demographic data, nutritional assessment, albumin, complications, rate of complete remission (CR), rate of early mortality (EM) and length of hospital stay (LOS).

RESULTS: 32 patients (18 men, 14 women) were included; median age 31 years (range 15–62); 16 with myeloid leukemia, 13 with lymphocytic leukemia and 3 with hybrid leukemia. The median of body mass index (BMI) was 25 kg/m2 (18.3–41.7); triceps skinfold (TSF) 16.5 mm (7–42); albumin 3.24 g/dL (1.98–4.18). At diagnosis, the prevalence of malnutrition was 59% according to the SGA (moderately and severely malnourished) and increased to 90% on day 30, (p<0.05). We obtained initial CR in 22 patients (69%), the mortality rate due to post-chemotherapy aplasia (EM) in the prospective cohort was 9%. Patients with deterioration of the nutritional status had longer LOS in comparison with those patients well nourished (15.61 vs. 9.68 p<0.05). They also needed antibiotics for a longer time (16 vs. 10 days, p< 0.05). Nutritional status was not associated with CR rate or EM rate.

CONCLUSIONS: The high incidence of malnutrition at the time of diagnosis and during chemotherapy indicate the importance of nutritional status evaluation. Malnutrition in AL is caused by chemotherapy side effects and/or the disease itself, during this period intensive supportive care, including nutritional therapy is required.
The Clinical Features and the Efficient Therapy of 66 Acute Promyelocytic Leukemia Patients with High Initial White Blood Cell Count.
Authors: Qiu Hui-ying , Xue Meng-xing , Sun Ai-ning , Liu Yue-jun , Wu De Pei
Published: October 2019

Objective: To investigate the clinical features and the efficient therapy of acute promyelocytic leukemia(APL) with high initial white blood cell(WBC) count.

Methods: The clinical features of 66 newly diagnosed APL patients with high initial WBC and 152 patients without high initial WBC were retrospectively analyzed. Additionally, 66 patients with high WBC were divided into different groups and the therapeutic effects were compared according to the different induction therapy.

Results: The early death rate and the incidence of disseminated intravascular coagulation(DIC) and retinoic acid syndrome(RAS) were 30.3%,57.6% and 31.8% respectively for the group with initial high WBC, those were higher than the indexes of 7.2%,38.1% and 21.05% respectively for the group without initial high WBC (p <0.05),Whereas CR rate (63.6%) for the group with initial high WBC were lower than that (88.2%) for the group without initial high WBC (p <0.05). 61 of 66 patients with high WBC accepted induction therapy: 31 patients treated with all-trans retinoic acid(ATRA) alone ,21patients treated with the combination of ATRA and arsenic trioxide (ATO),9 treated with ATO alone, the early death rates were 27.3%,14.3%,55.6% respectively, CR rates were 67.7%,81.0%,44.4% respectively, the result showed there were the lowest rate of early death and the highest rate of CR in the combination group. Of 61 patients, 41 cases were given low-dose chemotherapy, the CR rate was 73.2%,total death rate was 19.5%, that of 20 patients without chemotherapy were 45.0%,55.0% respectively. The differences between two groups had statistical significance(p<0.05).

Conclusions: Compared with APL without high initial WBC, APL with high initial WBC has lower CR rate, higher early death rate, higher incidence of DIC and RAS .The combination of ATRA,ATO and chemotherapy is the most efficient therapeutic approach to APL patients with high initial WBC, which can significantly reduce the early death rate and improve the CR rate.
Addition of ATRA to the Maintenance Protocol Did Not Improve Disease-Free Survival: Results of the Russian APL Trial.
Authors: Parovichnikova Elena N. , Savchenko Valeri G. , Isaev Valentin G. , Shuravina Elena N. , Demidova Irina A. , Olshanskaya Julia V. , Misiurin Andrey V. , Kulikov Sergey M.
Published: October 2019

Russian Leukemia study group is presenting the final analysis of the APL-06.01. randomized clinical trial. The aim of the study was to identify the efficacy of ATRA substitution of each other chemotherapy course while the 2-yrs maintenance after three induction/ consolidation 7+3 with daunorubicin dose of 180 mg/m2 per course and ATRA for 30 days while the first induction. The patients were randomized for two types of maintenance - (I type) rotation of 5-days ARA-C (100 mg/m2 bid) combined with daunorubicin (45 mg/m2 for 2 days, till the total dose of 650 mg/m2) or 6-MP (50mg/m2 p.o.5 days) or CHP (800 mg/m2 i.v.1 day) or (II type) rotation of the same courses of chemotherapy with 5-days 45 mg/m2 ATRA course. The II type of maintenance treatment contained twice less chemotherapy. The intervals between courses were 4 weeks From July, 2001 till January, 2006 114 APL patients from 26 centers were enrolled in the study. In 95% of pts APL was confirmed by cytogenetics or PCR. 102 patients were included in the analysis. CR rate was 85%, ED rate -15%, 4-yrs OS -68,3%, DFS - 77,4%, CCR - 89,5%. Suvival analysis according to randomization did not demonstrate statistical differences, but showed a trend (p=0,06) towards better DFS and CCR in patients maintened with chemotherapy only: 85,2% and 94,3% (I type) and 77,7% and 85,6% (II type). In the multivariant analysis only one factor was defined as statistically significant - the number of the patients randomized by the participating centers. OS and DFS were much higher in the centers randomized 6 and more patients comparing with less than 6 patients: 82,9%and 90,7% vs 48,8% and 63%, respectively (0,003). 1. So we can suggest that maintenance with ATRA alternating with chemotherapy is less effective than chemotherapy only. Whether addition not substitution of ATRA to chemotherapy maintenance will change the results will be checked in the next study.2. The experince of the centers in APL treatment is a cruicial point for survival.
Salvage Therapy with Standard Dose Cytarabine Is Appropriate for Patients with Acute Myelogenous Leukemia Refractory to Front-Line Therapy with Hypomethylating Agents.
Authors: Borthakur Gautam , Kantarjian Hagop , Garcia-Manero Guillermo , Pierce Sherry , Estey Elihu
Published: October 2019

Treatment without cytarabine (ara-C) is increasingly being used as initial therapy for acute myelogenous leukemia (AML) particularly in patients (pts) deemed ineligible, usually on the basis of age, for standard ara-C-containing therapy; prime examples are the hypomethylating agents decitabine (DAC) and 5-azacitidine (Aza). This practice raises the question whether it is useful for patients whose disease fails to respond to hypomethylating agents to receive ara-C as salvage. Thirty-five pts treated from 2001 to 2006 failed to respond to initial therapy with decitabine/azacitidine. Twenty-five pts subsequently received ara-C as salvage and 10 received other salvage regimens. We compared their complete remission (CR) rates (Table 1). Although more pts who received non ara-C salvage had poor prognosis karyotype, CR rates in pts with standard-risk karyotype receiving ara-C and non ara-C salvage therapy were 7/20 (35%) vs. 0/6 (0%) respectively. Although this difference was not significant (p=.15) the 95% confidence limits for the true difference, computed so as to account for the small sample size, was [−.69, .11] indicating as much evidence for a true difference of 58% as for a true difference of 0% since zero and 58% are equidistant from the midpoint of the confidence interval (−0.29). Although future improvements in results with non ara-C containing salvage therapies will impact decision, the results suggest that it is worthwhile to give pts who fail initial therapy with decitabine or azacitidine, standard dose ara-C salvage provided the pts do not have −5/−7 (CR rate 0/5).

Table 1 Induction/salvage Patients Median age −5 / −7 CR DAC or Aza/ara-C 25 67 5 (20%) 7 (28%) DAC or Aza/no ara-C 10 71 4 (40%) 0
Feasibility Study of Temozolomide Maintenance Therapy in Elderly Patients with AML Following Induction Therapy with High Dose Mitoxantrone and High Dose Ara-C.
Authors: Ponce Doris , Katragadda Sreedhar , Liu Delong , Rasul Muhammad , Ahmed Nasir , Ahmed Tauseef , Seiter Karen
Published: October 2019

With intensive chemotherapy, approximately 50% of elderly patients with AML achieve a complete remission, however essentially all will relapse within 2 years. To date, there is no standard effective postremission therapy for these patients, and most elderly patients tolerate multiple courses of intensive chemotherapy poorly. We previously demonstrated that temozolomide has significant antileukemic activity in patients with relapsed/refractory acute leukemia (Seiter K, et al JCO 2002). In that study 9/20 patients treated had a significant reduction in bone marrow blasts, and 4 patients had a formal complete response. Currently we are evaluating the feasibility and efficacy of temozolomide maintenance therapy administered after one cycle of intensive induction chemotherapy in elderly patients with newly-diagnosed AML. Induction consists of ara-C, 3 gm/m2 daily for 5 days, and mitoxantrone, 80 mg/m2 times one on day 2. Patients achieving complete remission then receive temozolomide 200 mg/m2/d for 7 days every 5 weeks until progression. To date, 34 pts have been treated: median age 69.5 (range 61–84); M/F: 21/13; prior AHD: 47%; cytogenetics: good: 1, intermediate: 24, poor: 7, insufficient: 2. Response to high-dose ara-C/high-dose mitoxantrone induction: CR: 56%, failure: 32%, toxic death: 12%. Two pts expired in remission unrelated to temozolomide: pneumonia, 1 pt, C. difficile colitis: 1 pt. Of 19 patients in CR, 16 received temozolomide maintenance. The median number of temozolomide cycles was 2 (range 1–14). 3 pts stopped temozolomide after 1 cycle (1 refused further therapy, 1 had stem cell transplant, 1 died from C. difficile colitis). Treatment was well tolerated and was administered in the outpatient setting. The median duration of remission is 6.8 months (95% CI: 4.5–9.1 months), and median survival is 5.5 months (95% CI: 3.7–7.3 months) for all patients and 10.1 months (95% CI: 4.9–15.2 months) for patients achieving CR. Temozolomide maintenance is feasible in patients with AML in CR. Further accrual is continuing to assess if remission duration and survival will be improved compared to historical controls.
Tumor Necrosis Factor-α Enhances DMSO-Induced Differentiation of HL-60 Cells through Activation of ERK/MAPK Pathway.
Authors: Park Jinny , Lee Young-Rae , Noh Eun-Mi , Han Myung-Kwan , Ahn Jung Ryul , Kim Young Sang , Park Se Hoon , Cho En Kyung , Shin Dong Bok , Lee Jae Hoon , Kim Jong-Suk
Published: October 2019

The differentiation of leukemic cells into mature cells is a major strategy for treatment of leukemia. Many trials for more efficient differentiation of leukemic cells have been performed using single uses of differentiation inducers including interferon-α (IFN-α), interleukin-4 (IL-4), tumor necrosis factor-α (TNF-α), and dimethyl sulfoxide (DMSO) or their combinations. Here we show that combination of TNF-α with DMSO enhances HL-60 cell differentiation. TNF-α enhanced DMSO-induced expression of CD11b and increase of G1 phase in the cell cycle, which are hallmarks for HL60 cell differentiation. Inhibition of ERK pathway, but not NF-κB pathway, abolished the enhancement of DMSO-induced HL-60 differentiation by TNF-α. These results suggest that TNF-α enhances HL-60 cells towards induction of differentiation by DMSO through activation of ERK/MAPK signaling pathway.
High Dose Simvastatin Has No Beneficial Effect on Markers of Bone Turn-Over in Multiple Myeloma.
Authors: Sondergaard Teis E. , Pedersen Per T. , Andersen Thomas L. , Lund Thomas , Garnero Patrick , Delaisse Jean-Marie , Plesner Torben
Published: October 2019

Background: Bone degradation in multiple myeloma (MM) is a result of increased bone degradation by osteoclasts that is not compensated for by bone forming osteoblasts. Ideally new drugs used for treatment of MM should target not only the myeloma cells but also the imbalance between bone resorption and bone formation. Statins have been shown to inhibit myeloma cell proliferation and induce apoptosis in vitro. Furthermore statins have been shown to stimulate osteoblasts and inhibit osteoclasts both in vitro and in animal models. Statins are normally used at doses around 20–80 mg/day, but in order to reach serum concentrations that can match the in vitro experiments MM patients were treated with 15 mg/kg/day of Simvastatin (HD-Sim) divided in two daily doses in this study. This high dose has previously been found to be safe for MM patients (Haematologica 2006, 91,542–545)

Patients and methods: Six patients with advanced MM have been included in this pilot study, 4 males and 2 females with an average age of 68 years and an average duration of disease of 43 months. The patients were treated with 2 cycles of HD-Sim for seven days followed by a break of 21 days in a 4-weeks cycle. Two of the patients were treated with bisphosphonates during the study, and 4 had previously been treated with bisphosphonates. Endpoints are change in concentrations of markers of osteoclast activity (TRAP) or bone resorption (CTX, NTX, ICTP) or markers of bone formation (Osteocalcin and PINP). Cholesterol, OPG and DDK-1 were also measured.

Results: Two patients completed the protocol with two cycles of HD-Sim at full dose, 2 patients were reduced to 7.5 mg/kg/day simvastatin in cycle 2 due to nausea and diarrhea and 2 patients left the protocol after 3 weeks (deaths not related to high dose simvastatin). All patients experienced gastrointestinal toxicity grade 1–2. Myalgia and other muscular symptoms grade 1–2 were reported by 5 patients but were not associated with an increase in creatin kinase. TRAP and NTX activity in serum increased for all 6 patients during the seven days of treatment with HD-Sim indicating that bone resorption may have been stimulated rather than inhibited. The other markers of bone resorption and the bone formation markers showed no change. All patients responded with a significantly reduced level of cholesterol in serum. None of the patients showed any reduction in free monoclonal light chains or monoclonal proteins in serum during treatment with HD-Sim and 2 of the 4 patients completing the protocol showed progression of diseases.

Conclusion: This pilot study of HD-Sim in advanced MM has been terminated due to lack of response and evidence from two markers of osteoclast activity (TRAP) and bone resorption (NTX) that HD-Sim may be harmful rather than beneficial in MM.
Bortezomib in Combination with Dexamethasone and Subsequent Thalidomide for Newly-Diagnosed Multiple Myeloma in Chinese Patients.
Authors: Cai Zhen , Zheng Weiyan , Wei Guoqing , Ye Xiujin , He Jingsong , Li Li , Wu Wenjun , Lin Maofang
Published: October 2019

Background: Bortezomib-dexamethasone-thalidomide has been reported to be effective in newly-diagnosed multiple myeloma (MM) with an overall response rate of 92% and a CR rate of 18% (Alexanian et al, Hematology12(3):235–239, 2007), but this regimen has not been reported in Chinese patients. We now report our experience with this combination.

Objectives: To investigate the efficacy and safety of bortezomib in combination of dexamethasone plus subsequent thalidomide as primary treatment for MM.

Patients and Method: Between June 2006 and August 2007, 11 consecutive newly-diagnosed patients with symptomatic MM were treated with bortezomib at 1.3 mg/m2 IV on days 1, 4, 8 and 11, dexamethasone at 20 mg/m2 IV daily on the day of bortezomib and the day after. All patients received daily oral thalidomide that was escalated from 100 mg to 200 mg. Seven of 11 patients were male and 4 were female. Median age was 57 years (range 47–86). Seven of 11 patients were stage 2 according to the International Staging System, 4 out of 11 patients were stage 3. Eleven patients received a median of 2 cycles of therapy (range 1–6). The Blade criteria were used for response evaluation. Toxicities were evaluated according to the NCI Common Toxicity Criteria version 3.

Results: Nine out of 11 patients (82%) achieved PR and 2 (18%) achieved CR; therefore the overall response rate was 100%. With a median follow-up duration of 5 months (1– 14 months), no patients died. Grade 3–4 toxicities included fatigue (3/11), thrombocytopenia (3/11), diarrhea (3/11) and orthostatic hypotension (2/11). Grade 2 neuropathy occurred in 3 out of 11 patients, herpes zoster occurred in 3 out of 11 patients. Routine anticoagulation or anti-thrombosis was not used. There was no DVT/PE in 11 patients.

Conclusion: Our preliminary experience indicated that bortezomib-dexamethasone-thalidomide is highly effective in newly-diagnosed MM. Grade 3 and 4 toxicities were rare after median 2 cycles of therapy. The relative lower rates of neuropathy and DVT/PE in this report with Chinese MM patients are being cautiously observed.
Hepatitis - Associated Aplastic Anaemia: Epidemiology and Treatment Results Obtained in Europe. A Report of the EBMT Aplastic Anemia Working Party.
Authors: Locasciulli Anna , Bacigalupo Andrea , Oneto Rosa , Bruno Barbara , Marsh Judith , Tichelli Andre’ , Socie’ Gerard , Passweg Jakob
Published: October 2019

Background. Most cases of acquired severe aplastic anaemia (SAA) are not associated with any other disease and are therefore classified as idiopathic. Hepatitis-associated aplastic anaemia (HAA) has been well described as a separate entity amongst the acquired aplastic anaemias. Aim of this study was to describe the epidemiology of HAA and to analyze treatment results obtained in Europe in the last 15 years.

Patients and methods. We evaluated 198 HAA out of 3684 AA patients reported to the SAAWP Registry between 1990 and 2005. Median age at diagnosis was 15 years (range 1–87) .132 were male and 66 female. Year, month and season of diagnosis were analyzed, as well as all variable related to the type and outcome of first line therapy.

Results.Prevalence of HAA amongst all acquired AA registered in Europe from 1990 to 2005 was 5.4% (198/3684).The majority of patients were male (66%), and median age was 15 years. The virus causing the hepatitis was unknown in most cases (94.5%) while only 11 / 198 (5.5%) were related to Hepatitis B or A virus infection. No significant differences were found in the prevalence according to the year, month or season of diagnosis. No geographic clusters could be identified. Seventy three HAA patients were treated with immunosuppression (IS) and 125 underwent Stem cell Transplantation (SCT): these were compared with 1539 and 2045 non-HAA patients given IS or SCT. The median age was significantly younger in the HAA as compared to the non-HAA patients (15 versus 20, p<0.0001). The median interval between diagnosis and treatment in IS and SCT was 8 and 67 days respectively. Median follow-up for surviving or deceased patients was 1356 and 155 days respectively. The actuarial survival at 10 years after IS was 66% for HAA and 52% for non-HAA (p=0.4). HLA identical sibling SCT produced survival of 83% in HAA and 74% in non-HAA patients (p=0.1); when the donor was unrelated, survival was respectively 63% and 52% (p=0.5). After stratification for stem cell source, actuarial survival was 83% vs 72% (p=0.06) for bone marrow, and 68% vs 57% (p=0.5) for peripheral blood.

Conclusions. The incidence of HAA in Europe is currently 5.4%, comparable with published data, and seems to be distributed homogeneously over the years, months and geographic areas. Results after IS or SCT in HAA patients are comparable to those obtained in idiopatic acquired non-HAA, treated in the same period in Europe. Therefore, HAA should be managed according to current guidelines for idiopathic aplastic anemia.
Response to Bortezomib Therapy for Multiple Myeloma (MM) in Actual Clinical Practice: Preliminary Effectiveness Findings of an International Observational Study.
Authors: Delforge Michel , Katodritou Eirini , Zervas Konstantinos , Sargin Deniz , Hulin Cyrille , Linderholm Mats , Verrou Evgenia , Poon Vicki , Dhawan Ravinder
Published: October 2019

Background: Bortezomib (VELCADE®) is indicated for the treatment of MM in patients (pts) who have received at least 1 prior therapy. The electronic VELCADE Observational Study (eVOBS) is designed to evaluate the clinical and outcomes benefits of bortezomib in actual clinical practice. The study is open for enrollment between October 2006 and December 2008 in Belgium, France, Greece, Russia, Spain, Sweden, and Turkey with 3-year follow up. This report includes preliminary outcomes in pts with at least 4 months of data as of July 2007.

Methods: Adults were eligible for study if they were scheduled to initiate bortezomib within the approved indication. All bortezomib dosages and concomitant treatments were permitted, except investigational therapies. Data on concomitant therapies, treatment response, and safety were collected prospectively during bortezomib therapy. Due to the non-interventional nature of the study, no predefined response criteria were mandated; response criteria could include M-protein, EBMT, SWOG, or others as defined by the investigator.

Results: A total of 86 pts with at least 4 months of data were included in this analysis. Median age was 61 yrs, and 50 (58%) pts were male. Median interval since diagnosis was 3 yrs. The number of previous therapies for MM was 1, 2–3, and ≥4 for 38%, 44%, and 11% of pts, respectively. Demographic and clinical characteristics of the initial participants were similar to those of the participants in the prospective controlled phase 3 APEX trial (Richardson, N Engl J Med, 2005:352; 2487–98). Most pts (61%) received bortezomib with dexamethasone. Adverse events (AEs) were reported in 61 (71%) pts, including Grade ≥3 AEs in 38% and Grade ≥4 AEs in 9%. AEs were treatment-related in 45% of patients and treatment-limiting in 9%. Presently, 72 of 86 pts have been evaluated for response, of whom 54 completed 4 or more cycles. Response rates are shown in the table. Updated data will be presented at the meeting.

Best Response(n = 72) Complete response (CR) 5 (7) Near complete response (nCR) 9 (13) Partial response (PR) 30 (42) Minimal response (MR) 9 (13) Stable disease (SD) 6 (8) Progressive disease (PD) 13 (18) Overall response (≥PR) 44 (61) Overall response (≥MR) 53 (74)

Conclusions: In this preliminary analysis of data from a prospective, observational study of actual clinical practice, most pts received bortezomib in combination with 1 or more other therapies for MM. Overall response in actual clinical practice was at least as common as previously reported with bortezomib monotherapy. Response rates and safety data from actual clinical practice demonstrate that bortezomib-containing regimens are effective and well-tolerated in the treatment of MM.
Simultaneous Haploidentically Mismatched Stem Cell Transplantation from Multi-Donors (MDT) in Humans Offers Better Engraftment as Compared to Single Haploidentical Donor.
Authors: Shapira Michael Y. , Resnick Igor B. , Gesundeheidt Benjamin , Samuel Simcha , Drey Liliane , Verkholevsky Yuri , Slavin Shimon , Or Reuven
Published: October 2019

Introduction: stem-cell-transplantation (SCT) aims to combine tumor cytoreduction and host replacement with donor cells. For patients with no matched sibling available for allogeneic SCT (approximately 70% of patients) an alternative option is SCT from a relative carrying only one identical HLA haplotype (haplo-SCT), which may provide a donor for almost every patient at optimal timing. In the small number of multi-cord blood transplants reported, it was found that usually only one unit engrafted faster, while the others were rejected. We hypothesized that multi-donors may improve the engraftment following haplo-SCT. Patients and methods: Fifteen ultra high-risk patients (median age 20y, 9–52, mostly after 1–2 previous SCTs and 13 of them with refractory disease) were included. Conditioning regimen was fludarabine based. All grafts were T cell depleted.

Results: All 14 evaluable patients engrafted but one of them later rejected the graft. The time for neutrophil engraftment (0.5 & 1.0x109/L) was fast, 11d (9–24) and 12d (10–28) respectively. Platelets engrafted even earlier (20 & 100x109/L), 10d (8–45) and 14d (12–32) respectively. Three patients demonstrate continuous bi-donor hematopoiesis (follow up 72, 128 and 168d) while 11 patients converted to single donor hematopoiesis within a median of 21d (14–39). Comparison of MDT patients to an earlier patient group receiving a similar protocol with a single donor (SDT) showed better granulocyte and platelet engraftment (p<0.001 for both, figures 1 and 2). Additionally, no primary graft failure occurred in patients undergoing MDT, while in SDT it occurred in 9/38 patients (p<0.001). One patient in-which leukemia persisted throughout the conditioning, leukemic cells disappeared upon the conversion to single donor chimerism. Despite major blood type differences, no hemolysis related side-effects were noted. Eight patients developed GVHD (mostly grade 1/2).

Conclusion: MDT is safe and induces fast and durable engraftment of both myeloid and thrombocyte lineages in patients undergoing haplo-SCT. Further studies should follow this report.

Figure Figure Figure Figure
Endothelial Dysfunction in Autologous Hematopoietic Stem Cell Transplantation.
Authors: Diaz-Ricart Maribel , Carbo Carla , Palomo Marta , Fernandez-Aviles Francesc , Rovira Montse , Escolar Gines , Carreras Enric
Published: October 2019

Introduction: Intensive conditioning regimen and allo-reactivity seem to be the main causes of the endothelial dysfunction observed after allogeneic HSCT. This dysfunction may be the origin of some early complications after HSCT. To evaluate if a less intensive conditioning without allo-reactivity phenomena produces endothelial dysfunction we analyzed a group of patients receiving autologous HSCT.

Methods: Blood samples were collected from patients (n=10) before receiving BEAM treatment (pre), and at days 0, 7, 14 and 21 after the HSCT. Soluble markers of endothelial damage (von Willebrand Factor [VWF], sE-selectin, sVCAM1, sICAM1, TNFalphaRI and II, MMP9) were measured in plasma samples by ELISA. Expression of the adhesion receptors E-selectin, VCAM1 and ICAM1, on endothelial cells (EC) cultured in the presence of a pool of sera, obtained at the different time points during the BEAM treatment, was analyzed by immunocytochemistry. Changes in the expression of VWF in the extracellular matrix generated by these EC were evaluated by using the same technique. Leukocyte adhesion on EC monolayers under flow conditions was also studied. The effect on cell cycle and apoptosis was also explored.

Results: Plasma levels of VWF and sICAM1 increased progressively after the HSCT, peaking on day 14 (84% and 118%, respectively, vs pre; p<0.01), although no increments were detected for sVCAM1 and sE-selectin. TNFalphaRI and RII increased from 0 (pre value) to 5, 12, respectively, and MMP9 from 0 (value at day 7) to 300ng/ml. Exposure of cultured EC to patients sera caused an increased expression of all adhesion receptors studied on the EC surface and of VWF on their extracellular matrix, more notable at day 14 and specially for VCAM1. Leukocyte adhesion on EC cultured with sera from day 14 showed a 6-fold rise compared to controls. Although no signs of apoptosis were detected, cell growth was accelerated at day 14.

Conclusions: The increases in the markers of endothelial inflammation detected both in the ex vivo and in the in vitro approaches suggest that there is endothelial damage associated with autologous HSCT. This damage seems to be more important at day 14 after HSCT. A better understanding of the mechanisms involved deserves further investigation.
Phase 1 Human Trial of Autologous Mesenchymal Stem Cell Transplantation for the Treatment of Decompensated Cirrhosis.
Authors: Alimoghaddam Kamran , Mohamadnejad Mehdi , Bagheri Mohahamd , Bashtar Maryam , Ghannati Hossein , Bonab Mandana Mohyeddin , Ghavamzadeh Ardeshir , Malekzadeh Reza
Published: October 2019

INTRODUCTION: Liver transplantation is the standard treatment in decompensated cirrhosis; however, it has several limitations including high cost, and several complications. Recent animal works have shown that mesenchymal stem cell (MSC) infusion through the rat tail vein can lead to regression of liver fibrosis. But there has been no human trial of autologous MSC transplantation (Tx) in cirrhosis. The aim of this study was to evaluate safety and feasibility of MSC transplantation in decompensated cirrhosis. In a recent study, infusion of CD 34+ (e.g. hematopoietic stem cells) through the hepatic artery was used to treat cirrhosis. However, we infused MSCs (e.g. non-hematopoietic stem cells) through a peripheral vein.

AIMS & METHODS: After protocol approval by the ethic committee, four patients with decompensated cirrhosis were included. 100 ml of their bone marrow was aspirated and MSCs were cultured. After two months culture, 10 to 70 million MSCs obtained which 95% of the cells were viable (as assessed by trypan blue method) and 95% of the cells were morphologically and immunophenotypically MSCs. The MSCs were infused through a peripheral vein. Outcomes were changes in liver function tests, MELD score, and liver volume (as measured by CT volumetry) 5 months after Tx. During follow up, patients’ medications (e.g duretics, or propranolol) remained the same as before MSCs Tx.

RESULTS: Four patients (1 male, 3 female) aged 30 to 56 were included. The etiology of cirrhosis was autoimmune hepatitis or cryptogenic. The follow up for the first patient has been completed, and other 3 patients have 2.5 to 4 months of follow up. In first patient, PT (18.5 to 13 seconds), and albumin (2.9 to 3.8 g/dL) normalized, and MELD score was decreased from 16 to 11. Her liver volume increased from 495 to 815 ml. In patient 2, PT decreased from 20.1 to 17.5. In patient 3, PT decreased from 19.1 to 15.7 seconds, and MELD score decreased from 17 to 14. Patient 4 has not yet been shown any significant improvement in liver function tests. The procedure was safe, and there was no any adverse effect.

CONCLUSION: MSCs Tx is a novel approach to treat cirrhosis. For the first time, we showed that this procedure is safe and feasible, and signs of improvement found in some of our patients are promising.
NK-Cell Reconstitution after HLA-Identical Blood and Marrow Transplantations for CML: The Recovery of NKG2D Is Inversely Correlated with NKG2A and It Promotes the GVL Effect.
Authors: Tong Juan , Liu Huilan , Geng Liangquan , Sun Zimin , Tang Baolin , Wu Zhentian , Qiao Chunlan , Chen Xin , Qian Bing , Song Kaidi
Published: October 2019

Natural killer (NK) cell alloreactivity is reported to mediate strong graft versus leukemia (GVL) effect in patients after allogeneic stem-cell transplantation. NKG2D receptors recognize human MHC class Ichain related A and B (MICA/B) and UL16-binding protein 1∼4(ULBP 1∼4) on target cells, thereby regulating NK cell activity. To examine the recovery of NKG2D, NKG2A and other receptors expression by NK cells, we used flow cytometry to evaluate samples from 11 chronic myeloid leukemia patients and their donors in the year following unmanipulated HLA completely matched peripheral blood stem cells plus bone marrow transplantation. Peripheral blood mononuclear cells from patients and their donors were tested in standard 51Cr release assays against cultured K562 targets to determine the cytotoxicity of the NK cells in the same intervals. There is no mismatched immunoglobulin-like receptor (KIR) ligand in both GVH and HVG direction. The reconstitution of KIR2DL1 (CD158a) after this transplantation protocol was very slow and these receptors didn’t reach normal value in the year and KIR2DL2 (CD158b) was much better. The NKG2D increased and the NKG2A decreased quickly at the same time after engraftment, and used linear regression analysis we demonstrated that NKG2A recovery was inversely correlated with NKG2D recovery in the year following transplantation. The ratio of NKG2D/NKG2A was directly associated with the capacity of NK-cell cytotoxicity. Thus, the reconstitution of NKG2D makes contribution to the recovery of the NK cytotoxicity. These results reveals that the NK cells generated after HLA matched blood plus bone morrow transplantation of CML patients are promoted at an immature state characterized by specific phenotypic features and enhanced functioning, having potential impact for immune responsiveness and transplantation outcome.
Development of a PML-RARα-IL-2 Recombinant Plasmid DNA for APL.
Authors: Zhou Yubing , Hu Gang , Cen Dongzhi , Yang Lijian , Chen Shaohua , Li Yangqiu
Published: October 2019

Specific active immunotherapy with DNA vaccines is one of the best approaches to eradicate minimal residual disease in cancer patients, when appropriate tumor-specific antigens are identified for this disease. However, little data are regarding the DNA vaccines in acute leukemia, recent reports has shown that a human PML-RARα breakpoint-drived DNA vaccine can prevent APL in mice model, but the host immune response were weakly, due to the weak immunogenicity. In order to improve the effect of DNA vaccine for acute promyelocytic leukemia (APL) therapy, we develop a vector coexpressing PML-RARα gene fused to human Interleukin- 2 (hIL-2) gene. PML-RARα fusion gene segment and the full-lenght hIL-2 gene were amplified from NB4 cells or Jurkat cells. Both PCR products were cloned into PIRES plasmid respectively to construct a recombinant plasmid PML-RARα-IRES-hIL-2. The recombinant plasmids were then transfected into K562 or A549 cells respectively. The expression of the PML-RARα/hIL-2 mRNA and protein in transfected cells were identified by RT-PCR, dot blotting, ELISA and Western-Blot respectively. By in vivo assays, BALB/c mice were vaccinated at 6–8 week of age with a total of 200 μg DNA in normal saline, injected into two sites in the quadriceps muscles on day 0, 7 and 21. The plasmid containing the same sequence of PML-RARα gene and blank plasmid served as controls. Two weeks after the final DNA boost, both PML-RARα/hIL-2 mRNA and protein, serum INF-γ and anti-NB4 cells specific cytotoxicity of splenocytes following 7 days of stimulation in vitro with freeze thawing NB4 cells and recombinant human IL-2 were assessed by ELISA and LDH assays. The results showed that the sequence of the fragments inserted in multi-clone site (MCS) A and MCS B of PIRES plasmid were absolutely correct by double restriction enzyme cutting analysis (Sal I/Not I) and sequence analysis, the PML-RARα/hIL-2 mRNA and protein could be identified in transfected K562 or A549 cells and in mice quadriceps muscles. The level of serum INF-γ and cytotoxicity of splenocytes against NB4 cells from immunized mice was significant increased than that from control groups. Our results indicated that the recombinant plasmid expressing PML-RARα and hIL-2 was successfully constructed, which can induce more effective immune response and anti-APL cells effect in animal models. The data suggest that effective vaccination approaches should be possible against APL. It could be farther used in the research as PML-RARα DNA vaccine for APL.
Potent Anti-Leukemic Activity of a Cationic Lipid-DNA Complex.
Authors: Herse Zachary , Chang Stella , Liggitt Denny , Fairman Jeffery , Claxton David F.
Published: October 2019

Cationic Lipid and DNA Complex (CLDC) is a chemically defined preparation of lipids and non-coding bacterial plasmid DNA able to activate antigen presenting and immune effector cells and thereby enhance cellular immune responses. We tested this preparation subcutaneously (SC) in two models of transplantable murine leukemia: 32D -(bcr-abl)p210 (leukemogenic in C3H/HEJ animals) and WEHI-3B cells (leukemogenic in BALB-C mice.) In multiple experiments using the 32Dp210 model a single SC dose of CLDC (200 ul) prevented or significantly delayed death from leukemia when delivered between -1 and 16 days from leukemic challenge. Specifically, control animals died at 28 days (SD=2.73) of leukemia whereas 20/24 similarly leukemia challenged but CLDC treated animals were alive >71 days (p<0.0001 − 4/20 animals died of leukemia). Animals dying of leukemia had swollen spleens and leukemic infiltration of spleen and liver on histopathological evaluation. Similarly Balb-C animals challenged with WEHI-3B cells died at a median of 30 days unless treated with CLDC (1 dose day 6 − 4/5 alive >day 40 p=0.0017). A green fluorescence protein (GFP) expressing subclone of 32Dp210 allowed quantitation of leukemia in peripheral blood (PB). PB leukocytes were 20% GFP positive on day 19. After CLDC on day 19, day 21 PB showed only 2% GFP. Animals receiving 32D-p210-GFP and treated with CLDC and CD8 antibody on day 19 showed higher GFP in followup and died earlier (median day 35) than animals treated with CLDC alone (>42 days) but later than animals receiving no treatment after leukemic challenge (median 23 days, p=0.0002.) See Figure below. We conclude that CLDC shows potent anti-leukemia activity in two murine models of AML. Antibody experiments suggest that CD8 positive effector cells contribute to this effect. Additional experiments are maturing and will be reported. This preparation has promise for activity in human acute leukemias.

Figure Figure
Y-Chromosome Genes Strongly Expressed in Acute Myeloid Leukaemia.
Authors: Gallo Antonio , Bund Dagmar , Mindnich Rebecca , Buhmann Raymund , Schmetzer Helga , Adamski Jerzy , Kolb Hans-Jochem
Published: October 2019

In hematopoietic stem cell transplantation (HSCT) the gender of the donors has a substantial impact on the relapse rate, graft-versus-host reaction and overall therapeutic outcome. In several studies including our own HLA-haploidentical setting female to male transplantation seemed to be superior to other gender combinations, suggesting the dominance of Y-encoded minor histocompatibility antigens for graft-versus-leukemia reactions. In order to explore the therapeutic possibilities we compared the Y-chromosomal gene expression between isolated AML blasts (FAB M4/M5) and isolated CD14+ cells from healthy male donors. RNA was isolated and subsequently analyzed by low density arrays (LDAs), a real time-PCR based method. This way we could identify four Y-encoded genes found to be strongly up-regulated in the AML samples. Bcorl2, a pseudogene; pcdh11y, known to be involved in acquisition of apoptotic resistance in prostate cancer; tgif2ly and vcy, both putative transcription factors. The genes were cloned in plasmids and transferred in 293 cells. The expression of these proteins was analyzed by western blot. Biostatistical soft ware was used to predict the proteasomal cleavage site and binding capacity to HLA-A 0201 of these peptides. Five peptides were found to bind strongly to HLA-A 0201; they are now studied for recognition by female T cells. In summary, the genes identified might be involved in the immune recognition of AML blasts and other tumor cells. These peptides may help to identify and select T memory cells specific for AML blasts.
Ganoderma lucidum Polysaccharides Can Induce Human Monocytic Leukemia Cells into Dendritic Cells with Immunotolerogenic Function.
Authors: Chan Wing Keung , Cheung Christopher , Law Helen Ka Wai , Lau Yu Lung , Chan Godfrey Chifung
Published: October 2019

Previous studies demonstrated Ganoderma lucidum polysaccharides (GL-PS), a form of bioactive β-glucan can stimulate the maturation of monocyte-derived DCs (Mo-DCs). However, the question of how leukemic cells especially in monocytic lineage respond to GL-PS stimuli remains unclear. We hypothesized that GL-PS may also have an effect on leukemic monocytic cells In this study, we used human dendritic cell (DCs) in vitro culture model with leukemic monocytic cell-lines THP-1 and U937 as cell sources. We treated the cells with purified GL-PS (100 μg/mL) or GL-PS with GM-CSF/IL-4 (cocktail). GL-PS alone induced proliferative response on both cell-lines with more markedly on U937. But it induced THP-1 cells to differentiate into cells with typical DC morphology. Additive maturation effect can be shown when THP-1 cells were stimulated with the cocktails shown by significant increase in HLA-DR, CD40, CD80 and CD86 which express in normal peripheral blood Mo-DCs. The GL-PS THP-1 DCs had similar antigen-uptake ability as the Mo-DCs positive control. However, we found that GL-PS THP-1 DCs could not induce allogeneic T cell proliferation in the one-way mixed lymphocyte reaction as Mo-DCs or THP-1, suggesting GL-PS THP-1 DCs were immunotolerogenic in nature. Our data indicated that the GL could induce monocytic leukemic cell proliferations as well as differentiate particular leukemic cells to DCs with immunosuppressive action. The possible clinical impact has to be carefully investigated.