bluuebunny/crossref_metadata_2025 huggingface dataset:
A Single Center Analysis over a Ten Year Period on Transplant Related Mortality (TRM), Early Relapse (ER) and Failure of Blood Cell Reconstitution (FBR) in High-Risk (HR) Cancer Patients (PTS) Undergoing Autologous PBSCT (auto-PBSCT): Frequency, Outcome and Prognostics.
Authors: Engelhardt Monika , Doeing Carsten , Ihorst Gabriele , Wider Dagmar , Finke Juergen
Published: April 2020

The outcome of hr cancer pts undergoing intensified chemotherapy (CTx) with risk factors, such as advanced disease, incomplete remission, tumor resistance and/or prior clinical complications can be dismal and remains a challenge. The prognostic factors for determining TRM, ER and FBR in hr cancer pts undergoing auto-PBSCT, allogeneic (allo)-PBSCT or intensified CTx have not yet been identified. Here, we analysed 50 cancer pts who died within three months of auto-PBSCT or had primary FBR, with a total of 796 pts undergoing auto-transplants over a 10 year period (6/93–5/2003) at our center. Median patient (pt) age at transplant was 52 years (y; range; 18–75), with underlying lymphoma, solid tumors or leukemias in 30, 14 and 6 pts, respectively. The disease stage in 90% (n=45) was advanced, bulky disease was present in 52%. Median performance status of all pts was 70%. An elevated LDH (median 365U/l) at diagnosis was present in 78% of pts. Treatment before auto-PBSCT consisted of a median number of 7 CTx cycles and radiation in 32% of pts. A median number of 4.1x10e6/kg bw PBSCs were transfused, with total CFUs (/1.5e5) of 128. Median platelet counts before PBSCT were 98x10e9/L, with persisting BM involvement in 38% and hypocellularity in 46% of pts. Transplant complications, leading to TRM, ER or FBR were observed in 18, 25 and 7 pts, respectively and resulted in early death in all pts, except one with CML and FBR, who was salvaged with back-up PBSCs. Thirteen pts were examined post-mortem and showed hypo- vs normocellular BM in 8 vs. 3 pts, respectively. Of note, FBR was observed in older pts (59 y), who were mostly heavily pretreated and had AML or CML (4 pts). They had been retransfused with lower CD34+ cells (2.9x10e6/kg), showed lower BFU-Es, CFU-GMs and GEMMs (30, 40, 3, respectively) and low platelet counts (27x10e9/L) before PBSCT due to active underlying disease (71%) and/or BM hypocellularity (71%). During post-transplant cytopenia all pts with FBR showed serious infections. These results suggest that the above risk factors are associated with TRM, ER and FBR. LDH and low platelet counts were significantly different between the three groups. In vitro culture data (cell expansion, CFU, LTC-IC) correlated with our clinical results. With a TRM of 2.26% and FBR in 0.88% in this hr pt cohort, auto-PBSCT is in general a very safe procedure. Nevertheless distinct risk factors can be determined and need to be considered before autotransplantation. Our results may also be valid for allo-SCT and intensified dose-dense chemotherapies.
Evaluation of Body Mass Index (BMI) as a Prognostic Factor among Patients with Relapsed/Refractory Hodgkin’s Lymphoma (R/R-HL) Undergoing Autologous Bone Marrow/Peripheral Stem Cell Transplantation (Auto-BM/PSCT).
Authors: El-Khoury Christian , Kojouri Kiarash , Pant Shubham , Sylvester-Kohrt Nancy , Selby George B. , Ozer Howard , Kamble Rammurti , Kharfan-Dabaja Mohamed A.
Published: April 2020

Background: High-dose chemo(radio)therapy (HDC) followed by auto-BM/PSCT is considered an acceptable therapeutic option for patients (pts) with R/R-HL. Various adverse prognostic factors have been previously described in this setting, including resistance to salvage therapy, active disease at the time of transplant, decreased performance status, B symptoms, extranodal disease, and complete remission duration of less than 1 year. On the other hand, overweight state (defined as BMI > or = 28) was associated with reduced overall and event-free survival among pts who received HDC followed by auto-BM/PSCT for non-Hodgkin’s lymphoma. The prognostic significance of excess body weight among pts with R/R-HL who undergo HDC followed by auto-BM/PSCT has not been reported.

Objective: To evaluate the relationship between excess body weight and overall survival (OS) in pts with R/R-HL who receive HDC followed by auto-BM/PSCT.

Methods: A retrospective, single institution study of consecutive pts with R/R-HL who received HDC followed by auto-BM/PSCT at the University of Oklahoma over the last 19 years (1985– 2004) was performed. BMI was calculated as [weight (kg)/height2 (m)]. Overweight state was defined as BMI > or = 28. OS was defined as the time period in months from the day of transplant until death, or the last of day of follow-up. Survival curves were estimated according to the Kaplan-Meier method, and were compared between the two groups (control group [BMI < 28] and overweight group [BMI > or = 28]) with the use of the log-rank test. Median survivals of the two groups were also compared by using Wilcoxon-Mann-Whitney test. A two-tailed P-value of < 0.05 was considered statistically significant. SAS® sofware (version 8.0; Carry, NC) was used for statistical analysis.

Results: 66 pts (40 males, 26 females) were identified, who were 13–69 years old (median 29 years) at the time of transplant. 40 pts (61%) died during follow-up, 4 pts (6%) were lost to follow-up at 15, 27, 58 and 103 months after transplant, and 22 pts (33%) are alive at the time of this analysis (7/30/04), 2–187 months (median 68 months) after transplant. 65 pts were included in this analysis (BMI could not be calculated for one pt). OS for the control group (n1= 42, 26/42 died) was 30 months (95% CI: 20–69 months) and for the overweight group (n2=23, 14/23 died) was 42 months (95% C.I. 8.0- N/A months). Overall survival did not differ between the two groups (p-values for log-rank and Wilcoxon-Mann-Whitney tests, 0.8109 and 0.7367, respectively).

Conclusion: Excess body weight is not a prognostic factor in patients with R/R-HL who undergo HDC followed by auto-BM/PSCT. A prospective study to evaluate and confirm our findings is warranted.
The Efficacy and Long-Term Toxicity of Intravenous Busulfan and Cyclophosphamide Conditioning Regimen for Stem Cell Transplantation in Advanced Lymphoma.
Authors: Santos Edgardo S. , DeCesare Teresa , Goodman Mark , Rosenblatt Joseph D. , Pereira Denise L. , Fernandez Hugo F.
Published: April 2020

Background: Although busulfan (Bu) in combination with cyclosphosphamide (Cy) is a commonly used conditioning regimen for the treatment of patients with hematologic diseases, its toxicity profile when administered intravenously (IV) as well as efficacy remains unclear, especially in the setting of autologous transplant for lymphoma.

Objective: To determine toxicity and the efficacy of the IV Bu/Cy conditioning regimen in patients who underwent autologous peripheral stem cell transplantation (APSCT) for the treatment of lymphoma.

Methods: A retrospective analysis was performed in 37 patients (21 male; 16 female) who underwent APSCT and received IV Bu/Cy regimen between January 2000 and May 2004.

Results: The median age was 50 years old (range: 20–68). Forty-seven percent of the patients were heavily pretreated (≥ 3 previous treatment, range 2–5). The distribution of lymphoma was: diffuse large B-cell lymphoma (DLBCL, 23), follicular lymphoma (8), mantle cell lymphoma (MCL, 5), and T-cell lymphoma (1). All patients received IV busulfan 3.2 mg/kg and cyclophosphamide 120 mg/kg, and were evaluable for toxicity. The patients engrafted (ANC ≥ 500 μ/l) at a median of 11 days (range: 9–14), and sustained platelet count (≥ 20,000 μ/l) at a median of 11 days (range: 8–42). The most common toxicities encountered were grade 1–2 including nausea, vomiting, stomatitis, esophagitis, and diarrhea. Grade 3–4 toxicities included hepatic veno-occlusive disease (HVOD, n=2), SIADH (n=1), and atrial fibrillation (n=2). Treatment related mortality (TRM) was seen in 3 patients due to HVOD (n=2) and septic shock (n=1). Twenty patients are still alive and 18 have no evidence of disease progression, with a median follow up of 17 months (range: 2–54 months). Kaplan-Meir actuarial OS at 24 months was 55.3% with a 47.3% DFS. Median event-free survival was 24 months (range 2–52).

Conclusions: IV Bu/Cy is a well-tolerated conditioning regimen with acceptable TRM, when compared with other regimens in heavily pretreated patients. The risk for HVOD is low. Further prospective trials are necessary to evaluate the role of Bu/Cy regimen in autologous stem cell transplant for lymphoma.
Immunotherapy Immediately Following Transplantation for Multiple Myeloma: Early Recovery of Aggressive Cytotoxic Cells and Improved Immune Recovery.
Authors: Bengtson Elizabeth , Wu Jia-Yan , Fitzmaurice Thomas , Cornell Cornelius J. , Ely Pamela , Gautier Marc , Hill John , Lowrey Christopher H. , Meehan Kenneth
Published: April 2020

Autologous peripheral blood stem cell transplantation (APBSCT) is effective for the treatment of multiple myeloma (MM), however the majority of patients relapse. Recent evidence shows an improved outcome if absolute lymphocyte count is increased immediatedly following APBSCT in patients with hematologic malignancy. We designed a phase II trial evaluating a dose escalation of SQ IL-2 and standard dose GM-CSF post-transplant for myeloma patients. Patients (n=18) received melphalan 200mg/m2 with GM-CSF 250mg/m2/day beginning on day 5. IL-2 began on the day of transplant and continued 5 days per week for 4 weeks. Peripheral blood samples were obtained at baseline (pre-transplant) and every week for 4 weeks post-transplant and evaluated using flow cytometry for cell subsets using antibodies directed against CD3, CD4, CD8, CD25 and CD56. Control patients (n=11) consisted of MM patients who received melphalan 200mg/m2 with either G-CSF or GM-CSF without IL-2. IL-2 at a dose of 1 X 106IU/m2/d was not tolerated in 2 of 6 patients due to grade 4 fatique and diarrhea (n=1) and grade 4 supraventricular arrhythmia (n=1). A dose of 6 X 105 IU/m2/d was well tolerated by 12 patients. Level 3 or greater toxicities included nausea (n=5), diarrhea (n=3), anorexia (n=11), mucositis (n=9). Engraftment of neutrophils occurred on day 12 and platelets on day 18. The LOS was 21 days in IL-2 treated cohort and 18 days in control group. Absolute lymphocyte counts on days 10–15 were increased an average of 152% in the IL-2 cohort (range 25–390%) over the control group. At day 21, there was a marked increase in the number of CD3+CD8+ cytotoxic T cells (43 %, +/− 7.5%) , CD56 +NK cells ( 78.3 %, +/− 9.1), and CD8+CD56+ NKT cells (36.8 %, +/− 9.8%) compared to patients’ baseline levels. Cytotoxicity of Day #21 PBMNC in the IL-2 cohort was strikingly increased at 28.1% (18.5 – 29.2%) compared to 5.5 % (3.59 – 6.52%) at baseline when tested against a human myeloma cell line (RPMI 8226) using chromium release assays (E: T ratio = 100:1). These results demonstrate a tolerable regimen of immediate post-transplant immunotherapy with marked increase in the number and function of early cytotoxic effector cells. The enhanced immune recovery may translate into an improved outcome.
Restoration of B Cell Development after Lentiviral Transduction of CD34+ Hematopoietic Cells from RAG-1 Deficient Patient.
Authors: Lagresle-Peyrou Chantal , Charneau Pierre , Hue Christophe , Mollier Karine , Andre-Schmutz Isabelle , Bonhomme Delphine , Hacein-Bey-Abina Salima , de Villartay Jean-Pierre , Fischer Alain , Cavazzana-Calvo Marina
Published: April 2020

Patients lacking expression of either RAG-1 or RAG-2 suffer from a Severe Combined Immuno-Deficiency (SCID) disease characterized by an early block in T and B lymphocytes differentiation leading to the absence of both mature lymphocyte subsets. This disease accounts for about 20% of SCID and the only curative treatment is hematopoietic stem cell transplantation, usually successful when an HLA-genoidentical donor is available. In the absence of such a donor, the success rate decreases along with the degree of HLA disparity between donor and recipient. Ex-vivo gene therapy of hematopoietic stem cells can be considered as an alternative treatment as a selective advantage of transgene-expressing cells is expected. Moreover, constitutive expression of only one of the two RAG proteins should not be harmful as concomitant expression of both genes is required for the recombination activity.

We used a lentivecteur containing the RAG-1 cDNA transgene as a therapeutic vector to transduce bone marrow CD34+ cells obtained from RAG-1 deficient patients. The transduced cells were injected into N0D-SCID mice previously irradiated (3Gy) and treated with an anti-TMb1 antibody. Ten weeks after transplantation, in all treated mice, 35±15% of the bone-marrow cells express the human CD45 marker. In this population, 24±2% co-express CD19 and IgM demonstrating that B cell differentiation capacity has been restored. We also detected some CD33+ cells attesting the presence of human myeloid progenitors cells. Altogether, these results suggest that both lymphoid and myeloid precursors have been transduced and demonstrate that gene transfer into hematopoietic cells can reconstitute B cell development in vivo.

Our data support the hypothesis that gene therapy could represent a possible alternative to bone marrow transplantation in RAG-1 deficient SCID disease.
Sustained Long Term Engraftment and Transgene Expression of Peripheral Blood CD34+ Cells Transduced with Advanced Lentiviral Vectors in the Preence of ABC Inhibitors.
Authors: Piacibello Wanda , Tesio Melania , Gammaitoni Loretta , Gunetti Monica , Pignochino Ymera , Aglietta Massimo
Published: April 2020

Because mobilized peripheral blood (MPB) represents an attractive source of cells for gene therapy, we investigated the ability of advanced-generation lentiviral vectors (LV) to transfer the Enhanced Green Fluorescent Protein (EGFP) gene into CD34+ cells isolated from MPB in culture conditions that allowed expansion of transplantable human HSC. So far, a few studies reported transduction of MPB cells with LV pseudotyped with VSV-G. However, the critical issue remains whether primitive, hematopoietic repopulating cells have indeed been transduced.

In vitro and in vivo experiments provided here show that MPB CD34+ cells can be effectively transduced by advanced generation LV: they do not lose their proliferation potential, self renewal and multilineage differentiation ability, moreover transgene expression is maintained. Following 48 hour-exposure to FL+TPO+SCF+IL6 and overnight incubation with vector particles, MPB CD34+ cells were further cultured for up to six weeks. During this period of culture the CD34+ cells population, the CD34+/GFP+ fraction and the CFC output were maintained.

Transduction efficiency of NOD/SCID repopulating cells (SRC) was assessed by serial transplants into NOD/SCID mice. Primary mice transplanted with transduced cells showed high level of engraftment (11,5 % CD45+ cells): within the CD45+ cells 11,4% expressed GFP (n=15). Moreover, mice transplanted with transduced, week-1 expanded cells showed higher levels of human engraftment than those transplanted with transduced, non-expanded cells (range from 9.25 % to 10.5 % for transduced basal cells; range from 14, 3 % to 38,19 % for transduced 1-week expanded cells). Engraftment was multilineage, with GFP+/lineage+ cells. Serial transplants were performed with transduced, week 1-expanded cells. Secondary engraftment levels were 1.6 %; 19.3 % of human cells were GFP+ (n=3). Moreover, levels of human engraftment were higher in mice injected with transduced, 1- week expanded cells (3.5 %) rather than in those injected with transduced, non expanded cells (1.2 %). Engraftment was multilineage with GFP+/lINEAGE+ cells.

These results show that LV efficiently transduced HSC, that maintain proliferation and self-renewal ability.
Use of RNA Interference to Inhibit Integrin Subunit alphaV-Mediated Angiogenesis.
Authors: Graef Thorsten T. , Steidl Ulrich U. , Fenk Roland R. , Rohr Ulrich U. , Nedbal Wolfgang W. , Haas Rainer R. , Kronenwett Ralf R.
Published: April 2020

Inhibition of angiogenesis is a promising approach for the treatment of solid tumors, inflammatory diseases and different hematological malignancies such as multiple myeloma. One of the central molecules in capillary formation during angiogenesis is the integrin alphaVbeta3 (aVb3), therefore aVb3 is a potential target molecule to inhibit angiogenesis. The aim of this study was to inhibit alphaV-mediated angiogenesis in vitro using RNA interference (RNAi) technology as well as antisense oligodeoxyribonucleotides (asON). We used synthetic small interfering RNAs (siRNA) and asON directed against the alphaV chain of aVb3 to inhibit integrin expression. Five siRNAs were selected on the basis of a systematic alignment of computer-predicted secondary structures of target mRNA and on the basis of current recommendations for siRNA oligonucleotide design. In parallel, 3 asON were examined. They had the sequence of the antisense sequence of 3 of the siRNAs molecules, respectively. SiRNAs, asON and respective control sequences were transfected into human umbilical vein endothelials cells (HUVEC) using lipofection. Following stimulation by phorbol 12-myristate 13-acetate (PMA), two siRNAs showed a dose- and time dependent inhibition of PMA-induced aV-mRNA and -protein upregulation as assessed by real-time RT-PCR and flow cytometry. At a concentration of 25 nM a 100% (SD: 4.9%) inhibition of aV upregulation was observed, whereas transfection of the respective asON sequences resulted in a 63% (SD: 6.1%) inhibition of aV upregulation at 25nM. To evaluate the anti-angiogenic potential of siRNAs in comparison to asON a cell culture model of human angiogenesis based on the co-cultivation of endothelial cells and dermal fibroblasts was used. Transfection of the most efficient siRNA sequence at a concentration of 50nM resulted in an inhibition of total length of capillary-like tubules by 48.7% (SD: 3.6%) in comparison to 21.8% (SD: 9.8%) by the respective asON sequence treated cultures.

In conclusion, siRNAs can successfully be selected on the basis of computer-predicted secondary structures. In comparison with asON having the same sequence as the antisense strand of the respective siRNA the siRNA-mediated inhibition of aV expression showed a stronger inhibition of capillary tube formation in an angiogenesis in vitro assay. Therefore, siRNAs are useful tools for functional aV knock-down experiments and might be a therapeutic alternative for antagonists which bind directly to the integrins aVb3 or aVb5.
Prevalence of Antiphospholipid Anitbodies in Idiopathic Venous Thromboembolism.
Authors: Paramsothy Koushi , Wells Philip S.
Published: April 2020

The presence of antiphospholipid antibodies (APLA), which include lupus anticoagulant (LA) and anticardiolipin antibodies (ACA), increases the risk of venous thromboembolism (VTE). Although, the association demonstrated between LA and VTE is strong, that between ACA and VTE is weak with the exception of high titers. This suggestion has been that this may be related to the induction of ACA by acute illness. This may be the explanation for the widely varying frequency of ACA positivity in patients with acute VTE in the current medical literature. The prevalence of LA in patients with acute VTE is more consistent but the value of testing for ACA and LA at presentation of acute VTE remains unknown. We routinely test all idiopathic VTE patients at presentation for LA and ACA. It had been our impression that most positive results revert to normal with subsequent testing. We sough to test this hypothesis. The objective was to determine the prevalence of abnormal LA and ACA results and the frequency of subsequent normalization in patients presenting with a first idiopathic venous thromboembolic event. A retrospective chart review was conducted at the Thrombosis Unit at the Ottawa Hospital. 278 charts of unrelated, consecutive patients with idiopathic VTE were reviewed. 232 patients with single idiopathic venous thromboembolic events with documented LA and ACA results were included. The patients were divided into two groups based on the time between the acute VTE and initial APLA test. The frequency of abnormal LA and ACA on initial and repeat tests was determined in each group. 170 and 177 patients were screened for LA and ACA within one month of the acute VTE. On initial tests, LA and ACA were detected in 14.1% and 7.9% of patients, respectively. However, LA and ACA were present in only 1.2% and 1.7% of patients, respectively, on subsequent tests. In patients screened more than one month after an acute VTE 3.8% of the 52 patients had evidence of LA on initial testing and all were positive on repeat tests. ACA were present in 8% of patients on initial tests and in 6% (75% of the initial positive results) of patients on subsequent tests. A high frequency of positive LA and ACA tests results is observed when these tests are performed at the time of presentation of acute VTE. Repeat testing beyond one month from presentation demonstrates most of the results return to normal suggesting false positive results are common at presentation. However, when the tests are performed one month or longer after the acute VTE, the frequency of positive APLA test is much lower and false positives are uncommon. Screening for APLA should not be performed until at least one month after the diagnosis of idiopathic.
Effect of Chemotherapy on the Distress and Quality of Life of Cancer Patients.
Authors: Hussain Mirsa B. , Sarita Gangadharan P. , Devi Nandakumar , Thomas Bejoy C. , Rita Krishnan , Ramdas Kunnambath , Rajan Balakrishna , Pandey Manoj , Narayanan Geetha N.
Published: April 2020

Introduction: The deteriorating effects of chemotherapy on cancer patients are well documented, as is the need and impact of psychosocial interventions in improving quality of life (QOL). In India, cancer centres have a very high patient load and providing quality treatment and achieving good survival is still the first priority. Published studies from India on QOL in cancer patients are far and few, current ongoing studies are even fewer. The present study is on effect of chemotherapy on QOL, and prevalence of psychological morbidity in the Indian context.

Patients and Methods: The study sample consisted of 117 cancer patients undergoing chemotherapy with curative intent as single modality or as part of the multi modality regimen. Nearly 30% of the sample had lymphoproliferative disease, 17% had haematological malignancies, 45% had solid tumours, and the rest had multiple myeloma. The assessments were carried out using the Distress Inventory for cancer version 2 (DIC V2), Functional Assessment of Cancer Therapy-General (FACT-G), and the Hospital Anxiety and Depression scales (HADS).

Results: Older patients (>=47 years) had significantly greater spiritual distress, and difficulties in activities of daily living, but had better emotional well being compared to younger patients (<47 years). Patients from lower income strata had significantly greater distress levels and poorer QOL than those in the upper income category. Those with advanced disease had significantly poorer scores on most of the QOL subscales. More than 20% of the patients were found to have significant psychological morbidity ranging from mood disturbances (12%), to anxiety (3%) and depression (5%). Though chemotherapy did not significantly influence anxiety and depression, it was noted that male patients had significantly higher depression rates than females.

Conclusions: The study was intended to measure the existence of psychological morbidity within the cancer patients. Older patients, those from lower incomes groups, and those with advanced disease had significantly greater levels of distress and significantly poorer QOL scores. Depression rates were significantly higher in male patients than in female patients. This shows the need to have psychiatric assessment and intervention among patients undergoing cancer chemotherapy.
Hematopoietic Growth Factors Reduce Side Effects of Cancer Treatment: Results of Two Comprehensive Meta-Analyses.
Authors: Bohlius Julia , Engert Andreas
Published: April 2020

Background: Hematopoietic growth factors such as G-CSF/GM-CSF and erythropoietin (EPO) are used in clinical practice to reduce side effects of cancer treatment but also to improve tumour response and overall survival. We conducted two meta-analyses on the effects of hematopoietic growth factors in the treatment of malignant diseases a) to determine the effectiveness of G-CSF and GM-CSF to i) prevent infection, ii) improve tumor response and overall survival in lymphoma patients undergoing standard chemotherapy and b) to assess the effectiveness of EPO to i) prevent anemia, ii) improve tumor response and overall survival in cancer patients.

Methods: The Cochrane Library, Medline, Embase and smaller databases were searched. Randomized controlled trials comparing G-CSF/GM-CSF or EPO versus placebo/no treatment were included in the reviews. Both study arms had to receive identical antineoplastic treatment and identical additional supportive care. We included full-text and abstract publications as well as unpublished data. Data were meta-analysed using fixed and random effects models.

Results: Twenty one trials with 2,794 adults were included in the EPO meta-analysis. Use of EPO significantly reduced the relative risk of red blood cell transfusions (RR 0.67; 95% CI 0.62 to 0.73). For participants with baseline hemoglobin below 10 g/dL hematological response was observed more often in participants receiving EPO (RR 3.60; 95% CI 3.07 to 4.23). There was inconclusive evidence whether EPO improves tumour response (random effects: RR 1.21; 95% CI 0.92–1.59) and overall survival (unadjusted data: HR 0.84; 95% CI 0.69 to 1.02). 12 trials with 1,823 randomized patients were included in the G-CSF/GM-CSF meta-analysis. Compared with no prophylaxis, G-CSF/GM-CSF significantly reduced the relative risk for infections (RR 0.74; 95% CI 0.64–0.85). There was no evidence for G-CSF/GM-CSF to decrease infection related mortality (RR 1.37; 95% CI 0.66–2.32), to improve complete response (RR 1.02; 95% CI 0.94–1.11) or overall survival (HR 1.00; 95% CI 0.86–1.16).

Conclusions: There is consistent evidence that the administration of EPO and G-CSF/GM-CSF reduces the side effects of antineoplastic therapy, such as risk for blood transfusions (EPO) and infections (G-CSF/GM-CSF). However, there is inconclusive evidence whether the administration of hematopoietic growth factors in these clinical settings also improve tumor response and overall survival.
Detection of Cytokeratin-19 in Peripheral Blood of Breast Cancer Patients: Results from a Systematic Review/Meta-Analysis.
Authors: Soares Heloisa P. , Santucci Rodrigo , Kumar Ambuj , Djulbegovic Benjamin , Glasberg Joao , Del Giglio Auro
Published: April 2020

Background: The identification of occult micrometastatic disease is potentially important in management of breast cancer (BC) patients. Therefore, detection of cytokeratin 19 (CK-19) and others molecular markers using reverse transcriptase polymerase chain reaction (RT-PCR)-based techniques to detect micrometastases could have significant implications in prevention, early detection and treatment of breast cancer.

Objective: To assess the diagnostic specificity and sensitivity of detection of CK-19 in the peripheral blood (PB) of BC patients as well as to assess its relation to prognostic factors and implication on survival.

Methods: We conducted a systematic review/meta-analysis of all studies that analyzed CK-19 in the PB of BC patients. We searched MEDLINE, LILACS and Cochrane Database of Systematic Reviews (last search June 2004). Studies that had at least 10 BC patients and controls and evaluated the presence of CK-19 in PB using RT-PCR were included in this review. Diagnostic sensitivity was defined as the percentage of detection of CK-19 in patients with histological proven diagnosis of breast cancer. The specificity was defined as the percentage of negative tests for CK-19 in the control group (healthy individuals or non breast cancer patients).

Results: Out of 1842 relevant studies that were identified, 26 met our inclusion criteria, enrolling 2398 patients (1176 treatment and 1222 controls). Results showed that PCR had an overall diagnostic sensitivity of 0.39 (95% CI: 0.36–0.42) and diagnostic specificity of 0.83 (95% CI: 0.76–0.87) regardless of the breast cancer stage. Six studies researched the association between CK-19 and well established prognostic factors (i.e. positive axillary lymph nodes, hormonal receptors, tumor size); however data were not available for quantitative synthesis. Survival was poorly reported making impossible to generate any conclusion about the implication of detection of CK-19 on prognosis. When studies were critically appraised, we also identified many methodological weaknesses in the design, conduct and the analysis of these studies (e.g. use of multiple control arms, use of non-consecutive patients, different timing of sample collection, inclusion of all stages of breast cancer and differential work-up in the study & control groups) hence resulting in biased findings due to selection, verification, detection and spectrum bias.

Conclusion: The existing body of evidence regarding the assessment and prognostic value of CK-19 using RT-PCR techniques is poor and needs ample improvement before the initiation of its clinical application.
The Role of Race on Survival after Alternative Donor Hematopoietic Progenitor Cell Transplant for Pediatric Acute Leukemia: Provocative Single Center Data.
Authors: Margolis David A. , Eapen Mary , Carreras Jeanette , Talano Julie-An , Browning Meghen , Casper James
Published: April 2020

Allogeneic blood or bone marrow transplant (BMT) can be a curative treatment for many children and adolescents with acute leukemia. With advances in unrelated donor transplant, others and we have shown that unrelated donor BMT can have similar survival to matched sibling BMT. There are several reports describing outcomes after matched related donor transplantation among various ethnic groups. Thus far, there are no published studies comparing outcomes among ethnic groups after alternative donor transplantation. Anecdotally, however, there have been concerns regarding outcomes among racial and ethnic groups, especially African-Americans. In order to address this question, we utilized our institutional database to analyze survival among children and adolescents receiving an alternative donor BMT at Children’s Hospital of Wisconsin from 1988-present. We compared survival in Caucasians and African-Americans undergoing unrelated donor and mismatched related donor transplantation (including haploidentical donors). One hundred and twenty four Caucasians underwent matched and mismatched unrelated donor transplantation compared to 11 African Americans. The 2-year probabilities of overall survival were significantly better for Caucasians at 53% (95% CI 44–62) than for African Americans, 18% (95% CI 2–45), p=0.01. Fifty-four Caucasians and 9 African Americans received mismatched family donor transplantation. Corresponding probabilities of overall 2-year survival were 38% (95% CI 25–51) and 30% (95% CI 5–64), respectively. Interestingly, our data show no statistically significant difference in survival after mismatched related donor transplantation between the Caucasian and African-American cohorts. Our data should be interpreted cautiously as the number of African Americans transplanted at our institution is few. Additionally, our analysis is limited by our inability to adjust for disease status at transplantation, HLA disparity and other known risk factors that may impact survival. Nevertheless these observations from a single institution cannot be ignored and warrant further analysis in a larger cohort such that outcomes after transplantation may be adjusted appropriately for relevant risk factors. We believe that a national database/registry study will have the numbers necessary to answer the questions that need to be asked regarding outcomes with alternative donor transplantation in the African-American population. We also believe that as cell processing and supportive care technologies improve mismatched family member transplantation outcomes, these advances could have a significant impact in improving leukemia-free survival for African-American children and adolescents.
Third Autotransplant for the Management of 98 Patients among 1358 Who Had Received Prior Tandem Autotransplants: Benefit Apparent When 2nd to 3rd Transplant Interval Exceeds 3 Years.
Authors: Lee Choon Kee , Barlogie Bart , Fassas Athanasios , Anaissie Elias , Zangari Maurizio , Rasmussen Erik , Tricot Guido
Published: April 2020

Tandem autotransplants have recently been reported in the IFM 94 randomized trial to confer more durable event-free survival (EFS) and overall survival (OS) than a single transplant. In the Arkansas program, scheduled tandem transplants have been part of Total Therapy 1 (TT 1) and Total Therapy 2 (TT 2) protocols since 1989 for newly diagnosed patients; this strategy was also applied for the management of previously treated patients. Among a total of 1358 patients receiving two prior autologous transplants at our institution (median interval from 1st to 2nd transplant, 4 mos), 98 received a further autologous transplant. In a search for features associated with favorable post 3rd autotransplant survival, pre-transplant 1, 2 and 3 features, especially the presence of cytogenetic abnormalities (CA) and standard laboratory features such as, B2M, CRP, LDH, albumin, hemoglobin and creatinine were examined along with the time interval from 2nd to 3rd transplant and whether a 3rd transplant was applied to rescue TT 1 or TT 2 front line or other patients receiving tandem transplants as salvage therapy. Remarkably, the presence of CA at any of the indicated time points had no impact on survival after a 3rd autotransplant usually with melphalan 200mg/m2. According to multivariate analysis, the 2nd to 3rd transplant interval (time-dependent covariate analysis) (HR 8.8, p=.002) and hypoalbuminemia < 3.0g/dL (HR 3.2, p<.0001) were independently important. Thus, post 3rd tx survival was superior among the 41 patients with a post-2nd tx survival >3 yr and albumin ≥ 3.0g/dL (median, 21 mos; 5 yr OS 20%), with an intermediate outcome noted for the 34 patients with one of these favorable features present (median, 10 mos; 3 yr 20%) while the worst outcome was noted in the 13 patients who were both hypoalbuminemic and had received their 3rd transplant within 3 yrs of the 2nd transplant (median, 3 mos, no survival beyond 12 mos). Our data indicate that meaningful survival can be obtained with a 3rd autotransplant maneuver in approximately 50% of patients who have derived meaningful survival from a 2nd transplant of at least 3 years and have a preserved albumin concentration as an indication of their overall performance status.

Figure Figure
Expression of Endomucin, a CD34-Like Sialomucin, Marks Definitive Hematopoietic Stem Cells throughout Development.
Authors: Maeda Azusa , Iwama Atsushi , Eto Koji , Ema Hideo , Kitamura Toshio , Vestweber Dietmar , Nakauchi Hiromitsu
Published: April 2020

In order to identify cell surface molecules specific to hematopoietic stem cells (HSCs), a modified signal sequence trap was applied to mouse bone marrow (BM) CD34− c-Kit+ sca-1+ lineage− (CD34−KSL) cells which is highly enriched for HSCs. Among the identified genes, mRNA expression of Endomucin, an endothelium-specific gene encoding a CD34-like sialomucin, appeared highly specific to CD34-KSL HSCs.

To further investigate the expression of Endomucin, we generated two rat anti-mouse Endomucin monoclonal antibodies that recognize different epitopes (AE2D4, AE7F2). Taking advantage of these and another monoclonal antibody, V7c7 (1999, Blood, 93; 1; 165), detailed expression analysis was performed. Endomucin expression was largely confined to lineage markers-negative (Lin−) cells. Approximately 7 % of Lin− cells were Endomucin-positive. Cells strongly expressing Endomucin represented 30% of c-kit+ sca-1+ cells. Gating out CD34+ cells from Lin− Endomucin+ population resulted in high yield of KSL cells. High correlation between Lin− Endomucin+CD34− cells and KSL cells was confirmed by in vivo bone marrow transplantation. When Lin− cells were fractionated by their expression of CD34 and Endomucin, only Lin− Endomucin+CD34− cells contributed to long-term repopulation (LTR), and as few as 100 cells were enough to obtain engraftment. Furthermore, the majority of CD34−KSL cells were Endomucin+, and again, only CD34−KSL-Endomucin+ cells had LTR activity. These data indicate two facts: 1) A single positive marker, Endomucin can substitute for c-kit+ sca-1+, 2) All LTR -HSCs express Endomucin.

We then analyzed the expression of Endomucin during embryonic development of the hematopoietic system. Definitive HSCs arise from the hemogenic endothelium lining the wall of the dorsal aorta in embryonic aorta-gonads-mesonephros (AGM) region, then seed to the fetal liver. E10.5 AGM CD45− cells were segregated into subpopulations by their expression of Endomucin and CD41, an early marker of embryonic hematopoiesis. In vitro coculture system with a stromal cell line, OP9, was applied to detect the ability of hematopoietic potential. Hematopoietic activity was exclusively found in the CD41+Endomucin+ population, that represents 24% of CD41+ cells. Taken together, these data indicate that Endomucin marks both embryonic and adult HSCs, providing a novel useful cell surface marker for definitive HSCs throughout development.

Figure Figure
Significant Differences in the IGVH and BCL6 Mutation Status in Aggressive B-Cell Lymphomas with and without MYC Breakpoints.
Authors: Pott Christiane , Trautmann Heiko , Harder Lana , Kneba Michael , Siebert Reiner ,   on behalf of the MMML-Network
Published: April 2020

Somatically mutated IGVH regions are a hallmark of germinal center (GC) B-cells. Moreover, aberrant somatic hypermutation (SHM) of oncogenes, like changes in the 5′ non-coding region of the BCL6 gene occurring in 75% of DLBCL, are a mechanism of oncogene activation independently of chromosomal translocations.

Large scale mutational screening using DHPLC and direct sequencing was applied to determine the somatic hypermutation status of clonal IGVH as well as several oncogenes like BCL6 and MYC in a series of more than 250 aggressive B-cell lymphomas included in the Deutsche Krebshilfe funded network “Molecular Mechanisms in Malignant Lymphoma”. Mutation patterns were correlated with the results of molecular cytogenetic analyses. MYC breakpoints detected by FISH clearly differentiated two groups of aggressive B-NHL with significantly different VH mutation status. MYC-positive lymphomas by FISH carried VH genes with a mutation frequency significantly lower compared to aggressive lymphomas lacking these features (median 4.8% vs.11.,0%, p<0.0001). Similarly, the mutation frequency of BCL6 was significantly lower in MYC-positive than in MYC-negative lymphomas (median 0.13% vs 0.25% p=0,020). We observed a bias in VH gene usage in both groups with an overrepresentation of VH4 (40% in both groups) and VH3 gene (24% in MYC-positive, 40% in MYC-negative). The group of MYC-positive lymphomas was heterogeneous with regard to the pattern of chromosomal aberrations. (IGH, MYC, BCL2, BCL6, MALT1 and REL loci). Lymphomas with IG-MYC fusion lacking breakpoints in BCL2, BCL6, MALT1 or REL loci as well as non-MYC associated IGH translocations displayed a median IGVH mutation frequency of 4,8% vs. 12,2% in those cases with additional chromosomal translocations (p=0.0060).

Molecular classification of aggressive B-cell lymphomas according to MYC breakpoints distinguishes subgroups with significant differences in the VH and BCL6 mutation frequencies.
Expression of Midkine(MDK) and Sensitivity to Anti-Leukemic Drugs in Pediatric Acute Lymphoblastic Leukemia (ALL).
Authors: Zeng Weihua , Chen I-Ming , Tibbitts Justin , Kang Huining , Ar Kerem , Atlas Susan , Willman Cheryl L.
Published: April 2020

Midkine (MDK), a heparin-binding growth factor, is highly expressed in pediatric ALL, neuroblastomas, and various solid tumors. Previous gene expression profiling studies conducted in our laboratory on a cohort of 220 children with B precursor ALL treated on Children’s Oncology Group (COG) clinical trials have shown that higher levels of MDK expression in leukemic blasts are strongly associated with continuous complete remission (CCR) and that MDK is a powerful prognostic predictor of outcome (Wilcoxon ranksum test, p=0.000001), particularly in patients with central nervous system (CNS) involvement (Wilcoxon ranksum test, p =0.009). To explore the role of MDK in hematopoiesis and leukemogenesis and to further explore MDK as a potential novel target for ALL therapy, we examined MDK mRNA expression in three stages of normal B cell development in healthy controls and compared it to expression levels in leukemic blasts from children with B-ALL. Quantitative RT-PCR assays showed that MDK mRNA levels in CD19+ cells from peripheral blood mononuclear cells (PBMNC) were significantly higher than those in CD34+ or CD19+ cells from bone marrow (BM) (Mann Whitney test, p=0.0025 and 0.0018, respectively). ALL leukemic blasts samples expressed significantly higher levels of MDK than those in CD19+ cell in the BM of healthy controls (Mann Whitney test, p=0.0043). Similar results were obtained by ELISA detection of MDK protein in plasma of ALL patients and healthy controls. We have also investigated the relationship of MDK expression and the chemo-sensitivity of ALL cell lines. Using a lentiviral system, we have successfully over-expressed or knocked down MDK gene expression in the REH cell line containing the t(12;21). Unexpectedly, MTS assays revealed that MDK expression is inversely associated with sensitivity to L-asparaginase and doxorubicin, but not to vincristine and cytarabine, in REH cells. Higher levels of MDK were associated with chemo-resistance. Finally, we tested the effects of MDK expression on proliferation and apoptosis in REH cells and determined that although MDK expression was not associated with REH cell proliferation in vitro, it protected REH cells from doxorubicin-induced apoptotic cell death. Taken together, these results suggest that expression levels of MDK in ALL is inversely associated with in vitro drug sensitivity to specific anti-leukemic agents; consistent with observations in solid tumor models. It has been suggested that 4β1- and 6β1-integrins bind directly to MDK and are involved in MK-dependent cell migration. Further studies of the role of MDK signaling in vivo in the complex marrow/adhesion microenvironment and the effects on ALL survival and chemo-sensitivity and resistance are in progress.
Two Distinct Subsets of dic(9;12)(p12;p11.2) among Children with B-Cell Precursor Acute Lymphoblastic Leukemia (ALL): PAX5-ETV6 and ETV6-RUNX1 Rearrangements: A Report from the Children’s Oncology Group.
Authors: Gastier-Foster Julie M. , Carroll Andrew J. , Ell Denise , Harvey Richard , Chen I-Ming , Ketterling Rhett , Meloni-Ehrig Aurelia , Opheim Kent E. , Patil Shivanand , Pettenati Mark , Rao Kathleen , Wu ShiQi , Heerema Nyla A.
Published: April 2020

The dic(9;12)(p12;p11.2) has been described as a rare cytogenetic abnormality in pediatric precursor B-cell ALL. Initial studies suggested that the rearrangement is associated with a favorable outcome, and recent studies demonstrated the presence of a PAX5-ETV6 fusion gene was associated with this cytogenetic abnormality. Twenty cases with a cytogenetic dic(9;12) were identified in the Children’s Oncology Group (COG) cytogenetics databases. FISH analysis with the ETV6-RUNX1 (TEL-AML1) probes was done on 12 of these samples. Five cases were positive for fusion, indicating a cryptic t(12;21)(p13;q22), and also had loss of the ETV6 probe from the chromosome 12 not involved in the t(12;21). Seven cases were negative for fusion and had loss of an ETV6 signal, although one of the latter had a diminished ETV6 signal identified. To determine whether both PAX5-ETV6 and ETV6-RUNX1 rearrangements occurred in some patients, a diagnostic sample from each patient was analyzed by RT-PCR for the PAX5-ETV6 and ETV6-RUNX1 fusion genes. Primers from exon 3 of PAX5 and exon 3 of ETV6 were used for the PAX5-ETV6 analysis and from exon 5 of ETV6 and exon 4 of RUNX1 for the ETV6-RUNX1 analysis. Of the 20 cases, only 8 were RT-PCR positive for the PAX5-ETV6 fusion with the above primers; however, an additional 2 were RT-PCR positive with alternate primers, and all 10 of these were negative for the ETV6-RUNX1 fusion by RT-PCR. Of the remaining 10 patients, 9 were RT-PCR positive for the ETV6-RUNX1 fusion, including all of the ETV6-RUNX1 cases positive by FISH. The gene rearrangement associated with the dic(9;12) in these cases is not known. One patient was negative for both fusions by RT-PCR, negative by FISH for ETV6-RUNX1 rearrangement, yet had loss of an ETV6 signal. No cytogenetic differences could be seen between the 2 groups, either in the appearance of the dic(9;12) or in the other abnormalities identified. These results demonstrate the presence of two mutually exclusive dic(9;12) rearrangements in pediatric ALL; one associated with ETV6-RUNX1 rearrangement and one resulting in PAX5-ETV6 fusion. Both PAX5-ETV6 and ETV6-RUNX1 rearrangements are associated with a favorable prognosis. However, molecular analysis of the dic(9;12) patients must be performed to determine whether the dicentric chromosome results in PAX5-ETV6 fusion or whether the case has ETV6-RUNX1 fusion.
A Two-Gene Classifier for Predicting Response to the Farnesyltransferase Inhibitor Tipifarnib in Acute Myeloid Leukemia.
Authors: Raponi Mitch , Lancet Jeffrey E. , Fan Hongtao , Dossey Lesley , Lee Grace , Gojo Ivana , Feldman Eric J. , Gotlib Jason , Morris Lawrence E. , Greenberg Peter L. , Wright John J. , Harousseau Jean-Luc , Lowenberg Bob , Stone Richard M. , De Porre Peter , Wang Yixin , Karp Judith E.
Published: April 2020

Currently there is no method available to predict response to farnesyltransferase inhibitors (FTI). We analyzed gene expression profiles from the bone marrow of patients from a phase 2 study of the FTI tipifarnib (R115777, ZARNESTRA), in older adults with previously untreated acute myeloid leukemia (AML). The RASGRP1:APTX gene expression ratio was found to predict response to tipifarnib with the greatest accuracy. This two-gene ratio was validated by quantitative PCR in the newly diagnosed AML cohort. We further demonstrated that this classifier could predict response to tipifarnib in an independent set of 54 samples from relapsed or refractory AML, with a negative predictive value and positive predictive value of 92% and 28%, respectively (odds ratio of 4.4). The classifier also predicted for improved overall survival (154 vs 56 days, p = 0.0001), which was shown to be independent of other prognostic factors including a previously described gene expression classifier predictive of prognosis. Therefore, these data indicate that a two-gene expression assay may have utility in categorizing a population of AML patients who are more likely to respond to tipifarnib.