bluuebunny/crossref_metadata_2025 huggingface dataset:
Off-Label Use of Recombinant FVIIa: Clinical Characteristics That May Influence Outcomes.
Authors: Saito Yoriko , Healy Brian , Kaufman Richard M. , Dzik Walter H.
Published: April 2020

Recombinant activated factor VII (rFVIIa) is increasingly being used in off-label settings. To understand patient characteristics that may affect outcomes in patients who receive off-label rFVIIa, a retrospective review of off-label rFVIIa use from 1/1999 through 3/2005 was conducted at two tertiary-care academic hospitals. Through a hospital IRB-approved protocol, patient charts were examined for all 117 consecutive patients who received off-label rFVIIa. Overall 30-day mortality was 75/117 (64%; 95% CI [0.55 - 0.72]). Overall incidence of new renal dysfunction (serum creatinine>1.3mg/dL for females, >1.5mg/dL for males) or worsening renal dysfunction (>50% increase in creatinine in patients with pre-existing renal dysfunction) was 31/108 (28.7%; 95% CI [0.21 - 0.38]). Thromboembolic complications were documented in 9/117 (7.6%; 95%CI [0.04 - 0.14]). Multiple clinical characteristics were recorded and the Fisher’s exact test was used to determine which characteristics were associated with either an increased (or decreased) frequency of renal dysfunction within 72 hours of rFVIIa administration; or with an increased (or decreased) frequency of 30-day mortality. The characteristics that showed statistically significant association in univariate analysis are listed in the table. The incidence of renal failure was significantly increased over baseline (29%) for patients with either one of 2 characteristics (rows g,h). The incidence of death was significantly increased over baseline (64%) for patients with any of 6 clinical characteristics (a – f) and decreased compared to baseline patients with 3 characteristics (i–k). Characteristics which showed no statistically significant association for either outcomes were: intracranial hemorrhage, subdural hematoma, gastrointestinal bleeding, post-procedural bleeding, trauma, surgical intensive unit, hx of solid tumor, hx of coronary artery disease, hx of atrial fibrillation, hx of diabetes mellitus, gender and age. This analysis helps identify covariant clinical features to be considered when selecting a matched control group to examine patient outcomes resulting from off-label rFVIIa use.

Patient Clinical Characteristics n Renal Dysfunction; Odds Ratio [95% CI] Death; Odds Ratio [95 % CI] *p<0.005; **p<0.05; †p>0.05; ¶One-sided Odds Ratio is reported because no patient survived. a Sepsis 23 0.49 [0.11–1.68]† >4.73 *,¶ b DIC 15 0.16 [0.004–1.2]† >2.67 *,¶ c BMT/Stem Cell Transplant 15 0.64 [0.11–2.7]† >2.67 *,¶ d Heme malignancy 21 0.41 [0.07–1.6]† 14.9 [2.2–632.6]* e Medical ICU 36 1.2 [0.45–3.2]† 10.2 [2.8–55.2]* f Pre-existing renal dysfunction 53 0.42 [0.16–1.1]† 4.7 [1.8–12.7]* g Liver disease/Ped liver transplant 23 3.19 [1.02–9.78]** 0.54 [0.19–1.5]† h Cardiac/thoracic surgery 18 3.1 [0.95–9.9]** 0.86 [0.28–2.9]† i Anticoagulant-related bleeding 19 0.64 [0.11–2.7]† 0.19 [0.06–0.62]* j Surgical prophylaxis 6 0.48 [0.01–4.6]† 0 [0–0.32]*
The Target Receptor Siglec-3 (CD33) Is Expressed on AML Stem Cells in a Majority of All Patients with AML.
Authors: Hauswirth Alexander W. , FLorian Stefan , Krauth Maria-Theresa , Schernthaner Gerit-Holger , Selzer Edgar , Wacheck Volker , Printz Dieter , Fritsch Gerhard , Sotlar Karl , Sperr Wolfgang R. , Valent Peter
Published: April 2020

The cell surface antigen Siglec-3 = CD33 is becoming increasingly important as target of therapy in acute myeloid leukemia (AML). In particular, a conjugate consisting of the humanized CD33 antibody P67.6 (gemtuzumab) and the cytostatic drug calicheamicin has been developed for clinical use and was found to work as an effective antileukemic agent (Mylotarg®) in patients with CD33+ AML. In normal myelopoiesis, expression of CD33 is restricted to advanced stages of differentiation, whereas primitive stem cells do not express CD33 (Siglec-3). In line with this notion, CD33-targeting therapy is a non-myeloablative approach. In the present study, we asked whether leukemic stem cells in patients with AML express CD33. For this purpose, a multicolor-staining technique was applied in eleven patients with AML. Leukemic stem cells were defined as CD34+/CD38−/CD123+ cells. In all patients in whom the majority of myeloblasts expressed CD33 (=CD33+ AML, n=8), the AML progenitor cells reacted with the CD33 antibody P67.6. Repopulation experiments utilizing NOD/SCID mice confirmed that the AML stem cells in these patients reside within the CD33+ subpopulation of leukemic cells. Moreover, AML stem cells (CD34+/CD38−/CD123+ cells) highly purified (>98% purity) from patients with (CD33+) AML by cell sorting, were found to express CD33 mRNA in RT-PCR analyses. To demonstrate that AML stem cells can also reside within the CD33-negative fraction of the AML clone, we purified CD33-negative cells in a patient with AML in whom a majority of leukemic stem cells were found to lack CD33. In this particular patient, the CD33-negative cells were found to repopulate NOD/SCID mice with leukemias in the same way as the entire leukemic clone did. The CD33 antigen was neither detectable on CD34+/CD38− cells in the normal bone marrow nor on leukemic stem cells in patients with CD33-negative AML. In summary, our data show that leukemic stem cells in patients with CD33+ AML frequently express the target receptor CD33. This observation is in favor of novel treatment concepts employing CD33-targeting antibodies (Mylotarg®) in acute myeloid leukemia.
Complex Translocation (8;16;21); a New Variant of t(8;21), with t(13;22) in Acute Myeloid Leukemia.
Authors: Gupta Shiphali , Brody Judith , John Veena , Koduru Prasad
Published: April 2020

The translocation (8;21)(q22;q22) is a common recurrent chromosome aberration present in 10% to 15% of all acute myeloid leukemias. In approximately 3% of all cases with t(8;21), a variant of t(8;21) involving chromosomes 8,21 and other chromosomes is present. We report here a case of AML-M2 with a new complex translocation (8;16;21)(q22;p16.3;q22) associated with additional abnormality t(13;22)(q22;q13) not described before. Fluorescent in-situ hybridization analysis with probes for ETO and AML1 genes, and probes for the subtelomeric regions of chromosome 16 demonstrated an ETO-AML1 fusion signal on der(8). The 5′ region of ETO gene with chromosome arm distal to translocation breakpoint had moves to der(16) while region of AML1 probe proximal to translocation brealpoint stayed on der(21). The signal for the subtelomeric probe for 16p was present on der(21). Use of painting probe for chromosome 13 confirmed t(13;22). In a hematopoietic cell the expression of chimeric AML1- ETO protein resulting from t(8;21) plays a key role in leukemic transformation by targeting AML1-CBFB transcription factor complex, an essential regulator of genes required for normal hematopoietic cell development, by dominant negative effect on normal AML1 protein. However, the expression of AML1- ETO does not block normal differentiation of stem cells. Additional mutational events must occur in hematopoietic progenitor cell to block normal path of differentiation. Although clinical features of variant translocation is same as cases with conventional t(8;21), prognostic implications of variant (8;21) are yet to be established. Chromosome region 16p13 has been involved in clinically important translocations with poor prognosis such as t(8;16)(p11;p13) and t(11;16)(q23;p13) and may have similar implications. Translocation (8;21) is frequently accompanied by additional chromosome abnormalities i.e. loss of sex chromosome, trisomy 8 and structural abnormalities of 9q. Additional reports on secondary genetic alterations accompanying t(8;21) are needed to understand their cumulative effect. Chromosome region 16p13, 13q22 and 22q13 may harbor putative genes influencing normal path of differentiation.
Study on Gene Exppression Profiling Associated with Prognosis in AML (M2a).
Authors: Meng Fan Yi , Tang Jiaming , Ma Wenli
Published: April 2020

Objective: to explore genes related to the prognosis of AML (M2a).

Methods: 6 newly diagnosed AML were involved in this experiment and were divided into 2 groups. Group A: 3 AML (M2a) patients with continuous complete remission (CCR1) less than 6 months; Group B: 3 AML (M2a) patients with CCR1 more than 12 months. Bone marrow mononuclear cells were separated and mRNA was purified and labeled with Cy3 and Cy5 respectively, which were used to hybridize against the Agilent human 1B 60mer oligonucleotide microarrays.

Results: in the 20173 genes tested, 22 genes were found to be expressed differently between these two groups, among them 10 genes were up-regulated in group A, and 12 genes were up-regulated in group B.

Conclusion: with microarray assay, 22 genes including APP were found to be differently expressed in AML (M2a) treated with standard chemotherapy. These genes may be early indicators for the diagnosis as well as prognosis of the refractory AML.
Intrathecal Administration of Rituximab for the Treatment of Primary Central Nervous System B-Cell Lymphoma.
Authors: Fernandes Flavo B. , Silla Lucia , Friedrisch Jõao R. , Brusius Carlos
Published: April 2020

Background: Patients with a primary central nervous system (CNS) lymphoma are very difficult to be treated. Regarding elderly patients, the side effects of the established strategies are considerable, particularly those caused by radiotherapy. Although with poor long term results, surgery, high dose chemotherapy and focal radiotherapy have been advised for these patients. Thus, new therapeutics strategies are needed. We report two cases of primary CNS B-cell lymphomas treated with rituximab intrathecally (ith). Case 1: In September 2003, a 59-year-old female with headache, paresthesia and weakness of the right limbs presented at our clinic. In August 2003 a brain biopsy of a parenchymal lesion was performed, vasculitis and cerebritis were found. The patient’s condition progressively worsened during the next 3 months and she developed a right-side hemiparesis. A PCR test to ascertain the presence of Mycobacterim tuberculosis in brain tissue was positive and rifampicin was prescribed. The patient presented at our clinic in September 2003, and in December 2003 she underwent another brain biopsy and CD20+ diffuse large B-cell lymphoma was diagnosed. Chemotherapy with high-dose methotrexate (MTX) 3.5 g/m2 iv, MTX 12 mg ith and vincristine 1.4 mg/m2 was scheduled every 15 days for a total of 5 cycles. The patient experienced a partial remission. In February 2004, rituximab 25 mg ith was administered on days 1–2 and days 8–9. There was no toxicity during, or after the rituximab infusions. In March, after a total dose of 200 mg, there was a complete radiological remission of the lymphoma and a consistent clinical improvement. In June 2004, the lymphoma relapsed, affecting the same area of the brain and in July 2004 a re-treatment protocol consisting of MTX 8g/m2 and high dose cytarabine was started, without response. The patient died of sepsis, secondary to mucositis and pancytopenia in August 2004. Case 2: a 67-year-old male with a primary malignant B lymphoma CD20+ of the posterior fossa which relapsed after two surgeries, high dose chemotherapy and focal radiotherapy with lymphomatous meningitis. Complete remission was obtained after one course of high dose MTX (4g/m2) and intrathecal MTX, intercalated with three courses of systemic (350 mg/m2) and intrathecal rituximab (25mg/d - 8 days). The patient presented hepatotoxicity after the first dose of MTX iv. The relapse free survival is seven months until now and the patient is physically and intellectually fully recovered. No major side effects were observed with the administration of rituximab intrathecally.

Conclusion: Intrathecal administration of rituximab had no toxicity and was well-tolerated. Further studies are necessary to establish the best dose and schedule to achieve an improvement in the therapeutic response.
The Bisphosphonate, Incadronate, Inhibits Cell Growth of HTLV-I-Infected T-Cell Lines and Primary ATL Cells by Inhibiting Mevalonate Pathway.
Authors: Ishikawa Chie , Okudaira Taeko , Tomita Mariko , Mori Naoki
Published: April 2020

Bisphosphonates have been used for the treatment of hypercalcemia associated with malignancies and osteoporosis. Recent reports have suggested that bisphosphonates may also exert direct antitumor effects on several cancers, in addition to their therapeutic antiresorptive properties. Adult T-cell leukemia (ATL) is a fatal malignancy of T lymphocytes caused by infection with human T-cell leukemia virus type I (HTLV-I), and remains incurable. ATL is associated with osteolytic bone lesions and hypercalcemia, which are major factors in the morbidity of ATL. Therefore, the search for anti-ATL agents that have antitumor and antiresorptive effects is warranted. The aim of this study was to determine the effect of bisphosphonate, incadronate, on HTLV-I-infected T-cell lines and primary ATL cells. Incadronate prevented cell growth of HTLV-I-infected T-cell lines and primary ATL cells, but not of uninfected T-cell lines and normal PBMCs. Incadronate induced S-phase cell cycle arrest and apoptosis in HTLV-I-infected T-cell lines. Treatment of HTLV-I-infected T-cell lines with cell-permeable substrates for mevalonate pathways, geranylgeraniol and farnesol, prevented incadronate-mediated growth suppression. Incadronate also prevented the prenylation of Ras proteins. These results demonstrate that incadronate-induced growth suppression is the result of inhibition of the mevalonate pathway. On the other hand, incadronate did not affect NF-κB and AP-1 pathways. Importantly, treatment with incadronate reduced tumor formation from an HTLV-I-infected T-cell line, HUT-102, when these cells were inoculated s.c. into severe combined immunodeficient mice. These findings indicate that incadronate may become a novel molecular therapeutic class for treatment of ATL.
Immunotherapy with Recombinant Xenogeneic Vascular Endothelial Growth Factor as a Vaccine Combined Low-Dose Adriamycin Induces Synergistic Antitumor Efficacy in EL4 Lymphoma Model.
Authors: Niu Ting , Liu Ting , Jia Yong-qian , Wen Yan-jun , Tian Ling , Liu Ji-yan , Wu Yang , Hu Bing , Wei Yu-quan
Published: April 2020

We explored the antitumor efficacy of a strategy using recombinant Xenopus laevis vascular endothelial growth factor (xVEGF) as a xenogeneic protein vaccine, which we have demonstrated to have effective antiangiogenic activities in several tumor models, in combination with low-dose adriamycin in lymphoma model. EL4 lymphoma model was established in C57BL/6 mice. Mice were randomly divided into four groups: combination therapy (xVEGF plus adriamycin), xVEGF treatment alone, adriamycin treatment alone, and normal saline (NS) groups, and received relevant treatments. Tumor growth, survival rate of tumor-bearing mice, and potential toxicity of regimens above were investigated. Anti-VEGF antibodies and anti-VEGF antibody-producing B cells (APBCs) were detected by Western blot analysis and enzyme-linked immunospot (ELISPOT) assay, respectively. In addition, microvessel density (MVD) within tumor tissues and tumor cell apoptosis were determined by immunohistochemistry and TUNEL staining, respectively. Our data showed that combination therapy inhibited tumor growth and improved survival of tumor-bearing mice significantly, and complete regression of tumor was observed in 3 of 10 mice in combination group in EL4 lymphoma model.Survival rate of mice was significantly higher in combination group than that in xVEGF alone group,in adriamycin alone group (P<0.05),or that in NS group (P<0.01). Remarkably, the combination treatment resulted in not only significant antiangiogenic effects but also promotion of tumor cell apoptosis. The calculated indexes of combination therapy showed synergistic relationship in terms of inhibition of tumor growth, antiangiogenesis, induction of tumor cell apoptosis. In addition, anti-VEGF antibodies and APBCs were found in mice treated with either xVEGF vaccine alone or combination treatment. No marked toxicities were found in the treated mice. In summary, these findings may provide an effective combination approach for lymphoma therapy in the future.
A Pilot Trial of Imatinib, Low-Dose Cytarabine (ara-C) and Idarubicin (Ida) in Patients (pts) with Chronic Myeloid Leukemia (CML) in Myeloid Blastic Phase (BP).
Authors: Quintas-Cardama Alfonso , Kantarjian Hagop , Ravandi Farhad , Garcia-Manero Guillermo , O’Brien Susan , Wierda William , Faderl Stefan , Cortes Jorge
Published: April 2020

Although imatinib is the single most effective agent in CML BP, only approximately 30% to 50% of pts achieve a hematologic response, and it is complete (CHR) and durable in 5% or less. Regimens including high-dose ara-C and daunorubicin are associated with remission rates of 20% to 25% in CML BP, a median survival of 3 months, and significant morbidities. Imatinib is additive or synergistic in vitro with ida and ara-C, particularly in imatinib-resistant cell lines. We thus investigated the efficacy of the combination of imatinib 600 mg daily, ara-C 10 mg daily subcutaneous, and Ida 12 mg/m2 IV every 14 days in pts with CML in myeloid BP. Nineteen pts were treated, including 6 who had BP as their first manifestation of CML. Seventeen pts had failed prior imatinib as a single agent and had received a median of 5 prior therapies (range, 1 to 9). Median age was 54 yrs (range, 27–81) and the median WBC was 28.6 x 109/L (range, 3.7 to 240.5), peripheral blood blasts 18% (range, 0 to 82), and bone marrow blasts 36% (range, 2 to 84). Four pts had extramedullary disease. Pts were treated for a median of 10.5 weeks (range, 4 to 124), receiving a median of 2 cycles (range 1 to 28). Fourteen (74%) pts responded, including 9 (47%) who achieved CHR, and 5 (26%) who returned to chronic phase. Four of the pts who achieved CHR also had a cytogenetic response (CGR) (3 complete, 1 minor). Responses lasted for a median of 10 weeks (range, 2 to 89 weeks). Two of 5 pts who failed to respond had a transient hematological improvement (>50% reduction of blasts in peripheral blood). Six pts were able to receive allogeneic BMT (4 in CHR, 1 in CP and 1 still in BP). Treatment was well tolerated and administered on an outpatient basis. Grade 3–4 non-hematological toxicities consisted of fatigue (n=4), vomiting (n=1), bone pain (n=1) and hyperbilirubinemia (n=1). Sixteen (84%) pts required brief hospitalization for neutropenic fever (n=8), bleeding (n=7), pneumonia (n=6), cellulitis (n=2) and CNS disease (n=2). Thirteen pts died: 4 after BMT and 9 due to disease progression. Among these, 4 had achieved CHR and 2 complete CGR. The median survival was 23 wks and 1 year survival rate 26%. In conclusion, the combination with imatinib + low-dose ara-C + Ida is an outpatient, well tolerated regimen that can induce responses in patients with CML BP who have failed imatinib and can be considered as an option to improve patient’s condition, and as a bridge to allogeneic BMT.
Coordination of Intrinsic, Extrinsic and Endoplasmic Reticulum-Mediated Apoptosis by Imatinib Mesylate Combined with Arsenic Trioxide in Chronic Myeloid Leukemia.
Authors: Zhang Ji , Du Yanzhi , Wang Kankan , Fang Hai , Li Junmin , Xiao Dakai , Zheng Peizheng , Chen Yulong , Fan Huiyong , Zhang Qinghua , Fang Jing , Auffray Charles , Chen Saijuan , Chen Zhu , Zhang Ji
Published: April 2020

A treatment strategy that combines arsenic trioxide (ATO) with the tyrosine kinase inhibitor imatinib mesylate (STI571, Gleevec) appears to induce more cell apoptosis than STI571 alone in chronic myeloid leukemia (CML). To understand the mechanisms underlying the synergistic action of these agents, we applied cDNA microarrays, component plane presentation integrated self-organizing maps (CPP-SOM) and methods of protein biochemistry to study proapoptotic and apoptotic activities in programmed cell death induced by STI571, ATO and the combination of the two agents in detail. Numerous features with temporospatial relationship were revealed, indicating the coordinated regulation of molecular networks from various aspects of proapoptotic and apoptotic activities in CML. In consequence, STI571 induces the intrinsic pathway of cell apoptosis, whereas ATO induces the ER stress-mediated pathway of cell apoptosis, and the combination of the two agents induces the intrinsic, extrinsic and ER stress mediated pathways of cell apoptosis, which results in a more effective and efficient induction of programmed cell death in CML. The potential impact of this study on the development of more sophisticated therapy in human malignancy can be substantial.
Characterization of Cancer Stem-Like Cells in a Novel STI571-Resistant Chronic Myeloid Leukemia Cell Line.
Authors: Song Yongping , Fang Baijun , Zhang Yanli , Wei Xudong , Lu Lulu , Zhu Xinghu , Wang Ping , Li Yufu , Gao Qunli , Wang Guiye , Qiu Lugui
Published: April 2020

To characterize a novel chronic myeloid leukemia (CML) cell line and to further elucidate the mechanisms of resistance to STI571, a novel K562 cell line (K562/VP16) was achieved after exposure of the K562 cells in gradually increasing concentrations of VP16 over a period of several months. A small subpopulation (K562/VP16 SP) that was capable of excluding Hoechst 33342 in the K562/VP16 cell line was isolated by flowcytometry sorting. The rest of the K562/VP16 cells were classified as non-SP K562/VP16. The mechanisms involved in K562/VP16 SP cells that became resistant to STI571 were studied. We found that the levels of Bcr-Abl and Abl proteins were similar in the K562 cells and in non-SP K562/VP16 and K562/VP16 SP cells. The MDR-1 gene expression of the 170 kDa P-gp was detected in K562/VP16 non-SP and K562/VP16 SP cells but not in K562 cells. The expression levels of P-gp in the two K562/VP16 cell lines were similar. Furthermore, there were no mutations of Abl in the K562/VP16 SP cells. Compared with non-SP K562/VP16, the K562/VP16 SP cells were more resistant to STI571, and many multidrug resistance inhibitors could hardly reverse this resistance. In addition, in vivo study showed that the K562/VP16 SP cells induced tumorigenesis in mice, while the K562/VP16 non-SP cells failed to do so. These results suggest that a small side population K562/VP16 SP cells in K562/VP16 cell line, had high resistance to STI571 treatment and more tumorigenic than the K562 cells and it may represent the cancer stem cells of the K562/VP16 cell line.
BCR Gene Breakpoint and Type of BCR/ABL-Transcript in Children with Chronic Myeloid Leukemia.
Authors: Suttorp Meinolf , Viehmann Susanne , Claviez Alexander , Martiniak Yvonne , Harbott Jochen
Published: April 2020

The hallmark of chronic myeloid leukemia (CML) is the reciprocal translocation t(9;22)(q34;q11) which creates a hybrid BCR/ABL gene that is transcribed into a chimeric mRNA. In the 8.5-kb mRNA either exon b2 or exon b3 of the major breakpoint cluster region (M-bcr) is coupled to C-ABL exon 2. These two junction variants (b2a2 or b3a2) differ in size by 75 nucleotides and can be selectively amplified by semi-nested PCR. A controversy exists whether or not the type of transcript and/or the breakpoint site exerts an influence on the clinical course of CML in adult patients (Mills KI 1991; Foiretos T 1993; Melo JV 1996). CML is a rare disease in childhood contributing only 2 – 3 % of all pediatric leukemia cases. This may be the reason why there are only scant data from small series of children with CML giving information on the transcript variant present at diagnosis. During an 11 year period (1995 – 2005) 203 pts younger than 19 years (median age: 11.4 yrs; 101 boys, 102 girls) with Philadelphia-chromosome positive CML were treated within the multicenter GPOH-trial “CML-paed”. 85% of the pts were diagnosed in chronic phase (CP). Treatment was performed by hydroxyurea +/− interferon and pts were scheduled for stem cell transplantation (SCT) from an HLA-matched family donor within 6 months after diagnosis and from an unrelated donor within 12 months. Bone marrow and/or peripheral blood specimen for molecular analysis were collected at diagnosis and during follow-up examinations at 3 – 6 months intervals.

RT-PCR with analysis of the breakpoint and transcript type was performed in 154 pts (77.4 % of the total study population). 133 pts exhibited a Major-BCR rearrangement, 5 pts a minor-BCR rearrangement and in 16 pts both rearrangements were present. In pts with M-BCR either 55 pts expressed the b2a2 transcript (37 %) or the b3a2 transcript (37 %), respectively. Both types of transcripts were found either simultaneously or alternately during follow-up examinations in 38 pts (25 %). One pt showed an atypical rearrangement which is awaiting further classification.

No statistical significant differences (p > 0.05) could be found with respect to hematological parameters at diagnosis (white cell count, platelets count, percentage of basophils in peripheral blood), clinical findings (liver size, spleen size) and disease status (CP versus advanced phase). The impact of either transcript upon the course of the disease (e.g. duration of chronic phase) was not analyzed because 70 % of the study population was transplanted early within 1 year after diagnosis.

We conclude that in contrast to some reports in adults the type of BCR breakpoint seems to have no impact in pediatric pts with CML.
Development of a BCR/ABL Real-Time Quantitative RT-PCR Assay and Correlation with an Existing Qualitative Test.
Authors: Howe John Greg , Crouch Jill , Smith Brian R.
Published: April 2020

Introduction: With the recent introduction of imatinib (Gleevec) as a therapeutic agent for chronic myeloid leukemia (CML), the need for quantitation of the level of tumor cells after treatment has become essential. Our lab has for a number years tested for CML using a sensitive qualitative nested RT-PCR assay for BCR/ABL. We wanted to develop a quantitative test that retained the sensitivity of our qualitative test and contained an initial screen to prevent unnecessary quantitative testing as well as develop quantitative tests for the various breakpoints.

Methods: The new test is divided into three parts. A real-time multiplex qualitative RT-PCR screen containing specific primers for the p190, p210 and p230 BCR/ABL forms and a common labeled probe. If a patient sample is positive in the screen, the original nested RT-PCR test is performed to determine which BCR/ABL form is present. This is followed by a real-time quantitative RT-PCR test for the identified BCR/ABL translocation. Subsequent follow up tests require only the specific quantitative test.

Results: Comparing negative results from 82 patient (various clinical presentations) samples tested by the previous qualitative test with the new qualitative screen showed completely correlation. Out of 31 CML patients studied, both test strategies gave the same result for 20 positive and 13 negative samples. Patients with multiple samples collected over time showed complete correlation between the two assays.

Conclusions: The new quantitative BCR/ABL testing strategy yielded results that correlated with the previous qualitative BCR/ABL assay.
High Dose Zevalin (90Yttrium Ibritumomab Tiuxetan) Treatment with PBSC Support in Refractory-Resistant NHL Patients: Preliminary Results of a Phase I/II Study.
Authors: Vanazzi Anna , Ferrucci Pierfrancesco , Ferrari Mahila , Calabrese Liliana , Cremonesi Marta , Bartolomei Mirco , Paganelli Giovanni , Martinelli Giovanni
Published: April 2020

Therapeutic options are limited for resistant-refractory high-grade NHL pts not suitable to HDCT or for those pts relapsing after ABMT. Zevalin has been already demonstrated active in elderly pts with resistant-primary refractory DLBCL at dose of 0.4 mCi/kg, however duration of response is short. Increasing RIT dose intensity might improve efficacy. There are no data about the use of Zevalin at myeloablative dosage. We present feasibility and toxicity results of a phase I/II study of HD-Zevalin with PBSC support in resistant-refractory NHL pts. Three Zevalin dose-levels were fixed: 0.8, 1.2, 1.5 mCi/kg; from April 2004 to July 2005 12pts were enrolled, 4pts at each dose level. Median age was 66,5 ys (28–73), 83% male. All pts had stage III/IV at diagnosis; 7DLBCL; 3MCL, 1FL, 1transformed MZL. 7pts had received more than 3 previous CT regimens. All pts had received prior rituximab, 3pts RT, 6pts HD-CT. BM was negative in all pts before RIT. 1Week prior to Zevalin all pts underwent dosimetry and if no abnormal uptake was observed they received the planned dose. On Day −7 and 0 imaging and therapeutic doses were preceded by rituximab 250mg/mq. On Day13 pts were reinfused with PBSC previously harvested. On Day28 from reinfusion engraftment was considered to be delayed if WBC/ANC were<1.0x109/L or PLT<20x109/L. All pts engrafted promptly after PBSCT with median time to WBC/ANC>1.0x109/L and PLT>20x109/L of 19 (range 0–23) and 12 days (0–22) from PBSCT. PLT and ANC count nadirs were observed at 21 (0–24) and 25 (0–31) days after Zevalin with median values of 11x109/L(4–35) and 0.24x109/L(0.01–1.09). Median time to recovery from nadir was 16,5 days (4–175) for PLT and 17days (7–175) for ANC. 8pts required PLT transfusions, 4pts RBC transfusions; median number of PLT transfusions:1(1–4). No G-CSF was administered. No significant differences in terms of haematologic toxicity were observed among the 3 levels. No pulmonary, renal or cardiac toxicity was observed. 11/12 pts are now evaluable for response: 5pts experienced a CR (2 at 0.8mCi/kg, 2 at 1.2mCi/kg, 1 at 1.5mCi/kg), 1 pt receiving 1.5 mCi/kg a PR (ORR 50%); to date 4 pts maintain CR at 12, 11, 9 and 8 months after treatment.

Conclusion: Zevalin at dosage higher than MTD is feasible with PBSC support. Even at dosage 4 times higher than standard, HD-Zevalin could be safely delivered in elderly and heavily pretreated pts, including those who previously received HD-CT. Clinical activity and mild treatment-related toxicities suggest that HD-Zevalin is an interesting modality treatment to be further investigated as an alternative therapeutic options for resistant-refractory NHL.
Philadelphia Chromosome (Ph’) Negative, MLL-Rearranged AML Arising in a Patient Treated with Imatinib for CML.
Authors: Perl Anna Sechser , Perl Alexander E. , Robinson Blaine W. , Bagg Adam , Porter David , Tomczak Ewa , Nowell Peter C. , Carroll Martin , Felix Carolyn A.
Published: April 2020

Background: Imatinib has become standard front-line therapy for CML. In ~3% of patients treated with imatinib, abnormalities including trisomy 8 and/or monosomy 7 occur in Ph’ negative subclones and dysplasia occasionally is present, but transformation to AML is rare. We describe the first known case of AML with an MLL translocation during an imatinib-induced molecular remission of CML.

Methods: The Ph’ and the t(11;19)(q23;p13.1) were detected and monitored in sequential marrows using cytogenetic and FISH analyses. The BCR-ABL1 fusion transcript was traced by RT-PCR. The MLL translocation was identified and characterized in the AML blasts by Southern blot analysis, panhandle PCR-based methods and conventional PCR.

Clinical History and Results: The patient presented with chronic phase CML at age 53 and was treated with hydroxyurea for 6 weeks, IFN-α for 20 months and imatinib for 32 months. Complete cytogenetic response was achieved at 10 months. Molecular remission, as indicated by absence of the BCR-ABL1 transcript, occurred after 15 months of imatinib (37 months from CML diagnosis). Mild dysgranulopoiesis was noted 6 months after imatinib was started and increased on subsequent studies. After 28 months of imatinib (50 months from CML diagnosis), there was marked trilineage dysplasia and the karyotype showed t(11;19)(q23;p13.1) in cells that were Ph’ negative and negative for the BCR-ABL1 fusion transcript. FAB M5b AML was diagnosed four months later. The patient succumbed to AML despite aggressive management with chemotherapy and allogeneic stem cell transplantation. Southern blot analysis of the AML revealed two MLL bcr rearrangements. The reciprocal breakpoint junctions on the der (11) and der (19) chromosomes indicated a translocation of intron 8 of MLL and intron 1 of the known MLL partner gene ELL, which encodes a transcription elongation factor. The involved region of MLL was more 5′ than the secondary leukemia translocation breakpoint hotspot. The der (11) fusion transcripts joined MLL exon 7 to ELL exon 2, which is consistent with alternative splicing, and the der (19) fusion transcript joined ELL exon 1 to MLL exon 9.

Conclusions: Secondary leukemias with MLL translocations have been associated with topoisomerase II poisons, but not with the agents administered to this patient. The entity that we describe is distinct from blast crisis, in which Ph’ positive subclones evolve to acquire additional alterations. This case establishes that persistent clonal abnormalities and/or dysplasia in Ph’ negative cells following imatinib therapy may not be benign and may herald AML transformation. With effective and selective molecular eradication of the Ph’ positive clone, the emergence of leukemia with independent abnormalities may become more common. This is a highly concerning clinical complication to consider with BCR-ABL targeted agents.
Treatment of Imatinib-Sensitive and -Resistant Chronic Myelogenous Leukemia Cells with a Combination of Imatinib and Farnesyltransferase Inhibitors.
Authors: Radujkovic Aleksandar , Topaly Julian , Zeller W. Jens , Ho Anthony D. , Fruehauf Stefan
Published: April 2020

Farnesyl protein transferase inhibitors (FTIs) represent a new class of anticancer agents. We were interested whether the farnesyltransferase inhibitors L-744,832 and LB42918 alone and in combination with imatinib are active in imatinib-sensitive and -resistant CML cells. LB42918 was more potent with ED50 = 4 ± 1 μM (mean ± S.D., n = 3) in the BCR-ABL+ imatinib-sensitive cell line EM-3 vs. ED50 = 25 ± 2 μM for L-744,832 after 48 hrs of treatment and assessment by MTT assay. The growth of imatinib-sensitive K562 and LAMA84 cells was not measurably inhibited by L-744,832 doses up to 25 μM. Similarly, the growth of K562 was not affected by 25 μM of LB42918. However, for LB42918 in LAMA84 cells an ED50 value of 30 ± 11 μM could be determined. In the imatinib-sensitive cell line EM-3, combination index values (CI) obtained using the method of Chou and Talalay indicated synergistic effects following simultaneous treatment with imatinib and FTI (CI = 0.5 ± 0.2 and 0.7 ± 0.2 at ED75 for imatinib + L-744,832 and imatinib + LB42918 respectively). In the cell imatinib-sensitive and -resistant lines K562 and LAMA84 a trend to lower ED50 values of imatinib was determined when up to 16 μM of L-744,832 or LB42918 were added, indicating potentiation of imatinib activity. Annexin V / propidium showed a strong increase of the apoptotic cell fraction in all imatinib-sensitive and an increase in the imatinib-resistant cells treated for 48 hrs by the combination imatinib + L-744,832 or imatinib + LB42918 as compared to treatment with each drug alone. Growth inhibition of CFU-GM colonies of primary imatinib-sensitive CML cells obtained from 4 patients is stronger after treatment with different concentrations of the combination of both drugs than after monotherapy with either imatinib or L-744,832. On the basis of the observed potentiation effects FTIs may find their place as supplement for CML patients on imatinib treatment.
Additional Cytogenetic Changes and Prior Genotoxic Exposure Predict Unfavorable Prognosis in Myelodysplastic Syndromes and Acute Myeloid Leukemia with der(1;7)(q10;p10).
Authors: Ohyashiki Kazuma , Hsiao Hui-Hua , Sashida Goro , Ito Yoshikazu , Ohyashiki Junko H.
Published: April 2020

We analyzed 23 patients with myelodysplastic syndromes (MDS) and acute myeloid leukemia (AML) showing der(1;7)(q10;p10) (hereafter der(1;7)), to identify the exact predictive factor of this cytogenetic change. Eight (34.8%) patients, including 6 MDS and 2 AML patients, had a prior history of genotoxic exposure, especially radiation and/or antimetabolites. Patients with der(1;7) consisted of three groups; one-third of patients had a prior history of genotoxic agents, one-third had additional cytogenetic changes at the time of MDS/AML diagnosis without prior exposure history, and the remaining one-third had neither prior exposure history nor additional cytogenetic changes. The current study demonstrated that the poor outcome of MDS/AML with der(1;7) is due to the high frequency of associated risk factors, i.e., prior history of genotoxic exposure, the presence of additional cytogenetic changes, or both. Identification of prognostic disadvantage might be required for appropriate strategy in managing MDS/AML patients with rare der(1;7) abnormality.
Myelodysplastic Syndromes with Myelofibrosis May Be a Target for the JAK2 V617F Tyrosine Kinase Mutation.
Authors: Ohyashiki Kazuma , Aota Yasuo , Gotoh Akihiko , Miyazawa Keisuke , Kimura Yukihiko , Ohyashiki Junko H.
Published: April 2020

The JAK V617F tyrosine kinase mutation is responsible for development of the disease in chronic myeloproliferative disorders, including myelofibrosis (MF). Since MF is associated with various hematologic diseases as a terminal hematologic condition and is an important issue in managing patients, we searched for the V617F mutation in primary and secondary MF in various hematologic diseases using the sequence-specific primer-single molecule fluorescence detection assay (SSP-SMFD). Of the MF patients associated with acute myeloid leukemia and (n = 3), lymphoma (n = 3), and chronic myeloid leukemia (n = 3), none of them showed the mutation at the time of the diagnosis of MF. Approximately 40% of essential thrombocythemia (ET: n = 16) and primary MF (n = 4) showed the JAK2 V617F mutation, whereas 6 of 8 patients with polycythemia vera showed the mutation. Of note is that 2 of 6 patients with myelodysplastic syndromes (MDS) terminating in MF showed the mutation, while no MDS patient without MF (MDS: n = 3) had the JAK2 V617F mutation. Our study clearly demonstrated that MF in MDS patients is sometimes associated with JAK2 V617F mutation, while other underlying diseases developing MF may involve other pathways. Moreover, it permits speculation that clonal evolution in some MDS patients with the JAK2 V617F tyrosine kinase mutation may be responsible for secondary MF.
Phase II Study of Bortezomib in Waldenstrom’s Macroglobulinemia: Results of WMCTG Trial 03-248.
Authors: Treon Steven , Hunter Zachary R. , Matous Jeffrey , Badros A. , Joyce Robin M. , Mannion Brian , Advani Ranjana , Cook D. , Songer J. , Hill John , Kaden Bruce R. , Sharon D. , Steiss R. , Branagan Andrew R. , Patterson Christopher J.
Published: April 2020

In this multi-center phase II study, 27 patients with the consensus panel definition of Waldenstrom’s macroglobulinemia received bortezomib as a single agent. All but one patient had relapsed/or refractory disease, and the median number of prior therapies was 2 (range 0–3). Intended therapy consisted of 8 cycles of bortezomib at 1.3 mg/m2 on days 1,4,8,11. Twenty-six patients have completed therapy and are eligible for evaluation. The median age for this group was 62 (44–79 years) and baseline laboratories were as follows: serum IgM: 4,565 (1,067–9,330 mg/dl); bone marrow (BM) involvement 30% (1–95%); and hematocrit 34.9% (25.6–45.3%). Following a median of 6 cycles of therapy, serum IgM levels decreased in 25/26 patients to a median of 2,685 (125–5,650 mg/dl), while hematocrit rose in 20/26 patients to a median of 37.2% (30.9–46.3%). The overall response rate was 84.6%, with 11 and 11 patients achieving a minor (<25% decrease in IgM) and partial (<50% decrease in IgM) response, respectively, using consensus panel criteria. Post therapy bone marrow biopsies were not required for response determination, but were available for 8 patients who demonstrated a median decrease in bone marrow involvement from 37.5%(range 20–55%) to 17.5% (range 5–48%). However, in 3 of 8 patients discordance between serum IgM levels, bone marrow involvement and/or adenopathy was demonstrated including in one patient who demonstrated a decrease in serum IgM levels from 2,291 to 618 mg/dL whilst bone marrow involvement rose from 25% to 48%. With a median follow-up of 8 months, 10 patients have demonstrated progressive disease. In general, therapy was well tolerated with grade III/IV toxicities as follows: neutropenia (15%); sensory neuropathies (12%); thrombocytopenia (8%); pleural effusion (4%); neuropathic pain (4%); diarrhea (4%); infection (8%); anorexia (8%); and fatigue (4%). The results of these studies demonstrate that bortezomib is an active agent in the salvage setting for patients with WM, and may independent of direct tumor cell killing inhibit secretion of IgM in some patients. Moreover, BM biopsies and CT scans (if baseline adenopathy or splenomegaly were present) should be considered to assess response in WM patients receiving bortezomib. In view of these results, a phase II study of bortezomib in combination with dexamethasone and rituximab has been initiated in patients with WM.
A Case of Fulminant Myelodysplastic Syndrome Versus Acute Erythroleukemia Variant.
Authors: Mazzella Fermina M.
Published: April 2020

Case: A 73 year old male with no significant past medical history presented with chronic fatigue. A routine CBC revealed an H/H of 6.8/19, MCV 103, WBC 4,100 and platelet count 92,000. Reticulocyte count was 0.6%. Serum iron, B12 and red cell folate studies were within normal limits. A bone marrow aspiration and biopsy were performed.

Bone Marrow Findings: The bone marrow aspirate showed an M:E ratio of 1.8:1, with 33% erythrocytic elements. There was markedly left shifted erythropoiesis, with 57% of erythrocytic precursors being pronormoblasts. 1.2% blasts were present. Moderate erythrocytic dysplasia was present, with mild granulocytic and megakaryocytic changes. Vacuolated pronormoblasts were readily identified. No ringed sideroblasts were present. A diagnosis of MDS, NOS was rendered. No cytogenetics were obtained.

Follow-up: The patient was transfused 4 units of packed red cells. Due to the patient’s rapidly deteriorating condition, comfort measures only were provided. The patient died 2.5 weeks after the bone marrow procedure. No autopsy was performed. The cause of death was listed as MDS.

Discussion: This case is an extremely rare instance of left shifted erythropoiesis in the absence of the requisite number of erythrocytic elements for consideration of acute erythroleukemia (50%). By current definitions, this is an example of a fulminant MDS. However, this case does beg the question of whether or not pronormoblasts should be included in the assessment of malignant disorders of bone marrow.
Deletion of Tumor Suppressor Genes (TSGS) in Patients with Myelodysplastic Syndrome (MDS).
Authors: Galanopoulos Athanasios G. , Papadhimitriou Stephanos , Tsourveloudis Ioannis , Michalis Evridiki , Marinakis Theodoros , Tsakiridou Anastasia , Savvidou Ioanna , Xanthopoulos Vasilios , Theodoropoulos George , Gortzolidis George , Zomas Athanasios , Georgiakaki Maria , Anagnostopoulos Nikolaos I.
Published: April 2020

Background: The deletion of genomic sites harboring TSGs is known to be a powerful prognostic indicator in various chronic hematologic malignancies. However, the clinical significance of TSGs loss in MDS has not been thoroughly investigated.

Aims: To summarize our experience on the incidence and essential clinical correlates of TSGs deletions in MDS, providing preliminary results from the study of 27 patients.

Methods: The study included 17 men and 10 women (median age 74; range 38–84 years) with a documented diagnosis of MDS. The distribution of FAB subtypes was RA in 13, RARS in 3, RAEB in 6, RAEB-T in 4 and CMML in 1 case. According to the IPSS stratification, 5 patients were characterized as “low-risk”, 16 as “intermediate-risk” (11 as INT-1 and 5 as INT-2) and 6 as “high-risk” patients. The diagnostic bone marrow smears were studied with fluorescence in-situ hybridization (FISH) for deletions of chromosome regions 9p21 (p16/p14 and p15 genes), 9q34, 12p13 (TEL gene), 13q14 (RB1 gene D13S319 locus) and 17p13 (p53 gene).

Results: A deletion in at least one of the chromosome regions or loci studied was detected in 10 patients (37%). In 6 of them, more than one region locus was lost, making up a total of 20 deletions. The commonest finding was loss of the 17p13 region (5 cases; two of them homozygous), followed by 9p21 loss (5 cases; one of them homozygous). Overall, the presence of TSGs deletions was associated with complex karyotype, high IPSS score and risk of death (regardless of leukemic progression). Interestingly, deletion in at least one of the TSGs studied was found in 5 of the 17 patients presenting with normal karyotype. In 3 of these patients, the MDS progressed to acute leukemia. Two of the 5 patients died at one and six months from diagnosis of the MDS, without leukemic conversion.

Summary/Conclusions: Deletions of TSGs are not uncommon in MDS. Interestingly, we have found that certain TSGs, such as the p16/p14 and p15 genes at 9p21, the loss of which is mostly involved in the initiation or progression of lymploid neoplasms, are also frequently deleted in MDS. Overall, TSGs deletion in this small group of patients is apparently associated with other adverse biological and clinical features and poor outcome. Therefore, these preliminary observations justify the expansion of the study in a larger patient cohort, in order to clarify if the loss of certain TSGs is an independent prognostic factor in MDS and, thus, may be of help in the initial diagnostic approach and risk stratification of the patients.