bluuebunny/crossref_metadata_2025 huggingface dataset:
Elevated Serum Neutrophil Elastase but Not Proteinase 3 Is Present in Patients with Myeloid Leukemia.
Authors: Sergeeva Anna , Ono Yoko , Molldrem Jeffrey J.
Published: April 2020

Proteinase 3 (P3) and neutrophil elastase (NE) are abnormally expressed in the cytoplasm of myeloid leukemia, and we have shown that the HLA-A2-restricted peptide PR1 is processed and cross-presented from both proteins by antigen-presenting cells (APC) to cytotoxic T lymphocytes (CTL). PR1-specific CTL kills leukemia cells and contributes to cytogenetic remission in patients with chronic myelogenous leukemia. Both exogenous P3 and NE can bind to monocytes and P3 promotes DC maturation in vitro. Since high levels of circulating P3 were found in patients with the ANCA-associated systemic vasculitis Wegener’s granulomatosis, modulation of DC by P3 has been implicated in reversal of tolerance. The role of extracellular P3 and NE in reversing tolerance in patients with myeloid leukemia is unknown. We used an ELISA assay to assess P3 and NE level in the sera of patients with AML and CML. Serum NE level was highly increased in all 15 AML patients studied (1045±149 ng/ml, n = 15) and in all 15 CML patients (1013±128, n = 15), compared to a control group of healthy donors (54±13 ng/ml, n = 4; p = 0.0038 and p = 0.0014, respectively). In contrast, serum P3 was increased in only 5 out of 18 (28%) AML patients (47 ± 18, n = 18 ng/ml) and in 8 out of 28 (29%) CML patients (95 ± 44 ng/ml, n = 28) compared to a group of 14 healthy donors (10 ± 2.2 ng/ml; p = 0.09 and p = 0.06, respectively). There was no relationship between elevated serum NE and P3 levels (r=0.16, n=35). P3 level in untreated patients and healthy donors did not change within at least 13 months, although P3 decreased after allogeneic BMT in 5 out of 5 patients (p < 0.05) with high serum levels of P3 pre-BMT. Because P3 is expressed by myeloid progenitor cells in the bone marrow, we studied the concentration of P3 in bone marrow-derived serum (BMS). To our surprise, BMS P3 concentration in patients with myeloid leukemia (23 ± 9.5 ng/ml, n = 9) was similar to that of a control group of patients with non-myeloid hematological malignancies (27 ± 11.8, n = 5) (p = 0.08), and BMS P3 concentration was similar to sera P3 concentration in the same patient (p = 0.52). Interestingly, BMS P3 level correlated with the level of BMS GM-CSF (quantified by ELISA) in a small group of patients with AML and CML (r = 0.94, n = 9), suggesting that P3 secretion by leukemia cells may be downstream of GM-CSF. We also analyzed membrane expression of P3 and NE in 5 AML patients using flow cytometry and found that P3, but not NE, was expressed on circulating cells in 3 of 5 patients. This unique expression of P3 and NE suggests that circulating NE might be the predominant source of cross-presented PR1 antigen by APC or myeloid leukemia cells.
Global Proteomics in AML: Using SELDI-TOF MS on Diagnostic Samples To Identify Spectra and Protein Biomarkers Indicative of Prognosis.
Authors: Stenke Leif , Pernemalm Maria , Forshed Jenny , Pawitan Yodi , Lagergren-Lindberg Marita , Kanter Lena , Lehtiö Janne
Published: April 2020

With the object to identify key proteins related to treatment response in acute myeloid leukemia (AML), and to eventually elucidate new targets for therapy, we have investigated protein spectra from diagnostic AML samples. Surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF MS) was used to spot proteins in the molecular weight area of 2–40 kDa. The samples were applied to two different chip surfaces (CM10 and Q10) for selective protein binding, avoiding the necessity of extensive purification and separation procedures prior to analysis. Peripheral blood samples from a total of 67 acute leukemia patients at diagnosis were analyzed. The cells were collected, frozen and stored in biobanks between 1988 and 2003. This provided a good basis for correlating detected protein peaks to a large number of clinical diagnostic and therapeutic variables and outcomes. Among the patients, aged 18 to 86 yrs, response to induction therapy was observed with complete remission (CR) durations ranging between 12 and 3701 days. The duration of first CR was used as a dichotomic variable in order to compare protein spectra from groups of patients with “good” and “bad” response, respectively, to chemotherapy. A peak detection and annotation software was developed in-house1,2 and employed. The software will be presented and a comparison to commercially available MS-spectral analysis methods will be provided. Initial results from MS profiling of the two response groups showed 18 significantly (p>0.01) differently expressed peaks on anion exchange chips and 22 peaks on cation exchange surfaces. Ongoing work is focused on further development of the RS peak detection method, including the influence of normalisation and filtering of the SELDI data and supervised linear and non-linear modelling. In addition, identification of selected peaks and their relation to various clinical variables is in progress.
Gene Expression Profiling of Pediatric MLL-Rearranged Leukemia: Relation between Acute Lymphoblastic Leukemia, Acute Myeloid Leukemia, and Leukemic Cell Lines.
Authors: Ishii Masami , Tsutsumi Shuichi , Taki Tomohiko , Nishimura Kunihiro , Ge Xijin , Tsuchida Masahiro , Ohki Misao , Bessho Fumio , Sugita Kanji , Ishii Eiichi , Ichikawa Hitoshi , Aburatani Hiroyuki , Hayashi Yasuhide
Published: April 2020

Although most patients with acute lymphoblastic leukemia (ALL) having MLL rearrangements suffer poor treatment outcome, there exists a subgroup of patients with this disease who enjoy long-term disease-free survival. In order to verify our previously reported new classification of this type of leukemia predicting treatment outcome, we analyzed gene expression profiles of MLL-rearranged leukemic cell lines and acute myeloid leukemia (AML) samples along with the previously reported ALL samples. Hierarchical clustering and the weighted vote method showed that cell line samples formed a strong association with the poorer prognosis subgroup in the classification, and that AML samples had some relationship with the better prognosis subgroup. Highly expressed genes in the poorer prognosis subgroup combined with cell line samples compared with the better prognosis subgroup include genes related to tumor aggressiveness, such as SNCG, GRM4, and EDG4. These results support the significance of the previously described classification associated with prognosis. Large-scale clinical studies, therefore, may be worth trying on the basis of this classification.
Elevated Serum Ras Levels in Patients with LGL Leukemia.
Authors: Nelli Erin M. , Loughran Thomas P. , Leitzel Kim , Ali Suhail M. , Demers Lawrence , Carney Walter P. , Hamer Peter J. , Lipton Allan
Published: April 2020

Background: Ras is a GDP/GTP binding G protein that acts as a molecular switch converting signals from the cell membrane to the nucleus to regulate cell proliferation, differentiation, and protein synthesis. Activation of ras oncogenes has been identified in a variety of cancers, including 30% of patients with acute myelogenous leukemia (AML). Large granular lymphocyte (LGL) leukemia is a clonal T-cell lymphoproliferative disorder associated with chronic neutropenia, anemia, or autoimmune diseases, particularly rheumatoid arthritis. The purpose of this study was to evaluate serum ras levels in patients with LGL leukemia.

Methods: A novel ras p21 ELISA (Oncogene Science/Bayer HealthCare) employing two monoclonal antibodies was utilized to quantify ras levels in baseline serum obtained from 32 patients with LGL leukemia. A control group of 48 healthy subjects was also used to establish a cutoff for the upper limit of normal for serum ras levels. A 95% non-parametric cut-off was used to determine significant differences in the frequency of elevated serum ras levels in control vs. LGL leukemia patients.

Results: The median serum ras level in the 48 healthy control subjects was 125 pg/mL, with a 5% to 95% range of 50–422 pg/mL. The upper limit of normal for serum ras was defined as 422 pg/mL, as determined using the 95 % non-parametric cut-off. In the LGL leukemia patient group, 19 of 32 patients (59 %) had elevated baseline serum ras levels (median 484 pg/mL) (p=0.0001) when utilizing the healthy control upper limit of normal cut-off value.

Conclusions: The results of this study indicate that the majority of LGL leukemia patients have significantly elevated serum ras levels when compared to healthy controls. Serum ras should be evaluated as a potential biomarker in larger leukemia trials, especially for response to treatment with inhibitors of the ras signaling pathway.
Measurement and Clinical Relevance of Proteasome Enzymatic Activity in Plasma of Patients with Acute Lymphoblastic Leukemia.
Authors: Ma Wanlong , Kantarjian Hagop , O’Brien Susan , Jilani Iman , Zhang Xi , Zestrov Zeev , Estey Elihu , Ferrajoli Alessandra , Keating Michael , Giles Francis , Albitar Maher
Published: April 2020

The ubiquitin-proteasome pathway plays a major role in regulating proteins involved in cell cycle regulation and apoptosis. Three different types of enzymatic activity have been reported for proteasomes: chymotrypsin-like (Ch-L), trypsin-like (Tr-L), and caspase-like (Cas-L) (postglutamyl peptide hydrolytic-like). Various proteasome inhibitors affect each of the three activities differently and at different concentrations. For example, NPI-0052 inhibits Ch-L and Tr-L activities at lower concentrations than does bortezomib, while bortezomib inhibits Cas-L at lower concentrations than does NPI-0052. These enzymatic activities are usually measured in normal or tumor cells to monitor therapy with proteasome inhibitors. We developed fluorogenic kinetic assays using peptide-AMC (7-amino 4-methylcoumoran) substrates to measure Ch-L, Tr-L, and Cas-L activities in cell-free plasma. We evaluated plasma activities in 57 patients with acute lymphoblastic leukemia (ALL) and assessed their correlations with clinical behavior. Significantly (P < 0.001) higher Ch-L, Tr-L, and Cas-L activities were detected in patients with ALL (medians: 1.40, 2.06, and 2.04 pmol AMC/sec/mL, respectively) than in healthy volunteers (n = 42) (medians: 0.80, 0.74, and 0.81 pmol AMC/sec/mL, respectively). While there was no significant difference between Ch-L and Cas-L activities in healthy controls, there was a significant difference between the 2 activities in patients with ALL. Cas-L, Tr-L, and Ch-L all correlated positively with lactic dehydrogenase (P <0.01). However, ALL patients with Cas-L activity ≥2.7 pmol AMC/sec/mL had significantly longer survival (P = 0.01) than did patients with lower activity. The increased cell-free circulating proteasome activities most likely reflect the leukemic cells and may represent a marker not only for the disease, but also for monitoring therapy. These data also suggest that patients with ALL may benefit differentially from proteasome inhibitors depending on the specific therapeutic properties of the inhibitor.
Identification of Novel Recurrent Genomic Aberrations by Array-CGH in Adult Acute Lymphoblastic Leukemia.
Authors: Viardot Andreas , Schwanen Carsten , Goekbuget Nicola , Hoelzer Dieter , Dohner Hartmut , Wessendorf Swen , Schmid Mathias
Published: April 2020

In acute lymphoblastic leukaemia (ALL), genomic aberrations like translocations t(4;11) or t(9;22) are associated with an adverse prognostic impact and play an important role in the development of risk adapted treatment strategies. So far, chromosomal banding analysis is the standard cytogenetic procedure for routine diagnosis. Genomic array-comparative genomic hybridization (aCGH) has become a useful technique for genomic screening allowing a more precise delineation of small genomic aberrations. We analyzed 44 patients with ALL, which were treated within the GMALL trials, by aCGH analysis using a genomic 2,8K chip. In 27 patients, clinical data were available. Thirteen patients had a common-ALL with normal karyotype, 10 patients had a common-ALL/Pr-B-ALL with t(9;22), two patients had either pre-B-ALL or T-lineage leukaemia, respectively. Ten patients were classified as very high risk (t(9;22) positive), 2 patient as high-risk (pre-T-ALL; c-ALL with high leukocytes) and 15 patients were standard risk according to the risk stratification of the GMALL-studies. We found genomic aberrations in 30 out of 44 patients (68%). The most frequent aberrations were deletions on chromosome band 9p21 (10 cases; 23%), deletions on chromosome band 7q35 (7 cases; 16%); gains on chromosome X and losses on 7p (6 cases; 14% each), deletions on 13q14 (5 cases; 11%) and gains on 8q24 (4 cases; 9%). The consensus region of the 9p21 deletion was narrowed down to a size of 420 kilobasepairs. This region covers 3 different genes with potential tumor suppressor gene (TSG) function: p14ARF, p15INK4B/p10 and p16INK4A. An additional TSG gene - methylthioadenosine phosphorylase (MTAP) - was found in 9 out of these 10 cases. In 3 cases with 9p21 deletions, none of the patients expressed the p16INK4A protein using western blot analysis. Two patients where also negative for the MTAP protein, whereas another patient whithout a genomic deletion on MTAP remains positive. There was a tendency to a higher number of aberrations per case in the standard risk group in contrast to the t(9;22) positive group (2.1 vs. 1.0 aberrations per case). aCGH analysis is useful for the identification of recurrent genomic aberrations which are not detectable using standard cytogenetic techniques.
Familial Malignant Hematological Disorders: A Systematic Assessment in 216 Consecutive Patients with Acute Leukaemia.
Authors: Huiart Laetitia , Lafage Marina , Rabayrol Laetitia , Charbonnier Aude , Bourdeleau Philippe , Sainty Daniele , Coso Diane , Mozziconacci Marie Joelle , Arnoulet Christine , Stoppa Anne-Marie , Murati Anne , Lehucher-Michel Marie Pascale , Sobol Hagay , Esinger François , Vey Norbert
Published: April 2020

Purpose

Our objective was to describe the biological characteristics of acute leukemia (AL) patients with or without a familial form of malignant hematological disorders.

Population

The study population included all newly diagnosed cases of AL treated in the departments of hematology of Institut Paoli-Calmettes between January 2002 and December 2004. We identified 291 eligible patients, of which 216 (74.2%) provided consent to participate in the study and filled a short questionnaire collecting demographic, personal and familial medical data. A face to face interview was arranged for 185 patients (85.7%) and a pedigree compiled for each family and reviewed by 2 familial cancer consultants.

Results

Among the 185 patients with full data on personal and familial history of cancer, 34 (18.4%) had a strong familial history of cancer of which 16 (8.6%) presented a familial form of malignant hematological disorders (at least another case of hematological malignancy in the 1, 2 or 3 degree relatives). Seven families had at least 2 first degree family members with leukemia and 2 families had 5 relatives (1st, 2nd or 3rd degree) diagnosed with leukaemia.

Most index cases were diagnosed with AML in both groups (88.1%). Among familial forms, 7 AML (50%) were classified as FAB M1 or M2. No M3, M6 or M7 were identified. White blood cell count was higher than 30 G/l in 37.5% of familial form as compared to 23.5% in sporadic cases (NS). The mean circulating blast percentage was higher in familial forms (66.6% (SD= 35.5)) than in sporadic cases (36.7% (SD=32.1)) (p=0.001). None of the familial forms were consecutive to pre-existing myelodysplasic or myeloproliferative syndrome. Among familial cases with AML, 43% had normal cytogenetics (vs 36% in sporadic cases); none had a complex karyotype; one presented with a t(3;15)(p26;q11) as a mosaic, a translocation not previously reported. Complete remission (CR) rates after 1st induction and Overall Survival (OS) were similar in both groups (CR in both groups: 61.5% (n=112); OS: median: 16 months (95%CI = 10.2 – 21.8) in familial forms vs 23.9 months (18.8 – 28.9)).

Conclusion

According to our data, among the 15 900 new cases of AL diagnosed every year in the US, 1 370 may correspond to familial forms. This warrants awareness of clinicians who should systematically assess family cancer history. Familial AL tends to present as a proliferative form, with no prior hematologic malignancies and normal cytogenetics. This is consistent with pathogenesis pathways described in other familial cancer syndrome.
The Growth Inhibition and Apoptosis Induction of Acute Myeloid Leukemia Blast Population by the Blocking IGF-IR Pathway.
Authors: Sun Si , He Yanli , Wang Xingbing , Liu Wei , Liu Jun , Huang Shiang
Published: April 2020

The insulin-like growth factor-1receptor (IGF-1R) is overexpressed in a variety of tumors and has been associated with cancer development. Here, we analysis the IGF-IR expression on the bone marrow cells from 45 newly diagnosed patients with acute myeloid leukemia (AML) by flow cytometry. IGF-1R universally expressed on AML blasts and the leukemia cell line HL-60, did not show significant correlation with FAB subtypes. However, the bone marrow cells from AML patients with high myeloblast counts (>80%) generally showed brighter IGF-IR expressions, which indicated the IGF-IR pathway might play an important role for AML blast proliferation and survival. Indeed, blocking the IGF-1R pathway by neutralizing monoclonal antibodies could reduce the proliferation of HL-60 cells by 38.28% at 48 hr. This inhibitory effect on blast growth was observed in 4 of 5 AML samples. In the same IGF-1R blocking treatment, the apoptosis of HL-60 cells was significantly induced, resulting in apoptosis of 57% of the cell population with the measurement of Annexin V vs PI staining by flow cytometry. The control contained only 20% apoptotic cells. We also demonstrated that the blockade of the IGF-1R pathway inhibited the phophorylation of the PI3K pathway component Akt in HL-60 cells when cultured in a serum free system with a supplement of 50ng/ml exogenous IGF. Since PI3K pathway activation greatly contributes to the proliferation, survival and drug resistance of AML, it is of interest to study whether blockading IGF-IR could also inhibit the PI3K pathway in primary AML blasts and synergize other anti-leukemia agents to improve the therapeutic effectiveness.

Conclusions:

IGF-IR may play an important role in the proliferation and survival of the AML blast population; Blocking the IGF-IR pathway could significantly inhibit the growth of AML blasts and considerably induce the apoptosis of AML blasts; IGF-IR could become a critical molecular target in anti-leukemia drug discovery.
Downs Syndrome Patients with Acute Lymphoblastic Leukemia; an Intermediate Outcome with a High Infectious Morbidity.
Authors: Saleh Mahasen , Khairy Ashraf R. , El-Solh Hassan , Al-Mahr Mohammed , Al-Musa Abdulrahman , Al-Seraihy Amel , Al-Ahmari Ali , Belgaumi Asim F.
Published: April 2020

Unlike AML, children with Down syndrome and Acute Lymphoblastic Leukemia (DS-ALL) have been reported to achieve significantly lower rates of remission, with higher mortality and a poorer overall survival. To further study this, we conducted a retrospective review for all DS children who were diagnosed and treated for ALL at our institution from 1997 to 2006. 12 (1.8%) out of 645 children (age 0–14 years) diagnosed with ALL had DS. The median age at diagnosis was 55 months (range 23 – 108 months) and eight were male. The most frequent signs and symptoms were fever (n=11; 91.6%), bleeding tendency (n=8; 75%) and organomegaly (n=9; 83.3%). The median WBC count at diagnosis was 4.99×109/L (range 0.58–500×109/L). The median platelet count and hemoglobin were 255×109/L and 80.5g/L, respectively. None of the patients had CNS disease at diagnosis. Leukemia cell cytogenetics revealed only the additional chromosome 21 for all patients, except in one patient who had 48XY,+X,+21. All cases had a precursor B-cell phenotype, and a DNA index of 1.0. Patients received treatment according to the protocol utilized for intermediate risk ALL at our institution. A good early response to induction therapy by a day-14 bone marrow (BM) evaluation was seen in all patients, and complete remission (CR) was achieved by day 28 of induction for the 11 who completed induction chemotherapy. One patient died toward the end of induction due to severe sepsis. Infection was the most commonly encountered morbidity throughout the course of therapy. Six episodes of febrile neutropenia, two documented bacteremias and one disseminated invasive Aspergillosis were reported during the induction phase. In the post-induction period, 21 episodes of febrile neutropenia were reported, in addition to six documented bacteremias and two patients developed fungal infections, one with fungemia and the other with a nail bed infection. Four patients relapsed at a median time of 13 months from diagnosis. Relapses occurred in the BM for two, as isolated CNS in one, and as combined BM and CNS relapse in one. Seven patients continue in first CR. The overall survival and the relapse free survival at 3 years were 53.7% and 60% respectively.

Conclusion: Although DS ALL children do not present with poor-risk features (infancy, T-cell phenotype, adverse cytogenetics, CNS involvement) they tend to have an intermediate treatment outcome even if treated on relatively intensive protocols. Delays in appropriate therapy due to infectious morbidity could contribute to this sub-optimal outcome.
Itraconazole-Enhanced Vindesine Neurotoxicity in Adult Acute Lymphoblastic Leukaemia.
Authors: Chen Suning , Wu De Pei , Sun Aining , Qiu Huiying , Jin Zhengming
Published: April 2020

Severe neurotoxicity following treatment with itraconazole and vincristine has been reported in 25 patients. Here we presented two adult acute lymphoblastic leukemia (ALL) patients developed neurotoxicity after treated with itraconazole and another vinca alkaloids, vindesine. Case 1: A 20-year-old male was diagnosed as having ALL and achieved complete remission (CR) after induction chemotherapy with DVP. He was treated with Hyper-CVAD for consolidation. Because he has a history of pulmonary invasive fungal infection, antifungal prophylaxis with itraconazole solution was started. Nine days after itraconazole was started, some symptom and signs of paralytic ileus were presented. He was diagnosed to have paralytic ileus secondary to neurotoxicity caused by itraconazole and vindesine. After the itraconazole was stopped, his paralytic ileus was improved rapidly. Case 2: A 37-year-old female was diagnosed as having ALL. A diagnosis of B-cell/T-cell biphenotypic acute leukemia was established based on the results of flow cytometry analysis. She was commenced on induction chemotherapy as for ALL (dexamethasone, Idarubicin, and vindesine). The patient presented as pulmonary infection with fever, cough, and expectoration in day 9. Emperical antifungal treatment with itraconazole injection was initiated. Five days after itraconazole injection was started, some symptom of neurotoxicity including limbs anesthesia, and paralytic ileus were presented. After the itraconazole and vindesine were discontinued, these symptoms were resolved. Given the interaction of itraconazole with vinca alkaloids leading to severe and even potentially fatal toxicities, the combination use of these drugs should be avoided.
Phase I Study of Temozolomide and Cisplatin in Patients with Acute Leukemia.
Authors: Katragadda Sreedhar , Liu Delong , Halicka Dorota , Kempin Sanford , Ahmed Nasir , Ahmed Tauseef , Seiter Karen
Published: April 2020

We previously demonstrated that temozolomide has significant activity in patients with acute leukemia (Seiter K, et al JCO 2002). Cisplatin can increase the activity of temozolomide through depletion of MGMT, the enzyme that rescues cells from temozolomide toxicity. We performed a phase I study of temozolomide in 17 patients with relapsed or refractory acute leukemia. Median age: 53 (range 26–73); M/F: 9/8; Dx: AML: 12, ALL: 3, biphenotypic: 2; median # of prior regimens: 2 (range 1–7); duration of 1st remission: primary refractory: 8 patients, 1–6 months: 3 patients, 7–12 months: 4 patients, greater than 1 yr: 2 patients; cytogenetics: good: 2, intermediate: 11, poor: 4; prior AHD: 6 patients; Cisplatin (50–100 mg/m2) was given on day 1; temozolomide (200 mg/m2/d for 5–7 days) was administered daily beginning 4 hours after cisplatin. Cisplatin was administered in the inpatient setting. Twenty-one cycles were administered. To date, no dose limiting toxicity has been observed. The median number of hospital days was 10 (range 2–37). Neutropenic fever occurred in 11/21 cycles. Other grade 3 non-hematologic toxicity included orthostatic hypotension, syncope, and constipation in one patient each, and elevated bilirubin in 2 patients. Four patients had grade 2 elevations of creatinine that resolved with hydration. Other grade 1/2 non-hematologic toxicity included fatigue, nausea/vomiting, constipation, bradycardia, diarrhea, hypokalemia and hypomagnesemia. The median number of RBC transfusions was 3U (range 0–8); the median number of platelet transfusions was 3 (range 0–12). No complete remissions were seen. However significant antileukemic activity was seen: the median pretreatment WBC was 3.2 (range 0.3–78.3); the median nadir WBC was 0.4 (range 0.1–9.6). Four patients had a significant reduction in bone marrow blasts: 40 to 22%, 87 to 38%, 69 to 3%, and 87 to 5%, respectively. The three latter patients were treated at the highest dose level. Cisplatin and temozolomide was well tolerated. We are continuing to accrue patients at the highest dose level to further assess efficacy.
Pegylated Recombinant G-CSF after Sequential High Dose Cytarabine Based Induction Chemotherapy in AML - Analysis of Pharmakokinetics and Effect on Neutrophil Recovery of Pegfilgrastim.
Authors: Fiegl Michael A. , Hiddemann Wolfgang , Braess Jan
Published: April 2020

BACKGROUND: The use of hematopoetic growth factors after conventional induction chemotherapy in acute myeloid leukemia (AML) remains controversial. Yet from former studies with sequential high dose AraC (S-HAM) in relapsed and refractory AML it was shown that the use of G-CSF reduced the early death rate significantly in this clinical setting. Pegylated filgrastim (NEULASTA©, Amgen) has shown to have a sustained-duration effect compared to conventional filgrastim due to decreased renal clearance. This analysis focuses on pegfilgrastim pharmacokinetics and effect on neutrophil recovery in AML.

MATERIAL AND METHODS: The ongoing AML CG pilot trial uses sequential high dose AraC as accelerated dose dense induction therapy in AML. It comprises 66 % of the established HAM-HAM double induction regimen (HAM: AraC 3 g/m2 bid for 3 days and mitoxantron 10 mg/m2 for 3 days) given within 2 weeks instead of four. 6 mg of Neulasta© was applied subcutaneously to all patients with complete blast clearance on day 18 after start of S-HAM. Blood samples for measurement of Neulasta plasma levels were obtained before application and afterwards daily up to day 21. Plasma concentrations of pegfilgrastim were measured by ELISA technique. Leukocyte count was obtained daily.

RESULTS: Analyses were performed in 19 patients at two institutions. After injection, peak plasma levels with a mean of 174 ng/ml were achieved within 48 hours after injection and t ½ was 6 days. 7 of these patients received a second injection of Neulasta 10 – 15 days later due to insufficient neutropenic recovery. Here plasma levels of > 200 ng/ml were achieved. Effective plasma levels of Neulasta (> 2.0 ng/ml) were observed up to 14 days after injection. In those patients who remained neutropenic, Neulasta plasma levels were observed even longer (t ½ > 6 days).

Pegfilgrastim clearance was significantly correlated to neutrophil recovery (r=−0.91, p=0.001). Median time to leukocyte recovery of > 1 G/l was 9 days after injection of the first dose in all evaluable patients.

CONCLUSION: In our study, time to neutrophil recovery in primary AML was shortened to 26 days by use of an accelerated, dose dense regimen plus pegfilgrastim application as compared historically to standard double induction therapy (median time to neutrophil recovery 45 days).

Figure Figure
Comparison of Survival Times after Administration of Tipifarnib or Ara-C-Containing Therapy, to Older Patients with Newly-Diagnosed Acute Myeloid Leukemia.
Authors: Estey Elihu H. , Thall Peter F. , Wang X.-M. , Giles Francis , Karp Judith E.
Published: April 2020

Non-cytotoxic (“targeted”) therapies are finding increasing use as initial treatment of older patients with untreated AML. However, the effect of this practice on survival is unknown. We compared survival times in 140 patients age ≥ 65 given the non-cytotoxic agent tipifarnib (T) at 5 centers (C1–C5) not including M.D. Anderson (MDA) with survival in similarly-aged patients treated at MDA with intermediate-dose ara-C + idarubicin (IA, 124 patients) or + agents other than idarubicin (XA, 49 patients). Because there was no randomization between T and IA or XA comparisons were confounded by center effects (C1–C5 versus MDA). This motivated use of Bayesian sensitivity analyses accounting for hypothetical between-center effects as well as age, performance status, cytogenetics, and whether AML was de novo or secondary. These analyses suggested that, while the odds were 7 to 1 that T was associated with shorter survival than IA or XA, it was nonetheless plausible (95% credible interval for relative risk included 1.0) that survival with T was in fact longer than with IA or XA. More importantly, the magnitude of the differences in survival was sufficiently short that a case could be made for use of T given its greater convenience. More generally we suggest that single-arm trials administering experimental therapies to patients with AML should include interim comparison of survival times with those obtainable with standard therapy, and that randomized phase II trials to avoid treatment-trial confounding may be desirable.
No Significant Difference in Therapy Results between Acute Myeloid Leukemia Patients with Myeloblastic Bone Marrow Infiltration within the Range of 20–29% and over 29% at Diagnosis.
Authors: Grosicki Sebastian , Holowiecki Jerzy , Kyrcz-Krzemien Slawomira
Published: April 2020

Definition of acute myeloid leukemia (AML) includes a heterogenous group of disorders differing in cytogenetics, clinical symptoms and prognosis. A common feature is bone marrow infiltration with leukemic clone which is also present in peripheral blood and different organs.

Over thirty years ago the FAB (French-American-British) group defined the morpho-cytochemical classification of acute leukemias and proposed ≥30% leukemic blasts in bone marrow for a threshold between muelodysplastic syndromes and acute leukemias. In 2002 the WHO group of experts decided to decrease this border line to 20%.

The question concerning differences in results of particular treatment programs for AML patient subgroups with original blastic bone marrow infiltration within the range of 20–29% and ≥30% remains disputable.

To address this question we performed an analysis in 405 patients treated according to the PALG (Polish Adult Leukemia Group) protocol DAC and DA. At diagnosis there were 26 patients with myeloblastic bone marrow infiltration ranging 20–29% (group A) and 379 patients with infiltration ≥30% (group B). In 62,5% patients (n=10) of the group A AML was preceding by myelodysplastic syndrome.

In the time period 1999–2002 patients under study received as induction DAC-7 regimen: daunorubicin 60 mg/m2/d iv 1–3; cytarabine 200 mg/m2/d ci 1–7; cladribine 5 mg/m2 2h inf. iv d 1–5 or standard DA-7 regimen (the same regimen without cladribine). Patients achieving CR received two courses of subsequent intensive consolidation:HAM (HD AraC, mitoxantrone)HD AraC with or without cladribine in the DAC-7 or DA-7 arm, respectively.In the case of PR after the first induction course the same regimen was repeated. Post-consolidation therapy was in both arms comparable.

Complete remission (CR) rate was comparable in both populations and reached 73% in the group A and 70% in group B. There were also no statistical differences between overall survival (OS) and leukemia free survival (LFS) between both groups. OS after 5 years equals 34% in group A and 27% in droup B and LFS 42% and 27%, respectively (p=NS).

Our study proves that there are no differences in treatment results between AML patients with original bone marrow infiltration within 20–29% and ≥30% given the same treatment. It confirms that the revised WHO criteria are reasonable.
SU11657 Cooperates with Doxorubicin To Increase Survival of Leukemic Mice.
Authors: Sevcikova Sabina , Lee Brian D. , Kogan Scott C.
Published: April 2020

Acute promyelocytic leukemia (APL) is characterized by t(15,17) resulting in fusion of RARα to PML; in rare cases, RARα may be fused to other partners. The PML-RARα fusion initiates APL-like disease in mice, including a response to ATRA. FLT3, a receptor tyrosine kinase, is mutated in a large number of human AMLs and roughly 40% of cases of human APL. In our mouse model of APL, we previously introduced activated FLT3 (ITD mutation) into bone marrow of PML-RARα transgenic mice and observed rapid development of AML. This disease is transplantable into sublethally irradiated mice with onset of disease between 3–6 weeks, creating a system suitable for therapeutic study.

There has been considerable effort focused on the development of small molecule inhibitors of FLT3 as therapeutics for hematological malignancies, in particular, AML; some of these inhibitors are currently in clinical trials. SU11657 is a multi-targeted inhibitor of class III/V receptor tyrosine kinases, including FLT3. We previously observed a synergistic effect of SU11657 in combination with ATRA in our PML-RARα/FLT3 model. We set out to use this model to assess the effectiveness of anthracycline chemotherapy in combination with SU11657. This model is in vivo resistant to the maximum tolerated dose of doxorubicin (3 mg/kg/day).

We found that the median survival of untreated and doxorubicin-treated mice with PML-RARα/FLT3 leukemia was not significantly different (42 and 45 days, respectively). SU11657 increased the median of survival to 55 days, whereas the combination therapy increased median survival to 62 days (P=0.003). Neither agent alone or in combination significantly increased survival of mice with PML-RARα leukemia lacking activated FLT3. These results suggest that the use of targeted therapeutics, such as a FLT3 inhibitor, can overcome chemoresistance to traditional therapies in AMLs.
Inhibitory Effect of ssRNA40, a Toll-Like Receptor-8 Agonist, on the Proliferation of Acute Myeloid Leukemia Cells.
Authors: Wang Xingbing , Xiong Fang , Liu Jun , Huang Shiang
Published: April 2020

Acute myeloid leukemia (AML) is a malignant hematopoietic disorder with considerable impairment of the immune system. We have previously reported that the T-cell-mediated immunity in AML patients could be in part damaged by the increased prevalence of CD4+CD25high regulatory T cells (T-regs). Toll-like receptors (TLR), found on many immune cells, including T-regs, have been shown to be important molecules in both innate and adaptive immune responses. Recent studies demonstrated that synthetic and natural ligands for human TLR8 could completely reverse the suppressive function of T-regs and enhance antitumor immunity, which provides a possible application of TLR8 ligands in immunotherapy for a patient with AML. However, an important question is whether the TLR8 ligands could also have direct impacts on the AML cells. In this study, we first examined the expression of TLR8 on fresh leukemic cells from 11 AML bone marrow samples and a human AML cell line (U937) by using RT-PCR analysis. We found that U937 cells and the majority of the AML patients (8+/11) expressed TLR8. To further investigate the functional role of TLR8, the cell line U937 was stimulated with ssRNA40, a Toll-like receptor-8 agonist, and analyzed by the measurements of cell proliferation, apoptosis and co-stimulatory molecules (CD80 and CD86) expression. The proliferation of U937 was detected by cell counting kit-8 (CCK-8) and the apoptosis was examined by AnnexinV and PI. Interestingly, the proliferation of U937 cells could be inhibited in a dose-dependent manner, whereas apoptosis and co-stimulatory molecules expression were not significantly changed with ssRNA40 stimulation. In the same condition, the inhibitory effect of ssRNA40 was not revealed on the proliferation of K562 CML cell line, even though TLR8 was also expressed on the cells. This might suggest that a specific pathological pathway in AML was blocked by ssRNA40. The signaling pathway that can restrain AML cells from proliferation is currently being studied. In conclusion, the present data may offer a unique therapeutic potential for ssRNA40 as an inhibitor, by which we will be able to directly inhibit the proliferation of AML cells and simultaneously enhance the patients’ immune response against the malignant cells.
Utility of Bone Marrow Biopsy, Bone Marrow Aspirate, Flow Cytometry, and Molecular Testing in the Clinical Staging of Lymphoma: An Institutional Experience.
Authors: Dumont Randall J. , Fernandez Louis A.V. , Juskevicius Ridas
Published: April 2020

Lymphomas are a diverse group of solid tumors of lymphoid cells that are broadly subdivided into Hodgkin lymphoma (HL) and non-Hodgkin lymphoma (NHL) categories. In the Western world, lymphoma is the most common hematologic malignancy. Complete and accurate staging of the patient with lymphoma is essential in determining the extent of initial disease. Bone marrow biopsy (BMB) remains the gold standard for assessing bone marrow involvement by lymphoma. It allows morphological diagnosis as well as the use of immunohistochemistry in difficult cases. At the time of BMB, other specimens are often gathered for use in ancillary investigations that can aid in diagnosis and/or staging. These investigations include bone marrow aspirate (BMA), flow cytometry (FC), and molecular studies (M) such as PCR. The objectives of this study were to evaluate the performance characteristics of BMA, FC, and M relative to BMB in lymphoma staging, to determine how frequently ancillary testing is positive for bone marrow involvement with lymphoma while BMB is negative, as well as to determine the clinical significance of this situation. Retrospective analysis was performed on 294 lymphoma cases from 1997 to 2002 at a single adult tertiary care center. Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), pre-test probability (PreTP), post-test probability given a positive test result (PTP+), and post-test probability given a negative test result (PTP−) were determined. Chart review was performed to determine clinical significance of the situation when BMB is negative and one or more ancillary investigations are positive. The number of cases positive for lymphoma by BMB, BMA, M, and FC was 157,113, 109, and 111, respectively. The performance characteristics are indicated in Table I. Twelve cases in which one or more ancillary investigations were positive when BMB was negative were identified. Clinical management was not altered in these cases. In staging of lymphoma, BMB remains the gold standard for the determination of bone marrow involvement. When compared to BMB, ancillary investigations have a high specificity and PPV, but only moderate sensitivity and NPV. Ancillary bone marrow investigations appear to add little information to lymphoma staging, and may not be fiscally justified.

Table I. Performance Characteristics Of Ancillary Bone Marrow Investigations In The Staging Of Lymphoma Parameter Bone Marrow Aspirate Molecular FlowCytometry Abbreviations: PPV = positive predictive value; NPV = negative predictive; PreTP = pre-test probability; PTP+ = post-test probability given a positive test result; PTP− = post-test probability given a negative test result Sensitivity 71% 64% 68% Specificity 98% 93% 96% PPV 97% 92% 95% PreTP 53% 53% 53% PTP+ 97% 92% 95% PTP− 26% 31% 28%
Absence of Chlamydial Infection in Japanese Patients with Ocular Adnexal MALT Lymphoma.
Authors: Yakushijin Yoshihiro , Kodama Toshio , Takaoka Ikue , Tanimoto Kazushi , Bessho Hiroko , Yamanouchi Jun , Azuma Taichi , Narumi Hiroshi , Hato Takaaki , Hasegawa Hitoshi , Yasukawa Masaki
Published: April 2020

The extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue (MALT lymphoma) is a distinct type of B-cell lymphoma that develops in extranodal sites and usually has an indolent clinical course as a localized disease. Recent clinicopathological studies suggest a strong relationship between MALT lymphoma and inflammatory diseases of the epithelium such as autoimmune diseases and infections (Sjogren’s syndrome, Hashimoto’s thyroiditis, Helicobacter pylori infection, and etc.), because chronic antigen stimulation has been causally linked to the development of lymphoproliferative diseases in B cells. MALT lymphoma grows in extranodal lymphoid tissue and exists in epithelium tissues, and it is often difficult to distinguish malignant lymphoid tissue from benign inflammation. However, this is only one of the possible causes; recent positive and negative correlations between ocular adnexal MALT lymphomas and chlamydial infections reported by different researchers indicate several different possible explanations. Here, we analyze twenty-three cases of ocular adnexal MALT lymphoma whose monoclonality was confirmed by immunoglobulin heavy chain gene rearrangement and/or the analysis of cell surface antigens for evidence of expected causative factors, using both ELISA analysis to detect antibodies and specific PCR gene amplification followed by Southern blot analysis. Our series of patients did not show any serological relations to the Epstein-Barr virus, the Hepatitis C virus, or to Chlamydophila psittaci. Two cases had positive serum antibodies for autoimmunity, another two cases had positive antibodies against Chlamydia trachomatis. None of our cases showed the presence of chlamydial 16S rRNA or 16S-23S spacer rRNA genes (C. psittac, C trachomatis, C. pneumoniae, and C. felis) after PCR amplification. These results indicate that the inflammatory agents are still unknown in our series of ocular adnexal MALT lymphomas, and that some types of chlamydial infections are not associated with ocular adnexal MALT lymphoma in southern regions of Japan.
Ki-67 Expression Level Is Not a Significant Prognostic Factor in Korean Diffuse Large B-Cell Lymphoma.
Authors: Sym Sun-Jin , Lee Sung Sook , Kim Min Kyoung , Kim Shin , Suh Cheolwon
Published: April 2020

The cell proliferation rate of tumors from patients with non-Hodgkin’s lymphomas has been shown to correlate with survival in previous studies. The aim of this study was to evaluate the independent prognostic value of Ki-67 expression level and the significance of cell proliferation rate as an outcome predictor. To this end, we reviewed the hospital case records and histopathological materials of 92 patients diagnosed with diffuse large B-cell lymphoma in a single center. The cell proliferative rate was expressed as percentage of Ki-67 positive cells (Ki-67 labeling index). All patients were available Ki-67 expression level. High Ki-67 labeling index was defined as nuclear Ki-67 expression in ≥ 70% of malignant cells. A cutoff point of 70% was used because this divided the group of cases into two subgroups of similar size (Ki-67 labeling index < 70, n = 46, Ki-67 labeling index ≥ 70, n = 46). The median age was 57 years (range, 15 to 83 years). The median follow-up time was 14 months. Sixty-nine (75%) patients were treated R-CHOP regimen initially and 13 (14%) patients were CHOP regimen. In a univariate analysis, age (p =.003), performance (p =.001), serum LDH (p =.020) were significant prognostic factors for overall survival. But Ki-67 had no statistical significance (p =.554). Multivariate analysis used Cox’s multivariate regression model revealed that serum LDH (relative risk estimate 6.7; 95% confidence interval, 1.5, 30.5), age (relative risk estimate 7.0; 95% confidence interval, 1.9, 26.4) were independent prognostic factors for overall survival. Taking all the results into consideration, it seems that Ki-67 expression level has no significance as outcome predictor in Korean diffuse large B-cell lymphoma.
SCFβ-TRCP Mediates Ubiquitination and Degradation of the Erythropoietin Receptor and Controls Cell Proliferation.
Authors: Verdier Frederique , Meyer Laure , Deau Benedicte , Forejtnikova Hana , Dumenil Dominique , Lacombe Catherine , Mayeux Patrick
Published: April 2020

Control of intensity and duration of erythropoietin (Epo) signalling is necessary to tightly regulate red blood cells production. After Epo stimulation of erythroid cells, 2 types of signal are transduced via the Epo receptor (Epo-R): positive signals involved in survival and proliferation, and negative signals involved in signal arrest. We have recently shown that the ubiquitin/ proteasome system plays a major role in the control of Epo-R signalling duration and desensitisation processes. Indeed, after Epo stimulation the Epo-R is ubiquitinated and its intracellular part is degraded by the proteasome, preventing further signal transduction. The remaining part of the receptor, together with associated Epo is internalised and degraded by the lysosomes (Walrafen et al 2005 Blood, 105, 600-608). Our aim was to identify the E3 ubiquitin ligase involved in Epo-R ubiquitination. The Epo-R contains a putative β-Trcp binding site in its intracellular domain. Interestingly, this putative binding sequence is located in a region of the Epo-R that is deleted in erythroid progenitors from patients with familial polycythemia. We show that β-Trcp is responsible for Epo-R ubiquitination upon Epo stimulation.

After Epo stimulation, β-Trcp binds to the Epo-R and this binding is dependent on Jak2 activation. Mutation of the Ser 462 residue of the Epo-R, located in the consensus β-Trcp binding site abolished β-Trcp binding, Epo-R ubiquitination and EpoR cleavage by the proteasome. Activation of the mutated Epo-R is prolonged in comparaison with Epo-R WT and BaF3 cells expressing this mutated receptor unable to bind β-Trcp are hypersensitive to Epo. Whether the removal of the β-Trcp binding site contributes to the hypersensitivity to Epo in familial polycythemia is currently under study.]