bluuebunny/crossref_metadata_2025 huggingface dataset:
Adult Acute Lymphoblastic Leukemia and Cytogenetic Abnormalities in Different Racial and Ethnic Subgroups.
Authors: Tamkus Deimante , Jajeh Ahmad , Berko Ebenezer , Osafo David , Griza Decebal , Kovarik Paula , Catchatourian Rose
Published: October 2019

Inferior survival of African American and Hispanic patients with acute lymphoblastic leukemia (ALL) has been reported. The reason for this is unclear. A retrospective analysis was conducted to see if abnormal cytogenetics account for the differences. We have analyzed cytogenetic studies of 39 adults (16 year and older) with newly diagnosed ALL who were consecutively treated at John Stroger Hospital of Cook County between 1997 and 2005. The study population included 13 (33%) African Americans, 18 (46%) Hispanics, 6 (16%) Caucasians, and 2 (5 %) other ethnic background adults. Male to female ratio was 2:1. Mean age at diagnosis was 26 (range between 16 and 71 years). A clonal cytogenetic abnormality was detected in 27 patients, and 12 patients had normal karyotype. Patients with t (9;22)(q34;q11), BCR-ABL + by FISH, t (4;11)(q21;q23), +8, −7 cytogenetic abnormalities were assigned to an unfavorable cytogenetic group. This group was composed of 14 patients. None of our study patients had favorable cytogenetics: del (12p), t(12p), t(14q11-q23), inv(14)(q11;q32) or t(10;14)(q24;q11). Remaining 25 patients with normal karyotype (n=12) or miscellaneous cytogenetic abnormalities (n=13) were classified as normal risk group. 54 % of African Americans had unfavorable cytogenetics, compared with 33 % Caucasians and 17 % Hispanics. Translocation t (9;22) alone or in association with other cytogenetic abnormalities was most commonly seen. 9 patients had single, and 18 had multiple chromosomal changes. Hispanic group had more complex cytogenetic changes when compared with the African American or Caucasian groups. Unfavorable cytogenetic abnormalities may account for inferior survival in African Americans, but other factors such as compliance or pharmacogenetics should be evaluated, especially in the Hispanic patient population.
The Wnt/β-Catenin Pathway Promotes Growth and Survival of Acute Lymphoblastic Leukaemia (ALL) Cells In Vitro.
Authors: Khan Naveed I. , Bradstock Kenneth F. , Bendall Linda J.
Published: October 2019

Wnt signalling is known to play an important regulatory role in self-renewal of haematopoietic stem cells as well as development of normal lymphocyte progenitors. Recently this pathway was implicated in a number of haematological malignancies including AML, CML, MM and CLL. However its role in acute lymphoblastic leukaemia is not clear. We have previously shown that primary pre-B ALL cells and cell lines express various Wnt family members, receptors and downstream transcription factors. We extended these studies further to elucidate a functional role of Wnt signalling in leukemic cells. Exposure to Wnt3a, a ligand inducing the canonical Wnt signal cascade, resulted in stabilisation of beta-catenin and its nuclear translocation in all growth factor independent pre-B cell lines (n=3), stromal-dependent cell line (n=1) and primary cases (n=3) examined. Wnt3a treated pre-B ALL cell lines and a patient sample were more resistant to apoptosis induced by serum deprivation (viability on day 4, 51.3±18.4% Wnt3a treated vs 29.5±19.9% control treated, p-value <0.05). 3H-thymidine incorporation studies revealed 4.0±1.7 fold increase in proliferation which was associated with increased cell cycle entry as determined by Ki-67 staining. Cell cycle analysis confirmed an increase in the proportion of cells in S and G2M phases of cell cycle. These effects were associated with increased transcriptional activity of toposiomerase II-alpha as assessed by semi-quantitative PCR and protein expression. Taken together our results indicate that Wnt-dependant β-catenin signalling pathway is active in pre-B ALL cells and promotes their growth and survival in vitro.
Epidemiology of Adult Acute Lymphoblastic Leukemia in Manitoba, Canada: Does This Approximate Clinical Trial Data?.
Authors: Dao Vi , Rubinger Morel , Bow Eric J. , Schacter Brent A. , Shore Tsiporah B. , Woloschuk Donna , Demers Alain , Sun Xiaoyun , Seftel Matthew D.
Published: October 2019

Background: Outcomes of unselected adult ALL patients (pts) have not been well studied at a population level. Whether such pts behave differently to those in formal clinical trials is unclear. We developed a uniform therapeutic protocol as the provincial standard of care in Manitoba, a province with 1.2 million people, for adult pts since 1989 (“ALL89-1”). The aim of this retrospective study was: to ascertain the local epidemiology of adult ALL; to analyze the efficacy and toxicity of the ALL89-1 regimen and any other regimens given to pts in a population-based setting.

Methods: All pts aged ≥18 years (yrs) diagnosed with ALL in Manitoba between 01/1989 and 11/2004 were identified via the provincial cancer registry. A review of computerized and paper charts was conducted. We focused on demographics, diagnostic clinical characteristics, therapy, treatment related toxicities, and whether stem cell transplantation (SCT) was performed.

Results: There were 81 pts with a median age of 44yrs (range, 18–90). 53 (65%) pts were male. 58 pts received ALL89-1, 12 received other regimens and 11 patients received palliation only. Thirteen pts were Philadelphia chromosome positive (Ph+). Complete remission (CR) rate after one cycle of induction therapy was 74% for all pts, and 74% for ALL89-1 pts. At a median follow-up of 56 months (range 7–265), estimated overall survival (OS) and leukemia free survival (LFS) at 5 years was 26% (95% Confidence interval [C.I.], 15–36) and 24%( 95% C.I. 14–35). The 5 year OS and LFS for the ALL89–1 group was 23% (95% C.I. 11–35) and 20% (95% C.I. 8–32). There were 13 treatment related deaths in the 58 ALL 89-1 pts, 4 of whom died post SCT. In multivariate analysis for all 81 pts, attainment of CR post induction chemotherapy independently predicted improved survival (hazard ratio 0.23, 95% C.I. 0.11–0.5). In contrast, age > 55yrs, Ph+ status, and high WBC at diagnosis were not significant survival predictors. Palliative pts survived for a median of 10 days (range, 0–53) from diagnosis.

Conclusions: This comprehensive province-wide study of adult ALL pts treated at a single cancer centre shows that durable leukemia free survival is possible, but for only a small subgroup. These results appear inferior to results of published clinical trials. The reasons for the relatively poor outcomes require further analysis in this and other jurisdictions.
Myeloid Blastic Cells with MLL-ELL Translocation Can Convert Their Morphology into the Fibroblastoid Cells.
Authors: Tashiro Haruko , Noguchi Mitsuho , Shirasaki Ryosuke , Goto Moritaka , Kawasugi Kazuo , Shirafuji Naoki
Published: October 2019

Objective: It has been reported that acute leukemia with the fusion product of MLL-related molecule is determined to be the poor prognosis. To make clear one of its biological characters, we cultured AML blasts with t (11; 19) (q23; p13.1), which produced fusion protein of MLL and ELL, and the morphological changes as well as their DNA, and RNA were examined.

Method: Bone marrow cells were obtained from two patients with MLL-ELL fusion gene after obtaining informed consent. There were over 90% blastic cells in bone marrow. Mononuclear cells were obtained with Ficoll (SG. 1077), and were cultured in DMEM with 10% FCS in CO2 incubator. The morphology of the cells was observed for 3 months. Also, DNA was extracted, and MLL-ELL translocation was analyzed with genomic Southern blotting with probe x which contains BamHI-digested 0.9 kb fragment from MLL cDNA. RNA was extracted with GTC method, and 1st strand cDNA was synthesized with oligo-dT primer, and PCR was performed with MLL common primer and ELL antisense primer. The reaction of DNA sequencing was performed with BigDye Terminator Cycle Sequencing kit (Applied Biosystems), and analyzed with ABI Prism 3700 DNA analyzer.

Result: After one month of the cultures, fibroblastoid cells were expanded without any blastic cells onto them. Southern analysis, RT-PCR study, and DNA sequence revealed that the fusion cDNA product with MLL and ELL was observed in fibroblastoid cells.

Discussion: AML blasts with MLL-ELL fusion gene can convert their morphology into the fibroblastoid cells. The fibroblasts have been reported to be resistant to the chemotherapeutic agents. This morphological conversion may contribute one of the causes of poor prognosis in AML patients with MLL-ELL fusion gene. We now characterize precisely these fibroblastoid cells in point of the expression of the specific proteins, and the functions including the binding and proliferating capability to the non-adherent blastic cells.
Platelet-Derived Factor VIII (FVIII) Corrects the Murine Hemophilia A Phenotype Even in the Presence of FVIII Inhibitors.
Authors: Shi Qizhen , Wilcox David A. , Fahs Scot A. , Cooley Brian C. , Desai Drashti , Weiler Harmut , Morateck Patricia A. , Montgomery Robert R.
Published: October 2019

Inhibitory antibody development is not only the most severe and important complication of FVIII infusion in hemophilia A patients, but also a major potential concern in patients following hemophilia A gene therapy. We have developed a transgenic model in which human B-domain deleted FVIII (hFVIII) expression is controlled by the platelet-specific αIIb promoter (2bF8) and results in FVIII storage together with VWF in platelets. The transgenic mice were then bred into FVIII knockout mice (FVIIInull) so that the only FVIII in peripheral blood is derived in platelets. The bleeding phenotype was corrected in the 2bF8/FVIIInull transgenic mice, even though FVIII was not detectable in plasma. We further hypothesize that platelet-derived FVIII in a complex with VWF will 1) protect FVIII from inactivation by circulating FVIII-specific antibodies and 2) release FVIII in sufficiently high local concentration at sites of vessel damage so as to overcome immediate inactivation by FVIII inhibitors. To address our hypothesis, we used three strategies -1) an acute model infusing plasma from highly immunized FVIIInull mice, 2) immunization of 2bF8 transgenic mice with human FVIII, and 3) a model using transplantation of spleen cells from highly immunized mice into sub-lethally irradiated 2bF8 transgenic mice. Thirteen mice were infused with plasma containing 48-64 BU and 12 survived following tail clipping using a 1.59 mm template. FVIII-immunized transgenic mice (given adjuvant) generated anti-FVIII inhibitory antibodies as high as 350 BU after a single immunization. The level of platelet-FVIII and the platelet number in these mice were not significantly different from pre-immunization levels and 13 of 15 survived tail clipping. In the 3rd group, spleen cell transplantation resulted it titers of >5000 BU and 8 of 12 transgenic mice survived tail clip. In contrast, only 2 of 7 wild type mice survived tail clip with inhibitor titers >5000 BU after spleen cell transplantation. None of 9 untransplanted FVIIInull mice survived. These results demonstrate that platelet-derived FVIII is protected from inhibitor inactivation and correct the phenotype of murine hemophilia A even in the presence of a high-titer anti-FVIII antibody. This approach may be useful not only for preventing inhibitor formation in hemophilia A gene therapy but also for gene therapy in the case of pre-existing inhibitors.
Investigation into the Pro-Apoptotic and Pro-Inflammatory Properties of 6-Thioguanine and 6-Mercaptopurine.
Authors: Bruneau Johanna C. , Jenkins P. Vincent , O’Marcaigh Aengus , Smith Owen P.
Published: October 2019

The thiopurines, 6-mercaptopurine (6MP) and its analog 6-thioguanine (6TG), have considerable anti-leukaemic activity. 6MP has contributed to the cure of tens of thousands of children with acute lymphoblastic leukaemia (ALL) with little in the way of significant toxicity and hence it is now established as the purine nucleoside of choice in maintenance therapy of childhood ALL. This is despite the fact that 6TG may indeed be a better alternative to 6MP as it has been shown to be at least one log more active in vitro than 6MP and in a recent large U.S. ALL Trial, isotoxic comparisons of 6MP and 6TG showed 6TG to have fewer CNS events. However, like in a similar recent U.K. study there was increased toxicity noted in this U.S. Trial, namely the emergence of an inflammatory response-like syndrome affecting the liver resulting in sinusoidal occlusion (SOS) with hepatomegaly, thrombocytopenia, ascites, and elevated bilirubin and liver transaminases. Although these purine nucleosides have been in widespread use for more than 50 years, their exact cytotoxic mechanism of action remains to be elucidated with incorporation of the nucleoside analogs into DNA, resulting in apoptosis, and by the inhibition of de novo purine synthesis leading to metabolic arrest being the two main contenders. The mechanism of the inflammatory insult causing SOS with 6TG is also poorly understood. In order to identify the differential effects of 6-TG and 6-MP in vitro, we measured apoptosis via the TUNEL assay, viability using the WST-1 assay, and the secretion of inflammatory cytokines (TNF-a and IL-8) by ELISA in a panel of human leukaemia cell lines: THP-1 (acute monocytic leukaemia), Jurkat E6.1 (acute T lymphocytic leukaemia), 697 (acute pre-B lymphocytic leukaemia), as well as the hepatocyte cell line HepG2 after treatment with either of the two drugs. There was a significant (p≤ 0.01) increase in apoptosis in THP-1and 697 cells following treatment with 6-TG but not 6-MP, although there was a significant decrease in viability observed with both agents (p≤ 0.01). In contrast, a significant (p≤ 0.01) increase in apoptosis was observed in Jurkat E6.1 cells after treatment with both 6-TG and 6-MP. Neither apoptosis nor viability was significantly affected by either 6-TG or 6-MP in HepG2 cells. IL-8 secretion was significantly (p≤0.01) increased by 6-TG and 6-MP in THP-1 and HepG2 cells, however there was no corresponding increase in TNF-a levels. Neither agent had a significant effect on the secretion of either TNF-a or IL-8 in 697 or Jurkat E6.1 cells. These results show that there are different apoptotic and pro-inflammatory responses to these drugs most likely reflecting their efficacy and toxicity profiles
Amphotericin B and Itraconazole as Empirical Antifungal Therapy in Children with Acute Leukemia with Neutropenic Fever.
Authors: Kim Sun-Young , Park Jong-Sun , Jeong Nak-Gyun , Jeong Dae-Chul , Cho Bin , Kim Hack-Ki
Published: October 2019

Purpose: Fungal infections are one of the important causes of morbidity and mortality in patients with hematologic malignancies. Amphotericin B (ABV) and itraconazole (ITZA) have been used as the standard empirical antifungal therapy in neutropenic patients with acute leukemia who have persistent fever that does not respond to antibiotic therapy. ABV is an antifungal drug associated with side effects such as fever and chills, symptoms which may be mediated by pro-inflammatory cytokines such as interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α). We assessed modulation of these pro-inflammatory cytokines as well as the anti-inflammatory cytokines (IL-4, IL-1Ra) by ABV and ITZA.

Methods: From October 2004 to February 2005, a total of 30 episodes from acute leukemia patients with febrile neutropenia were analyzed for this study. They were randomly allocated to receive intravenous ABV or ITZA for 14 days. Clinical responses were evaluated at the completion of therapy, and cytokine IL-1β, TNF-α, IL-4, and IL-1Ra were measured for determination to know the correlation between two antifungal agents and inflammatory cytokines.

Results: Empirical antifungal agents were given to 37 patients (ABV 20, ITZA 17), and 30 patients (ABV 15, ITZA 15) were evaluable for efficacy. White blood cell and absolute neutrophil count in the group treated with ITZA increased early days of treatment, so the duration of neutropenia in ITZA group is shorter. Serum creatinine level is lower in ITZA group than in ABV group but this is not statistically significant. There was no significant difference in response rate between two groups. The IL-1β was increased in ABV treatment group and the ratio of IL-1Ra/IL-1β is markedly decreased in ABV treatment group while increased in ITZA group.

Conclusions: ITZA and ABV have at least equivalent efficacy as empirical antifungal therapy in neutropenic children with acute leukemia. However ITZA is associated with significantly less toxicity in clinical and molecular aspects.
Pilot Study of the Feasibility of Assessing DNA Damage and Apoptosis Induced by Chemotherapy in Children with Acute Leukemias.
Authors: Halicka Dorota , Ozkaynak Fevzi M. , Huang Xuan , Traganos Frank , Darzynkiewicz Zbigniew
Published: October 2019

The aim of the present study was to explore whether early assessment of the extent of DNA damage in leukemic blasts and incidence of apoptosis induced during chemotherapy is feasible. The long term goal is to develop a rapid assay to assess the effectiveness of chemotherapy early in the course of treatment of leukemia. Mononuclear cells were isolated from blood samples of 7 ALL patients with peripheral blasts, 4 of whom were, at diagnosis, high-risk ALL while the remaining 3 were in relapse. The median age of the patients was 17 yrs (2–25 yrs) and all were entered into an anthracycline-containing drug regimen. Samples were collected:

prior to drug administration; and, one hour after completion of the drug infusion.

The extent of DNA damage was assessed by analysis of histone H2AX phosphorylation (γH2AX) occurring at sites of DNA double-strand breaks (DSBs), which could be detected immunocytochemically and measured by multiparameter flow cytometry in conjunction with DNA content. The incidence of apoptosis was estimated 1 h after drug administration and also after 24 h ex vivo culturing, by identification of cells with DNA fragmentation detected by the TUNEL assay. In patients with complete clinical response (6 of 7) at the end of the induction phase, an increase of γH2AX (post treatment - pretreatment mean) was detected 1 h after the administration of drug treatment. The apoptotic index in this group after 24 h ex vivo culture was 1–16%. In the only patient not responding to treatment - one of 2 relapsed ALL patients -, there was no induction of DSBs or increase in apoptosis.

Response to induction therapy γH2AX mean fluorescence in arbitrary units Apoptotic index ex vivo (%) post 24 h Complete response 8 16 Complete response 8 5 Complete response 5 1 Complete response 29 7 No response 0 0 Complete response 4 2 Complete response 4 Not estimated

Although the number of patients is very small in this pilot study, the results suggest that a cytometric assay that reveals the extent of DNA damage based on detection of H2AX phosphorylation may provide an early prognostic marker.
Induction of Complete Remission with 5-Azacytidine in a Patient with Acute Myeloid Leukemia Refractory to Intensive Chemotherapy.
Authors: Kuendgen Andrea , Graef Thorsten , Knipp Sabine , Hildebrandt Barbara , Czibere Akos , Huenerlituerkoglu Ali N. , Germing Ulrich , Haas Rainer , Gattermann Norbert , Kobbe Guido
Published: October 2019

Intensive chemotherapy achieves complete remission in about 75% of patients with acute myeloid leukaemia (AML). For patients refractory to intensive chemotherapy, prognosis is very poor and treatment options are limited. We report a case of AML which was refractory to induction chemotherapy as well as two salvage regimens. The patient then achieved CR through monotherapy with low-dose azacitidine.

The 57-year-old patient was admitted to our hospital with a diagnosis of AML M1. A bone marrow biopsy revealed a blast count of 88%. The karyotype was 48,XX,+8,+11 [4/20]. The patient received induction chemotherapy with idarubicin, cytarabine, and etoposide (ICE). Because her disease was refractory to treatment, with a bone marrow blast count of 66%, the patient received a salvage regimen with high-dose cytarabine, mitoxantrone, and all-trans retinoic acid (A-HAM). Still, AML blasts persisted, with a blast count of 52%. The patient then received FLAMSA (fludarabine, amsacrine, and high-dose cytarabine) as a second salvage regimen, but again failed to achieve remission (medullary blast count 50%). Pancytopenia persisted over a period of approximately 3 months (WBC <100/μl, Hb and platelets transfusion-dependent). We then decided to treat her with 5-azacitidine. During the first cycle, she received additional G-CSF because of fungal pneumonia. Azacitidine was given at a dose of 100mg/m2 subcutaneously for 5 days. Treatment was tolerated without side effects. The patient responded swiftly. After the first cycle, peripheral cell counts normalized and the peripheral blast count decreased to 1%. The bone marrow blast count was 3% after 3 cycles, and <1% after 5 cycles. Peripheral blood counts dropped only slightly during treatment cycles, and the patient required no further transfusions. Cytogenetic analysis, including FISH with a centromeric probe for chromosome 8, gave normal results. With the exception of the first cycle, azacitidine was administered in an outpatient setting. After completing the fifth cycle, the patient went on to receive allogeneic stem cell transplantation.

In the nineteen-seventies and -eighties, conventional (cytotoxic) dosages of 5-azacytidine showed some activity against AML, but did not achieve remission rates comparable to high-dose cytarabine. Treatment-related toxicity was considerable. Recently, low-dose azacitidine, supposedly acting as a demethylating agent, was approved for treatment of myelodysplastic syndromes in the U. S.. According to recent studies, methyltransferase inhibitors seem to achieve good response rates in patients with high-risk MDS. Cytogenetic remissions have been achieved in patients with poor risk karyotypes. Our case report confirms that 5-azacitidine can be effective in AML, even in patients who have a poor prognosis with conventional therapeutic approaches.
Attenuated Dose of Idarubicin for the Elderly De Novo Acute Myeloid Leukemia with Normal Karyotype.
Authors: Cheong June-Won , Kim Yuri , Park Sun Young , Park In Hae , Kim Jin Seok , Min Yoo-Hong
Published: October 2019

The incidence of acute myeloid leukemia (AML) increases with age. Because of poor performance status, co-morbidity and treatment-related side effects, a conventional dose chemotherapy containing anthracyclins may be toxic to the majority of elderly patients. In contrast, the administration of suboptimal dose of myelosuppressive chemotherapy could lead to an unsuccessful clinical outcome including lower complete remission (CR) rate. To evaluate the effect of attenuated dose of idarubicin, compared to the standard dose, on the clinical outcome and chemotherapy-related complications, we analyze the consecutive 32 elderly de novo AML patients (range, 60 – 74 years) with normal karyotype. Eleven patients received one cycle of conventional-dose remission induction chemotherapy (idarubicin, 12 mg/m2/day on days 1–3 and cytarabine 100mg/m2/day on days 1–7) (Group 1) and 21 patients received attenuated-dose idarubicn (8 mg/m2/day on days 1–3) and cytarabine (100mg/m2/day on days 1–7) (Group 2). Six cases (54.5%) in Group 1 and 12 cases (57.1%) in Group 2 had CR. The difference of CR between the two groups was not significant (P = 0.59). The intervals from the chemotherapy-starting date to the date of CR documentation were not also different between two groups (median 31.5 days on Group 1 vs 27.0 days on Group 2) (P = 0.29). The median number of transfusion requirement during the induction therapy was not different in the red blood cells (10 units, each) and platelets (16.5 units in Group 1 vs 18.0 units in Group 2; P > 0.05). Thirty patients received the recombinant human granulocyte colony-stimulating factor (G-CSF) three days after termination of chemotherapy. The duration of G-CSF administration was not different between two groups (P = 0.86). However, the frequency of septicemia and septic shock after induction chemotherapy was statistically significantly higher in Group 1 (54.5% and 9.5%, respectively) compared to that in Group 2 (36.3% and 0.5%, respectively) (P < 0.01). We also observed a higher incidence of clinically-documented invasive fungal infection in Group 1 (45.5%) compared to Group 2 (15.0%), although the difference was not statistically significant (P = 0.095). The incidence of other regimen-related toxicities including renal dysfunction, hepatic dysfunction and heart failure was not different between two groups. Overall survival and disease-free survival also were not different between the groups. In conclusion, the attenuated dose of idarubicin can be recommended for the remission induction chemotherapy for the elderly de novo AML patients with normal karyotype since it is associated with lower incidence of sepsis and septic shock with comparable CR rate.
Extended Study for Topotecan, Idarubicin, and Intermediate-Dose Cytarabine Combination Chemotherapy in Patients with Refractory or Relapsed Acute Myelogenous Leukemia and High-Risk Myelodysplastic Syndrome.
Authors: Kim Yuri , Cheong June-Won , Kim Jin Seok , Park In Hae , Park Sun Young , Lee Seung Tae , Min Yoo-Hong
Published: October 2019

The development of more effective therapy for patients with refractory or relapsed acute myelogenous leukemia (AML) and high-risk myelodysplastic syndrome (MDS) is a major concern in hematology. We previously reported a preliminary data that combination chemotherapy with topotecan, idarubicin, and intermediate dose cytarabine had a significant anti-leukemia activity and acceptable range of toxicity for the high-risk AML/MDS patients. In this study, we demonstrate an extended data for investigating larger number of patients with long-term follow-up of five years. The forty-seven patients were enrolled: 10 with primary refractory AML, 16 with AML in first relapse, 9 with AML in second relapse, and 12 with MDS-RAEB/RAEBT. Patients received as salvage therapy a single course of idarubicin 12 mg/m(2) IV bolus on days 1–3, cytarabine 1 g/m(2) over two hours q 12 hr on days 1–5, and topotecan 1.25 mg/m(2) over 24 hr on days 1–5. Median age of patients was 47 years (range 17–69 years). Patients with AML were categorized to high-risk (n=21), intermediate-risk (n=11), and low-risk (n=3) cyrogenetic group. All patients were evaluable for response: 23 (48.9%) achieved complete remission, 18 with AML (51.4%) and 5 with MDS (41.7%), respectively. The median remission duration and survival of patients with AML were 7 and 13 months, respectively. Median remission duration and survival of MDS patients was 10 and 15 months, respectively. Severe myelosuppression was observed in all patients, resulting in the fever or documented infections in 85.1% of patients. Median time to recovery of neutrophils > or =0.5 x 10(9)/l was 23 days (11–37 days) and for platelets > 20 x 10(9)/l 37 days (11–63 days). Reversible grade 3–4 toxicities included diarrhea (3 patients) and mucositis (13 patients). The relapsed AML patients (n=16) were grouped into four categories to stratify the probability of achieving complete remission (CR): group 1, first CR duration > or = 2 years and receiving first salvage treatment (S1); group 2, first CR duration 1–2 years and receiving S1; group 3, first CR duration 0–1 years and receiving S1; and group 4, first CR duration 0–1 years and receiving S2, S3, or S4 after failing S1. The response rate of group 4 (33.3%) was significantly lower than that of other groups (P < 0.05). We conclude that combination chemotherapy with topotecan, idarubicin, and intermediate dose cytarabine is an effective salvage chemotherapy for high risk AML/MDS patients, and the first CR duration is a significant predictor for response.
Does High Dose Cytosine Arabinoside Improves Disease Free Survival for Down Syndrome Acute Myelocytic Leukemia Patients?.
Authors: Saleh Mahasen , Khairy Ashraf , Al-Mahr Mohammed , El-Solh Hassan , Al-Musa AbdulRahman , El-Hassan Ibrahim , Belgaumi Asim F.
Published: October 2019

Acute myelocytic leukemia (AML) in Down Syndrome (DS) children is characterized by a young age of onset (< 2 years), a low white blood cell count and high frequency of Megakaryocytic leukemia. DS children with AML have higher disease free survival (DFS) rates as compared to non DS AML patients. Previous studies have suggested that intensification chemotherapy may not be necessary for the treatment of DS children with AML. The objective of this study was to clarify the effectiveness and toxicities of using high dose Cytosine Arabinoside (HD AraC) intensification in the treatment of DS AML. Clinical data for children (<14 years) with DS AML, diagnosed between September 2000 to May 2005, were retrieved from the hospital data base. Patients were divided into two groups; Group A patients received chemotherapy containing HD AraC, while Group B patients did not. A total of 15 patients were included, eight in Group A and seven in group B. The median age at diagnosis was 22 months (A=23 months, B=22 months). The two groups were matched regarding their clinical and laboratory parameters. There was no significant difference in DFS between groups A and B, 75% and 85% respectively (P = 0.82) at a mean observation period of 42.9 months for group A and 23.12 months for group B. The median time to relapse was 6 months for group A and 8 months for group B. The overall treatment related toxicity was higher in Group A patients but achieved only borderline significance (P = 0.06). However, when toxicity was assessed separately for induction and post induction phases of chemotherapy there were significantly more infectious events (17 v. 2; p=0.0006) in the post induction phase which includes HD AraC intensification in Group A. Even when only serious infections (bacteremia, fungal infection, sepsis) were included in the evaluation this difference persisted (7 v. 1; p=0.0339), with less toxicity for Group B patients. No such difference was noted between the two groups during induction chemotherapy. In conclusion the use of HD AraC in post-induction intensification phases for DS AML children does not improve DFS and is associated with more treatment related toxicity.
Retroviral Vector Integration Deregulates Gene Expression but Has No Consequences on the Biology and Function of Transplanted T Cells.
Authors: Recchia Alessandra , Bonini Chiara , Magnani Zulma , Urbinati Fabrizia , Sartori Daniela , Muraro Sara , Ciceri Fabio , Bordignon Claudio , Mavilio Fulvio
Published: October 2019

Retroviral vectors have raised safety concerns for the genotoxic risk associated with their uncontrolled insertion into the human genome. The occurrence of T-cell leukemia in patients treated by gene therapy for X-SCID has been correlated with insertional activation of a T-cell proto-oncogene. However, no such event has been reported in other pre-clinical or clinical trials, suggesting the existence of disease- and/or transgene-specific risk factors in the X-SCID case. We have analyzed the integration pattern of a retroviral vector expressing a suicide gene (HSV-TK) and a surface marker (ΔLNGFR) in T-lymphocytes from patients treated with donor lymphocyte infusion (DLI) after hematopoietic stem cell transplantation for therapy of acute and chronic leukemia. The suicide gene allows to control graft-versus-host disease, a major complication of DLI, while preserving anti-tumor and antiviral activities. Proviral integrations were analyzed by sequencing the vector-genome junctions in T-cell DNA obtained from 4 different patients up to 9 years after treatment. Over 85% of proviral integrations occurred within transcription units, with a preference for first introns and upstream promoter regions. Over 75% of intragenic integrations were in genes active in T cells at the time of transduction, and ~4% occurred at recurrent genomic locations (hot spots). Microfluidic real-time PCR in RNA extracted from individual T-cell clones harboring proviral integrations in proximity of transcription start sites showed that one fifth of these integrations affect the expression of nearby genes. However, transduced T-cell populations maintain remarkably stable gene expression profiles, phenotype, biological functions and immune repertoire in vivo, with no evidence of clonal selection up to nine years after administration. Analysis of transduced cells before and after transplantation indicates that integrations interfering with normal T-cell function are more likely to lead to clonal ablation than expansion. Despite the potentially dangerous interactions with the T-cell genome, retroviral integration has therefore little consequences on the safety and efficacy of T-cell transplantation.
Granulocytic Sarcoma in Erythroleukemia (M6).
Authors: Abbasi Elham , Mims Martha P. , Udden Mark M.
Published: October 2019

Granulocytic Sarcomas are extramedullary tumors composed of immature myeloid cells, these tumors can be seen in myeloid malignancies such as chronic myelogenous leukemia, myelodysplastic syndrome and acute myelogenous leukemia. They can be seen at the time of diagnosis of the myeloid malignancy, prior to diagnosis, or after treatment as a first sign of relapse. We report here the case of a young woman who presented with a 6 month history of a slowly enlarging nasal mass. Biopsies of the lesion had been inconclusive until a CBC revealed abnormal blasts in the peripheral blood. Subsequently bone marrow biopsy revealed more than 59% erythroid precursors with a preponderance of immature erythroblasts and 33% myeloblast consistent with AML (M6). Reanalysis of the nasal biopsy demonstrated positive staining for CD68, Myeloperoxidase, Lysozyme, and CD117, but negative for CD34, suggestive of granulocytic sarcoma. The patient received induction chemotherapy with idarubicin and cytarabine and achieved bone marrow remission along with complete resolution of the nasal mass. To our knowledge this is the first report of a granulocytic sarcoma in association with AML M6.
Phase I/II Study of Clofarabine Combined with Standard Dose Cytarabine as Remission Induction Therapy of De Novo AML in Elderly Patients ≥60 Years.
Authors: Foran James M. , The Angeline S. , Devetten Marcel , Meleth Sreelatha , Worrell Jeff , Dixon Pam , Sekeres Mikkael A.
Published: October 2019

Despite significant advances in the treatment of acute myeloid leukemia (AML) in younger adults, there has been little progress in the treatment of older patients (age≥60 years), who comprise the majority of those with the disease. Clofarabine is a purine nucleoside analog with single agent activity in patients with relapsed AML. In addition, clofarabine potentiates Ara-C cytotoxicity in vitro through increased intracellular Ara-CTP accumulation, making this an attractive combination.

A phase I/II study has therefore been initiated combining clofarabine with standard dose cytarabine (100mg/m2/day x 7) in pts age ≥60 years with de novo AML. The starting dose of clofarabine was 30mg/m2/day x 5 beginning on day 2 (Dose level I). Patients were accrued in cohorts of 3–6 to establish dose limiting toxicity (DLT); cohort expansion at the maximum tolerated dose (MTD) is planned in phase II using a Simon 2-stage design. Detailed plasma and intracellular pharmacokinetics were performed during induction therapy with Ara-C alone (day 1), and following the addition of clofarabine (day 2) to determine the effect of clofarabine on intracellular Ara-CTP accumulation. Pts with residual AML on d14–21 restaging bone marrow (BM) biopsy were eligible to receive Re-Induction with 5 days of clofarabine & Ara-C. Those achieving complete remission were also eligible to receive 1–2 cycles of consolidation with Ara-C (d1–5) & clofarabine (total 3 cycles of planned therapy).

Dose limiting toxicity was encountered at dose level I (see Table 1). 2/4 pts achieved CR, in 1 case with residual cytogenetic abnormality, and there were 2 treatment-related deaths from infxn (culture neg sepsis, n=1; Candida tropicalis, n=1). In the latter case (pt 4), BM aplasia was achieved, but the pt died on d25 prior to hematologic recovery. In view of the DLT, the protocol has therefore been amended to allow 25% dose de-escalation of clofarabine to Dose Level -I (22.5mg/m2/day x 5), and to limit eligibilty to pts age 60–75 yrs inclusive. Routine use of aggressive pre-hydration and antibiotic and antifungal prophylaxis is now mandated.

Clofarabine & cytarabine is a highly active induction regimen in older adults age ≥60 yrs with de novo AML, but has significant myelosuppressive and infectious toxicity. The study is proceeding in phase I at Dose Level -I to establish the MTD.

Phase I, Dose Level I PT AGE, GENDER FAB CYTOGENETICS F/U BM TOXICITY (Gr.III/IV) OUTCOME MLD - multilineage dysplasia; F&N - fever & neutropenia; CR - complete remission 1 66M M2 Diploid D21: residual AML renal, infxn Death 2 61M M2 Complex D14: aplastic F&N CR 3 69M M2 with MLD Intermediate Risk D21: recovering F&N CR 4 77F M2 Diploid D14: aplastic renal, infxn, capillary leak Death
Peripheral Blood Lymphocytic Profile in Patients with Diffuse Large B Cell Lymphoma and Follicular Lymphoma.
Authors: Cavallin Fernando , Vicario Giovanni , Manente Paolo , Di Gaetano Rosa , Tagariello Giuseppe
Published: October 2019

Diffuse Large B Cell Lymphoma (DLBCL) and Follicular Lymphoma (FL) are the most common adult low-grade non Hodgkin’s lymphomas. The influence of these diseases in peripheral blood lymphocytes is not well defined. Indeed the lymphocytic arrangement can be altered on account of the leukaemic form (although it slightly ever occurs); on the other hand the cause of occasional anomalies can be the involvement of the immune system against neoplasm. In order to contribute to the knowledge of these conditions we have analysed, at diagnosis, the lymphocytic immunophenotype in peripheral blood of 61 subjects: 27 were affected by DLBCL, average age 68, and 34 by FL, average age 61 years. Therefore we quantified the number of lymphocytes and evaluated essential markers, using flow cytometry, to define T, B, NK subsets by: CD3, CD4, CD8, CD19, SIgk, Sigl, CD56, and expression of CD11a molecule on T CD8. The absolute peripheral blood lymphocytes count presented a reduction in 51% and in 32% of the cases with an increase in 4% and in 3% of the subjects respectively considering DLBCL and FL. On the contrary T cells (CD3) had similar decrease, 33% and 32%, and different augmentation 15% and 3%. T cells ratio CD4/CD8 was under normal in 23% and in 12% of the patients but over normal in 12% and 29% always in DLBCL and FL. B cells (CD19) were reduced in 35% and in 12% of the subjects but increased in 8% and in 14%, whereas clonal restriction was present in 8% and in 20% of the components of the two groups. Natural Killer lymphocytes (CD56) were under normal in 12% and in 6% of bthe cases but over in 40% and 20%. Finally CD11a was over-expressed in 87% and in 68% of the patients of the respective pathologies. After selecting patients aged over 60 years, following four parameters that showed a significant variability was obtained: 1) lymphopenia in 50% of the cases in both groups; 2) similar results 11% and 15% about clonal restriction; 3) increase of the NK population 42% and 30% in DLBCL and FL; 4) very high over-expression of CD11a on T CD8 of 90% and 80%. Therefore DLBCL and FL are lymphoproliferative diseases where there is an important subtraction of lymphocytes, particularly in elderly people, from peripheral blood (perhaps because of accumulation in lymphnodes). These lesions present clonal restriction of B cells only in few cases (confirming the low known leukaemic form) while Natural Killer population are well represented especially in DLBCL. The over-expression of CD11a is the most altered parameter and seems almost a typical marker of these diseases above all in over 60 years subjects. Consequently if rarely happens that a leukaemic form of DLBCL and FL are found by flow cytometry however immunological defined alterations are very frequent in most of the cases of old patients.
WNT6 mRNA Is Overexpressed in Variuos Non-Hodgkin’s Lymphoma Cases with No Apparent Relation to Cyclin D1 Level.
Authors: Swoboda Pawel M. , Gos Monika , Rymkiewicz Grzegorz , Blachnio Katarzyna , Walewski Jan , Janik Przemyslaw
Published: October 2019

A casual role for Wnts in the development of some human cancers seems to be established. Binding of Wnt to its receptor results in β-catenin stabilization and in consequence, translocation to the nucleus, where it activates the specific target gene’s expression including CCND1 (cyclin D1). Abnormal levels of β-catenin may therefore contribute to neoplastic transformation by causing accumulation of cyclin D1. Overexpression of cyclin D1, as a result of the t(11;14)(q13;q32) translocation, which places cyclin D1 gene under the enhancer of immunoglobulin heavy chain is considered the hallmark of mantle cell lymphoma (MCL) but high levels of cyclin D1 have also been observed with various levels in some cases of non-MCL lymphoproliferative disorders not involving t(11;14), such as hairy cell leukemia (HCL), splenic lymphoma with villous lymphocytes (SLVL), prolymphocytic leukemia (PLL) and some atypical CLL cases. Little is known about potential Wnt signaling in mature lymphocytes or lymphoid malignancies. Recent finding, that the Wnt pathway regulates the proliferation and survival of normal B lymphocytes, together with WNT16 overexpression in leukemic cell lines and high expression of five other Wnts including WNT6 in CLL suggests that Wnt family members can contribute to tumorgenesis in the immune system. In the current study we evaluated the relationship between Wnts and cyclin D1 mRNA expression level in NHL patients. WNT6 and CCND1 mRNA expression level normalized to 18 sRNA was evaluated by RT-PCR on 200 biopsy samples taken from 150 lymphoma patients. In parallel, samples underwent pathological and flow cytometry examination. WNT6 mRNA was detected in all specimens tested. The 18 sRNA/WNT6 ratio ranged from 1 to 650. Most of the CLL specimens were among those with high WNT6 mRNA expression, but no apparent correlation between the level of WNT6 mRNA expression and cyclin D1 status was found so far.
Clinical and Endoscopic Presentation of Primary Gastric Lymphoma: An Italian Multicenter Study.
Authors: Patti Caterina , Zullo A. , Tedeschi L. , Di Raimondo F. , Hassan C. , Recine U. , Bibas M. , Miedico A. , Romanelli A. , Vinci M. , Costa A. , Restivo G. , Mistretta A. , Montalbano L. , Linea C. , Marrone C. , Caruso L. , Chiarenza A. , Bonanno G. , Lizzani G. , Stella F. , Mirto S. , Morini S. , Cottone M. , Andriani Alessandro
Published: October 2019

Background. The stomach is the most frequent site of intestinal lymphomas. However, few data are available on the clinical-endoscopic presentation of gastric lymphoma as well as on possible differences in clinical pattern and endoscopic features between low-grade (LG) and high-grade (HG) lymphomas. In this study, we evaluated such aspects on consecutive primary gastric lymphoma patients observed in the last 12 years (1993–2004) in four Italian Hospitals (1 North, 2 Centre, 2 South).

Methods. Clinical, histological, and endocospic records of consecutive patients diagnosed with LG or HG gastric lymphoma were retrieved and accurately evaluated. Symptoms were categorized as “alarm” (anaemia/melaena/heamorrage, persistent vomiting, weight loss) or “no alarm” (epigastric/abdominal pain, heartburn, dyspepsia/bloating). The endoscopic findings were classified as “normal” (no macroscopic lesions) or “abnormal” (ulcer, erosions, nodular pattern, hypertrophic folds, polypoid mass). Statistical analysis was carried out by using the Chi squared test.

Results. During the study period, 143 patients with primary gastric lymphoma were detected. Overall, 61 patients were observed in the first 6 years and 82 in the last 6 years. The main results of the study are summarized in the table 1.

Conclusions. The incidence of primary gastric lymphoma seems to be increasing. The overall prevalence of alarm symptoms is quite low, and they may be absent in near 75% of LG lymphoma patients. Moreover, contrarily to HG, LG lymphoma may present as a normal endoscopic finding and it is more frequently associated with H. pylori infection. At diagnosis, HG lymphoma is more frequently detected in an advanced stage as compared to LG lymphoma.

Overall (143 patients) LG lymphoma (73 patients) HG lymphoma (70 patients) P value Age (mean ± SD) yrs 59.5 ± 14.2 59.4 ± 13.3 59.7 ± 15.1 0.4 Sex (M /F) 83/60 44/29 39/31 0.6 Alarm symptoms 57 (40%) 19 (26%) 38 (54%) 0.0009 Normal endoscopy 15 (10%) 15 (20%) 0 (0%) 0.0004 H. pylori infection 66 (73%) 47 (86%) 27 (39%) <0.0001 Stage (IA />IA) 78/65 58/15 20/50 <0.0001
Beta-2 Microglobulin and Soluble Interleukin-2 Receptor Serum Levels Correlates with Human Immunodeficiency Virus Associated Non-Hodgkins Lymphoma-Diffuse Large B-Cell Lymphoma (HIV-NHL-DLBCL).
Authors: McPherson Eugene , Ng J. , Hazel E. , Tassy P.
Published: October 2019

Beta-2-microglobulin (B2M) is a cell surface protein on many somatic cells including T and B lymphocytes and macrophages as a subunit of class I major histocompatibilty complex (MHC). Soluble interleukin-2 receptor (sIL-2R) is a T cell derived cytokine that induce proliferation of activated T cells, participates in differentiation of B cells and modulates macrophage function and phenotype. Both serum B2M and sIL-2R levels reflect immune-system activation and are elevated in patients with HIV-NHL-DLBCL. Combined detection of B2M and sIL-2R can be used to predict treatment response or failure in patients with aggressive HIV-NHL-DLBCL. When highly active antiretroviral therapy (HAART) chemotherapy is administered to refractoriness of elevated B2M and sIL-2R serum levels may be seen in HIV-NHL-DLBCL poor responders or failures. As sIL-2R levels decrease, B2M levels remain elevated in early stage HIV-NHL-DLBCL. Treatment of these patients with CHOP-rituximab and HAART both markers decrease. However, in late stage disease (stage III-IV, HIV-NHL-DLBCL based on DLBCL), B2M and sIL-2R remain significantly elevated and prognosis poor even after immune restoration of CD4+ T cell count and HIVRNA viral load approach normal.

CASE: We present one of several cases - a 38 year old HIV positive male with DLBCL of his sinus turbinates, had turbinectomy surgery with sIL-2R level of 617 U/mL (5,460.226 pg/ml) postop which gradually increased to 27,673.95 pg/ml in less than six months. The B2M level while on HAART decreased from 18.8 mg/L to 4.4 mg/L. His LDH, CRP, and GGT levels were elevated but the CD4+ T cell count was 26% and his HIVRNA viral load became undetectable. After CHOP-rituximab therapy sIL-2R decreased to 10,806 pg/ml from 27,674 pg/ml but slowly rose to 39,975.25 pg/ml and B2M increased to 19 mg/L. The CD19+ B cell count was zero and a CD4/CD8 ratio improve to 0.546.

CONCLUSION: B2M and sIL-2R serum level measurements provides data that when used together may offer a better management approach to these HIV-NHL-DLBCL patients. B2M and sIL-2R elevated refractory levels correlates with tumour burden and predicts poor prognosis in aggressive HIV-NHL-DLBCL.