bluuebunny/crossref_metadata_2025 huggingface dataset:
Recurrent Chromosome Abnormalities Define Non-Overlapping, Unique Subgroups of Tumors in Chronic Lymphocytic Leukemia Patients with Known Karyotypic Abnormalities
Authors: Jimenez-Zepeda Victor H , Chng Wee Joo , Braggio Esteban , Kay Neil , Leis Jose , Fonseca Rafael
Published: April 2020

Background B-cell chronic lymphocytic leukemia (B-CLL) is a well-defined clinical entity with heterogeneous molecular and cytogenetic features. Chromosome aberrations could be associated to specific CLL clinical features and outcomes and their impact on clonal evolution should have been addressed by using temporal analysis. In order to demonstrate the role of temporal analysis in CLL we conducted a comprehensive karyotypic survey of a large number of CLL cases with already known genetic aberrations to fully describe their meaning in terms of biological evolution.

Methods A total of 1749 karyotypes were retrieved from the Mitelman Database of Chromosome Aberrations in Cancer. A matrix depicting the 360-band human chromosome ideogram was created. Regions that were either lost or gained in more than 3% of the cases were retained and identified as recurrent imbalances. Early and late imbalances were defined according to the appearance on the complex karyotypes. Descriptive statistics were used to summarize the genetic abnormalities.

Results The median chromosome number and the median number of abnormalities per tumor (NAPT) were 46 and 1 respectively. (Figure 1 A and 1B) The most common abnormality seen was trisomy 12 which occurred in 29% (508 cases) followed by 13q14del (10.34%), 13q13del (6.63%) and add14q32 (6.63%). The temporal analysis revealed +12, 13q-, −17, 17p-, −Y and −X to be early imbalances (TO<5), followed predominately by a late loss with TO=5–10.5 (11q-). Hierarchical clustering suggested there are different groups including:

trisomy 12, 13q-, 11q-, and 6q-, which at least in this clustering analysis appeared to be mutually exclusive.

In summary, we can conclude that overall CLL is a neoplasia that shows remarkable chromosome stability even when only abnormal karyotypes are evaluated. Interestingly, clustering analysis suggests there are non-overlapping, unique subsets of CLL cases where trisomy 12 is the most common and along with 13q-emerged as early events on CLL clonal evolution.

Figure 1A. Chromosomes distribution in chronic Lymphocytes Leukemia karyotypes Figure 1A. Chromosomes distribution in chronic Lymphocytes Leukemia karyotypes Figure 1b. Chronic Lymphocytes Leukemia and Number of Abnormalities Per Tumor (NAPT) distribution Figure 1b. Chronic Lymphocytes Leukemia and Number of Abnormalities Per Tumor (NAPT) distribution Figure Figure
The Prognostic Significance of Positive Direct Antiglobulin Test in Chinese Patients with Chronic Lymphocytic Leukemia
Authors: Xu Wei , Li Jianyong , Cao Xin , Zhu DAN-Xia , Yao Lin , Liu Qiong , Qiao Chun , Wu Yu-Jie
Published: April 2020

Chronic lymphocytic leukemia (CLL) is the most common type of adult leukemias in the Western countries, however, infrequent in the Eastern. Autoimmune hemolytic anemia (AHA) is a complication in chronic lymphocytic leukemia (CLL). The direct antiglobulin test (DAT) may be positive at some time during the disease course in up to 35% of cases, but overt AHA occurs less frequently. The aim of the study was to explore the prognostic impact of positive DAT in Chinese patients with CLL and its correlation with other prognostic factors, including Binet stages, lymphocyte count in peripheral blood, lactate dehydrogenase (LDH), β2-microglobulin (β2-MG), IgVH mutation status, ZAP-70, CD38 and cytogenetic abnormalities. Out of the 80 Chinese patients with CLL, positive DAT was found in 21 (30.6%) cases. The incidence of positive was 12.5% in Binet A, 23.8% and 44.4% in Binet B and C, respectively. The incidence of positive DAT was significantly increased at Binet C, compared with Binet A (P=0.006), and the presence of higher LDH and β2-MG levels correlated strongly with positive DAT (P=0.006 and P=0.004, respectively). Patients with unmutated IgVH genes had higher incidence of positive DAT than did patients with IgVH mutations (P=0.042), and positive DAT was also associated with higher level of ZAP-70 and CD38 (P=0.004 and P<0.001, respectively). We also analyzed positive DAT in different cytogenetic subgroups. Higher incidence of positive DAT was found in patients with unfavorable cytogenetic aberrations (deletion in 17p13 or 11q22) in contrast to lower level in good risk cytogenetics (deletion in 13q as the sole abnormality) (P = 0.002). Positive DAT was associated with poor outcome. Survival analysis showed that the patients with positive DAT had significantly shorter OS (mean, 106.3 months) (95% CI, 74.7 to 137.8 months) than the patients negative DAT (mean, 151.5 months) (95% CI, 122.3 to 180.6 months) (P=0.024). Patients treated with fludarabine were not likely to remain DAT positive and to change from negative to positive (P=0.209). In conclusion, DAT status provides a new prognostic indicator and correlates with other clinical or laboratory prognostic factors, and might be applied for the assessment of prognosis in patients with CLL.
Serum Thymidine Kinase Level in Chinese Chronic Lymphocytic Leukemia and Its Correlation with Other Prognostic Factors
Authors: Li Jianyong , Xu Wei , Yu Hui , Qiu Hongxia , Qiao Chun , Wu Yu-Jie , Liu Qiong , Zhu Danxia , Miao Kourong
Published: April 2020

Objective To investigate serum thymidine kinase (TK) level in Chineses patients with chronic lymphocytic leukemia (CLL) and its correlation with other prognostic factors, including Binet stages, absolute lymphocyte count (ALC), lactate dehydrogenase (LDH), immunoglobulin heavy-chain variable region (IgVH) gene mutation status, ZAP-70 protein and CD38 expression level, and cytogenetic aberrations.

Methods Serum TK1 level in 39 CLL patients was detected by TK monoclonal antibody (Anti-TK mAb) and enhanced chemiluminecence (ECL). IgVH mutation status was detected by multiplex PCR and sequencing of purified PCR amplification products. A panel of monoclonal antibodies and multiparametic flow cytometry were employed to immunophenotype and determine the expression of ZAP-70 protein and CD38. A panel of probes and interphase fluorescence in stu hybridization (FISH) were used to detect cytogenetic aberrations including 6q-, 11q-, +12, 13q-,17p- and IgH translocation.

Results The level of TK1 was higher in CLL patients that in normal control (P<0.05). TK1 level was not signifigantly correlated with sex, age, Binet stages, CD38, and cytogenetic aberrations. Patients with higher level of ALC, LDH, ZAP-70 and unmutated IgVH genes had higher levels of TK1 than those with lower level of ALC, LDH, ZAP-70 and mutated IgVH genes (P=0.018, P=0.018, P=0.038 and P=0.030, respectively).

Conclusions Serum TK1 level significantly correlates with ALC, LDH, ZAP-70 and IgVH mutational status, and could be a predictive marker of IgVH mutation status. Serum TK1 might be applied for the assessment of prognosis in Chinese patients with CLL.
CD38 Expression above 20% Predicts Disease Progression in Chronic Lymphocytic Leukemia and Is a Useful Prognostic Marker Where Cytogenetic and Molecular Markers Are Not Available
Authors: Alipour Sina , Leitch Heather , Vickars Linda M , Foltz Lynda M , Galbraith Paul F , Leger Chantal S , Ramadan Khaled M
Published: April 2020

The prognostic significance of CD38 expression and the cut off value has not been fully investigated. As CD38 is readily available test in patients with chronic lymphocytic leukemia (CLL), we investigated its role in prediction of disease progression when a cut off value of 20% is used. Progression free survival (PFS) was defined as the time from diagnosis to first treatment or last follow up. An electronic database search of pts with CLL who presented at St Paul’s Hospital between 1969 and 2007 was performed. Among 465 pts with CLL, 161 pts (35%) had their CD38 expression measured by flow cytometry. CD38 expression and its association with other prognostic factors such as age, Rai stage, lymphocyte count at diagnosis, gender and other immunophenotypic makers were analyzed. Out of 161 pts, positive CD38 expression (>20%) was found in 36 patients (22%). Comparing the baseline characteristics of the CLL pts with CD38+ and negative disease, we found CD38 positivity more common in male pts than in female pts (p=0.03). Also patients with CD38 positive disease tend to present with more advanced stage disease (p=0.056). Progression free survival at 2, 5 and 10y for the CD38+ CLL pts was 89%, 61% and 41% respectively compared with 95%, 81% and 62% for the CD38 negative group (p=0.03). Univariate analysis revealed the following factors as significant or marginally significant for disease progression: CD38+ (p=0.03), male gender (p=0.07), Rai stage (p<0.0001), lymphocyte count above 20 ×109/l at diagnosis (p<0.0001), CD5 expression <10% (p=0.01). On multivariate analysis, only disease stage at diagnosis (p<0.0001) and CD38 expression above 20% (p=0.04) retained significant and were predictive for disease progression. We conclude that CD38 expression above 20% at the time of diagnosis can be prognostically useful and predicts for disease progression and along with Rai staging can provide inexpensive tool to follow and monitor patients with CLL.

Table: Characteristics of patients with CLL based on CD38 ≥20% and <20%

Parameter CD38 ≥20% (%) CD38<20% (%) p value* *for differences between the groups. Number 36 125 Sex: M/F (ratio) 22/14 (1.6:1) 66/60 (1.1:1) 0.03 Age above 60 y 21 (58) 81 (65) 0.2 Rai stage: 0, 1+2, 3+4 20, 11, 5 (55, 31, 14) 100, 17, 1 (80, 14, 0) 0.056 Lymphocyte count above 20×109/l 9 (25) 23 (18) 0.3 CD5 <10% 6 (16) 29 (23) 0.3

Fig: Progression free survival for patients with CLL based on CD38 expression, cut off level 20%. Fig:. Progression free survival for patients with CLL based on CD38 expression, cut off level 20%.
Nucleolar Stress and p53 Mediated Erythroid Failure in Diamond Blackfan Anemia
Authors: Sieff Colin , Lodish Harvey F.
Published: April 2020

The discovery that several ribosomal protein genes can be mutated in DBA suggests that ribosomal protein gene mutations may account for many or all cases of DBA, and focuses attention on the ribosome. While experiments in yeast and mammalian cells show that RPS19 depletion or mutation leads to a block in ribosomal RNA biosynthesis, this result does not explain why erythropoiesis is so severely affected in DBA. We hypothesize that during fetal development immature erythroid cells proliferate more rapidly than other lineages and therefore require very high ribosome synthetic rates to generate sufficient capacity for translation of erythroid specific transcripts that must take place before these unique cells enucleate. To test this kinetic hypothesis we measured RNA biogenesis in primary mouse fetal liver cells and reported previously that during the first 24 hours cell number increases 3–4 fold while, remarkably, there is a 6-fold increase in RNA content during the same period, suggesting that the cells accumulate an excess of ribosomal RNA (80% of measured RNA) during early erythropoiesis. Retrovirus infected siRNA RPS19 knockdown cells show reduced proliferation of FACS sorted GFP positive cells at 48 hours. Although the cell yield is reduced, the differentiation pattern of the surviving GFP positive cells is similar to that of the controls. While quantitative RT-PCR analysis shows that RPS19 mRNA is rapidly depleted, Western analysis during this time course does not show a deficiency of RPS19 protein. This suggests strongly that the proliferative defect is not due to insufficiency of RPS19 protein, and is more likely due nucleolar stress induced by the block in ribosome biogenesis. Molecular consequences could lead to redistribution of cell cycle proteins normally resident in the nucleolus with consequent p53 mediated cell cycle arrest and or apoptosis. To test this hypothesis we used a culture system that allows expansion without differentiation of immature cells in SCF, EPO, IGF-1 and dexamethasone. Under these conditions proliferation of siRNA expressing precursors is reduced with an increased proportion arrested in G0/G1 in the knockdown cells. Furthermore, p53 is increased in the knockdown cells. Taken together, these data suggest that RPS19 insufficient cells undergo a nucleolar stress response and erythroid cells proliferate poorly because of p53 mediated cell cycle arrest and apoptosis.
Treatment of Chronic Lymphocytic Leukemia with a Hypomethylating Agent Induces Expression of NXF2, An Immunogenic Cancer Testis Antigen
Authors: Dubovsky Jason A , McNeel Douglas G , Powers John J. , Sotomayor Eduardo M. , Pinilla Javier
Published: April 2020

Critical to success of active immunotherapy against cancer is the identification of immunologically recognized cancer-specific proteins with low tolerogenic potential. Cancer testis antigens (CTAs) in particular, fulfill this requirement as a result of their aberrant expression restricted to cancer cells and lack of expression in normal tissues bypassing tolerogenic mechanisms against self. Although CTAs have been extensively studied in solid malignancies little is known regarding their expression in chronic lymphocytic leukemia (CLL). Using a two-pronged approach we evaluated the immunogenicity of 29 CTAs in 22 patients with CLL and correlated these results to RTPCR data from CLL cell lines and patient cells. We identified IgG specific antibodies for one antigen, NXF2 and confirmed this response by ELISA and Western blot. We found that treatment of CLL with 5-aza-2′-deoxycytidine can induce expression of NXF2 that lasted for several weeks after treatment. Treatment also increased levels of MHC and costimulatory molecules (CD80, CD86, and CD40) necessary for antigen presentation. In addition, we identified other promising antigens such as NY-ESO-1 and MAGE which may have potential immunotherapeutic application. Our findings suggest that NXF2 could be further pursued as an immunotherapeutic target in CLL, and that treatment with demethylating agents could be exploited to specifically modulate CTA expression and effective antigen presentation in malignant B-cells.
Anti-Drug Antibody Responses to Lumiliximab Are Not Detected in Relapsed Refractory CLL Patients Treated with Lumiliximab in Combination with FCR in a Phase 1/II Study
Authors: Ranuio Julie , Estrellado Annalee , Harris Sarah , Shannon Megan , Subramanyam Meena , Hughes Steve , Woodworth James , Tangri Shabnam
Published: April 2020

Lumiliximab, an anti-CD23 monoclonal antibody that binds specifically to human CD23, is under investigation for the treatment of patients with relapsed refractory chronic lymphocytic leukemia (CLL). The clinical safety monitoring for Lumiliximab includes testing for the presence of anti-drug antibodies (ADAs) in patient serum samples. In order to determine if patients develop an ADA response to Lumiliximab, serum samples were collected at specific time points throughout a Phase I/II study (152–30) of Lumiliximab in combination with FCR in relapsed CLL. Patient sera from this Phase I/II study were tested in a standard bridging ELISA in which samples were added to a drug coated plate and ADAs were detected using conjugated drug and colorimetric detection. Validation results showed the sensitivity of the assay was 1000ng/ml using a purified polyclonal rhesus anti-Lumiliximab antibody in undiluted human sera. ADAs were not detected in any patients that had evaluable post treatment (Week 13, Week 25, Week 41, and Month 12 post first dose) serum samples in the 152–30 study. Over the last few years, there has been an effort in the immunogenicity field to develop ADA assays with increased sensitivity and to adapt formats that, in general, have increased drug tolerance, i.e. reduction of drug interference in the assay. Therefore, a new ADA assay was developed using Meso Scale Discovery (MSD) technology that has become the reliable and current method for detection of ADA responses and incorporates an acid dissociation step that has been shown in literature to improve drug tolerance. Using the new method the same evaluable samples from the phase I/II study were tested to determine if any previously unseen ADA responses could be detected. The samples were incubated with biotinylated and ruthenylated drug, added to a streptavidin coated plate and ADAs were detected using electrochemiluminescence. Validation results show that the newly developed MSD assay has sensitivity of 250ng/ml using a purified polyclonal chicken anti-Lumiliximab antibody in undiluted human sera. All the samples tested in the MSD assay (n=32) were negative despite greater sensitivity than the ELISA method and an acceptable drug tolerance at the time points samples were drawn. PK concentrations at the specified ADA collection times ranged between 0μg/ml and 114μg/ml whereas drug tolerance ranges from 1μg/ml to 200μg/ml depending on ADA concentrations. Drug interference is a major factor in not being able to detect antidrug responses, yet several of the corresponding post dose PK time points showed low to no drug levels and yet no anti-drug antibodies were detected. These data imply that Lumiliximab when administered in combination with FCR has a low immunogenicity rate.
Do Microenvironmental Factors Play Important Role in Supporting the Maintenance of Leukemic Stem Cells of Chronic Myelogenous Leukemia?
Authors: Cho Yeon Hee , Kim Dong Hwan , Jang Jun Ho , Kim Seok Jin , Kim Kihyun , Kim Won Seog , Jung Chul Won
Published: April 2020

Background : In the era of tyrosine kinase inhibitor(TKI) for chronic myelogenou leukemia(CML), it is becoming more important to develop the way to get rid of the leukemic stem cells(LSC), as there is frequent relapse when TKI therapy is stopped even when molecular remission is achieved. Some researchers reported macrophages and Wnt signaling in bone marrow microenvironment might play significant role in the development and the progression of CML.

Methods : We isolated CD34+CD38− LSC from bone marrow cells from 8 patients with CML chronic phase and also CD34+CD38− normal hematopoietic stem cell from 7 normal controls using magnetic cell separation system. CD34+CD38− cells were cultured on stromal cells for 5 weeks and transfered to methylcellulose media for long-term culture initiating cell(LTC-IC) assay. We checked whether there is a difference in LTC-IC capacity of LSC when cultured on leukemic or normal stromal feeder cells. CD45− stromal supporting cells were established by 3-week culture after 12Gy irradiation of CD34− bone marrow cells. We studied the expression of VCAM, ICAM, E-selectin in stromal cells and the presence of BCR-ABL transcript. Wnt and β-catenin, frizzled, Notch1 were analyzed in the LSCs. CD14+ cells were isolated and added in LTC-IC assay whether the presence of leukemic or normal CD14+ cells would make a change in the maintenance of LSCs.

Results : The number of CD34+CD38− cells in CML was in the range of 1.3 × 103 to 9.2 × 104, that we could do only two sets of 2 × 2 LTC-IC assay that statistical analysis was impossible. LTC-IC frequency of LSCs was about 5-times higher compared to that of normal stem cells irrespective of the kind of stromal cells used. CD14+ cell co-culture did not changed significantly the LTC-IC frequency both in normal and LSCs. Wnt 2B, 5A and 10B were highly expressed in LSCs irrespective of the stromal cells. β-catenin was also strongly expressed in LSCs compared to normal stem cells. Notch1 was expressed only in CD34+CD38− LSCs but not in normal stem cells. The kind of stromal cells or the addition of CD14+ cells in LTC-IC assay did not affected the Notch1 expression in LSCs.

Conclusion : CML CD34+CD38− LSCs seem to have more LTC-ICs and wnt and notch pathway may play important role in the maintenance of LSCs that could be therapeutic targets.
Heme Oxygenase-1(HO-1): A Promising Novel Target Molecule for Overcoming Imatinib Resistance in Chronic Myeloid Leukemia
Authors: Wang Jishi
Published: April 2020

Objective : HO-1 is a microsomal enzyme catalyzing the first, rate-limiting step in degradation of heme, HO-1 is a inducible isoform of HO, it can be strongly induced in response to cellular stress and diverse oxidative stimuli, including its substrate heme, Many studies have convincingly shown that HO-1 is a cytoprotective and antiapoptotic enzyme. the objective of this study was to investigate the influence on the K562 cell growth and apoptosis after hemin-induced HO-1 expression, and to investigate the influence on K562 cells and imatinib-resistant CML cells after ZnPPIX-induced HO-1 inhibition.

Methods: different concentrations hemin (0umol/l A20umol/l 30umol/l)was used respectively to induce HO-1 expression of cultured chronic myeloid leukemia cell line K562, then detected HO-1 mRNA expression under different time by RT-PCR, and MTT was used to detected the viability of K562 cells. In addition, we used STI571(2 μmol/L) deal with the hemin-induced cells, then confirm HO-1 protective effect against STI571 use MTT. Then ZnPPIX was used to inhibition HO-1 expression of K562 and imatinib-resistant cells, similarly, RT-PCR and MTT was used for analyzed.

Results:

The HO-1 mRNA was not tested when absence of hemin, 8h after treated with hemin of 20 μmol/L, we can test the HO-1 mRNA expression, and at 16h the expression is reach to the peak, 16h after treated hemin under different concentrations (10umol/l, 20umol/l, 30umol/l), we found the expression is in a dose-dependent manner. In the group of 10 umol/l and 20 umol/l, the survival of cells is significantly increased in comparison to the control and also have significantly difference in the two groups(p<0.05), in the group of 20 μmol/L, 16h to 48h after hemin-induced, the survival of cells presents a time-dependent manner. In the group of 10μmol/L and 20 μmol/L, exposure of K562 cells to STI571 resulted in a substantial decrease of cell viability in comparison to the STI571 single treatment group(p<0.05). ZnPPIX-induced HO-1 inhibition leads to induction of apoptosis in K562 cells, having significant difference with the control group(p<0.05). ZnPPIX-induced HO-1 inhibition can suppress the survival of imatinib-resistant cells(p<0.05).

Conclusion: our studies have shown that hemin-induced HO-1 gene expression may promote the proliferation of K562 cells, and can against the cell apoptosis. And we found hemin-induced HO-1 gene expression can protect K562 cells against STI571-induced apoptosis, ZnPPIX-induced HO-1 inhibition leads to decreased viability of imatinb-resistant CML cells. these all indicates HO-1 may represent a novel targeting in CML.
Detection of BCR-ABL Kinase Domain Mutations in Chronic Myeloid Leukemia Patients Treated with Tirosin-Kinase Inhibitors
Authors: Manrique Gonzalo , Bittencout Roberta , Pérez Verónica , Sholl Vanesa , Cappetta Monica , Zubillaga Noel , Hassan Rocio , Boschi Susana , Elizondo Victoria , Seuanez Hector , Zalcberg Ilana , Martinez Lem , Escuder M. Rosario Uri arte
Published: April 2020

Background. Point mutations in the kinase domain (KD) of the BCR-ABL are the most frequent mechanism of drug resistance in CML patients treated with kinase inhibitors (TKI). More than 80 mutations with different frequency and clinical significance have been reported. One of them, the T315I confers resistance to all TKIs available. The detection of mutations in KD allows early identification of high-risk patients and therefore guides clinical therapy decisions.

Aim. To assess the mutation status of a group of CML pts resistant to TKI from Uruguay (n=35) and Brazil (n=30).

Methods. KD mutation screening was performed by RT-PCR and direct sequencing according to Branford et al. (2002). Additionally, we developed a rapid, specific, sensitive and low cost allele specific (AS)-RT-PCR assay to identify T315I, using Branford’s KD amplification primers in combination with an allele specific primer for the T315I point mutation detection. BCR-ABL transcript levels were also measured by RQ-PCR according to international recommendations.

Results and Discussion. RT-PCR and direct sequencing analyses performed in all pts showed the presence of T315l mutation in 3/65 cases. Other 11 showed the alternative mutations Y253H (n=2), E450A, G250E (n=2), E459K (n=2), E450G, F317L (n=2) and E255K; and the remaining 55 showed no mutations in the ABL KD. All 65 samples together with cDNA from 15 non-resistant CML pts and 10 cDNA from non-CML were analyzed by AS-RT-PCR assay for T315l mutation in order to validate the method. T315l was identified in the 3 samples in which the mutation was previously detected by direct sequencing and in 1 pt that had been classified as KD mutation negative. This result was then confirmed by direct sequencing of the AS-PCR product. T315 was neither detected in samples positive for other mutations nor in samples of non-resistant CML and non-CML patients, supporting the specificity of the method. Assessment of the sensitivity of the AS-RT-PCR was performed on serial dilutions experiments using RNA from T315 positive pt into RNA from CML-T315l negative pt, showing that the T315I mutation was detectable to a level of 0.01 % by AS-PCR, while through direct sequencing method the sensitivity was 10–20%. The prevalence of mutations in our study was 15/65 (23%).

Conclusions. Our results showed that the AS-RT-PCR described here is a convenient and easy tool to be used in a clinical routine laboratory for rapid screening for BCR-ABL T315. This, together with direct sequencing, constitutes a suitable approach for CML resistance monitoring and therapeutic choice.
The Vast Majority of CML-CP Patients Treated with Dasatinib (70 mg BID) Who Progress after Long-Term Follow-up (24 mo) Remain in Chronic-Phase
Authors: Cortes Jorge , Mauro Michael , O’Brien Susan , Borthakur Gautam , Wierda William , Matloub Yousif , Sinha Ritwik , Liu David , Kantarjian Hagop
Published: April 2020

Patients with Chronic Myeloid Leukemia (CML) who develop resistance to imatinib have poor clinical outcomes. The disease stage at the time of imatinib failure is an important prognostic factor for this outcome. Specifically, although patients with advanced-stage CML, particularly those in blast-crisis at the time of imatinib failure, had a high rate of cytogenetic response to second-line tyrosine kinase inhibitors (TKIs), these responses were not durable in most instances. In contrast, patients who remained in chronic-phase (CML-CP) at the time of imatinib failure substantially benefited from second-line TKIs, achieving PFS and OS that appear favorable compared to historical data of patients treated with non-TKI therapeutic modalities. Because the prognosis for those in chronic-phase is significantly better than patients who have progressed to accelerated- or blast-phase, it is important to examine the clinical status of patients at the time of TKI failure. This study examined the clinical status of patients in the START-C trial (CA180-013) at the time of disease progression after imatinib and dasatinib therapy. Progression was defined as: loss of complete hematologic response (CHR), loss of major cytogenetic response (MCyR), increasing WBC count, development of accelerated- or blast- phase CML, or death due to any cause. Of 387 participants, 322 (83%) were progression-free at the 2-year follow-up. Of the 65 patients with any criteria for progression as defined above, 46 subjects had progressive disease (loss of MCyR = 14, loss of CHR = 14, increasing WBC counts = 8, development of accelerated-phase CML = 10). The remaining 19 patients died. Thus, only 10 out of 65 (15%) progression events were due to the development of advanced disease and 36 patients (55%) remained in chronic-phase despite a loss or lack of therapeutic response to imatinib and subsequent dasatinib. In summary, 2.6% (10/387) of all patients analyzed transitioned to accelerated-phase CML after first- and second-line therapy. Importantly, development of blast-crisis CML was also not observed in our study population. In a 5-year follow-up of the IRIS trial comparing imatinib to interferon as first-line therapy, a projected 17% of subjects relapsed and 7% developed advanced-phase disease after imatinib. Despite the fact that patients analyzed in our study sequentially failed both imatinib and dasatinib, most remained in chronic-phase after dasatinib failure and are expected to benefit from additional therapeutic options.
Vitamin B12 Deficiency in Patients with Chronic Myeloid Leukemia
Authors: Siddique Muhammad K , Sarwar Hafiz M. Y.
Published: April 2020

High vitamin B12 (cobalamin) levels have been documented in patients with untreated chronic myeloid leukemia. However, the literature about vitamin B12 deficiency in patients with chronic myeloid leukemia who are receiving imatinib is scarce. The objective of this study was to investigate the prevalence of vitamin B12 deficiency in patients with chronic myeloid leukemia. We retrospectively reviewed the medical records of 195 patients with chronic myeloid leukemia seen in the outpatient clinic of our institution between 2006 and 2007. The median age of patients was 35 (range 9 – 77); 118 (60%) patients were males and 77 (40%) were females. All patients included in the study were receiving imatinib. Macrocytic anemia at any given time during the course of treatment was documented in 61 (31%) out of 195 patients. Serum vitamin B12 and folate levels were measured in all patients with macrocytic anemia using chemiluminescent enzyme immunoassay. Among these 61 patients, 33 (54%) had low vitamin B12 levels (< 200 pg/ml), 12 (20%) had indeterminate levels (200 – 300 pg/ml) and 16 (26%) patients had levels > 300 pg/ml. Folate levels were within normal range (3 – 17 ng/ml) in all but two patients and both of these patients had folate levels < 3 ng/ml and vitamin B12 levels > 300 pg/ml. Among 195 study patients, 33 (17%) had macrocytic anemia with low vitamin B12 levels and 12 (6%) had macrocytic anemia with indeterminate vitamin B12 levels. We conclude that vitamin B12 deficiency is prevalent in our patients with chronic myeloid leukemia who are taking imatinib. This is of significance because our patient population is non-vegetarian. Serum vitamin B12 should be measured as a part of work up for anemia in patients with chronic myeloid leukemia. Correction of vitamin B12 deficiency in these patients could improve their tolerance to imatinib, which potentially is a myelosuppressive drug. In our study, measurements of serum methylmalonic acid (MMA) and homocysteine levels in patients with indeterminate vitamin B12 levels were not performed; although these measurements could have confirmed the diagnosis of vitamin B12 deficiency in this subgroup. We suggest that a similar study be conducted in a different patient population elsewhere.
Prognostic Factors in Chronic Myelomonocytic Leukemia: A Retrospective Analysis of 41 Patients
Authors: Chen Bobin , Xu Xiaoping , Ma Yan , Wang Xiaoqin , Lin Guowei
Published: April 2020

Purpose To investigate clinical characteristic and prognostic factors for chronic myelomonocytic leukemia (CMML).

Methods Retrospective cohort study was used in the study. Information for CMML patients was collected, including symptoms, CBC, results of bone marrow aspire and pathology, cytogenetic. All patients were followed up regularly. Analysis of survival and prognostic factors was performed by Kaplan-Merier cure, log rank test and Cox regression model.

Results Forty-one cases were diagnosed as CMML, including 27 male and 14 female patients. Median WBC was 13.7× 109/L. Five patients had leukocytopenia(1.92- 3.46×109/L). Median monocyte count in the peripheral blood was 2.13×109/L, but lower monocyte count (<1×109/L) occurred in 8 patients. All patients presented with bone marrow dysplasia, and most showed hyperplasia, except 3 cases. Abnormal chromosome was detected in 34% cases. Median survival time was 20 months, and there were no difference of survival duration between CMML-1 and CMML-2. Univariate analysis showed that age (> 60 yrs), neutrophil count (<2.0′109/L), lymphocyte count (<1.0′109/L), mature monocyte count (35′109/L) and anemia (Hb< 60g/L) were associated with poor prognosis for CMML. There was no statistical significance in LDH, gender, abnormal chromosome for survival time. Only lymphocyte count and neutrophil count in peripheral blood were independent prognostic factor for CMML after Multivariate analysis.

Conclusion CMML mainly occur in elderly patients. Although most patients have leukocytosis and monocytosis at diagnosis, few case shows leucopenia and monocytopenia. Median survival time for CMML is 20 months. Age, neutrophil, lymphocyte and monocyte count, severe anemia are related to CMML prognosis. Neutropenia and lymphopenia in peripheral blood are independent prognostic factor for CMML.
09:00–09:05Escalated Dasatinib Dose up to 60 Mg O.D. in Elderly Patients with Chronic Myeloid Leukemia in Late Chronic Phase Resistant to or Intolerant of Imatinib-12 Months Follow-up
Authors: Iuliano Francesco , Russo Antonio , Caroleo Maria Cristina , Conforti Filomena , Serra Anna
Published: April 2020

Background: Dasatinib (SPRYCEL®, formerly BMS-354825) is a novel, oral, multitargeted kinase inhibitor of BCR-ABL, SRC, and other kinases that is approximately 300 times more potent than Imatinib in vitro. Dasatinib has been shown to be effective and safe in pts with CML resistant or intolerant to Imatinib, As might be expected, older patients experienced more adverse events, both hematologic and non-hematologic with the standard dosage of 100 mg once daily. Data regarding tolerability in elderly pts are scanty The aim of this study is to test if escalated dasatinib dose up to 60 mg once daily is as effective as standard dose of 100 mg QD with a better toxicity profile in elderly patients with chronic myeloid leukemia in late chronic phase resistant to or intolerant of Imatinib.

Patients and Methods: As of July 2007, 4 eligible patients have been enrolled and treated :4F ; median age 77 y [range 73–82]; 1 imatinib-resistant, 3 imatinib-intolerant). Dasatinib was given at 20 mg once daily (QD) starting dose with dose escalation to 40 mg QD or 60 mg QD in pts tolerant or lacking response. Complete blood counts were obtained weekly for the first 12 weeks; bone marrow cytology and cytogenetics every 1 months, and molecular monitoring of BCR-ABL transcript levels by real-time qPCR every 12 weeks. The primary endpoint was toxicity. Median time from diagnosis of CML was 62 months (range 36 – 88). Prior therapy included interferon-alpha in 1/4. 50 percent of pts had 400 mg of prior imatinib. 25% had 600 mg. 100 % of pts received imatinib for >3 yrs. Best response to prior imatinib therapy was a CHR in 100, and partial (PCyR) cytogenetic responses in 75 % of pts. No BCR-ABL baseline mutations were found.

Results: The analyses with a median follow-up of 12 months (7–12) months show hematologic toxicity (grade 3 thrombocytopenia and leukopenia) at 60 mg OD in 2 out 4 pts. Dose interruptions occurred in 1/4 pts at 60 mg One pts was intolerant to dasatinib and switched to HU Patients received an average daily dose of 40 mg/day (range 20–60 mg). Non-hematologic toxicity consisted mainly of grade 1 diarrhea, headache 2 patients experienced grade II muscolo-skeletal toxicity and fatigue at 60 mg/day.) 100% pts had a CHR, and 50% a PCyR.

Conclusions: Dasatinib demonstrated substantial hematologic activity in elderly patients with late CP-CML. Despite the maximum dose reached was 60 mg QD only the 75% of pts were able to continued the treatment. because of toxicity. The novel TKIs therapies constitute an important therapeutic challenge in this particularly subset of patients when compared with younger patients
Hepatitis B Virus Reactivation and Severe Hepatitis during Chemotherapy with Fludarabine In a Patient with Leukemia
Authors: Türkkan Emine , Pekün Fügen , Atay Didem Y.
Published: April 2020

It is generally accepted that seroconversion of hepatitis B virus (HBV) surface antigen (HBsAg) to an antibody to HBsAg (HBsAb) indicates clearance of HBV. The reverse seroconversion to hepatitis B virus infection has been sporadically described in hematological patients receiving cytotoxic therapy and can be associated with the development of acute hepatitis. Here we report a 14-year-old male patient who developed HBV reactivation after intensive chemotherapy containing fludarabine for second bone marrow relaps of acute lymphoblastic leukemia. Fludarabine is a chemotherapatic agent that used for the treatment of relapsed acute leukemia in pediatric patients. He was HBsAg(−)/anti-HBs(+) and with normal ALT levels before intensive chemotherapy (fludarabine, dexamethasone, GCSF, alexan)for treatment of second bone marrow relaps. 6 weeks after completing two cyclus of chemotherapy with fludarabine, the patient developed transaminitis and evidence of active HBV infection (the patient had test results positive for HBsAg, negative for HBsAb, and positive for HBV DNA). the serum levels of aminotransferases were increased progressiveley. Lamivudine was administered immediately after confirming the HBV reactivation, which considerably improved jaundice, aminotransferase levels and serum HBV DNA titers after one month. The patient was able to resume the chemotherapy under continuis lamivudine treatment. This case indicates the need for physicians to be aware of fludarabine-related HBV reactivation, closely monitor and detect reactivation early, even in an HBsAg-negative patient.
Pre-Transplant Hyperferritinemia Is Associated with Low Incidence of Chronic Gvhd, High Relapse Rate and Impaired Survival in Patients with Hematological Malignancies after Allogeneic HSCT
Authors: Hagglund Hans Gustaf , Lorenz Fryderyk , Remberger Mats , Wahlin Bjorn , Wahlin Anders
Published: April 2020

Background: Iron overload is common after allogeneic hematopoietic stem cell transplantation and may cause liver damage. High pre-transplantation serum ferritin levels have been reported to be associated with impaired survival post transplantation in patients with acute myeloid leukemia or myelodysplastic syndrome.

Design and Methods: We performed a retrospective study of 309 adult patients with haematological malignancies who underwent allogeneic hematopoietic stem cell transplantation after myeloablative (51%) or non-myeloablative conditioning (49%). The aim was to determine the incidence of hyperferritinemia and iron overload and the effects of these conditions on graft versus host disease, relapse and survival in patients with blood disorders who receive allogeneic stem cell grafts.

Results: For overall survival and for relapse-free survival, only high pretransplantation ferritin (>400ug/l) and advanced stage of disease were significant in multivariable analysis. High pretransplantation ferritin and advanced stage of disease were also significant for relapse risk at 3 years, but ferritin was not significant for non-relapse mortality. In multivariate analysis, ferritin, age, type of donor, and previous transplantation were significant factors for cGVHD at one year.

Conclusions: Hyperferritinemia was associated with lower risk of cGVHD and increased risk of relapse and death, but not with increased non-relapse mortality. The lower rate of cGVHD among patients with high ferritin levels can be explained by an immunosuppressive effect of ferritin, reducing graft versus host disease and graft versus leukaemia effect, resulting in increased risk of relapse and impaired chance of survival. Therefore, these conditions might be best explained by an immunosuppressive effect of ferritin and not by a direct toxic effect.
Neurological Complications Following Allogeneic Haematopoietic Stem Cell Transplantation
Authors: Tan Yamin , Huang He , Luo Yi , Shi Jimin , Cai Zhen , Lin Maofang
Published: April 2020

Allogeneic hematopoietic stem-cell transplantation (Allo-SCT) has become the standard of care for a number of malignant and non-malignant disorders. Neurological complications associated with these procedures remain frequent. These complications result from a variety of causes. Known risk factors include the underlying disorder, therapies used for immunosuppression, especially the use of cyclosporine A or FK506, and toxicities from radiation or chemotherapy, but other risk factors might also be of importance. We retrospectively analyzed neurological complications of 152 patients (range 10–49 years old) who underwent Allo-SCT in our transplantation center between August 2005 and July 2008. In all, there were 95 (62.5%) cases of unrelated donor transplantation, 54 (35.5%) cases of sibling donor transplantation, and 3 (2.0%) cases of haploidentical transplantation. Post-SCT neurological complications were seen in 16(10.5%) patients. They were seen in 4 cases after related donor SCT and in 12 cases after unrelated donor SCT. Neurological symptoms occurred between day −4 day and +512 day after transplantation. The complications included 4 cases with relapsed leukemia of central nervous system (2 in ALL, 1 in CML, 1 in AML), 3 cases with encephalorrhagia, 3 cases with epilepsy, 2 cases with demyelinating disease, 2 cases with encephalitis, 1 cases with multiple myositis,1 cases with peripheral neuritis. Seven cases died of neurological complication, including 4 with relapsed leukemia of central nervous system and 3 with encephalorrhagia. Other patients were cured. High dose gamma globulin conbined with Methylprednisolone were used in demyelinating disease. Antiepileptic agents were used in epilepsy. Cyanocobalamin was used in peripheral neuritis. In conclusion, neurological complications commonly occurred in Allo-SCT, and encephalorrhagia might be the main indication that needs intensive care. Moreover, central nervous system leukemia and demyelinating disease, multiple myositis should have clinical interests.
The Relationship of JAM2 Polymorphisms and Graft-Versus-Host-Disease (GVHD) after Allogeneic Stem Cell Transplantation
Authors: Park Eunkyung , Park Sung-Hyo , Lim Kyu-Hyoung , Song Ji Ae , Kim Inho , Yoon Sung-Soo , Park Seonyang
Published: April 2020

Introduction: The junctional adhesion molecule 2 (JAM2) gene plays a key role in tight junction formation in epithelial and endothelial cell. We studied JAM2 polymorphism in GVHD and non-GVHD patients after allogeneic stem cell transplantation.

Methods: We tested 3 sites of JAM2 gene - rs2276220 (G/A), rs963103 (T/C), rs974680 (G/A) - and studied the possible link between the JAM2 polymorphism and the risk of GVHD. We studied 251 patients who had treated with allogeneic stem cell transplantation. The three-SNP block study was performed with rs2276220-rs963103-rs974680 haplotypes. The JAM2 gene was screened for polymorphisms by SNaPShot method. We used SAS version 8.1 (SAS Institute Inc., Cary, NC, USA) for statistic analysis. The odds ratio and p-value was adjusted with multiple logistic regression analysis about age, gender, stem cell source, disease status at the time of transplantation, and conditioning regimen.

Results: The minor allele C of rs963103 (T/C) was dominant in patients with acute GVHD (Odds ratio (OR) 2.918, 95% confidence interval (CI) 1.178~7.233; p=0.0207). This phenomenon was distinct in gut GVHD (OR 1.684, 1.019~2.784; p=0.0419). The minor allele A of rs2276220 (G/A) was dominant in patients with infection after allogeneic stem cell transplantation (OR 6.058, 1.266~28.979; p=0.0241). The overall survival (OS) was associated with rs2276220 (hazard ratio (HR) 0.887, 95% CI 0.633~1.242; p=0.0346) and rs974680 (HR 0.969, 0.763~1.23; p=0.0193). The progression-free survival (PFS) was associated with rs2276220 (HR 0.761, 0.544~1.064; p=0.0095) and rs974680 (HR 0.551, 0.274~1.108; p=0.0382). In three-SNP block study which was performed with rs2276220- rs963103-rs974680 haplotypes, ATA (HR 0.844, 0.645~1.105; p=0.0016) and GTA (HR 5.568, 1.686~18.384; p=0.0048) was associated with OS. ATA (HR 0.796, 0.559~1.133; p=0.0039) and GTG (HR 1.339, 0.834~2.149; p=0.0353) was associated with PFS.

Conclusions: The JAM2 gene polymorphism rs963103 was associated with acute GVHD especially with gut GVHD after allogeneic stem cell transplantation; with higher risk in carriers of the C allele. The JAM2 gene polymorphism rs2276220 was associated with infection, OS, and PFS after allogeneic stem cell transplantation; with higher risk in carriers of the A allele.
Obesity Is Associated with An Increased Risk of Infectious Diseases after Unrelated Bone Marrow Transplantation
Authors: Fuji Shigeo , Kim Sung-Won , Yoshimura Ken-ichi , Akiyama Hideki , Okamoto Shin-ichiro , Sao Hiroshi , Takita Junko , Kobayashi Naoki , Mori Shin-ichiro
Published: April 2020

(Background) Both obesity and malnutrition have been considered to be risk factors for complications after bone marrow transplantation (BMT). To elucidate the impact of pre-transplant body mass index (BMI) on the clinical outcome, we performed a retrospective cohort study with registration data from the Japan Marrow Donor Program (JMDP).

(Results) From January 1998 to December 2005, 3935 patients received unrelated BMT through JMDP, and 3827 patients for whom pre-transplant height and weight data were available were included. Patients were stratified according to pre-transplant BMI values (low BMI: BMI<18, n=295; normal BMI: 18≤BMI<25, n=2906; overweight: 25≤BMI<30, n=565; obese: 30≤BMI, n=61). In a univariate analysis, pre-transplant BMI was associated with a significantly higher risk of grade II-IV acute graft-versus-host disease (GVHD, P=0.03). Other factors which was associated with higher incidence of grade II-IV acute GVHD was HLA allele disparity, GVHD prophylaxis with CSP compared with TAC and donor age (≥40). Multivariate analysis showed that pre-transplant BMI tended to be associated with an increased risk of grade II-IV acute GVHD (P=0.07). Obesity was associated with an increased risk of infectious diseases compared with normal BMI [odds ratio (OR) 1.9; 95% confidence interval (CI) 1.1–3.2; P=0.02]. Progression-free survival was 54%, 52%, 56% and 47% in low BMI, normal BMI, overweight and obese, respectively in 1 year after BMT, and there was a trend that 1-year non-relapse mortality was higher in obese group (low BMI 29%, standard BMI 31%, overweight 32%, obesity 39%).

(Conclusions) This study shows the correlation between pre-transplant BMI and posttransplant complications. Although BMI strongly depends on multiple factors, the effect of obesity on clinical outcome, as suggested here, should be evaluated in a prospective study.