bluuebunny/crossref_metadata_2025 huggingface dataset:
ERG, EVI1 and PRAME Are Relevant Markers of Treatment Response and Survival, and They Could Be Useful for Improving the Risk-Stratification in Citogenetically Normal Acute Myeloid Leukemia (CN-AML)
Authors: Santamaría Carlos , Chillón Carmen , García-Sanz Ramon , Pérez Cristina , Ramos Fernando , de Coca Alfonso García , Alonso Jose- Maria , Giraldo Pilar , Bernal Teresa , Queizán José-Antonio , Rodríguez José Nicolas , Fernández-Abellán Pascual , Barez Abelardo , Peñarrubia María-José , Amigo Mari Luz , García-Sancho Alejandro Martín , Pozas Miguel Angel , Balanzategui Ana , Díaz-Mediavilla Joaquin , San Miguel Jesús F. , González Marcos
Published: April 2020

Several mutations and aberrant gene expression have been proposed as prognostic factors in cytogenetically normal acute myeloid leukemia (CN-AML), but few studies have analyzed the clinical value of several genes in a large group of CN-AML patients. In 121 de novo CN-AML included into the Spanish PETHEMA therapeutic protocols, we evaluated FLT3 and NPM1 mutations and BAALC, ERG, EVI1, MLL-PTD, MN1, PRAME, RHAMM and WT1 RNA levels. Ninety-one patients achieved CR with standard induction therapy while 20/111 (16.5%) were refractory to treatment. This latter subgroup showed higher ERG (media 1.0±0.8 vs. 0.6±0.6;p=0.012) and lower PRAME (media 28.8±52.9 vs. 1640.6±6102.0;p=0.014) levels than no-refractory patients. Moreover, high ERG expression (defined by median value) was associated with shorter OS at 2-years (38% vs. 61%;p=0.012) and RFS at 2-years (38% vs. 60%;p=0.005). By contrast, high PRAME levels (defined by 75th percentile) was related to longer OS (61% vs. 48%;p=0.029) and RFS (74% vs. 43%;p=0.016). Additionally, high EVI expression (defined by 75th percentile) was associated with a poorer OS (37% vs. 56%;p=0.049). The same results in OS were observed in patients with high BAALC RNA levels (median value; 41% vs. 62%; p=0.049). Interestingly, ERG, EVI1 and PRAME markers improved the current prognostic classification of CN-AML based on FLT3 and NPM1 status. Therefore, intermediate risk patients (FLT3-ITD/NPM1mut and FLT3wt/NMP1wt) with low ERG, low EVI1 or high PRAME achieved long OS and RFS, similar to the favorable FLT3wt/NPM1mut subgroup. By contrast, the opposite expression subgroup (high ERG, high EVI1 or low PRAME) showed OS and RFS similar to the adverse FLT3-ITD/NPM1wt subset. These results allowed identify clearly two risk-subgroups for OS and RFS, based on three molecular markers (FLT3, NPM and one of the 3 proposed genes). In conclusion, we demonstrated that ERG, EVI1 and PRAME are relevant prognostics markers in CN-AML and they could improve their risk-stratification.
Analysis of γH2AX and NHEJ Signaling as Molecular Determinants for GOSensitivity in AML
Authors: Hååg Petra , Lindberg Marita Lagergren , Kanter Lena , Lewensohn Rolf , Stenke Leif , Viktorsson Kristina
Published: April 2020

Most patients with acute myeloid leukemia (AML) display an initial response to high dose chemotherapy but a majority develops resistance and relapse within 1–2 years. The targeted drug gemtuzumab ozogamicin (GO) has shown promising clinical effects. Consisting of a monoclonal CD33 antibody coupled to a toxin, calicheamicin, GO enters the cell after binding to the CD33 antigen, preferentially expressed on myeloid cells. Calicheamicin is then released from the antibody and causes multiple signaling effects resulting in cell death. We have previously shown that GO-induced apoptotic signaling in AML cells involves activation of the Bcl-2 proteins Bax and/or Bak. In this study we aimed to clarify if altered DNA repair or DNA damage signaling are molecular determinants of GO-sensitivity in AML. Moreover, attempts were made to connect the upstream DNA double stranded break (dsb) signaling to GO-induced activation of Bax and Bak. Therefore, we analyzed GO-induced DNA damage signaling in GO responsive (HL60) and resistant (KG1a) AML cells. We can show that GO-resistant cells display a higher basal activity of γH2AX and DNA-PKcs and hence a higher DNA repair capacity. Moreover, by treating AML cells with clinically relevant doses of GO we have observed an increased level of both γH2AX and phosphorylated DNA-PKcs which indicates the prevalence of DNA dsbs. This activation was observed in HL60 as well as in KG1a cells, in vitro. Thus, our data suggest that resistance to GO is not due to lack of DNA dsbs induction, but is rather influenced by the DNA dsbs signaling to the downstream apoptotic components. Key activators downstream of DNA-PK but upstream of Bak/Bax, including c-Abl, caspase-2, Bid, Bad, Bim, Bcl-2 and Bcl-xL, are therefore currently being analyzed. In conclusion, we show that GO induced DNA dsbs in both GO sensitive and GO resistant AML cells in vitro, and that resistance to GO treatment is at least in part due to factors downstream of DNA-PK but upstream of Bak/Bax activation.
Prevalence and Clinical Characterization of CD20 and CD22 Expression in Pediatric Precursor B-Cell Acute Lymphoblastic Leukemia
Authors: Rayburg Melissa S , Marmer Daniel , Mo Jun , McMasters Richard , Smolarek Teresa , Lampkin Beatrice , Perentesis John
Published: April 2020

Immunophenotypic classification of leukemia has important therapeutic and prognostic implications. In B-cell malignancies, CD20 and CD22 also represent therapeutic targets. CD20 expression in adult patients with B lineage ALL has been associated with a poor prognosis. The data are conflicting in the pediatric population and may be impacted by the type of therapy employed. A recent study of pediatric patients treated in consecutive St Jude Children’s Research Hospital Total Therapy ALL regimens observed excellent outcomes and no prognostic significance for CD20 expression (Jeha et al. Blood2006, 108:3302–3304). We retrospectively analyzed 50 consecutive patients aged 4 months to 28 years with precursor-B ALL treated with contemporary risk-adapted BFM-based ALL regimens for whom flow cytometric, genetic, and early response data were available. Cases were defined as positive for CD20 and/or CD22 expression if surface expression was identified in more than 20% of leukemic blasts. We found that CD22 was expressed at high levels (68–99%) in all patients evaluated. CD20 expression was positive in 27 (54%) of patients. CD20 expression did not correlate with known NCI prognostic features, including presenting white blood count or age. All 3 patients with BCR-ABL translocation ALL were CD20 positive. Consistent with previously published data, neither of the 2 patients with MLL-AF4 translocation were CD20 positive. There was no association of CD20 expression with trisomy 4/10/17 or TEL-AML1 status. We did not observe an association between CD20 expression and rapid early bone marrow response to therapy at day 8 or 15; 47/50 patients were in remission at day 29. At a median follow-up time of 48 months 46/50 patients were alive without relapse. These limited data do not suggest a strong association between CD20 expression and known prognostic features or early treatment response in pediatric precursor-B ALL treated with contemporary BFM therapy platforms. However, our findings of frequent expression of CD22 on precursor-B ALL blasts from children supports its consideration as a target for immunotherapy approaches in high risk or relapsed disease.
T-Cell Large Granular Lymphocytic Leukemias/Proliferations and Clinical Implications in a Community Practice
Authors: Mathews Malcolm (Mack) , Gale Peter F. , Silva Marco A.C.P. Da , Bailey Teresa W
Published: April 2020

Clonal T-Cell large granular lymphocytic leukemias/proliferations were diagnosed in eighteen patients in the community of Kingsport, TN, from 2000–2008, based on peripheral blood and bone marrow morphology, flow cytometry, and T-cell receptor gene rearrangement studies. These clonal expansions were associated with anemia, neutropenia, and thrombocytopenia. Some patients required no treatment, while others responded to weekly methotrexate. Associated disease entities included myelodysplastic syndrome, non-hodgkins lymphoma, multiple myeloma, mast cell disease, febrile illness, and rheumatoid arthritis. Acquired autoimmune hemolytic anemia developed in one case. Anemia was a common presenting feature. Splenomegaly was noted in several cases. Clonal rearrangement of T-cell receptor gamma chain genes was confirmatory in all cases. Chronic disease with uncomplicated cytopenias was noted in the majority. Diagnosis should be suspected in patients with neutropenia and anemia with a relative (and at times an absolute) peripheral blood large granular lymphocytosis. Clonality can be assessed with molecular studies, and therapy is reserved for symptomatic disease or transfusion dependency.
A Case Report of Aggressive Adult Neuroblastoma Mimicking Acute Leukemia with Fulminant Course and Fatal Outcome
Authors: Panovska Irina , Ivanovski Martin , Trajkova Sanja , Pivkova Aleksandra , Genadieva-Stavric Sonja , Hadzi-Pecova Liljana , Latifi Arif , Trajkov Dejan , Spiroski Mirko , Cevreska Lidija
Published: April 2020

Neuroblastoma is an embryonic neoplasm of the sympathetic nervous system derived from the primitive neural crest cells. It is the second most common extra cranial malignant tumor of childhood and the most common solid tumor of infancy, comprising 8 to 10 percents of pediatric cancers. Forty percent of patients who present with clinical symptoms at diagnosis are under 1 year of age, and less than 5% with clinical symptoms are over the age of 10 years. The tumor very rarely occurs in adult patient (defined as 20 years of age and older) and usually has an indolent course with poor survival. Up until June 2008, no more than 65 neuroblastomas in adults had been reported in the literature. Majority of the reports describe an indolent and longer course of the disease among adults with different biological characteristics when compared to children, but ultimately poor outcome regardless of the initial stage of the disease. Here, we report a case of aggressive adult neuroblastoma mimicking acute leukemia with fulminant course and fatal outcome. The patient, a previously healthy 38 years old Caucasian male, presented at our department with a 4 months history of hip pain, headaches, malaise, weakness, persistent diarrhea, poor appetite and weight loss. On admission he was febrile, prostrated, pale, fixed in bed. His abdomen was distended with present bowel movements. The liver and spleen were 3 cm and 5 cm below the costal margins respectively. No abdominal mass was palpable. Neurological examination showed right Horner’s syndrome, reduction of the superficial sensitivity on the Th1 and Th2 level and loss of the tendon-muscle reflexes of the lower limbs. He had paraplegia, indicating possible spinal cord compression. The complete blood counts revealed pancytopenia and presence of 11 % peculiar circulating blastoid cells. The LDH was 3552U/l. Under suspicion of acute leukemia bone marrow aspirate and biopsy was performed. Immunophenotyping of the bone marrow mononuclear cells showed presence of around 50% CD45negative, CD56 and CD9 positive cells, indicating a possible involvement of the bone marrow with neuroblastoma. A routine second opinion from a reference pathologist was also interpreted as neuroblastoma and confirmed with positive immunohistochemical staining for NSE, chromogranin, synaptophysin and negative for LCA. A work-up for neuroblastoma was done and investigations confirmed disseminated stage 4 neuroblastoma. In spite of the appropriate treatment with intensive chemotherapy regimen, the patient failed to improve and died of progressive disease. Our case report illustrate the usefulness of including CD56 marker in flow-cytometry acute leukemia assays, especially in cases when CD45 expression is absent, as initial screening tool for recognizing cases of neroblastoma which simulate acute leukemia.
Differential Expression of VEGF Receptors on Human T Cell Subsets
Authors: Lee Stephen A , Corless Nichole L , Lee Dean A.
Published: April 2020

Vascular endothelial growth factors (VEGF) and their receptors (VEGFR) deliver important signals that controlling the angiogenesis of growing, injured, and malignant tissues. Expression of VEGF receptors has been described on a variety of normal and malignant leukocytes, and presumably plays a role in their trafficking to injured tissue. Differential expression of neuropilin-1 (NRP-1) has been described as a marker of regulatory T cells (Treg) in mice, and may be expressed on human thymocytes, but expression on human Tregs has not been confirmed. Moreover, earlier reports of Treg-specific markers in humans were based on the CD4+CD25+ subset, which is not as reliable for Treg identification as it is in mice. We used cell sorting to obtain pure populations of human Tregs (using the more stringent criteria of CD4+CD25+CD125−) and T helper cells (Th) (CD4+CD25−CD127+). We compared expression in these subsets of VEGFR-1, -2, -3, and NRP-1 and -2 using flow cytometry and quantitative RT-PCR, and NRP-1 splice isoforms by RT-PCR using isoform-specific primers. We were not able to identify any NRP-1 expression in Tregs by any of these methods. In contrast, there was marked differential expression of VEGFR-2 (Flk-1) and -3 (Flt-4), both of which exhibited up to 16-fold higher expression in Tregs than in Th. These findings suggest a role for VEGF in recruiting Tregs to sites of tissue growth as a mechanism for blunting potential autoimmunity to neoexpressed antigens, and thus may also explain the accumulation of Tregs in tumors.
Simultaneous Phenotype-Specific Telomere Length Measured by Flow-FISH Using Thermally Stable Quantum Dots
Authors: Kapoor Veena , Hakim Fran , Rehman Najibah , Gress Ronald E , Telford William G
Published: April 2020

The Flow-FISH technique used to measure telomere length by incorporation of fluorescein-labeled oligonucleotide probes complementary to telomeres repeat sequences has been extended to permit simultaneous multiple phenotypic analysis within a single sample. The necessary thermal stability of the fluorescent labeling needed for phenotyping cell populations can be achieved using quantum dots instead of conventional organic fluorophores, which are usually damaged during the high-temperature hybridization treatment and lose their fluorescence, limiting their usefulness for phenotypic analysis. In this study, quantum dots and conventional fluorophores were compared for their ability to survive the 82oC hybridization step necessary for PNA probe annealing. Quantum dots preserved their fluorescence following heat treatment and gave good signal-to-noise ratios when used to detect lymphocyte and monocyte markers (CD3, CD4 and CD14). This was in contrast to phycobiliproteins and low molecular weight fluorochromes such as fluorescein and the Alexa Fluor dyes, which decreased considerably in fluorescence following hybridization. Since addition of fluorescent immunophenotyping complicated an assay already heavily dependent on uniform technique, internal reference standards (previously reported by Lansdorp) were particularly critical in this technique. We used aliquoted frozen calf thymocytes (CT) as an internal standard, determining their telomere length and incorporating them into each sample to monitor tube to tube and day to day assay variability. The telomere length of the CTs and of CD3+CD4+, CD3+CD4− and CD14+, were determined and then corrected by normalization to the known CT telomere length. To validate the overall method, we then measured age dependence on telomere length for monocyte and lymphocyte subpopulations in young, middle-aged and older donors. Results were consistent with results previously reported using traditional Southern blotting and Flow-FISH with no immunphenotyping, suggesting that incorporation of immunolabeling did not adversely affect this technique. This method allows for the rapid, accurate and simultaneous determination of the telomere lengths of different cell populations within a single sample when little amount of sample is available.
Characterization of Circulating Monoclonal B Lymphocytes in Patients with HCV Infection
Authors: Ohtsubo Korenori , Okamura Takashi , Sata Michio , Kawaguchi Takumi , Morishige Satoshi , Takata Yuka , Oku Eijiro , Imamura Rie , Seki Ritsuko , Hashiguchi Michitoshi , Osaki Koichi , Yakushiji Kazuaki , Kanaji Taisuke , Yoshimoto Kohji , Ueno Takato
Published: April 2020

Hepatitis C virus (HCV), being lymphotrophic as well as hepatotrophic, has been reported to induce B-cell proliferative disorders such as mixed cryoglobulinemia and B-cell lymphoma. To investigate the association between HCV and B-cell proliferation, we evaluated the incidence and characteristics of monoclonal B cells in the circulating blood of HCV-infected cases relative to controls with non-HCV hepatic diseases. We first evaluated the surface immunoglobulin κ:λ light-chain ratios of the circulating B (CD19+) cells in 240 HCV-infected cases and 150 controls. Light chain restriction (κ:λ ratio >3:1 or < 1:2) was detected in 7 cases with HCV (2.9%) (Table 1), but was never detected in the controls (p<0.05). As these monoclonal B cells were not identified morphologically, they were analyzed for CD5/CD20 expression. None of them showed the so-called chronic lymphocytic leukemia (CLL)-phenotype cells. The clonal and normal B cells did not differ significantly in their intensity of CD5 expression (Figure 1). The B-cell clonality was confirmed in all 7 cases by polymerase chain reaction (PCR) analysis of the immunoglobulin heavy-chain (IgH) gene rearrangements and the t(14;18) fusion gene was detected in one case. The loss of clonality was observed in 2 cases treated with interferon and in one case treated with splenectomy. The longitudinal study is required to determine whether these circulating monoclonal B cells progress to lymphoproliferative disorders or not.

Table 1. The clinical data of the 7 HCV-infected cases with monoclonal H cells

Case No Age Sex WBC count./ul Lymphocyte count./ul CD19+ cells in lymphocytes. % Light chain κ/λ. CD5+B cell. % Cryoglobulin IFN therapy 1 78 M 2,800 710 26.46 κ 33.59 17.45 (+) (−) 2 65 M 3,400 720 5.41 λ 0.23 15.93 (−) (+) 3 84 M 5,700 3,520 46.68 κ 11.31 6.8 (+) (−) 4 74 F 3,300 1,750 15.01 λ 0.037 46.31 (−) (−) 5 65 F 2,500 580 34.46 κ 8.07 40.17 (+) (−) 6 72 F 2,000 940 25.4 λ 0.198 38.77 (−) + 7 66 M 4,200 1,380 9.73 λ 0.41 3.53 (+) (−)

Figure 1. The three groups did not differ significantly in the frequency of CD5+ B cells (mean±SEM: group1, 24.14±6.55; group2, 30.27±1.69; group3, 28.53±1.93). Figure 1. The three groups did not differ significantly in the frequency of CD5+ B cells (mean±SEM: group1, 24.14±6.55; group2, 30.27±1.69; group3, 28.53±1.93).
Ultimate Outcomes of Patients with Recurrent Diffuse Large B Cell Lymphoma Who Do Not Respond to Second-Line Chemotherapy
Authors: Elstrom Rebecca , Martin Peter , Ostrow Katya , Barrientos Jacqueline , Furman Richard R. , Ruan Jia , Schuster Michael , Shore Tsiporah , Chadburn Amy , Coleman Morton , Leonard John P.
Published: April 2020

Background: Patients with diffuse large B cell lymphoma (DLBCL) who relapse after or are refractory to initial therapy can in some cases experience long term disease free survival after second-line chemotherapy alone or followed by autologous stem cell transplantation. The major predictor of outcome for patients undergoing autologous transplant is response to prior (second-line) chemotherapy. Patients not responding to second-line regimens may receive third-line or subsequent chemotherapy regimens in hopes of achieving response and proceeding to transplant, but available outcome data from this group have been limited.

Methods: We used pathology and treatment records to identify patients with relapsed or refractory DLBCL (excluding transformed lymphoma) at the Weill Cornell Medical Center for whom data on response to second-line chemotherapy could be determined. An online social security database verified survival. Median overall survival (OS) was calculated by the Kaplan-Meier method.

Results: One hundred eight patients with relapsed or refractory DLBCL who underwent second line chemotherapy were identified, and adequate treatment records were available for 74 of these patients. Chemotherapy consisted of ifosfamide-containing regimens in 60 patients (81%), platinum-containing regimens in 61 (82%) and included rituximab in 29 (39%). Forty-seven patients (64%) achieved at least a partial response (R), while 27 patients (36%) did not respond (NR). Median OS from second therapy was 4 months (range 1–61) in the NR group, and was projected at 66 months (range 2–106) in the R group. Only one patient in the NR group (4%) survived for greater than one year. Twenty-four NR patients underwent third-line therapy, of whom 5 of 19 with available response data achieved a clinical response (5 died prior to response assessment) and 3 underwent autologous transplantation. Two patients progressed within 3 months of transplantation, with OS from transplantation of 4 months, while the third patient is alive in remission at 59 months after transplant. Neither longer duration of first remission (HR 1.00, p=0.85) nor choice of subsequent aggressive chemotherapy over less intensive approaches (HR 0.81, p=0.65) conferred a survival benefit in patients who had not responded to second-line therapy.

Conclusion: Patients with recurrent DLBCL who do not respond to second-line chemotherapy have poor outcomes, with only rare patients achieving extended survival following subsequent chemotherapy. Clinical trials of novel therapeutic regimens should be prioritized as management strategies for these patients.
Analysis of Long-Term Outcome and the Risk of Second Malignancies in Hodgkin Lymphoma Survivors
Authors: Cedrych Ida , Blamek Slawomir , Deptala Andrzej , Skladowski Krzysztof
Published: April 2020

Background: Therapy of Hodgkin lymphoma (HL) can contribute to delayed impairments of health, which some of them (e.g. secondary cancer) can be fatal.

Aim: The purpose of this retrospective study was to evaluate long-term outcome of HL survivors and the risk of second malignancies.

Methods: The medical report data analysis of 170 patients (pts) with HL treated in one oncological center in Poland between 1979–2005 has been performed. There were 88 females and 82 males, median age 30.5 years (range 16–72). Among the patients there were 22 pts (12.5%) in CSI, 95 pts (55.8%) in CSII, 26 pts (15.4%) in CSIII and 27 pts (15.8%) in CSIV. Half of the 170 pts presented B-symptoms. Chemotherapy (ABVD, MOPP/ABV or MOPP regimen) was given to 155 (93.5%) pts, and radiotherapy alone (mantle-field or involved-field) was given to 11 pts (6.5%). Chemoradiotherapy received 105 pts (61.8%).

Results: CR achieved 136 pts (80%), PR 11 pts (6.4%), and progression of HL was observed in 23 pts (13.5%). With a median follow-up of 242 months (range 0–439), there were 37 relapses (21.7%), and 25 deaths (14.7%), usually due to HL progression (20 pts), and also because of secondary cancer (4 pts = 2.4%) and coronary heart disease (1 patient). Five year DFS and OS were 80% and 90% respectively. There was no statistical difference between ABVD and MOPP/ABV regimen efficacy (p=0.94). Late impairments of health were revealed in 78 pts (45.9%), and there were: secondary malignancies, amenorrhea/infertility, skeletal lesions, depressive syndromes, pulmonary fibrosis, polyneuropathy, hypothyroidism, cardiovascular disease, and gastro-duodenal peptic ulcer. Nine pts (5,3%) developed secondary malignancies, i.e.: 2 melanomas, 2 skin basal-cell carcinomas, 1 osteosarcoma, 1 thyroid carcinoma, 1 non-small cell lung cancer, 1 colon carcinoma, 1 cervical carcinoma, and 1 prostate carcinoma. A median time from the end of the treatment to the development of secondary cancer was 114 months (range 28–434). The risk of secondary malignancies development among pts treated with either initial chemotherapy alone or combined modality therapy was similar, and did not reach statistical significance (p=0.20).

Conclusions: Chemotherapy or chemoradiotherapy of HL provides freedom from the disease in more than 3/4 of our patients. In the group of fairly young patients such treatment caused a substantial amount of variety of health impairments including secondary malignancies. Despite the high rate of HL curability our study indicates the need of a careful, long term medical surveillance of HL survivors.
Study on Treatment of Lymphoma-Associated Hemophagocytic Syndrome with Fludarabine Combined with Methylprednisolone and Globulin
Authors: Wang Yini , Wang Zhao , Feng Cuicui , Chen Xi
Published: April 2020

Hemophagocytic syndrome (HPS) is a life-threatening condition of severe hyperinflammation caused by the uncontrolled proliferation of activated lymphocytes and histiocytes secreting high amounts of inflammatory cytokines. Lymphoma-associated hemophagocytic syndrome (LAHS) clinically featured by histiocytic proliferation, fever, hepatosplenomegaly, cytopenia, deranged liver function and frequent coagulopathy. The immediate aim of therapy in LAHS is suppression of the severe hyperinflammation, which can be achieved with immunosuppressive/immunomodulatary agents and cytostatic drugs. In this study, we diagnosed 11 cases of LAHS according to HLH-2004 diagnostic criteria in Beijing Friendship Hospital during the period from June 2005 to Jan 2008, and collected informations about the patients with LAHS: primary diseases, manifestions, laboratory findings and survival. 2 cases large B cell lymphoma, 1 case mantle lymphoma, 4 cases extranodal NK/T cell lymphoma, 3 cases peripheral T cell lymphoma and 1 case angioimmunoblastic T-cell lymphoma were included in this research. The median age was 49 years, and the range was 33–74 years. 7 out of 11 were female and 4 were male. All the 11 patients received regimens containing fludarabine (25mg/m2/d, d1–5), methylprednisolone (2mg/kg/d, d1–14, 1mg/kg/d, d15–28) and immunoglobulin (0.5g/kg/d, d1–14) after finally diagnosed. 7 of the 11 patients were markedly improved after treatment and then received lymphoma chemotherapy. The median survival time had reached to 1.4 year. The other 4 aggravated. This outcome was much better than what A-Reum Han had reported (Ann Hematol2007; 86: 493–498). It suggests that fludarabine will be a promising agent in the therapy of LAHS.
Thalidomide: An Active Agent in AILT Lymphoma
Authors: Henderson Ross , Simpson David , Hanna Merit , Chunilal Sanjeev
Published: April 2020

Angioimmunobalstic T cell lymphoma (AILT) is a relatively rare subtype of lymphoma, accounting for approximately 1 – 2% of all cases of NHL. The pathogenesis is thought due to clonal expansion of follicular T-helper cells which subvert normal germinal centre function. It is characterised by systemic disease and significant immune dysregulation with frequent atypical infections and autoimmune disease. It has a poor prognosis with a median survival of less than 3 years using CHOP or CHOP-like regimens. Dose intensification, or anthracycline use, appears to be ineffective. Thalidomide is an immunomodulatory agent with isolated case reports of activity in this disease. We report 3 cases of AILT where thalidomide was effective at inducing or providing a sustained remission.

Case 1 was an 86 year old woman who presented with systemic symptoms, widespread lymphadenopathy and a polyclonal increase in immunoglobulins. AILT was diagnosed from both lymph node biopsy and bone marrow biopsy. Symptoms improved only temporarily with prednisone. She commenced thalidomide at 200mg daily with improvement in symptoms and resolution of lymphadenopathy and after 2 months the dose was reduced to 100mg daily. She continued the thalidomide at the same dose and remained in remission until her death from unrelated causes 4 years later

Case 2 was a 72 year old man who initially presented with autoimmune haemolytic anaemia and red cell aplasia which was unresponsive to steroids, intravenous immunoglobulins (IVIG), cyclosporine and splenectomy. He then developed cervical and axillary lymphadenopathy, biopsy of which showed AILT. Bone marrow examination showed no evidence of AILT. He commenced thalidomide at 150mg daily with resolution of the haemolysis and red cell aplasia within 12 days and reduction in all lymphadenopathy. He remains in remission 3 months after commencing thalidomide and is currently on 100mg daily.

Case 3 was a 57 year old woman presented with ITP with platelets < 10, systemic symptoms, widespread lymphadenopathy and hepatosplenomegally. AILT was diagnosed from lymph node biopsy and bone marrow biopsy. She commenced on prednisone and IVIG for the ITP, then proceeded to CHOP chemotherapy. Her first cycle was complicated by Pneumocystis jirovecii infection. Following the chemotherapy, her systemic symptoms subsided and the lymphadenopathy and organomegaly resolved. She commenced thalidomide at 100mg daily and has remained in remission 2 months later.

There are now several case reports in the literature indicating that thalidomide is effective in AILT which is reflected by the cases reported here. Given the poor prognosis of AILT using conventional chemotherapeutic regimens, such as CHOP, prospective clinical trials using thalidomide, or the newer derivatives such as lenalidomide, are warranted.
Skeletal Disorders as Late Complications in Hodgkin Lymphoma Survivors
Authors: Cedrych Ida , Blamek Slawomir , Deptala Andrzej , Skladowski Krzysztof
Published: April 2020

Background: Long-term complications of musculoskeletal system are rare in adult Hodgkin lymphoma (HL) survivors, thus is uncertain whether these complications are treatment-related.

Aim: The goal of this study was to analyze risk, frequency and type of skeletal damage in HL survivors.

Methods: The study comprises of a retrospective analysis of 170 patients (pts) suffered from HL, and treated in an oncological center in Poland between 1979–2005. There were 88 females and 82 males, median age 30.5 years (range 16–72). According to the Ann Arbor clinical staging system there were 22 cases (12.5%) in CSI, 95 cases (55.8%) in CSII, 26 cases (15.4%) in CSIII and 27 cases (15.8%) in CSIV. Fifty six percent of the patients presented B-symptoms, and 60% had bulky disease. Vast majority of cases were diagnosed as classical HL, with histologic subtypes of nodular sclerosis grade 1 in 45%, nodular sclerosis grade 2 in 14%, mixed cellularity in 21%, lymphocyte depletion in 5.2%, and lymphocyte-rich in 7%. However 5.8% of the cases were unclassified by a histopathologist. Chemotherapy with a median of 6 cycles/patient was given to 93.5% of cases (ABVD – 29.4%, MOPP/ABV – 37.6%, MOPP – 24.1%, other – 2.25%). Chemoradiotherapy was given to 61.8% of pts. Radiotherapy alone (mantle-field or involved-field), with a median dose of 40 Gy (range 21–46.8) received 6.5% of pts. For statistical analysis we used Kaplan-Meier method and log-rank test

Results: With a median follow-up of 242 months (range 0–439) a skeletal disorder (SD) was detected in 27 (15.8%) of the patients. Median time to identify SD was 29.5 months. Skeletal disorders were diagnosed as follows: osteoarthritis (11 pts), avascular (aseptic) necrosis of bone (3 pts), osteoporosis (3 pts), osteopenia (3 pts), rheumatoid arthritis (2 pts), vertebral osteochondritis (2 pts), idiopathic (nontraumatic) subluxation of wrist (1 patient), fibrosis of vertebral body (1 patient), and inflammation of nucleus pulposus (1 patient). In the statistical analysis there was no relationship between the risk of SD and: sex, bulky disease, B symptom, involvement of extralymphatic organ, histologic subtype, radiotherapy, chemotherapy, and chemotherapeutic regimen. However, we found an association of SD risk with clinical stage. The difference between CSI and CSIII was statistically significant (p = 0.0235). There were no differences between CSII and CSIII (p = 0.0564), as well as CSIII and CSIV (p = 0.0787).

Conclusions: In our group of adult HL survivors skeletal disorders caused substantial amount of long-term complications. We found no relationship between treatment modality and the risk of a skeletal disorder, despite high median dose of radiotherapy and extensive use of alkylating agents. The association between skeletal disorder and more advanced clinical stage is difficult to explain, however it might suggest a possible role of initial tumor burden and excessive crosstalk between cancer and reactive immune/inflammatory cells.
Successful Treatment of Primary Refractory Bulky Diffuse Large B-Cell Lymphomas with Concomitant Radio-Chemotherapy Followed by Autologous Stem Cell Transplantation : A Single Center Experience
Authors: Couturier Marie-Anne , Champeaux-Orange Elise , Senecal Delphine , Lissandre S.éverine , Benboubker Lotfi , Barillot Isabelle , Calais Gilles , Colombat Philippe , Gyan Emmanuel
Published: April 2020

Diffuse large B cell lymphomas (DLBCL) with bulky presentation display a poor prognosis because of a substantial rate of primary refractory disease. Second and third lines of standard chemotherapy associated with rituximab do not always achieve a complete response (CR) Autologous stem cell transplantation (ASCT) provides the best progression-free survival when patients are transplanted in CR. Concomitant radio-chemotherapy (CRC) has been used in the treatment of solid carcinomas, and may be useful in the treatment of locally refractory DLBCL to obtain a better tumor control with a minimal risk of systemic progression. We report 5 retrospective cases of patients (less than 60 years old) treated at our institution with CRC for bulky DLBCL with localized refractory disease assessed by CT and/or PET-Scan. All 5 patients presented at diagnosis with at least one adverse prognostic factor : 4 with elevated LDH, 4 with stage III or IV. The largest diameter of the bulky sites ranged between 70 and 160 mm. All 5 patients experienced locally refractory disease after one (2/5) or two (3/5) lines of immuno-chemotherapy, at mediastinal (2/5) or abdominal (3/5) sites. They were treated by 3 cycles of rituximab (375 mg/m2 on d1), CDDP (30 mg/m2 d1–3) and etoposide (120 mg/m2 d1–3) at 21d intervals and concomitant radiotherapy delivered to the residual disease sites, administrated in 2 Gy fractions once daily to a total dose of 30 Gy, beginning on d21. For one patient, CDDP was replaced by carboplatin because of previous renal toxicity and another received standard DHAP together with radiation therapy as a second-line treatment. The whole procedure was feasible, with no grade III or IV toxicities. One patient died of early disease progression outside the irradiation field. Four patients out of 5 achieved a complete response and were subsequently transplanted. With a follow-up ranging between 2 and 75 months, all 4 patients are alive and free of disease as well as of long-term toxicity. These preliminary results suggest that CRC deserves further investigation as an option for the management of localized refractory DLBCL.
Diffuse Large Cell Lymphoma Transformed from Rosai-Dorfman Disease
Authors: Jendiroba David , Sampaio Marcus Magnus , Gonçalves Márcia Vilela , Rodrigues Juliana Alves , Jendiroba Daniella Barqueti , Cabanillas Fernando
Published: April 2020

INTRODUCTION: Rosai-Dorfman disease, also known as Sinus Histiocytosis with Massive Lymphadenopathy, is an almost rare benign entity which mimics non-Hodgkin’s lymphoma, and is characterized by enlargement of lymph nodes. Knowing the difference between these two diseases is critical for the adequate therapeutic approach. The most common presentation of the disease is within the cervical basin, and most of the times, lymph node dissection and removal of their chains is enough for cure.

PATIENT AND METHODS: We describe a 23-yo white woman with past history of persistent cervical lymphadenopathy, who underwent 4 surgical procedures to remove left cervical nodes, diagnosed as Rosai-Dorfman disease within the last 3 years. When relapse after the 4th surgery happened, the patient came to our Service and sought for a 2nd opinion consultation.

RESULTS: Pathology reviews were done on the previously collected material, corroborating Rosai-Dorfman disease, except for the review of the slides of the last material, which showed inconclusiveness. Histopathology and immunohistochemistry were then repeated for the last surgical specimens at 2 different Pathology labs, and both confirmed diffuse large cell lymphoma. Patient started on chemo with CHOP 6×, and showed a partial response up to the 3rd cycle, when the tumor started progressing during the 4th cycle. Chemo was then changed to ESHAP, which was well-tolerated and promoted CR after 6 cycles. The patient is currently in CR and asymptomatic.

CONCLUSION: Rosai-Dorfman disease is a proliferative reaction to an unknown, unidentified pathogen or specific cause, which leads to lymph node enlargement and extra nodal involvement in 25% of the cases. Some studies have attempted to correlate the presence of klebsiella as a proliferative stimulus present in this disease, but there is not enough evidence to confirm this hypothesis. Interestingly, it may seem that Rosai-Dorfman is a middle step between a truly benign process and malignancy itself because, not only other cases of lymphoma transformed after Rosai-Dorfman’s disease have been reported, but also extra nodal cancer following extra nodal presentation of the disease. Although many clues point to its involvement in the process of carcinogenesis, more clinical and molecular evidence are necessary to tie the link between Rosai-Dorfman disease and cancer.
Potent Anti-Cancer Activity of Anti-NTB-a Monoclonal Antibodies in Preclinical Leukemia and Lymphoma Models
Authors: Korver Wouter , Zhao Xiaoxian , Singh Shweta , Pardoux Cecile , Barman Ishita , Aguilar Gerard , Zhao Jingsong , Hsi Eric , Abo Arie
Published: April 2020

NTB-A is a CD2-related cell surface protein expressed on lymphoid cells including B-lymphocytes from Chronic Lymphocytic Leukemia (CLL) and lymphoma patients. We have generated a series of mAbs against NTB-A and assessed their therapeutic potential in preclinical models. Selected mAbs to NTB-A were further tested in functional Complement Dependent Cytotoxicity (CDC) and Antibody Dependent Cellular Cytotoxicty (ADCC) assays in cell lines and B lymphocytes freshly isolated from CLL and lymphoma patients. Potent cytotoxic activity was demonstrated against B cells isolated from CLL patients and B lymphoma cell lines. Chimeric anti-NTB-A mAbs demonstrated anti-tumor activity equal to rituximab against CA46 human lymphoma xenografts in nude mice at a low dose. In a chimpanzee safety study, a single dose of lead anti-NTB-A mAb 994.1 resulted in near-complete depletion of peripheral lymphocytes while having a minimal effect on T cell activation. Taken together, these results demonstrate NTB-A as a promising target with an acceptable safety profile for immunotherapy of leukemia and lymphomas.
Fatty Acid Synthase and Diffuse Large B Cell Lymphoma
Authors: Uddin Shahab , Hussain Azhar , Bavi Prashant , Al-Kuraya Khawla
Published: April 2020

Diffuse Large B-cell Lymphoma (DLBCL) is considered to be the most common type of lymphoma in adults, accounting for 30–40 percent of cases of non-Hodgkin’s lymphoma. The incidence of DLBCL is about 40–60% in Saudi Arabian population. The reason of high rate for DLBCL in Saudi Arabian population is not known, however recent studies suggests that differences in molecular signature as compared to western population accounts for this incidence. Although patients with DLBCL are potentially curable with combination chemotherapy, the disease proves fatal in approximately 50% of patients. The cause of most DLBCLs remains unknown; however several studies suggest that dysregulated survival/apoptosis or defective repair pathways have been implicated. Many human cancers, particularly those with a poor prognosis express high levels of fatty acid synthase (FASN), a key metabolic enzyme linked to the synthesis of membrane phospholipids in cancer cells. Over-expression of FASN can be largely attributed to activation of phosphatidylinositol-3′-kinase (PI3K)/AKT pathway. However, the role of FASN in the pathogenesis of DLBCL has not been elucidated. Therefore, in this study, we investigated the role of FASN in a large series (301) of DLBCL patient samples and a panel of DLBCL cell lines. Using immunohistochemistry, FASN was detected in appreciable number of DLBCL tumors and was strongly associated with the expression of p-AKT protein. We next examined the effect of C-75, a synthetic slow binding inhibitor of FASN activity on DLBCL cell lines (SUDHL4, SUDHL5, SUDHL8 and OCI-LY19) in vitro and found that C-75 treatment inhibits growth and induces apoptosis in all 4 DLBCL cell linesused in the study. We show based on in vitro studies employing a variety of experimental tools using different FASN inhibitors, FASN siRNA and AKT siRNA that FASN exert its oncogenic action in DLBCL cells via activated AKT. Our data show that inhibition of FASN leads to de-phosphorylation of p-AKT and it’s down stream effectors, FOXO1 and GSK-3. This, in turn leads to activation of the intrinsic apoptotic pathway by, initially causing conformational changes of the Bax protein leading to changes in the mitochondrial membrane potential and release of cytochrome c into cytosole. This causes activation of caspases-9 and -3 and cleavage of PARP. zVAD-fmk, a universal inhibitor of caspases prevents caspase-9 and -3 activation and abrogates apoptosis induced by C-75 treatment. Finally, C-75 treament of DLBCL cell lines causes down-regulation of the inhibitor of apoptosis proteins, XIAP, cIAP1 and Survivin. In summary, data presented here demonstrate a significant correlation between the expression of FASN and active AKT in DLBCL and indicate that inhibition of PI3K/AKT signaling synergize the FAS inhibitors to induce apoptosis in DLBCL cell lines with constitutively active AKT. This may have significant clinical implications. Therefore, FASN has become a promising target for anti cancer drug development.
Multikinase Inhibition by Sorafenib in B and T NHL Cell Lines: Increasing Cytostatic Efficacy after Prolonged Exposure
Authors: Chapuy Bjoern , Schuelper Nikolai , Kaehler Elisabeth , Truemper Lorenz H. , Wulf Gerald
Published: April 2020

Introduction: A substantial fraction of patients with relapsed or primarily refractory Non Hodgkin Lymphoma (NHL) can still not be cured with current chemotherapy regimens. Therefore, we evaluated the efficacy of tyrosinkinase inhibition by sorafenib against NHL cells in vitro.

Methods: Phosphorylation of RAF/MEK/ERK pathway members as well as of PDGF and VEGF receptors was analyzed by western blot with phospho-antibodies in a panel of cell lines, i.e. aggressive B-NHL (Balm3, Karpas422, Raji, Ramos, SuDHL4), mantle cell lymphoma (Granta, Jeko, Mino), and T cell lymphoma (CEM, HPB-ALL, Jurkat, Molt4). Cell viability under sorafenib exposure was tested by tetrazolium-based (MTT) assays and the proapoptotic and antiproliferative mechanisms were evaluated by annexin V and cell cycle assays, respectively. Clonogenic potential after long-term (10 days) exposure to sorafenib was analyzed by colony formation in inhibitor-free semisolid media.

Results: Sorafenib reduced cell viability at clinically relevant concentrations with IC50 between 3.7 and 10.9 μM in seven cell lines, whereas four cell lines (Balm3, CEM, HPB-ALL, Granta) were less susceptible requiring IC50 of >15 μM. Most sensitive were the diffuse large B cell lines, SUDHL4 and Karpas422, and the Burkitt cell lines, Ramos and Raji. Induction of apoptotic death through an annexin V positive intermediate was documented in the responding cell lines (range 20–90% at 10.9 μM) after short-term exposure (24 through 72 h). After prolonged exposure (>10 days), apoptosis increased significantly, with proportions of annexin V positive cells at 7.3 μM above 75% in all cell lines except Jeko (39% ± 1.2%). In the remaining cells, we found cell cycle arrest to occur, leading to a complete loss of in vitro clonogenicity after cessation of sorafenib exposure in all cell lines (7.3 μM). Activation of the RAF/MEK/ERK-pathway was inhibited to variable degrees in all cell lines. Intensity of RAF and ERK phosphorylation inhibition alone, however, did not predict susceptibility to sorafenib after short-term exposure, suggesting a heterogeneous and time-dependent pattern of sorafenib targets to be involved.

Conclusions: Sorafenib has significant in vitro efficacy against B and T cell lymphoma cells, especially after prolonged exposure.
ITF2357, a Novel Histone-Deacetylase Inhibitor, Is Effective against Peripheral T-Cell Lymphomas in Vivo
Authors: Piccaluga Pier Paolo , Rossi Maura , Esposito Aurora , Tazzari Pier Luigi , Ricci Francesca , Nicoletti Giordano , Landuzzi Lorena , Fanti Stefano , Ambrosini Valentina , Leoni Flavio , Mascagni Paolo , Gazzola Anna , Lollini Pier Luigi , Zinzani Pier Luigi , Pileri Stefano A.
Published: April 2020

Background. Recently, gene expression profiling (GEP) indicated histone-deacetylases (HDAC) as potential therapeutic targets in peripheral T-cell lymphomas (PTCL) not otherwise specified (NOS), the commonest PTCL type. Consistently, phase II trials demonstrated the efficacy of some HDAC inhibitors (HDACi), including SAHA, which was approved for cutaneous T-cell lymphomas (CTCL) treatment.

Aims and methods. We investigated the anti-tumour effects of ITF2357 (Italfarmaco, Italy), a novel hydroxamic acid HDACi, on PTCL primarily-cultured cells and cell lines (HH and FEDP), and in a xenografted mouse-model of CTCL.

Cultured cells were incubated with different dosages of ITF2357 and SAHA (ranging from 0.5 to 2.5 mM). Cell viability, assessed by trypan-blue exclusion assay, cell-cycle progression, assessed by bromodeossiuridine assay, and apoptotic rate, determined by flow-cytometry analysis of annexin-V binding populations were determined at 48, 72 and 120 hours. Nude mice, injected with HH cells, received ITF2357 (10–20mg/Kg, per os) for 14 days. Micro-PET scan was adopted for disease measurement and treatment response evaluation. Finally, GEP of cell lines exposed to ITF2357 and SAHA were generated to elucidate their mechanisms of action.

Results. Cell viability of HH cells treated with ITF2357 ranged from 50% (0.5 mM, at 48 h), to <10% (0.5–2.5 mM, at 72–120 h), in comparison to untreated cells. Differently, cell viability of HH cells treated with SAHA ranged from 80% (0.5 mM, at 48–120 h) to 25% (2.5 mM at 48 h). Analogue effects were documented in FEDP and primarilycultured PTCL cells. Conversely, viability of normal T-lymphocyte was not significantly affected. Interestingly, exposure to ITF2357 was associated to G0/G1 cell-cycle arrest and apoptosis induction. Finally, ITF2357 determined significant reduction of tumoral masses and survival benefit in a xenografted mice-model inoculated with HH cells.

Conclusion. Taken together, these data demonstrate that ITF2357 is effective against PTCLs ex vivo and in vivo, by nominating it for clinical evaluation in this setting.