bluuebunny/crossref_metadata_2025 huggingface dataset:
Treatment of Primary Cutaneous Marginal Zone B-Cell Lymphomas with Intralesional Rituximab
Authors: Domingo Domenech Eva , Salas Queralt , Servitje Octavio , Climent Fina , Pomares Helena , Lucas Ana , Aguilera Carmen , Comai Alessandra , Mercadal Santiago , Gonzalez Barca Eva , Sureda Anna
Published: October 2019

BACKGROUND

Primary cutaneous marginal zone B-cell lymphomas (PCMZL) are low-grade lymphomas with an indolent course, but with a high rate of skin relapses. Surgical excision, local radiotherapy, or combinations of both are the first-line treatment in solitary or localized disease. Therapeutic options in cases of multifocal disease or relapse, include steroids or intravenous Rituximab in monotherapy or in combination with intravenous or oral chemotherapy. The effectiveness of intralesional Rituximab (ILR) in primary cutaneous B-cell lymphomas has been described in a small multicenter series of patients.

OBJECTIVE

To evaluate the effectiveness and safety of ILR in patients with PCMZL.

METHODS

In our center ILR is used since 2010 as fist-line treatment for patients with PCMZL with single facial lesions, multifocal disease that cannot be irradiated or in relapsed disease. The treatment regimen consist of a course of three injections of 10 mg (days 1, 3 and 5) in a single week at monthly intervals. Oral paracetamol and dexclorfeniramine were administrated 30 minutes before the procedure. Courses of ILR were monthly repeated until response or in cases of relapse. The primary outcome measure of the study was the rate of remissions (complete and partial remissions).

RESULTS

Twelve patients with PCMZL have been treated so far, 50% as first-line. The relapsed-group had previously received a median of 3 previous treatments (range, 1-6) including radiotherapy in 5 cases, surgery in 1 case, intravenous chemotherapy in 2 cases, and topic steroids in 2 cases. Clinical characteristics of the 12 patients are shown in the table below.

A total of 42 courses of ILR have been completed. All patients responded: a complete therapeutic response was achieved in 10 patients, whereas a partial response was observed in 2. No cases of progression or relapse in the site of ILR treatment were documented.

Four patients experienced transitory mild perilesional erythema that did not require medication and resolved in less than 4 hours. No cases of significant adverse events occurred and none of the patients suffered severe bacterial or opportunistic infections.

CONCLUSIONS

In our preliminary experience, ILR is an effective and safe therapeutic approach for selected PCMZL patients with single facial lesion, multifocal disease than cannot be irradiated, or in relapsed cases. To date, this is the largest experience with ILR of a single center.

Disclosures

Gonzalez Barca: Roche: Speakers Bureau; Janssen: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Gilead: Speakers Bureau.
Drug Shortage: The Impact of Substituting Mechlorethamine in Stanford V for Hodgkin's Lymphoma
Authors: Monteiro Luis P , Gaspar Catia L , Fevereiro Margarida , Tome Ana L , Almeida Nuno , Monteiro Alexandra , Sousa Aida B.
Published: October 2019

Introduction: the shortage of some anticancer drugs has forced their substitution in established combination chemotherapy regimens, but the replacement may have critical effects on the results of treatment and need careful evaluation. Since 1996 we treat Hodgkin's lymphoma (HL) with the Stanford V (SV) regimen, which achieves the same results as the more widely employed ABVD regimen with the benefits of a shorter duration (12 weeks) and lower cumulative doses of bleomycin and doxorubicin. When mechlorethamine became unavailable in 2008, we substituted cyclophosphamide at a dose of 650 mg/m2 for mechlorethamine 6 mg/m2. In 2012 the Pediatric Hodgkin Lymphoma Consortium, which had adopted the same policy, suggested in a retrospective analysis that the modification could lead to an inferior event-free survival (EFS). To assess the impact of the substitution, we retrospectively compared our results with the original SV (SVo) and the modified SV (SVm).

Methods: we evaluated the 265 patients consecutively treated in our center with SV between Nov/96 and Dec/14. We compared the pre-treatment characteristics, the complete remission (CR) rate, EFS and overall survival (OS) of the 184 patients who received SVo to those of the 81 patients treated with SVm. Radiotherapy indications and supportive measures were unchanged between the 2 periods.

Results: median age of the 2 groups was identical: 31 (16-69) for SVo and 31 (15-72) for SVm. No differences were found in histologic subtypes (nodular sclerosis 76/74%), frequency of B symptoms (36/47%, p=0.09), proportion of advanced stages (32/33%) or International Prognostic Score (≥3 in 20/13%, p=0.22). The CR rate was 89% for SVo and 86% for SVm. With a median follow up of 9 years for the SVo group and 4.6 years for the SVm group, there were no significant differences in EFS (76/73%, p=0.66) and OS (89%/85%, p=0.63) at 5 years.

Conclusion: the impact of forced substitutions in components of combination chemotherapy regimens due to drug shortage must be carefully scrutinized, notably in first line protocols for curable diseases. In the case of HL treated with the SV regimen, the present study is the first, to our knowledge, to evaluate the impact of replacing mechlorethamine by cyclophosphamide in adults. We found no difference in OS nor, unlike the pediatric study, in EFS. The substitution can be considered safe, since the modified version of SV achieves the same favorable results as the original SV.

Disclosures

No relevant conflicts of interest to declare.
NR4A3 Suppresses Lymphomagenesis In Vitro and In Vivo by Induction of Similar Patterns of Pro-Apoptotic Genes like NR4A1
Authors: Deutsch Alexander JA , Rinner Beate , Pichler Martin , Fechter Karoline , Greinix Hildegard T. , Wang Chenguang , Beham-Schmid Christine , Neumeister Peter
Published: October 2019

The nuclear orphan receptors NR4A1 and NR4A3 have been demonstrated as cooperating tumour suppressor genes leading to rapid development of acute myeloid leukaemia (AML) in double knock-out mice. In humans, their expression is reduced in leukemic blasts in AML patients. Furthermore, NR4A1 and NR4A3 hypoallelic mice develop preleukemic myelodysplastic/myeloproliferative disorders with progression to AML in some cases. Recently we published a comprehensive study of NR4A nuclear receptor expression levels in lymphoid neoplasms that revealed a marked reduction of NR4A1 and NR4A3 in the majority of patients with B-cell chronic lymphocytic leukaemia, with follicular lymphoma, and with diffuse large B cell lymphoma. Interestingly, functional characterization demonstrated that NR4A1 induces apoptosis of aggressive lymphoma cells in vitro and suppresses tumour growth in a xenograft mouse model.

Since the role of NR4A3 in aggressive lymphomas is unknown, we aimed to investigate its etiopathogenic function in these tumors. Low expression of NR4A3 was associated with poor survival in aggressive lymphoma patients. Experimentally, induction of NR4A3 expression by inducible ectopic expression in a variety of lymphoma cell lines led to a significantly higher proportion of apoptotic cells as demonstrated by DNA cleavage, Annexin V staining and increased caspase 3/7 activity. To test the tumor suppressor functions of NR4A3 in vivo, the stably transduced SuDHL4-lymphoma cell line was xenografted in the NOD-SCID-gamma (NSG) mouse model. In this system NR4A3 expression abrogated tumor growth in the NSG mice, whereas vector control and uninduced cells formed massive lymphoid tumors. Pharmacological activation of NR4A3 by Thapsigargin and BF175 resulted in a NR4A3 dependent induction of apoptosis in vitro. To dissect differential transcriptional activity of NR4A3 and NR4A1,both factors were separately over-expressed in four different aggressive lymphoma cell lines followed by semi-quantitative mRNA expression analysis of intrinsic and extrinsic apoptotic genes. NR4A1 or NR4A3 over-expression caused apoptosis by induction of BAK, Puma, BIK, BIM, BID and Trail to the same degree.

In summary, our data suggest that NR4A3 possesses tumor suppressive function in aggressive lymphomas by pro-apoptotic transactivation and that NR4A3 is functional redundant to NR4A1 in aggressive lymphomas.

Disclosures

No relevant conflicts of interest to declare.
Dnmt3a Haploisufficiency Cooperates with Oncogenic Kras to Promote an Early-Onset T-Cell Leukemia
Authors: Chang Yuan-I , Kong Guangyao , Zhang Jing , Ranheim Erik A.
Published: October 2019

Recent whole genome/exome sequencing efforts in myeloid malignancies identified that mutations in DNA methyltransferase 3A (DNMT3A) are prevalent in acute myeloid leukemia (AML). In addition, DNMT3A mutations are also identified in various T cell malignancies. Of note, DNMT3A mutations are typically heterozygous and some WT DNMT3A functions thus remain in this state. However, the predominant DNMT3A R882 mutations, which locate in the catalytic domain, seem to inhibit the methyltransferase activity of the remaining WT DNMT3A due to its dominant-negative function (Yang L, Rau R, Goodell MA, Nat. Rev. Cancer 15: 152-165, 2015). COSMIC database analysis reveals different prevalence of DNMT3A R882 mutations in various hematopoietic malignancies. Approximately 60% of DNMT3A mutations in AML are R882 mutations, while the frequency of R882 mutations drops to ~40% in myelodysplastic syndrome (MDS) and myeloproliferative neoplasm (MPN). In contrast, the frequency of R882mutations is less than 25% in T-cell acute lymphoblastic leukemia (T-ALL). The significantly different frequencies of DNMT3A R882 mutations in AML versus T-ALL inspired us to investigate whether downregulation of DNMT3A regulates malignancies of different lineages in a dose-dependent manner.

We previously showed that Dnmt3a-/- promotes MPN progression in KrasG12D/+ mice and ~1/3 compound mice develop AML-like disease (Chang et al. Leukemia 29: 1847-1856, 2015). Here, we generated KrasG12D/+; Dnmt3afl/+; Mx1-Cre mice to determine how Dnmt3a haploisufficiency affects KrasG12D/+-induced leukemogenesis. After pI-pC injections to induce Mx1-Cre expression, primary KrasG12D/+; Dnmt3a+/- mice died quickly as primary KrasG12D/+ mice; the survival rates of these two groups of animals were not significantly different. However, in a competitive transplant setting, recipients transplanted with KrasG12D/+; Dnmt3a+/- bone marrow cells displayed a significantly shortened survival than recipients with KrasG12D/+ cells. Moreover, all of the recipients with KrasG12D/+; Dnmt3a+/- cells developed a lethal T-ALL without significant MPN phenotypes, while ~20% of recipients with KrasG12D/+ cells developed MPN with or without T-ALL. This is in sharp contrast to the recipients with KrasG12D/+; Dnmt3a-/- cells, in which ~60% developed a lethal myeloid malignancy (MPN or AML). Our data suggest that in the context of oncogenic Kras, loss of Dnmt3a promotes myeloid malignancies, while Dnmt3a haploisufficiency induces T-ALL. This dose-dependent phenotype is highly consistent with the prevalence of DNMT3A R882 mutations in AML versus T-ALL in human. We are currently investigating the underlying mechanisms.

Disclosures

No relevant conflicts of interest to declare.
GDPT Versus CHOP in Newly Diagnosed Peripheral T-Cell Lymphoma: A Prospective Randomized Controlled, Open-Label Study (No.NCT01664975)
Authors: Li Ling , Duan Wenjing , Young Ken H. , Li Zhaoming , Zhang Lei , Fu Xiaorui , Li Xin , Sun Zhenchang , Zhang Xudong , Yan Jiaqin , Nan Feifei , Chang Yu , Lin Li , Zhang Mingzhi
Published: October 2019

Background: Peripheral T-cell lymphoma is a distinct lymphoid neoplasm with aggressive course and poor outcome. Optimal treatment strategies for peripheral T-cell lymphoma have not been well defined. We compared the efficacy and safety of GDPT and CHOP regimens for patients with newly diagnosed peripheral T-cell lymphoma in a prospective randomized controlled and open-label clinical trial (No.NCT01664975).

Methods: All eligible patients with newly diagnosed peripheral T-cell lymphoma had measurable disease with an ECOG performance status ≤ 2 and adequate organ function. GDPT or CHOP chemotherapy were randomly assigned to patients. Patients in arm GDPT received intravenous gemcitabine (0.8g/m2) in 30 min on days 1 and 8, cisplatin (25mg/m2) on days 1-3, and oral prednisone (60 mg/m2) on days 1-5, thalidomide (200 mg) until the end of the whole chemotherapy. Patients in group CHOP received intravenous cyclophosphamide (750 mg/m2), doxorubicin(50 mg/m2) and vincristine (1.4 mg/m2, maximum 2 mg) on day 1, and oral prednisone (60 mg/m2) on days 1-5. Each cycle was repeated six times every 3 weeks. Efficacy was evaluated every two cycles. The primary endpoint was to evaluate the efficacy assessed by progression-free survival. Secondary end points included response rate and overall survival.

Results: Between July 2010 and June 2016, 103 patients allocated into two groups randomly, of whom 52 were treated with GDPT therapy and 51 were treated with CHOP therapy. Patient characteristics were well balanced within the two arms of treatment at enrollment (Table 1). The 2-year progression-free survival (PFS) and overall survival (OS) rates were better in GDPT group than that in CHOP group (57% versus 35% for 2-year PFS, P=0.0035; 71% versus 50% for 2-year OS, P =0.0001) (Figure 1). Complete remission (CR) rate and overall response rate (ORR) of GDPT group were higher than that in CHOP group (52% versus 33%, P =0.044 for CR rate; 67% versus 49%, P =0.046 for ORR) (Table 2). Adverse effects of chemotherapy was hemocytopenia predominantly in both arms. No differences were observed between the two arms in terms of grade 3/4 myelosuppression, digestive tract, hepatic, renal, cardiac or neurological toxicity (Table 3). Acute toxicity was moderate, tolerable and well managed in both arms.

Conclusions: GDPT chemotherapy resulted in significant improvement in PFS and OS compared with CHOP chemotherapy and side effects of chemotherapy was well tolerated for newly diagnosed peripheral T-cell lymphoma patients. Therefore, GDPT is a promising new regimen as potential first-line therapy against peripheral T-cell lymphoma.

Disclosures

No relevant conflicts of interest to declare.
Long-Term Follow-up of Primary Bone Diffuse Large B-Cell Lymphoma Treated with m NHL-BFM-90
Authors: Gabeeva Nelly G. , Zvonkov Eugene E. , Morozova Anna K. , Koroleva Daria A. , Kovrigina Alla M. , Savchenko Valery Grigorievich
Published: October 2019

Background: Primary bone lymphoma (PBL) accounts for about 5% of extranodal lymphomas, the most common type are Diffuse Large B-cell Lymphoma (PBL-DLBCL). R-CHOP chemotherapy combined with or no radiation therapy can induce 5-year overall and progression free survival about 63% and 60% for localized-stage and only 41% and 35% for advanced stage and with adverse factors (AF). Due to the rarity of the disease the optimal treatment strategy still remains unknown. Further research is thus needed to increase treatment efficacy for patients with advanced stage PBL-DLBCL and with AF.

Aims: To evaluate the efficacy of mNHL-BFM-90 program in adult patients with advanced stage PBL-DLBCL and with AF.

Methods: Twenty three previously untreated pts with PBL-DLBCL underwent mNHL-BFM-90 treatment between May 2007 and July 2014; mean age 41 years (range 17-65); age ≥60 years 3 pts (13,6%); M\F=14\9. Tumor stage according to the Ann-Arbor classification: stage I (involvement of 1 bone) - 7 pts (30,4%), II (involvement of regional lymph nodes) - 3 pts (13%), IV (multiple bone involvement) - 13 pts (56,6%); stage >I 16 pts (69,5%). All pts with stage I had large-size tumors (>6sm) with soft tissue contact invasion. LDH elevation was observed in 10 pts (43,5%), B-symptoms in 15 pts (65%), large-size tumors - 20 pts (87%). The most common tumor sites were the skull bones, vertebrae, and femur. All patients received treatment according to the mNHL-BFM-90 intensive program This program was modified in the following way: adriamycin was administered on the 3rd day of course AA at a dose 50 mg/m2; methotrexate was administered on the 1st day of course C at a dose 1.0 g/m2 for 12 hours. All patients underwent from 4 to 6 courses. Twelve (52%) pts received rituximab on day 0 prior to each course of therapy. Five pts received radiotherapy, all pts received intrathecal prophylaxis of central nervous system (CNS) involvement.

Results. Complete remissions were achieved in 23 pts (100%). Relapses were observed in three patients in 6, 9 and 40 months after completion of treatment. From 2 pts with early relapse, one was diagnosed as «double-expression lymphoma» (MYC expression >70%, BCL2 >60%), both pts proliferation index Ki-67 accounted 100%. All 3 pts received salvage therapy and died from progression of disease.

With a median follow-up of 69 months (range 17-110) both progression-free and overall survival of 23 pts constituted 87%. Grades 3, 4 hematological toxicity was observed in all pts. No deaths related to mNHL-BFM-90 treatment occurred. No second malignancies were observed.

Conclusion The mNHL-BFM-90 demonstrated high efficacy and acceptable toxicity in patients with advanced-stage PBL-DLBCL and with AF.

Figure Figure.

Disclosures

No relevant conflicts of interest to declare.
Impact of Treatment Center Characteristics on Early Death Rates in Patients with Newly Diagnosed Acute Promyelocytic Leukemia: Analysis of Data from the SEER Program
Authors: Park Jae H. , Devlin Sean , Lynch Charles , Schwartz Ann , Keegan Theresa , Gonsalves Lou , Hermit Kim , Green Michael , Douer Dan , Tallman Martin S.
Published: October 2019

Background: Acute promyelocytic leukemia (APL) is a unique subtype of AML with a high cure rate of 80-90% when treated with all-trans retinoic acid (ATRA)-based therapy. However, early death at presentation or during induction therapy due to hemorrhage remains the major cause for treatment failure. Using the Surveillance, Epidemiology, and End Results (SEER) data, we have previously reported an early death rate of 17.3% (Park J et al. Blood 2011;118:1248). We hypothesized that the early death may be associated with the expertise and availability of blood products at treatment centers and undertook a retrospective cohort study.

Methods: APL patients diagnosed between 1992 and 2009 were identified using SEER 13 data. Patients with available survival and treatment center information were eligible for the study. Early death was defined as death within one month of diagnosis. The treatment center information was obtained from the American Hospital Association database and included the following characteristics: number of hospital beds (<200, 200-500, >500), presence of intensive care unit (ICU), presence of trauma center (as a surrogate indicator for availability of on-site blood products), and the teaching status (nonteaching, minor, major). Logistic regression was used to compare early death by selected hospital characteristics.

Results: The full-set of pre-specified treatment center information were available from five SEER 13 cancer registries (Iowa, Utah, Detroit, Connecticut, Hawaii); from these registries, we identified 507 eligible patients ofwhom [154 patients (30%)] experienced an early death. Data on bed size, teaching status, ICU presence and trauma center were available in 460, 460, 445 and 434 patients, respectively. Most APL patients were treated at centers with ³200 hospital beds (n=345), dedicated ICU (n=442), trauma center (n=287) and at teaching hospitals (n=349) (see Table 1). Of the treatment center characteristics, analysis revealed that the presence of trauma center was statistically significantly associated with decreased odds of early death (odds ratio: 0.59, 95% CI: 0.39-0.91; p=0.02).

Conclusions: Early death remains as the major cause of treatment failure in APL. To the best of our knowledge, this is the first and largest study reporting an association of treatment center characteristics with survival in APL, and suggests a prompt referral of APL patients to a large treatment center may further improve the outcome of these patients.

Disclosures

Park: Novartis: Consultancy; Baxalta: Consultancy; Juno Therapeutics: Research Funding; Amgen: Consultancy. Douer:Pfizer: Consultancy; Shire: Consultancy, Speakers Bureau; Spectrum: Consultancy; Jazz Pharmaceuticals: Honoraria; Gilled Sciences, Inc: Consultancy.
CCND1 Mutations Increase Protein Stability and Promote Ibrutinib Resistance in Mantle Cell Lymphoma
Authors: Mohanty Atish Ranjan , Sandoval Natalie , Das Manasi , Amin Hesham M. , Marcucci Guido , Pillai Raju , Weisenburger Dennis D , Rosen Steven T. , Pham Lan , Ngo Vu N.
Published: October 2019

Mantle cell lymphoma (MCL) represents 6-8% of non-Hodgkin's lymphoma with a median survival of 3-5 years. CyclinD1 (CCND1) chromosomal translocation t(11;14) is the hallmark of this neoplasm, which leads to its deregulated expression. Recent knock down studies of CCND1 in MCL have shown that its role is not only limited to promote cell cycle progression but also for maintaining genomic stability. Knock down of CCND1 results in increased DNA damage in MCL cells and makes them more susceptible to chemotherapy. However, CCND1 is not only prone to frequent translocation but also to somatic mutations. Genomic studies have identified recurring somatic mutations in the coding region of CCND1 in approximately 30% of MCL cases. The hot spot of these mutations is concentrated in exon 1 of CCND1. However, the functional consequence of these mutations is not known. We generated the most common CCND1 mutations (E36K, Y44D or C47S) by site directed mutagenesis and overexpressed these mutants in JEKO-1, UPN-1 or Z-138 MCL cell lines. Here, we show that these single mutations, as compared to the wild type (WT), increased CCND1 protein levels in MCL cell lines and primary tumors. Mechanistically, mutations Y44D or C47S increase the stability of CCND1 protein through attenuation of threonine-286 phosphorylation, which is a prerequisite for export of CCND1 from the nucleus to the cytoplasm, followed by proteolysis through the ubiquitin-proteasome pathway. But the two mutants have stronger affinity to interact with the threonine-286 phosphorylating enzyme glycogen synthase kinase 3 beta ,whereas interaction with cyclin-dependent kinase 4 remains unaffected. Furthermore we observed that these two mutants are preferentially localized in the nucleus compared to WT, as determined by cell fractionation and immunofluorescence assays. In order to understand the relevance of stable CCND1 protein in MCL, we studied the response of WT or mutant CCND1-expressing MCL cell lines to ibrutinib, an FDA- approved Bruton tyrosine kinase inhibitor used for MCL treatment. We observed that forced expression of WT or mutant CCND1 increased the resistance of JEKO-1 cells to ibrutinib. The Y44D mutation sustained the resistance to ibrutinib at supraphysiologic concentrations (5-10 μM). Furthermore, primary MCL tumors with CCND1 mutations expressed a more stable protein and were resistant to ibrutinib than WT cases. These findings uncover new mechanisms for MCL cells to increase CCND1 protein level through somatic mutations, which may provide a survival advantage to the lymphoma cells in developing resistance against the emerging ibrutinib therapy.

Disclosures

Pillai: Trillium Therapeutics: Research Funding. Pham:Vyripharm Biopharmaceuticals: Research Funding.
Histone-Modifying Gene Mutations Are Related to Tumor Progression and Response to Histone Deacetylase Inhibitors in Patients with Peripheral T-Cell Lymphoma Not Otherwise Specified
Authors: Ji Mengmeng
Published: October 2019

Objective

Histone modifications are critically involved in pathogenesis of peripheral T-cell lymphoma not otherwise specified (PTCL-NOS).Histonedeacetylaseinhibitors (HDACIs) exhibited potential anti-lymphoma activity through restoring normal chromatin modification patterns. Here we detected histone-modifying gene mutations in PTCL-NOS and correlated them with tumor progression and therapeutic response to different HDACIs.

Methods

Mutations of important lymphoma-associated histone-modifying genes (MLL2, TET2, CREBBP, EP300, SETD2 and EZH2) were detected by targeted sequencing in tumor samples of 105 newly diagnosed patients with PTCL-NOS and compared with clinical parameters. Moreover, T-lymphoma cell lines bearing representative mutations were treated with four clinically available HDACIs (valproic acid, VPA;suberoylanilidehydroxamic acid, SAHA;Romidepsin,Romi; andChidamide,Chid).

Results

A total of 65 histone-modifying gene mutations were identified in 39 of 105 PTCL patients (Figure 1A). As illustrated in Figure 1B, somatic mutations in MLL2 (encoding H3K4-specific methyltransferase) were most frequently observed (20.0%), followed by mutations of TET2 (13.3%), EP300 and CREBBP (encoding H3K18 acetyltransferase, 7.6% and 6.7%, respectively), and SETD2 (encoding H3K36 methyltransferase, 7.6%). No EZH2 mutation was detected. Clinically, mutation-positive PTCL patients presented with shorteroverall survival than patients without mutations (P=0.0038, Figure 1C).

Based on the crystal structure of each gene (Figure 1D), most mutations were located on SET domain of MLL2 (p.5397-5513), HAT domain of EP300 (p.1287-1663) and SET domain of SETD2 (p.1550-1667), which could lead to loss of H3K18ac, H3K4me3 and H3K36me3 expression, respectively.

Meanwhile, Jurkatcells were treated with VPA, SAHA, Romiand Chidfor 48h and the results showed that all the HDACIs increased the expression of EP300/CREBBP, as compared to the untreated group. Interestingly, VPA and SAHA specifically upregulated SETD2 expression, while Romi and Chid upregulated MLL2 expression (Figure 2A). No obvious change of TET2 expression was found during HDACI treatment. Thereafter,Jurkat cells were transfected with MLL2 mutant (p.V5389M), EP300 mutant (p.H1377R) and SETD2 mutant (p.R1598_). Compared with the WT, mutant reduced the level of H3K18ac, H3K4me3 and H3K36me3 expression, respectively. All the HDACIs restored H3K18 hypo-acetylation induced by EP300 mutant. VPA and SAHA restored H3K36me3 expression inhibited by SETD2 mutant, whileRomi andChid restored H3K4me3 expression inhibited by MLL2 mutant (Figure 2B-2D).

Conclusion

Our study provided the mutational spectrum of histone-modifying genes in PTCL-NOS. HDACIs targeted differently histone H3 acetylation or methylation modulated by the mutations, suggestive their distinct therapeutic efficiency in PTCL-NOS.

Figure. Figure. Figure. Figure.

Disclosures

No relevant conflicts of interest to declare.
The Syk Inhibitor TAK-659 Prevents Splenomegaly and Tumor Development in a Murine Model of EBV-Associated Lymphoma
Authors: Cen Osman , Kannan Karuppiah , Huck Jessica J. , Yu Jie , Zhang Mengkun , Gordon Leo I , Longnecker Richard
Published: October 2019

Epstein-Barr virus (EBV) is associated with the development of several cancers including Burkitt's Lymphoma, Hodgkin Lymphoma, Nasopharyngeal and Gastric Carcinomas. EBV-encoded Latent Membrane Protein 2A (LMP2A), a BCR mimic, is expressed in cancer cells and is essential for transformation, migration and inhibition of differentiation. LMP2A/λ-MYC double transgenic mice develop splenomegaly and lymphoma much faster than mice that express Myc only. LMP2A induces cancer phenotype by recruiting and activating BCR signaling components such as Lyn and Syk kinases to the SH2 motifs found on its N terminal cytoplasmic domain. In this study, we explored inhibition of Syk kinase activity with a novel Syk/FLT-3 inhibitor TAK-659, in lymphomas from these transgenic mice to assess SYK inhibition as a potential therapeutic strategy for EBV-associated malignancies.

The cell lines developed from LMP2A/Myc transgenic mice derived lymphomas showed higher basal pSYK levels than Myc lymphoma cell lines. TAK-659 treatment resulted in early apoptosis within 8hrs in LMP2A/Myc lymphoma cells in comparison to 24 hrs in Myc lymphoma cells. In both pre-tumor and tumor transfer models in vivo, compared with placebo and Myc tumors, LMP2A/MYC mice treated with TAK-659 did not develop splenomegaly and tumor development was totally inhibited in these mice indicating SYK signaling is essential for tumor development in LMP2A/Myc mice. In addition, metastasis of tumor cells into BM was abrogated. TAK-659 killed tumor cells but not host cells in spleen and tumors.

Taken together our data show that TAK-659, even in low nanomolar concentrations, inhibits pSYK and induces apoptosis in LMP2A/Myc tumor cells. It also inhibits splenomegaly and tumor development in our autochthonous and syngeneic tumor transfer models of EBV-associated lymphoma by targeting tumor cells while sparing non-tumor cells. Therefore, TAK-659 may provide an effective therapeutic option for the treatment of EBV-associated malignancies and should be explored further in clinical trials.

Disclosures

Kannan: Millennium Pharmaceuticals Inc., Cambridge, MA, USA, a wholly owned subsidiary of Takeda Pharmaceutical Company Limited: Employment. Huck:Takeda Pharmaceuticals International Co, Cambridge, MA: Employment. Yu:Takeda Pharmaceuticals International Co, Cambridge, MA: Employment. Zhang:Takeda Pharmaceuticals International Co, Cambridge, MA: Employment. Gordon:Northwestern University: Patents & Royalties: Patent for gold nanoparticles pending.
The Role of MYC and BCL-2 Double Expression the in DLBCL Patients with Advanced Stage and Elevated LDH Who Received Upfront ASCT after R-CHOP
Authors: Kim Yu Ri , Yoon Sun Och , Kim Soo-Jeong , Cheong June-Won , Lee Hyewon , Chung Haerim , Lee Jung Yeon , Jang Ji Eun , Kim Yundeok , Yang Woo-Ick , Min Yoo-Hong , Kim Jin Seok
Published: October 2019

Diffuse large B-cell lymphoma (DLBCL) with MYC and BCL2 protein concurrent expression, double- expressor lymphoma (DEL), by immunohistochemistry (IHC) was related with poor prognosis. To investigate the role of upfront autologous hematopoietic stem cell transplantation (ASCT) in double-expresser lymphoma, we retrospectively evaluated 44 DLBCL patients with advanced stage and elevated LDH who received upfront ASCT. We used the cut off value of BCL2 was 50% and c-MYC was 40%. All patients were treated with frontline rituximab plus cyclophosphamide, doxorubicin, vincristine and prednisone (R-CHOP) chemotherapy.

Median age of 44 patients was 52.0 years (range, 23 ~ 64 years). Two-year overall survival (OS) was 83.3% and progression free survival (PFS) was 76.6%. Double-expressor lymphoma (DEL) were detected 14 patients (31.8%). Two-year OS of the patients with DEL was 77.4%, non-DEL was 86.6%. Two-year PFS of the patients with DEL was 77.9% and non-DEL was 80.9%. Eleven (25.0%) patients were germinal center B-cell (GCB) subtype and 33 (75.0%) patients were non-GCB subtype. There was no survival difference between GCB and non-GCB subtype (Figure 1A, 1B). In the patients with GCB subtype, 2 (18.2%) patients were classified to DEL. OS and PFS were not different according to DEL. Among non-GCB subtype, the patients with DEL (n=12) showed the comparable OS and PFS compared to the patient with non-DEL (n=21). Two-year OS of DEL patients was 73.3%, non-DEL was 90.9%. Two-year PFS of DEL patients was 74.1 %, non-DEL was 82.6 (Figure 2A. 2B).

In conclusion, MYC and BCL2 concurrent expression did not show the poor outcome among the high risk DLBCL patients treated with upfront ASCT regardless of molecular classification. It will be need to investigate the role of ASCT in these group of patients.

Figure 1 Kaplan-Meier analysis of overall survival and progression survival in germinal center B-cell subtype versus non-germinal center B-cells subtype Figure 1. Kaplan-Meier analysis of overall survival and progression survival in germinal center B-cell subtype versus non-germinal center B-cells subtype Figure 2 Overall survival and progression survival of patients treated with upfront autologous hematopoietic stem cell transplantation according to the presence of double expression Figure 2. Overall survival and progression survival of patients treated with upfront autologous hematopoietic stem cell transplantation according to the presence of double expression

Disclosures

No relevant conflicts of interest to declare.
Tissue Mast Cell Leukemia
Authors: FRIEDMAN BEN I. , WILL JOHN J. , FREIMAN DAVID G. , BRAUNSTEIN HERBERT
Published: October 2019

A case of "tissue mast cell leukemia" in a 34-year old white woman is reported. The possible association with urticaria pigmentosa and myeloid leukemia is considered.
Cytochemical Studies of Glycogen Content of Lymphocytes in Lymphocytic Proliferations
Authors: MITUS W. J. , BERGNA L. J. , MEDNICOFF I. B. , DAMESHEK W.
Published: October 2019

The glycogen content of the lymphocytes of 44 cases of various lymphocytic proliferations was studied using a semiquantitative cytochemical technic. The lymphocytes of chronic lymphocytic leukemia and of lymphosarcoma (with one exception) showed greatly increased glycogen content. In acute lymphocytic leukemia and in one case of lymphosarcoma the glycogen content was low. The presence or absence of lymphosarcoma cells in the peripheral blood in cases of lymphosarcoma made no difference in the total score. The difference between the glycogen-positive "lymphosarcoma cell" and the glycogen-negative lymphoblast is apparently due to the greater maturity of the cytoplasm of the lymphosarcoma cell (which results from an asynchronism of nucleocytoplasmic maturation) and immaturity of the cytoplasm of the lymphoblast.

Three cases of benign lymphocytosis with atypical lymphocytes in the peripheral blood gave increased values. One case with atypical lymphocytes had normal values and two cases of lymphocytosis with morphologically normal lymphocytes also had normal values. It is concluded that the increase of glycogen in lymphocytic cells is due to an over-all proliferation of the lymphocytes and is not necessarily related to malignancy per se.
Studies of Megakaryocyte Glycogen. I. A Semiquantitative Method of Measurement: Effect of Phlebotomies in Young Adults
Authors: DANIELL HARRY W.
Published: October 2019

1. The distribution of glycogen and nonglycogen structures in megakaryocytes as demonstrated by the Hotchkiss technic is described.

2. A semiquantitative method for estimating the relative amounts of megakaryocyte glycogen in a bone marrow aspiration has been developed and is described. The result is expressed as the Megakaryocyte Glycogen Score (MGS).

3. The megakaryocyte glycogen scores on 25 aspirations from 14, healthy, young adult volunteers are presented and fall between 20 and 80, except in those subjects with recent acute blood loss or repeated phlebotomies over a period of years.

4. Subjects who have had repeated phlebotomies tend to have higher MGS than subjects without repeated phlebotomies. These higher values of MGS exist in the absence of abnormalities in the reticulocyte count, bone marrow differential count, platelet count, hemoglobin, hematocrit, wbc and differential, and rbc.

5. In one subject with a normal MGS, a 500 cc. phlebotomy was followed in one hour by a decrease in MGS. The MGS had increased above normal by four hours postphlebotomy and was still elevated 25 days after phlebotomy.

6. Implications of these observations are discussed.
Observations on Heinz Bodies in Normal and Splenectomized Rabbits
Authors: ROTHBERG HARVEY , CORALLO LEON A. , CROSBY WILLIAM H.
Published: October 2019

The disappearance from the circulation of Heinz bodies, produced by incubation of blood with phenylhydrazine, was studied in normal and splenectomized rabbits. In the intact animal, red cells damaged by this procedure are removed largely by the spleen. In both intact and splenectomized rabbits, the Heinz bodies are withdrawn from the circulation by removal in toto of the red cells containing them.
Vitamin B12 and Folic Acid Metabolism in Hookworm-Infected Patients
Authors: LAYRISSE MIGUEL , BLUMENFELD NORMA , DUGARTE IRIS , ROCHE MARCEL
Published: October 2019

Studies on the metabolism of B12 and folic acid were performed in patients with heavy hookworm infection and severe iron deficiency anemia, and in patients with light infection, noninfected patients and normal subjects.

Patients with heavy hookworm infection showed a marked decrease of the serum B12 as compared with normal subjects. Eight of 21 cases studied showed values of serum B12 below 100 µµg./ml.

Twelve of 13 patients with severe hookworm infection showed impairment of the pteroylglutamic acid intestinal absorption; however, none of them exhibited megaloblastic proliferation in the bone marrow. They all recovered with iron therapy alone. The patients with light infection and the noninfected patients with iron deficiency anemia did not demonstrate significant differences from the normal subjects studied.
The Pelger-Huët Anomaly in Three Families and Its Use in Determining the Disappearance of Transfused Neutrophils from the Peripheral Blood
Authors: ROSSE WENDELL F. , GURNEY CLIFFORD W.
Published: October 2019

1. Thirteen cases of Pelger-Huët anomaly occurring in three families are presented and discussed. The second recorded case of the anomaly in Negroes is included; the occurrence of familial hereditary elliptocytosis in this same family and the possible relationship to the Pelger-Huët anomaly are discussed.

2. Certain less well-known facts about Pelger-Huët anomaly are considered in relation to the cases presented.

3. The survival time of transfused neutrophils in the peripheral blood was investigated using the anomaly as a tagging device. Most of the cells were found to be absent from the peripheral blood stream in 6 to 8 hours, and none was found after 49.5 hours.
The Effect of Anemic Anoxia on the Cellular Development of Nucleated Red Cells
Authors: ERSLEV ALLAN J. , Ruiz Elva
Published: October 2019

The action of an anoxic stimulus on red cell production was studied in rabbits bled 20 ml./Kg., kept anemic for 20 hours and then reinfused with the previously removed blood. This 20-hour period of anemic anoxia was followed by a characteristic reticulocyte response, a response which was modified by nitrogen mustard or colchicine administered immediately after the 20-hour period of anemia, but was not influenced by anoxia or hyperoxia in the postanemic period. When mitotic division was arrested by colchicine during the 20-hour period of anemic anoxia, the onset of the reticulocyte response, though delayed by 1 to 2 days, was otherwise of characteristic magnitude.

These observations indicate that (1) the anoxic stimulus operates in the bone marrow by accelerating the differentiation of stem cells into pronormoblasts, and that thereafter (2) the maturation and multiplication of differentiated nucleated red cells proceed at fixed rates independent of the anoxic stimulus.
Normal Functions of the Spleen Relative to Red Blood Cells: A Review
Authors: CROSBY WILLIAM H.
Published: October 2019

During all the stages of a red cell’s life the normal spleen exerts a normal function. Eight of these functions have been considered: (1) erythropoiesis; (2) an effect upon red cell production; (3) an effect upon maturation of the red cell surface; (4) the reservoir function; (5) the "culling function"; (6) iron turnover and storage; (7) the "pitting function"; (8) destruction of old red cells.
Preserved Platelets: Their Preparation, Storage and Clinical Use
Authors: TULLIS JAMES L. , SURGENOR DOUGLAS M. , BAUDANZA PHILIPPA
Published: October 2019

A method is described for the isolation and preservation of human blood platelets. Their use in a platelet bank after storage of up to two years is described. The practical advantages to the recipient include small administration volume, freedom from pyrogenicity and immediate availability in the event of thrombocytopenic bleeding. The advantages of this technic to the hospitals include a saving of time, a saving of blood and the elimination of the frantic efforts of blood bank and laboratory personnel to produce large numbers of fresh platelet concentrates at the time of platelet needs.

The remarkable stability of preserved platelets is believed largely due to the method developed for their isolation from blood, wherein the cells are separated from the coagulation proteins with which they normally interact, prior to enzymatic degradation. The importance of a controlled atmosphere of CO2 for retention of clot retraction is presented. The in vitro and in vivo activities of the preserved cells are discussed. A hypothesis is suggested for the autocatalytic stimulation of new platelet production through temporary correction of a thromboplastic deficit by the transfusion of preserved platelets.
Immunologic Mechanisms in Heavily Irradiated Mice Treated with Bone Marrow
Authors: HOLLINGSWORTH J. W. , Perfetto Mary C.
Published: October 2019

1. Humoral antibody production has been studied in severely irradiated mice treated with isologous (same strain) or homologous (different strain) bone marrow.

2. The two methods of study involved functional end points of humoral antibody production as evidenced by in vivo lysis of rat erythrocytes or by regression of mouse leukosis E.L. 4 in histoincompatible mouse recipients.

3. Humoral antibody production was lost after irradiation and isologous marrow treatment, but recovered partially in two weeks and almost completely in four weeks.

4. Established immunity was not abruptly terminated after irradiation and treatment with either isologous or homologous marrow, although there was premature loss of immunity to rat erythrocytes by the irradiated, isologous marrow-treated mouse.

5. Permanent immunity could not be transferred by isologous marrow or spleen from immunized donors to irradiated recipients.

6. Treatment of mice histoincompatible to E.L. 4 leukosis with histocompatible donor bone marrow failed to establish rejection of the tumor.

7. These studies support the concept that humoral antibody production in irradiated, marrow-treated mice remains a function of the host rather than of the transplanted tissues.

8. These studies failed to clarify the conflicting evidence concerning the mechanism of the late illness that occurs after treatment of the irradiated mouse with bone marrow from a different strain or species.
Evaluation of Plasma Erythropoietic-Stimulating Factors in Anemic Uremic Patients
Authors: GALLAGHER NEIL I. , MCCARTHY JOHN M. , HART KATHLEEN T. , LANGE ROBERT D.
Published: October 2019

1. Plasma samples from certain chronically anemic humans have a humoral erythropoietic-stimulating factor.

2. Sixteen anemic uremic patients were studied to evaluate plasma levels of erythropoietic-stimulating factor. Fifteen of these patients showed no active erythropoietic-stimulating factors.
Effect of Phospholipid Fractions upon the Coagulation Defects in Patients on Long-Term Dicumarol Therapy
Authors: BOYLES PAUL W. , JONES RALPH , NICHOL E. STERLING
Published: October 2019

The coagulation defects of the blood of 30 patients after six months of dicumarol therapy have been studied. A preparation of brain or soybean phospholipids corrected the abnormal coagulation time and the thrombin generation test of patients on long-term dicumarol therapy. The deficiency in thromboplastin generation of these patients was not corrected.
Plasma Hemoglobin-Binding Capacity in Sickle Cell Disease
Authors: LATHEM WILLOUGHBY , JENSEN WALLACE N.
Published: October 2019

Studies of the capacity of plasma proteins to bind hemoglobin were made in patients with sickle cell anemia (SS), sickle cell trait (SA), hemoglobin C disease and in patients with hemolytic anemias. Hemoglobin binding was quantitatively normal in sickle cell trait, but was greatly reduced or absent in sickle cell anemia, hemoglobin C disease and in other hemolytic disorders. These alterations have been attributed to a reduction in the level of hemoglobin-binding proteins in circulating plasma. The mechanism of this reduction was not established, but the observed changes were correlated with the presence of increased hemolytic activity.

The binding of hemoglobin C and hemoglobin S by normal plasma was quantitatively normal.
Brief Note: Particle Size as a Factor in Thromboplastin Formation
Authors: BOYLES PAUL W.
Published: October 2019

Latex particles with diameters of 0.14 micron have been shown to be partially effective in the absence of platelets or phospholipids in the thromboplastin generation test.
Relation of Bone Marrow Findings to Serum Protein Changes in Lymphosarcoma, Chronic Lymphocytic Leukemia and Hodgkin’s Disease
Authors: ONAT ALTAN , COOPER TALBERT
Published: October 2019

In order to correlate the electrophoretic alterations in the serum proteins and the findings in the bone marrow, with particular reference to atypical lymphocytic forms and plasma cells, 14 cases of lymphosarcoma, six of chronic lymphocytic leukemia and six of Hodgkin’s disease were studied. No numerical relationship could be found between the degree of lymphoid atypia and the changes in serum globulin. A correlation, however, was apparent between the plasma cell concentration in the bone marrow and the changes in the serum gamma globulin, inasmuch as pronounced hypergammaglobulinemia occurred in all four cases of malignant lymphoma associated with bone marrow plasmacytosis (ranging from 2.8 to 12.8 per cent) and hypogammaglobulinemia of moderate to marked degree was present in five of the six cases of malignant lymphoma in which plasma cells were virtually absent in the bone marrow. When the findings of the fixed sections of the marrow are included, plasmacytosis was demonstrated in bone marrow in eight of 10 cases in which serum gamma globulins were elevated.

In two cases, serum globulins were qualitatively abnormal. In one, a case (case 15) of Hodgkin’s disease, a small fraction of electrophoretically homogeneous globulin was noted. In the other, case 5, in which the histologic diagnosis was lymphocytic lymphosarcoma, macroglobulins constituted 40 per cent of the serum proteins, and Bence Jones proteinuria was associated. Predominantly mature plasmacytosis of 12.8 per cent was present in the bone marrow. This case may be regarded as illustrative of the interrelationship between myeloma and lymphosarcoma and supports the view21 that, clinically and morphologically, transitional forms occur between lymphatic tumors and plasmacytomas.

On the basis of this material, it is suggested that in cases of lymphosarcoma, Hodgkin’s disease and chronic lymphocytic leukemia, alterations in the plasma content of the bone marrow, either as reactive plasmacytosis or as virtual absence of plasma cells, appear to be a major pathogenetic factor in the occurrence of hypergammaglobulinemia and hypogammaglobulinemia, respectively.

Hypogammaglobulinemia was observed in three of six cases of chronic lymphocytic leukemia; two of 14 cases of lymphosarcoma; and one of six classified as Hodgkin’s disease. Conversely, quantitative increases in the gamma globulin level occurred in none of the six cases of chronic lymphocytic leukemia; in six of 14 cases of lymphosarcoma (including two of three classified as lymphosarcoma, reticular cell type), and in three of the six cases of Hodgkin’s disease.
Studies in Leukemia. XII. Interference and Reactivation Experiments with Cell-Free Tissue Extracts of Leukemic Swiss Mice
Authors: SINKOVICS JOSEPH G. , SPURRIER WILMA A. , SCHWARTZ STEVEN O.
Published: October 2019

1. The virulence of the Swiss mouse leukemia agent can be enhanced by rapid brain to brain passages.

2. The existence of an occult agent in the tumor tissue is demonstrable by interference and reactivation experiments.

3. Tumor extracts inhibit the leukemogenic activity of brain homogenates of leukemic mice.

4. Mixtures of tumor extracts and heat-inactivated brain extracts, neither of which is capable of inducing leukemia alone, have leukemia-inducing activity.

5. A heat-labile inhibitor is probably responsible for the masking of the virus in tumor extracts.