bluuebunny/crossref_metadata_2025_split huggingface dataset:
Characteristics and Outcome Of Children and Young Adults With Early T-Precusor (ETP) ALL Treated On UKALL 2003
Authors: Patrick Katharine , Wade Rachel , Goulden Nicholas , Mitchell Chris , Rowntree Clare , Hough Rachael E , Vora Ajay J 
Published: October 2019

Abstract
Early T-precursor (ETP) ALL is a recently described poor prognostic sub-group of childhood T-ALL identified by a distinctive gene expression profile and immunophenotype (CD1a-, CD5 weak, CD8- and presence of myeloid or stem cell markers). It accounts for around 12% of T-ALL and is associated with a poor outcome in retrospective US and Italian studies with a 5 year event-free survival (EFS) of less than 50% on historical treatment protocols. We sought to determine the outcome for this sub-group on a contemporary MRD stratified treatment protocol, UKALL 2003.
As full immunophenotype data was not collected centrally, individual centres were asked to provide individual patient data retrospectively. Of 387 T-ALL cases in the trial, immunophenotype data to classify ETP status was not available for 148 patients. Of the remaining 239 cases, 35 (15%) were classified as definite/probable (CD8 and 1a neg with stem cell or myeloid antigen expression) and 17 (7%) as possible (CD8 neg and stem cell or myeloid antigen expression, 1a not available) ETP. The definite, probable and possible ETP groups had similar patient characteristics and outcomes and have, therefore, been analysed together. Compared to those without an ETP phenotype, ETP patients were more likely to present with a low (<50x109/l) white cell count (56% vs 26%, p<0.0001) but were less likely to be morphological rapid early responders at day 8/15 (63% vs 83% p =0.002) or MRD low risk at the end of induction (4% vs 27%, p = 0.008). Unlike previous reports, the risk of induction failure was not significantly higher in the ETP group (n= 2 vs 1 among non-ETP cases).
With a median follow-up of 4 years 10 months (range 1 year 4 months - 9 years), the ETP sub-group have a non-significantly inferior EFS (76.0% (95% CI: 64.0-88.0%) vs 84.6% (79.3-89.9%) at 5 years, p = 0.2) and OS (84.1% (73.9-94.3%) vs 90.9% (86.6-95.2%), p = 0.08) compared to the non-ETP sub-group, due primarily to a non-significantly higher relapse rate (17.7% (6.5-28.9%) vs 9.6% (5.1-14.1%), p = 0.1). Although MRD risk group was a determinant of outcome in the non-ETP group (p=0.05), its significance within the ETP group could not be reliably determined since only 2 patients were MRD low risk, of whom 1 suffered a relapse. Interestingly, of the 8 relapses in the ETP group, 7 were isolated marrow relapses and there was one isolated testicular relapse, whereas 12 of the 17 relapses in the non-ETP group involved the CNS (5 isolated and 7 combined).
Although ETP ALL is associated with an adverse outcome compared with non-ETP T-ALL, a 5 year EFS of 76% on a contemporary MRD stratified protocol is not sufficiently poor to warrant experimental treatment or first remission allogeneic transplant for the group universally.

Disclosures:
No relevant conflicts of interest to declare.
In Vivo Analysis of PML-RARA in a Humanized Mouse Model
Authors: Matsushita Hiromichi , Yahata Takashi , Sheng Yin , Nakamura Yoshihiko , Muguruma Yukari , Matsuzawa Hideyuki , Tanaka Masayuki , Hayashi Hideki , Sato Tadayuki , Damdinsuren Anar , Onizuka Makoto , Ito Mamoru , Miyachi Hayato , Pandolfi Pier Paolo , Ando Kiyoshi 
Published: October 2019

Abstract
Acute promyelocytic leukemia (APL) is a subtype of acute myeloid leukemia (AML) characterized by the formation of a PML-RARa fusion protein, which leads to the accumulation of abnormal promyelocytes. Xenograft mouse models with human leukemic cells have advantages for analyzing the human leukemias in vivo, especially for genetic analyses. However, human primary APL cells are difficult to engraft even in very severely immunodeficient mice, such as NOD/shi-SCID IL2Rg-/- (NOG) mice. In order to understand the mechanisms involved in human APL leukemogenesis, we established a humanized in vivo APL model using the transplantation of PML-RARA-transduced CD34+ cells from human cord blood into NOG mice. The expression of PML-RARa in the CD34+ cells disrupted the nuclear bodies in vitro. The clonogenic assay showed that PML-RARa inhibited the total colony formation, but favored the growth of myeloid colonies. When CD34+ cells with PML-RARA were transplanted, they proliferated in the NOG mice for more than three to four months after transplantation (in 24 out of the 34 mice). All 16 mice with more than 3,000 PML-RARA-transduced CD34+ cells were engrafted, while the engraftment was only detected in eight out of 18 mice when the cell density used for transplantation was less than 3,000 cells. These cells possessed abundant azurophilic abnormal granules in the cytoplasm, and some of them had bundles of Auer rods. They expressed CD13, CD33 and CD117, but not HLA-DR or CD34. In addition, the gene expression analysis revealed that these cells and human primary APL were clustered together among various types of AML, suggesting that these induced APL cells well recapitulated human primary APL. Similar to human primary APL, the induced APL cells possessed the ability for myeloid differentiation after treatment with all-trans retinoic acid in vitro and in vivo, and a very low potential for re-transplantation, which was similarly observed in both unsorted induced APL cells and the CD34- fraction. When human cord blood was fractionated before the PML-RARA transduction, the CD34+/CD38+ cells and common myeloid progenitors (CMP) in the CD34+/CD38+ cells led to the efficient development of APL in vivo. These findings demonstrate that CMP is a target for PML-RARA in APL, whereas the resultant CD34- APL cells may share the ability to maintain the leukemia.

Disclosures
No relevant conflicts of interest to declare.
CS-1 Re-Directed Central Memory T Cell Therapy for Multiple Myeloma
Authors: Wang Xiuli , Wong ChingLam W , Urak Ryan , Chang Wen-Chung , Budde Elizabeth E. , Brown Christine , Forman Stephen J. 
Published: October 2019

Abstract
Multiple myeloma (MM), a plasma cell malignancy, accounts for approximately 1 percent of all cancers and slightly more than 10 percent of hematologic malignancies in the United States. Approximately 20,000 new cases will be diagnosed this year and over 11,000 people will die from this disease. Current therapies for MM often induce remission, but nearly all patients eventually relapse and die. T-cell mediated anti-tumor therapies using genetically modify T cells with specific chimeric antigen receptors (CARs) have non-overlapping activity, toxicity and tumor resistance profiles compared to conventional chemotherapeutic agents.
The main challenge in designing a CAR T cell immunotherapeutic approach is identifying the best antigen for tumor targeting. CS-1 is a cell surface glycoprotein of the signaling lymphocyte activation molecule (SLAM) receptor family that is highly and selectively expressed on normal plasma cells and MM cells, with lower expression on NK cells and little or no expression on normal tissues. This unique expression pattern and proven clinical benefit of CS-1 monoclonal antibody for the treatment of relapsed MM makes CS-1 a good target for CAR T cell therapy.
Central memory T cells (TCM) from PBMC were isolated using a two-step process on the AutoMACS device to first deplete CD14+, CD45RA+ and CD25+ cells, then to positively select CD62L+ cells. These TCM undergo anti-CD3/CD28 bead stimulation and transduction with a lentiviral vector encoding CS-1 CAR containing a CD28 co-stimulatory domain and two mutations on IgG4 linker CH-2 portion to ensure enhanced potency and persistence after adoptive transfer. Gene modified CS-1 CAR T cells specifically lysed MM.1S, a MM cell line, in 4-hour 51Cr release assays and all the CAR+ cells expressed 107a upon co-cultured with the MM.1S cells. To investigate the potency of the CS-1CAR T cells, 2x106 MM.1S cells that were engineered to express GFP firefly luciferase were inoculated into NSG mice by tibia injection. 7 days post tumor engraftment, 1x106 CS-1 CAR T cells were intravenously injected into the tumor bearing mice. In contrast to untreated and mock cell treated mice in which tumor progressed rapidly systemically, single intravenous infusion of CS-1 CAR T cells induced dramatic tumor regression and significantly prolonged survival.
In addition to CS-1, CD44v6 and BCMA are antigens that have also been shown to be over-expressed on MM tumor cells. We therefore compared the two CARs with CS-1 CAR for their anti-MM activity. Based on our studies, targeting CS-1 resulted in the best efficacy (Figure 1) and would be an attractive strategy for development of a clinical trial.

Disclosures
No relevant conflicts of interest to declare.
Early and Transient Microchimerism Associated with Complete Remission after Adoptively Transferred Haploidentical NK Cells Against High Risk Myelodysplastic Syndrome and Refractory Acute Myeloid Leukemia
Authors: Björklund Andreas T , Carlsten Mattias , Schaffer Marie , Liu Lisa , Cooley Sarah A. , Miller Jeffrey S. , Watz Emma , Palma Marzia , Hansson Lotta , Wahlin Björn E , Möllgård Lars , Hagglund Hans , Blomberg Pontus , Ljungman Per T. , Hellström-Lindberg Eva , Ljunggren Hans-Gustaf , Malmberg Karl-Johan 
Published: October 2019

Abstract
Introduction: We here report from an ongoing phase I/II study of HLA-haploidentical NK cell therapy to patients with high-risk myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML) not eligible for standard therapies. The preparative regimen consisted of intermediate doses of Cyklophosphamide (Cy), Fludarabin (Flu) and titrated doses of total lymphoid irradiation (TLI). The trial design excluded systemic IL-2 treatment to avoid expansion of regulatory T cells and to test if in vivo expansion could be obtained without IL-2 support.
Patients:The first 12 patients were treated with Cy/Flu and an escalating dose of TLI (2 Gy and 4 Gy), followed by infusion of short-term IL-2 activated (16 hours) NK cells. Three patients received daily cyclosporine A after the conditioning. Three had relapsed, chemotherapy-refractory, primary AML, seven had secondary relapsed or refractory MDS-AML and two had high risk MDS with fibrosis.
Results: The treatment was well tolerated and no severe non-infectious toxicity could be observed in the patients. The endpoint of expansion (>100 donor NK cells/ul at day 14) was not reached, but six patients had positive microchimerism, NK cells of donor origin detectable by RT-PCR at day 7-14, that thereafter became undetectable within 7-14 days. Four of these six patients achieved complete remission (CR) whereafter they become eligible for and could proceed to allogeneic stem cell transplantation.
None of the patients with negative microchimerism obtained CR. Four patients died from progressive disease and three patients, with minor response and progressive disease, died in infections within three months of therapy.
Discussion: Although the long-term efficacy needs to be evaluated, the results suggest that a combined regimen with mild conditioning followed by NK cell therapy may induce remission in patients with chemo-refractory disease and provide a bridge to allogeneic stem cell transplantation. Notably, clinical responses were observed after only a minimal in vivo NK cell expansion and were independent on KIR-ligand mismatch.

Disclosures
Blomberg: VECURA: Employment. Hellström-Lindberg:Celgene: Research Funding.
Missense SNP rs3731249 Explains the CDKN2A Association with Childhood ALL and Shows Risk Allele Selection in Tumors with Somatic CDKN2A Alterations
Authors: de Smith Adam J. , Walsh Kyle M. , Hansen Helen M. , Barcellos Lisa F. , Chokkalingam Anand P. , Jenkins Robert B. , Wrensch Margaret R. , Wiencke John K. , Metayer Catherine , Wiemels Joseph L. 
Published: October 2019

Abstract
Little is known about the interaction between constitutive and somatic genetic variation in carcinogenesis. Genome-wide association studies (GWAS) of childhood acute lymphoblastic leukemia (ALL) have identified 6 genetic associations, including the single nucleotide polymorphism (SNP) rs3731217, located between the CDKN2A and CDKN2B genes. Somatic deletions of this chromosome 9p21.3 region are common in cancer, and occur in ~30% of childhood ALL.
We replicated the rs3731217 association with childhood ALL in Hispanics (P = 0.021) via a GWAS of 323 childhood ALL cases and 454 controls, using the Illumina OmniExpress SNP array. SNP imputation analysis across chromosome 9p21.3 revealed a stronger association at SNP rs3731249 (P = 6.4x10-4; OR = 2.77), a low-frequency missense variant in exon 2 (p.Ala148Thr) of CDKN2A (MAFcontrols = 2.1%). These associations were replicated in an additional 378 ALL cases and 536 controls using Taqman assays, where the missense variant again had a greater magnitude of effect (Prs3731249 = 6.5x10-3; OR rs3731249 = 2.00 versus Prs3731217 = 0.029; OR rs3731217= 1.38). We assessed the effect of rs3731249 on risk of other cancers with known chr9p21.3 GWAS associations, but observed no association between this variant and either adult glioblastoma or melanoma risk, suggesting an ALL-specific role for rs3731249.
We hypothesized that cases heterozygous for rs3731249 may show allelic imbalance, with preferential loss of the wildtype (WT) allele when hemizygous chr9p21.3 deletions occur in leukemic cells. To test this, we developed a novel method termed “SMART-ddPCR” (Somatic Mutation Allelic Ratio Test using Droplet Digital PCR). Concentration of risk and WT alleles was measured by ddPCR using the rs3731249 Taqman assay in diagnostic bone marrow (i.e. tumor) DNA from 35 known heterozygotes. Allelic imbalance due to somatic CDKN2A alterations was identified in 17 cases, of which 14 had a higher risk:WT allele ratio (P = 0.006, 1-sided binomial significance test), thus demonstrating preferential loss of WT rs3731249 and retention of the missense allele. Subclonal heterogeneity in CDKN2A copy number was also evident, suggesting that CDKN2A alterations are secondary events in leukemogenesis.
Preferential allelic imbalance towards the rs3731249 risk allele in ALL tumor DNA provides strong evidence of an important role for this missense variant in leukemogenesis. We show, for the first time, a direct relationship between constitutive and somatic genetic variants underlying ALL development, suggesting that such constitutive variants can provide an additional substrate for selection during tumor evolution.

Disclosures
No relevant conflicts of interest to declare.
Assessing the Value of Echocardiograms for Patients with Acute Myeloid Leukemia (AML) Receiving Anthracyclines
Authors: Seastone David J. , Nazha Aziz , Pophali Priyanka A , Ornstein Moshe C. , Carraway Hetty E. , Advani Anjali S. , Gerds Aaron T. , Kalaycio Matt E. , Maciejewski Jaroslaw P. , Sekeres Mikkael A. 
Published: October 2019

Abstract
Background
AML is a life-threatening disease that requires prompt administration of anthracycline-based induction chemotherapy, particularly in younger adults. Prior to treatment initiation, patients (pts) undergo routine echocardiography (echo) screening to assess cardiac ejection fraction (EF) as a measure of induction tolerability, despite American Society of Clinical Oncology and National Comprehensive Cancer Center guidelines which state that only pts with prior cardiac disease or exposure to cardiotoxic drugs or radiation therapy require such evaluation This is costly to the health-care system and may be unnecessary. In this study, we assess whether the use of echo prior to anthracycline-based therapy in AML has a major effect on pt outcomes.
Methods
We reviewed clinical data on AML pts treated with anthracycline-based induction chemotherapy regimens at our institution from 2002-2014. EF was obtained by 2D echo prior to induction chemotherapy initiation. Charlson comorbidity index was used to evaluate comorbidities prior to therapy. A cox proportional hazard model was used to analyze the impact of baseline echo on overall survival, with p-values <.05 denoting significance.
Results:
Of the 120 pts included, 60 (50%) were female. At diagnosis, median age was 57 years (range, 23-85), white blood cell count (WBC) 8.9 x 109/L (0.3-227), hemoglobin 9.1g/dL (4.7-13.8), platelets 85 x 103 (8-277), bone marrow blasts 48% (14-95%). Cytogenetic risk categories per CALGB 8461 criteria: 12 pts (10%) were favorable, 63 (53%) indeterminate, and 41 (34%) unfavorable. Charlson morbidity index was > 1 in 23 pts (19%) and 50 (38%) were current or former smokers. All pts received induction chemotherapy with either idarubicin (38%), daunorubicin (38%), or mitoxantrone (23%) combined with cytarabine (7+3). In 116 pts (97%), the baseline echo showed a left ventricular ejection fraction (LVEF) >50% (+/- 5). In the 4 remaining pts (3%), the EF was < 40%. In a multivariable analysis controlling for age, WBC, cytogenetic risk group, Charlson comorbidity index, and smoking exposure, LVEF was not associated with overall survival (HR 1.05 with 95% CI 0.99-1.12 and p=.09). Among the 4 pts with EF<40% (age 57-71 years), all received full dose anthracycline and only the 71-year-old patient experienced congestive heart failure during treatment. Six pts (5%) died during induction therapy (4 weeks) and none of these had an EF <40%.
Conclusion
Among AML pts who received anthracycline-based induction chemotherapy, results from echocardiography did not impact outcome in >99% in this cohort. It is not clear that echocardiography, particularly in younger pts, provides value, and may delay treatment initiation.

Disclosures
No relevant conflicts of interest to declare.
Left Ventricular Ejection Fraction Measurement and Doxorubicin-Based Chemotherapy in Patients with Diffuse Large B-Cell Lymphoma
Authors: Enns Deborah L , Aboulafia David M 
Published: October 2019

Abstract
Purpose:
Doxorubicin-based chemotherapy (DOX) is commonly administered to patients with diffuse large B-cell lymphoma (DLBCL). Prior to chemotherapy, left ventricular ejection fraction (LVEF) is routinely measured to assess left ventricular dysfunction. While LVEF screening is recommended by many national regulatory bodies, evidence supporting the usefulness of LVEF measurement prior to administering DOX is lacking. Our goal was to perform a retrospective analysis of patients with DLBCL to establish (1) how often LVEF was measured prior to administering DOX, and (2) whether the chemotherapy regimen was modified based on LVEF values. As cumulative doses of doxorubicin greater than 400 mg/m2 have been associated with an increased risk of congestive heart failure (CHF), we also determined the incidence of CHF in patients with DLBCL who did receive DOX.
Patients and Methods:
We identified 268 patients diagnosed with DLBCL at Virginia Mason Medical Center between 2001 and 2012 and collected the following data: age at diagnosis; stage of lymphoma; type of chemotherapy given; cumulative doxorubicin dose (mg/m2); LVEF status; and incidence of CHF or cardiac disease. We also compared the number of CHF risk factors between patients who did and did not have LVEF measured. Statistical analyses included a Fischer’s exact or Chi-squared test to compare study groups as well as the number of CHF risk factors. The level of significance was set at a P value of < 0.05.
Results:
LVEF was measured in 238 patients (89%) prior to initiation of chemotherapy. LVEF values were normal in 225 patients (95%) and low (< 50%) in 13 patients (5%). Of the patients with normal LVEF, 193 received DOX (86%), and of these patients, 14 developed CHF post-treatment (7%). For the 13 patients with low LVEF, 8 received DOX (62%) and 1 developed post-treatment CHF (13%). The remaining thirty patients did not have LVEF measured and none received DOX. Of the 268 patients studied, 176 are alive (66%) and 3 were lost to follow-up. The mean follow-up time was 43 months (range 3 d to 12.1 y). The mean number of CHF risk factors did not differ between patients who did and did not have LVEF measured (1.70 vs. 1.65, P = 0.87).
Conclusion:
Our results suggest that the decision to administer DOX was not directly affected by LVEF values. These findings challenge the existing policy of routinely screening patients with DLBCL with echocardiograms or MUGA scans prior to treatment.

Disclosures
No relevant conflicts of interest to declare.
Complement Component 3 (C3) Is a Mediator of Epithelial-Mesenchymal Transition (EMT)
Authors: Cho Min Soon , Hu Qianghua , Rupaimoole Rajesha , Sood Anil , Afshar-Kharghan Vahid 
Published: October 2019

Abstract
We have shown that complement component 3 (C3) is expressed in malignant ovarian epithelial cells and enhances cell proliferation in vitro and tumor growth in vivo. C3 is secreted by cancer cells into the tumor microenvironment and promotes tumor growth through an autocrine loop. To understand the mechanism of upregulation of C3 expression in malignant epithelial cells, we studied the transcriptional regulation of C3, and found that TWIST1, a major regulator of EMT, binds to the C3 promoter and regulates C3 transcription. Knockdown of the TWIST1 gene reduced C3 mRNA, and TWIST1 overexpression increased C3 mRNA.
TWIST1 promotes epithelial-mesenchymal transition (EMT) during normal development and in metastasis of malignant tumors. An important marker of EMT is a reduction in the surface expression of E-cadherin on cells facilitating migration and invasion of these cells. TWIST1 is a transcriptional repressor of E-cadherin; and because TWIST1 increases C3 expression, we investigated whether C3 is also a negative regulator of E-cadherin expression. We overexpressed C3 in ovarian cancer cells by stable transduction of lentivirus carrying C3 cDNA. Overexpression of C3 was associated with 32% reduction in the expression of E-cadherin resulting in enhanced migration ability of cells by 2.3 folds and invasiveness by 1.75 folds, as compared to control cells transduced with control lentivirus.
To investigate whether TWIST1-induced reduction in E-cadherin is C3-mediated or not, we studied the effect of TWIST1 overexpression simultaneous with C3 knockdown in ovarian cancer cells. Overexpression of TWIST1 alone resulted in 70% reduction in E-cadherin mRNA and this was completely reversed after simultaneous C3 knockdown in these cells.
To investigate the correlation between C3 and TWIST1 in vivo, we studied the co-expression of these two proteins in mouse embryos (physiologic EMT) and in malignant tumors (pathologic EMT). Given the role of EMT in embryogenesis we immunostained mouse embryos at different stages of development, using antibodies against TWIST1 or C3. Transverse section of 9.5-day post-coitum (9.5dpc) mouse embryos showed co-expression of TWIST1 and C3 in otocyst (ot) and hindbrain (hb) of neural crest. In the whole-mounted 11.5dpc mouse embryos, C3 and TWIST1 were co-expressed in limb buds.
Given the role of EMT in malignancy, tumors induced in mice after intraperitoneal injection of murine ovarian cancer cells were resected and immunostained for C3 and TWIST1 proteins. TWIST1 and C3 co-localized at tumor edges, where EMT and tumor cells migration occur. Taken together, these data provide evidence that TWIST1 regulates C3 expression, and C3 promotes EMT through E-cadherin.

Disclosures
No relevant conflicts of interest to declare.