bluuebunny/crossref_metadata_2025 huggingface dataset:
Propensity Score Matched Analysis of Risk for Malignancies in Sickle Cell Anemia
Authors: Afzal Amber , Siddiqui Nauman , Godara Amandeep , Khan Zarmina , Tuncer Hande H.
Published: October 2019

BACKGROUND:

The survival of sickle cell patients is limited as compared to the general population. The major morbidity and mortality comes from sickle cell complications including cardiovascular disease, renal failure, infections and thrombosis (Manci Br J Haematol 2013). However, literature is conflicting about risk for malignancies in this population and remains to be investigated by larger studies. Our retrospective study is aimed at studying the prevalence of different malignancies in a large cohort of hospitalized sickle cell patients in the year 2014 using the National Inpatient Sample (NIS).

METHODS:

Study Population:

We identified hospitalized sickle cell patients in NIS using ICD-9 codes (282.5,282.6X -282.6X) for year 2014. We also identified the various malignancies through ICD-9 codes and studied the prevalence of individual malignancies in hospitalized sickle cell patients (SCD/cases) compared to a matched cohort of hospitalized patients with no sickle cell disease (No SCD/controls).

Statistical Analysis:

The 1:N Case-Control Matching Macro (Parsons LS et al SUGI 29) was used to match cases with controls with respect to potential confounders including age, race, gender in the ratio of 1:4. Chi Square was used to determine the difference in the proportions of individual malignancies between the two groups. P value <0.05 was considered statistically significant. All analyses were performed using SAS 9.4.

RESULTS:

We identified 117,405 hospitalized sickle cell patients in the NIS database who were matched to 469,620 controls (Table 1). A total of 1890 malignancies were found in sickle cell cohort. The prevalence of malignancies in hospitalized sickle cell patients was 1.6% as compared to 4.6% in controls (OR 0.34; range:0.30-0.38). The prevalence of all studied malignancies including hematological malignancies was lower in hospitalized sickle cell patients as compared to a matched population of hospitalized patients with no sickle cell disease (Table 2).

CONCLUSION:

Our study showed a lower prevalence of both solid tumors and hematological malignancies in patients with sickle cell disease as compared to matched cohorts. Future prospective cohort studies are needed to evaluate the risk of malignancies in sickle cell anemia and identify if any protective mechanisms exist against the development of malignancy in this patient population.

Disclosures

No relevant conflicts of interest to declare.
Preclinical Development of CTX120, an Allogeneic CAR-T Cell Targeting Bcma
Authors: Dar Henia , Henderson Daniel , Padalia Zinkal , Porras Ashley , Mu Dakai , Kyungah Maeng , Police Seshidhar , Kalaitzidis Demetrios , Terrett Jonathan , Sagert Jason
Published: October 2019

Autologous CAR-T cells targeting BCMA have induced robust and durable responses in patients with relapsed/refractory multiple myeloma. However, autologous cell therapies face several challenges which will likely limit the number of patients that will have access to these therapies. These limitations include manufacturing failure rates, wait time and supply constraints in addition to other factors such as reimbursement. Allogeneic CAR-T cells can potentially overcome these access challenges, and may have several other advantages over autologous therapies. Allogeneic CAR-T cells are derived from robust healthy donor T cells through a batch manufacturing process, which may result in a highly consistent product with greater potency and enable better safety management. Here we show further development and preclinical data for CTX120, an allogeneic "off the shelf" CAR-T cell targeting BCMA. CTX120 is produced using the CRISPR/Cas9 system to eliminate TCR and MHC class I, coupled with specific insertion of the CAR at the TRAC locus. CTX120 shows consistent and high percent CAR expression from this controlled insertion and exhibits target-specific cytotoxicity and cytokine secretion in response to BCMA positive cell lines. CTX120 CAR-T cells retain their cytotoxic capacity over multiple in vitro re-challenges, demonstrating durable potency and lack of exhaustion. In mouse models of multiple myeloma, CTX120 showed typical CAR-T persistence and eliminated tumors completely, resulting in long-term survival as compared to untreated animals. These data support the ongoing development of CTX120 for treatment of patients with multiple myeloma and further demonstrate the potential for our CRISPR/Cas9 engineered allogeneic CAR-T platform to generate potent CAR-T cells targeting different tumor antigens.

Disclosures

Dar: CRISPR Therapeutics: Employment, Equity Ownership. Henderson:CRISPR Therapeutics: Employment, Equity Ownership. Padalia:CRISPR Therapeutics: Employment, Equity Ownership. Porras:CRISPR Therapeutics: Employment, Equity Ownership. Mu:CRISPR Therapeutics: Employment, Equity Ownership. Kyungah:CRISPR Therapeutics: Employment, Equity Ownership. Police:CRISPR Therapeutics: Employment, Equity Ownership. Kalaitzidis:CRISPR Therapeutics: Employment, Equity Ownership. Terrett:CRISPR Therapeutics: Employment, Equity Ownership. Sagert:CRISPR Therapeutics: Employment, Equity Ownership.
Role of Serial Positron Emission Tomography (PET) Scans in Indolent Lymphomas' Management - Interim Results from an Ongoing Prospective Study at King Fahd Medical City, Riyadh, Saudi Arabia
Authors: Alshehry Nawal Faiez , Ahmed Musab , Iq bal Shahid , Gill Atta Munawar , Dawiama Mohammad E , Butt Ahmed Ali , Albtoosh Belal , Motabi Ibraheem H. , AlGhamdi Mubarak S. , Tailor Imran Khan , Altaf Syed , Zaidi Syed Ziauddin A. , Raja Shanker
Published: October 2019

Background:

Indolent lymphomas (ind-LYM) are a subset of B-cell lymphomas, characterized by slow growth, protracted course without treatment and a tendency to reoccur after therapy. The management of ind-LYM is varied and evolving. To-date reliable tools for imaging based prognostication and treatment strategies are limited. We explored the utility of serial FDG PET scans (baseline (BL), interim/6 months (INT) end of therapy/1 year (EOT)), in managing ind-LYM).

Methods:

From an ongoing prospective study, examining the role of PET scan in ind-LYM at our institution, we performed an interim analysis of all patients hitherto enrolled in our study. Patients diagnosed as ind-LYM and having undergone serial PET scans from 2015 to date (N=40, males= 27, females =13) had median age of 59.68 ± 14.5 (28-82 at diagnosis. All PET scans were obtained per accepted protocols. SUVmax and Deauville scores (DS) were obtained from five target lesions. The average of composite SUV (cSUV) and composite Deauville scores (cDS) were computed for each patient. Statistical analyses (using t-test, mean delta change) were performed with the cSUV and cDS.

Results:

The types of ind-LYM were CLL/SLL (17), Follicular (15), mantle cell (3), marginal zone lymphoma (3), and 1 each of MALT and lymphoplasmacytic lymphoma. The data were analyzed for two arms of the study treatment chemotherapy arm (CHEMO-22/40) and Rituximab + observation arm (OBSR-18/40). At BL, patients on CHEMO had significantly higher cSUV and cDS compared to those who were on OBSR (SUV: 6.99 vs. 4.32 p-value 0.0124; cDS: 4.37 vs 3.71 p-value 0.0075 respectively). Although there was a delta change in the cSUV and cDS in both arms after INT and EOT, both treatment lines showed no significant difference in the SUV mean and DS (p-value: 0.754 and 0.5721, receptively). A 2-tailed T-test was used for delta change of cSUV. BL to INT cSUV was statistically significant (p-valve 0.05) and difference in cSUV was not significant for BL to EOT (p-valve 0.98). Difference in cDS was not significant between BL, INT and EOT.

Conclusion:

The higher cSUV/cDS at BL for the treatment arm may be due to disease profile at the time of presentation and/or selection bias. At EOT no difference in both estimates were noted. Both arms showed similar trends in delta change from BL-INT-EOT. Our findings suggest that SUV and DS at BL maybe an independent criterion for treatment selection in ind-LYM.

Disclosures

No relevant conflicts of interest to declare.
Multi-Dimensional Humanistic Care for the Kids with Hematological Malignancies in Guangzhou Women and Children's Medical Center (GWCMC)
Authors: Huang Haiying , Liang Zhimin , Huang Yongxian , Zhang Hui
Published: October 2019

Objective: To evaluate the effects of multi-dimensional humanistic care on kids with hematological malignancies and their parents and facilitate the effective nursing experience and professional nursing in such clinical care.

Methods: A total of 567 children with cancer and their parents were enrolled in this study. From January to December in 2017, multi-dimensional humanistic care was systematically implemented to provide care for all these patients and their family members. A warm and comfortable in-patient atmosphere was created with regular playing-games, health education meetings, and personal palliative care support were provided to all newly-diagnosed pediatric cancer patients and their parents who admitted to our department. In the meanwhile, case management were implemented by nurses. Children and parents in demand were referred to game therapists and social workers for visits, psychological counseling and emotional support. Financial assistance was also provided to care for those well-off families to support their systemic cancer treatment. Follow-up visits to hospitals were also offered.

Results: The implementation of multi-dimensional humanistic care measures significantly reduced the anxiety level of hospitalized children with cancer and their parents, which in turn improve their treatment compliance. Next, more easier caregiving ability and compliance were also recorded. The results of satisfaction survey showed that the satisfaction to nursing care rose from 85.6% in 2016 to 96.9% in 2017 after the implementation of multi-dimensional humanistic care in our hospital. The nurses' service attitude, medication guidance, hospitalization guidance, timely bedside services, and nurses' operating technology satisfaction also increased by 10.86%, 12.17%, 8.3%, 12.80%, and 12.15% respectively.

Conclusion: The multi-dimensional humanistic care effectively reduces the anxiety of kids with hematological malignancies and their parents, improves the caregiver's caring ability and compliance, and their satisfaction to nursing care. All these results point out the significance of systemic care services in the childhood cancer therapy, it is noteworthy to pour more attention and capacity to the clinics which may facilitate the cancer treatment.

Figure. Figure.

Disclosures

No relevant conflicts of interest to declare.
Cell Intrinsic and Extrinsic Mechanisms Mediate the Role of c-Myc Haploinsufficiency in Controlling HSC Fate
Authors: Sheng Yue , Yu Chunjie , Ma Rui , Qian Zhijian
Published: October 2019

It has been shown that loss of c-Myc leads to accumulation of hematopoietic stem cells (HSCs) and severe cytopenia as a consequence of a blockage of HSC differentiation. Here we report a role of c-Myc haploinsufficiency in regulating HSC quiescence and self-renewal. We showed that c-Myc haploinsufficient mice displayed decreased white blood count and number of lymphocytes with normal myeloid cell differentiation. The number of HSCs and hematopoietic progenitor cells (HPCs) were all decreased significantly in c-Myc haploinsufficient mice as compared with control mice. We found that c-Myc haploinsufficiency inhibited HSC self-renewal capacity, increased proliferation and decreased quiescence of HSCs in vivo. By transplantation assays, we showed that c-Myc haploinsufficiency has extrinsic and intrinsic effects on the maintenance of HSCs in vivo. Our study suggests that loss of c-Myc activity and reduced dosage of c-Myc have distinct effects on HSC functions.

c-Myc is a critical downstream mediator of the Wnt/b-catenin pathway. We showed that c-Myc haploinsufficiency is sufficient to prevent severe anemia in Apc heterozygous mice, and to significantly prolong the survival of Apc heterozygous mice. In addition, treatment of Apc haploinsufficient mice by a c-Myc inhibitor significantly reversed anemia in Apc-deficient mice.

By transplantation assay, we further demonstrated that reduced expression of c-Myc in the bone marrow niche is responsible for prevention of severe anemia in Apc-deficient mice. However, we found that reduction of c-Myc by loss of a single allele of c-Myc did not rescue defective self-renewal capacity of Apc haploinsufficient HSCs. Taken together, our studies indicate that c-Myc mediates the function of the Wnt/b-catenin signaling pathway in bone marrow niche but not in HSCs.

Disclosures

No relevant conflicts of interest to declare.
Survey of Compliance to Infection Control Measures and Its Impact on Healthcare of Patients in a Hematopoietic Stem Cell Transplant Unit
Authors: Yarlagadda Naveen , Krishnan Gayatri , Mohan Meera , Johnsrud Andrew , Wolfe Frankie , Kothari Atul
Published: October 2019

Background:

Isolation precautions are used in hospitals to interrupt the chain of transmission of nosocomial pathogens. There are several reports that isolation negatively impacts the care given to patients. Isolation protocol compliance has been reported between 40-70% in several studies.

Methods:

We conducted an anonymized survey of healthcare providers assessing the knowledge, practice and perceived impact of isolation precautions. The survey was conducted in a hematopietic stem cell transplant unit and a medical unit in an academic medical center. Surveys were given to nursing staff and resident physicians working on those units. Results were analyzed using chi-square test.

Results:

A total of 106 responses were received, 60 from nursing staff and 46 from resident doctors. Self- reported compliance to hand hygiene practices was 59.9%, this was significantly higher on leaving as compared to entering patient rooms amongst nurses (72% vs 50%, p=0.015). Overall adherence to isolation precautions was 60.3%, surprisingly nursing staff in the transplant unit had a significantly lower rate as compared to nurses in the medicine unit (44% vs 83%, p=0.025). 70.7% of respondents felt that patients on isolation precautions have to wait longer for non-emergent care. Only 56.6% of respondents felt that patients were educated about the reasons for isolation precautions; this number was significantly lower in residents as compared to nurses (41.3% vs 68.3%, p=0.005). 77.3% felt that patients in isolation are less satisfied in the care they received. 86.9% of resident physicians felt that isolation precautions helped reduce the spread of infection vs 68.5% of nurses (p=0.025). 82% of respondents expressed overall satisfaction with infection control practices.

Conclusions:

In our survey, we observed a low compliance of hand hygiene and isolation practices. While respondents felt that isolation adversely impacted patient satisfaction and care, they also opined that these practices help reduce the spread of infection. Surveys can be an important tool to assess knowledge and impact of infection control practices; and can be used to direct quality improvement initiatives.

Disclosures

No relevant conflicts of interest to declare.
Genomic and Immune Profiles of Multiple Myeloma Revealed By Whole Genome and Transcriptome Sequencing
Authors: Yoon Christopher J. , Song Hyojin , Song Seulki , Park Hyejoo , Shin Dong-Yeop , Hong Junshik , Kim Inho , Lee Hyun Jung , Kim Byung-Su , Lee Jung Lim , Yoon Sung-Soo , Ju Young Seok , Koh Youngil
Published: October 2019

Introduction:

We analyzed genomic profile in line with immune checkpoint molecule expression in multiple myeloma using whole genome sequencing (WGS) and whole transcriptome sequencing (WTS) data.

Methods:

We performed WGS of CD138+ selected tumor and the matched normal from saliva of multiple myeloma patients at 90-60x and 30x respectively from 37 patients. At the same time, we performed WTS using both tumor (CD138+) and immune milieu (CD138-) samples from these patients. Structural variations were called with Delly2 and Manta and the intersect of two call sets were used as the final SV calls. Copy number analysis was done with Sequenza and custom scripts. Somatic single nucleotide variants were identified with Mutect1 and Strelka and filtered against the panel of normal. RNA-seq was aligned using STAR algorithm and gene expression was quantified using RSEM algorithm. We extracted the expression of 21 immune checkpoint-related molecules (LAG3, PD-L1, IDO1, TIM3, CD27, CTLA-4, ICOS, TIGIT, PD-1, PD-L2, VISTA, and their ligands). To compare cell populations within a set of CD138-sorted MM samples, we used CIBERSORT algorithm to estimate predict fractions of leukocyte RNA.

Results & Discussion:

A total of 1,359 somatic structural variants were identified, with a median of 28 structural variant events per patient. The most common translocation event was t(4;14) (n=6, 16.2%). We identified novel recurrent deletions in CCSER1 (n=5, 13.5%) either deleting exon 10 (n=3) or exon 6 (n=2). CCSER1 has been proposed to a have a role in cell division with defects leading to cytokinetic defects. We observed an enrichment of whole-genome duplicated samples with CCSER1 deleted patients. We propose that CCSER1 deletion may have a role in whole-genome duplication events seen in multiple myeloma. Chromothripsis was also seen in 7 patients (18.9%). A total of 9 chromothripsis events were observed and chromosome 6 was most frequently involved (n=3). Recurrent mutation in IGKV4-1 chr2: 89185484 and 89185485 (p.Ala60) was observed suggesting dysfunctional antigen recognition in myeloma cells.

From immune checkpoint perspective, expression of Galectin-9, a ligand of TIM3, was outstandingly high in immune milieu suggesting potential role of TIM3 inhibitor in myeloma. On the other hand, VISTA was highly expressed in tumors. When we matched the transcriptomic profiles with genomic variants, NRAS G61R mutant showed high expression of TIM3/Galectin-9 axis suggesting characteristics immune milieu of RAS mutant myeloma.

Conclusions:

We could identify novel CCSER1 recurrent deletion and IGKV4-1 mutation in multiple myeloma using WGS. WTS immune milieu analysis suggest potential role of TIM3 inhibitor in myeloma, especially in RAS mutated cases.

Figure. Figure.

Disclosures

No relevant conflicts of interest to declare.
Lysine Histone-Induced Thrombocytopenia in Zebrafish Depends on Endothelial Von Willebrand Factor
Authors: Zheng Liang , Zheng X. Long
Published: October 2019

Thrombotic thrombocytopenic purpura (TTP), a potentially fatal blood disorder, is caused by severe deficiency of plasma ADAMTS13 activity, primarily resulting from acquired autoantibodies that inhibit ADAMTS13's ability to cleave von Willebrand factor (VWF). However, severe deficiency of plasma ADAMTS13 activity alone is often not sufficient to cause an acute episode of TTP; additional environmental or genetic factors may be required for the development of acute TTP. Here, we report a creation and characterization of a novel zebrafish model of TTP and use the model to identify a potential mechanistic link between an innate immunity and onset of TTP. Using CRISPR/cas9, we generated adamts13-/-,vwf-/-, and adamts13-/-vwf-/-zebrafish on a double transgenic (Fli1-eGFP/Gata1-dsRed) background in which erythrocytes, thrombocytes, and endothelial cells were uniquely labeled with an endogenously expressed fluorescent protein. Our results demonstrated that adamts13-/-zebrafish larvae (5-dpf) had an increased rate of developing occlusive thrombi in the caudal venues after FeCl3 injury; also, adamts13-/- adult zebrafish (3-4 months) exhibited ~27% reduction of their total or mature thrombocyte counts with a significantly increased number of fragmented erythrocytes; moreover, an intraperitoneal administration of a lysine-rich histone resulted in severe and more persistent thrombocytopenia with an increased mortality in adamts13-/-zebrafish than their wild-type littermate; finally, both spontaneous and histone-induced thrombocytopenia and thrombotic microangiopathy in adamts13-/-was eliminated when endothelial vwf was genetically deleted from zebrafish. Together, these results provide experimental evidences to support a mechanistic link of acute infection or inflammation to acute onset of TTP in patients lacking plasma ADAMTS13 activity.

Disclosures

Zheng: Alexion: Research Funding, Speakers Bureau.
Clinical Significance of Germ Line and Acquired Somatic Mutations of 30 High Frequency Genes in Hematologic Malignancies
Authors: Zhang Nanchen , Gong Yuping
Published: October 2019

Objective: To analyse germ line and acquired somatic mutations of 30 high frequency genes and explore their clinical significance in hematologic malignancies. Methods: A total of 254 patients with hematologic diseases (145 males and 109 females) from Jan 2017 to Apr 2018 in the Southwest China were included and the samples from bone marrow and oral mucosa were collected, and the gene mutations of germ line and acquired somatic cells were examined by the next-generation sequencing technology or PCR. The frequency and locus of germ line and acquired somatic mutations of 30 high frequency genes, as well as their clinical significance to the diagnosis and classification in hematologic diseases were analysed. Results: 1.The frequency and locus of germ line and acquired somatic mutations, and their clinical significance: no mutation was detected in 24 cases (9.4%), and a total of 659 genetic locus mutations were detected in 230 cases (90.6%), including 520 somatic cell mutations (78.9%) and 139 germ line mutations (21.1%). The results of gene mutations are listed in order: the number of acquired somatic mutation: the number of germ line mutation: common mutation locus: the type of hematological disease for the patients diagnosed in the West China hospital, as described in Table 1. 2.Characteristics of acquired somatic and germ line mutations: the genes of WT1, U2AF1, TP53, TET2, STAG2, RUNX1, PIGA, NRAS, KIT, IDH2, EZH2, DNMT3A, CBL and BCOR were dominated by acquired somatic mutations, while the genes of ZRSR2, JAK2 and BCORL1 were dominated by germ line mutations, the genes of SRSF2, SF3B1, SETBP1, PTPN11, NPM1, PHF6, KRAS, IDH1, FLT3-ITD, ETV6 and CEBPA were detected only in acquired somatic mutations, BCOR and ASXL1 have a similar proportion of acquired somatic and germ line mutation; only 42-4C>A of ZRSR2 mutation was existed in both somatic and germ line, however, the mutation sites of the acquired somatic and germ line of other genes have not been found to be existed at the same time. 3.The clinical significance of the acquired somatic and germ line mutation: It was found that the genes of TP53, SF3B1, EZH2 and DNMT3A tend to appear in MDS, the genes of ZRSR2, WT1, U2AF1, TET2, RUNX1, NRAS, NPM1, KRAS, KIT, IDH1/2, FLT3-ITD, CEBPA and BCOR tend to appear in AML, and the similar probability of the STAG2, PTPN11, PHF6, ETV6, CBL, BCORL1, ASXL1 and NF1 genes appear in AML and MDS, while the genes of PIGA tend to appear in PNH, and BCOR in AA. Conclusion: The acquired somatic and germ line mutations are significantly different in the frequency, mutation site and the type of hematological disease, and the further research will clarify their potential roles in the pathogenesis of hematological tumors, and have important clinical significance for the classification and prognosis of the hematological diseases.

Key words: gene mutation, somatic, germ line, hematologic malignancies

Table 1. Table 1.

Disclosures

No relevant conflicts of interest to declare.
Donor KIR3DL1/Receptor HLA-Bw4-80I Combination Reduce Acute Leukemia Relapse after Umbilical Cord Blood Transplantation without in Vitro T-Cell Depletion
Authors: Fang Xinchen , Sun Zimin , Liu Huilan , Xia Lijun
Published: October 2019

Objective: How to reduce relapse is still a puzzle in the treatment of acute leukemia patients with allogeneic umbilical cord blood hematopoietic cell transplantation (UCBT). Donor natural killer (NK) cell alloreactivity in UCBT can control leukemic relapse, However, NK cell function is calibrated through education, that is governed by human leukocyte antigens (HLAs), our study aim to investigate the Graft-resistant leukemia effect after transplantation of KIR3DL1 and its different ligand HLA-B subtypes(BW6, Bw4-80T, Bw4-80I).

Design and method: we retrospectivly analysed single center of 232 acute leukemia patients who underwent UCBT(myeloablative conditioning without ATG). The killer immunoglobulin-like receptor (KIR) genotype of cord blood donors was detected by SSP method, they all contains KIR3DL1. There were 104 cases of acute lymphoblastic leukemia, 112 cases of acute myeloid leukemia, 16 cases of myelodysplastic syndrome. 168 cases of transplant patients in remission period, and 64 cases of non-remission. There were 122 males and 110 females with a median age of 13 years (2-45 years). The patients were divided into three groups according to HLA-B locus, 1.patients had BW6 in both two HLA-B locus(group 1, N=86) , which means uneducated; 2.patients had HLA-Bw4-80T in the HLA-B locus(group 2, N=68), which means educated; 3.patients had HLA- Bw4-80I in the HLA-B locus(group 3, N=77), which means highly educated. The receptors were followed-up from July 31, 2012 to December 30,2017, in graft failure, neutrophil engraftment, platelet engraftment, acute graft-versus-host disease (aGVHD), chronic graft-versus-host disease (cGVHD), relapse, overall survival (OS), and leukemia free survival (LFS).

Results: In group 3 patients, donor KIR3DL1 and receptor HLA-Bw4-80I combinations that were predictive of highly educated were associated with significantly lower relapse (hazard ratio [HR], HR = 0.49; P =0 .004) and better LFS (HR = 0.68, p = 0.014), compared with the educated(group2) and uneducated (group3) group, with no significant increase in aGVHD and cGVHD incidence. While no difference was observed in the other index in the three group.

Conclusion : Alloreactivity of NK cells demonstrated reproducible patterns of strong or weak graft-versus-leukemia (GVL) effect against leukemia blast, which is depend on donor KIR3DL1/receptor HLA-B subtype combinations. Our data suggests that donor KIR3DL1/receptor HLA-Bw4-80I combination in UCBT may achieve superior graft versus leukemia effects, lower risk for relapse among patients with acute leukemia.

Keywords: KIR, Umbilical Cord Blood Transplantation, UCBT, GVL

Disclosures

No relevant conflicts of interest to declare.
Prognostic Value of Epstein-Barr Virus (EBV)-Induced Gene 3 (EBI3) in Diffuse Large B Cell Lymphoma
Authors: Zheng Hong , Huang Juan
Published: October 2019

Background: Epstein-Barr virus (EBV)-induced gene 3 (EBI3) as a subunit for heterodimeric cytokines IL-27 and IL-35, plays important roles both in regulation of T cell proliferation and Th1,Th2 and Th17 cells differentiation. EBI3 was closely related with tumor prognosis. Diffuse large B-cell lymphoma (DLBCL) is the most common lymphoid malignancy in adults with cure-rates of 40-60%. However, Relapse and refractory rate remain in 40% or so. In the present study, we aimed to detect the expression of EBI3 in DLBCL and to analyze its relationship with prognosis.

Methods: We retrospective reviewed medical records of 51 newly diagnosed DLBCL patients in Sichuan People's Hospital between January 2010 and December 2016. Immunohistochemical (IHC) assay was used to detect the expression of EBI3 and PD-1 in DLBCL. The expression of CD5, CD30, Bcl-2, Bcl-6, C-myc and Epstein-Barr virus (EBV)-encoded RNAs (EBERs) in tumor specimens from 53 patients was also detected by IHC. The Kaplan-Meier method with log-rank test was used for univariate analysis. Cox proportional hazards model was used for multivariate analysis.

Results: Of the 51 patients, 40 (78.4%) were EBI3-positive in tumor specimens. And PD-1 expression (39/40) was almost in parallel with EBI3 in tumor specimens. EBI3 expression was common in the non-germinal center B-cell-like (GCB) subtype than in the GCB subtype. Patients with EBI3 expression in tumor were more likely to be resistant to first-line chemotherapy when compared with the patients without EBI3 expression in tumor microenvironment (P <0.05). EBI3 expression in tumor microenvironment was correlated with PD-1 expression (r = − 0.20, P = 0.04). Only one patient with EBI3 expression was no PD-1 expression. No correlations were detected between EBI3 expression and the expression of EBER as well as other markers: ALK, CD5, c-Myc and CD30. The complete remission (CR) rates were 7.5% (3/40) and 71.4% in patients with and without EBI3 expression in tumor cells (P = 0.02). EBI3 expression in tumor cells was an independent risk predictor for ORR (P < 0.05).

Conclusions: Our results indicate that EBI3 associated with poor prognosis and EBI3 may be a potential therapeutic target and prognosis marker.

Disclosures

No relevant conflicts of interest to declare.
Evaluation of Elderly Patients with Hematological Malignancies By Two Different Geriatric Risk Scoring Systems: G8 and Flemish Version of the Triage Risk Screening Tool (fTRST)
Authors: Atilla Erden , Yalciner Merih , Kilic Ozlem , Muftuoglu Nurhayat , Ataca Atilla Pinar , Civriz Bozdag Sinem , Toprak Selami Kocak , Kurt Yuksel Meltem , Özcan Muhit , Gurman Gunhan , Ilhan Osman
Published: October 2019

Background: High burden of patients with hematological malignancies are diagnosed >65 years old. Since then, geriatric interventions can improve quality of life, compliance to therapy and overall survival. Geriatric screening is the best tool to access patients' prognosis. In this study, our aim is to identify our geriatric patients with different geriatric scoring systems to evaluate their prognostic value for overall survival (OS).

Patients and Methods: We included 127 patients (age>65 years) with hematological malignancies and assessed two geriatric screening tools, G8 and Flemish version of the Triage Risk Screening Tool (fTRST) during a hospital visit at diagnosis or follow-up. G8 is considered normal if the score is >14, high risk patients in fTRST are defined as patients scored ≥ 2. OS was described using Kaplan-Meier plots, and comparisons were made using log-rank and Wilcoxon rank sum tests.

Results: From 127 patients, 50 patients (39%) were newly diagnosed. The patient characteristics were defined in table. Patients under follow-up received median of 2 lines of treatment (range, 1-5); 11 patients (14%) treated with radiotherapy, 15 patients (19%) underwent autologous stem cell transplantation. Abnormal G8 score was detected in 120 patients (94%). High fTRST score was present in 72 patients (57%). Both abnormal G8 score and high fTRST score were detected more likely in follow-up patients (65% vs 40%, p=0.005).There was no statistical difference in 5-year OS in terms of abnormal G8 score or high fTRST score alone. In patients with both abnormal G8 and high fTRST scores had statistically significant lower 5-year OS compared to normal G8 and/or low fTRST scores (57% vs 85%, p=0.048) (figure).

Conclusion: In investigation of two screening tools-G8 and fTRST, statistically significant survival advantage was shown if both scores were normal.

Disclosures

Civriz Bozdag: MSD: Research Funding; NOVARTIS: Consultancy; TAKEDA: Consultancy. Özcan:Jazz: Other: Travel support; Roche: Honoraria, Research Funding; Takeda: Honoraria, Other: Travel payment, Research Funding; Janssen: Other: Travel Support, Research Funding; Celgene: Other: Travel support, Research Funding; Novartis: Research Funding; MSD: Research Funding; Abbvie: Other: Travel payment; BMS: Honoraria; Bayer: Research Funding; Jazz: Other; MSD: Other: travel support, Research Funding; Archigen: Research Funding. Ilhan:BMS: Speakers Bureau; Alexion: Speakers Bureau; Celgene: Speakers Bureau; Roche: Speakers Bureau.
Effects of Splenectomy before Transplantation on β-Thalassaemia Major with Matched Sibling and Unrelated Hematopoietic Stem Cell Transplantation
Authors: Chen Libai , Wen Jianyun , He Yuelin , Ruan Yongsheng , Li Chunfu , Wu Xuedong
Published: October 2019

Purpose:Splenectomy can improve the quality of life of patients with β-thalassaemia major to some extent, but also increase some unpredictable risk.In order to understand the effects of splenectomy before transplantation on hematopoietic stem cell transplantation reconstruction time, infection, graft versus host disease and hepatic veno-occlusive disease of matched sibling and unrelated hematopoietic stem cell transplantation, the following studies were conducted.

Methods:A retrospective analysis of 80 β-thalassaemia major patients in the southern hospital between January 2012 and December 2017 for matched hematopoietic stem cell transplantation, and 35 of them had a splenectomy before the transplant. Cox model adjustment, t test and χ2 test were used to compare the incidence of neutrophil implantation time, hemoglobin implantation time, platelet implantation time, infection, graft versus host disease and hepatic vein occlusion.

Results:Neutrophilic granulocytes and platelets were implanted more rapidly in patients who underwent splenectomy than those who did not (P < 0.05).There was no significant difference in the incidence of infection, graft versus host disease and hepatic vein occlusion.

Conclusion: For level of Ⅱ-Ⅲ β-thalassaemia major patients, hypersplenism can cause delayed reconstruction of hematopoietic stem cell transplantation, while before transplantation splenectomy to speed up the implantation of hematopoietic stem cell transplantation, and does not increase the incidence of complications after transplantation.

Disclosures

No relevant conflicts of interest to declare.
Bmi-1 Regulates Ex Vivo Self-Renewal of Human Erythroblasts
Authors: Olsen Jayme L. , Aeschlimann Judith , Westhoff Connie M. , Palis James
Published: October 2019

About twelve million units of red blood cells (RBCs) are transfused yearly in the United States. Ex vivo cultured RBCs could serve as a supplemental source to treat alloimmunized patients requiring chronic transfusions. The limited proliferative capacity of erythroid precursors is a major obstacle to generating sufficient numbers of RBCs to constitute even one unit of blood. We previously determined that erythroblasts derived from the murine embryo have a unique ability to self-renew extensively ex vivo, and that Bmi-1, a member of the polycomb repressive complex 1 (PRC1), is both necessary and sufficient to drive the extensive self-renewal of murine bone marrow-derived erythroblasts. Here, we tested the hypothesis that Bmi-1 regulates the ex vivo self-renewal of human erythroid cells. Bmi-1 overexpression increased the proliferative capacity of adult human peripheral blood mononuclear cell-derived erythroblasts, which normally have restricted ex vivo self-renewal, more than 10 billion-fold. Bmi-1-induced self-renewing erythroblasts (iSREs) retained an immature erythroid morphology and cell surface phenotype throughout culture. Chemical inhibition of Bmi-1 led to collapse of the culture, with a massive reduction in cycling cells and an increase in apoptosis. Taken together, these data indicate that Bmi-1 in iSREs is both necessary and sufficient to increase the self-renewal capacity of human erythroid precursors. Importantly, Bmi-1 overexpression does not interfere with the ability of the iSREs to mature into reticulocytes in vitro. Serological analysis of multiple blood group antigens demonstrated that iSRE-derived reticulocytes express the same antigens as the donor's RBCs. Bmi-1 can act as part of canonical or non-canonical PRC1 and different complex members have been associated with hematopoietic self-renewal versus differentiation. Expression studies of self-renewing human erythroblasts revealed that the non-canonical PRC1 member, RYBP, was expressed at higher levels than canonical PRC1 members. In addition, maturation of erythroid precursors is associated with down-regulation of Bmi-1 and non-canonical PRC1 members, as well as the upregulation of several canonical members. Knockdown of RYBP reduced the proliferative capacity of the iSREs, suggesting that Bmi-1 regulates erythroid self-renewal through non-canonical PRC1 interactions. Consistent with this hypothesis, preliminary studies inhibiting canonical PRC1 members led to an additional several trillion-fold increase in the ex vivo proliferative capacity of human iSREs compared to vehicle-treated control cultures. These data suggest that Bmi-1 regulates erythroid self-renewal through non-canonical PRC1. Increasing the ex vivo self-renewal capacity of human erythroblasts ultimately paves the way for the generation of sufficient numbers of cultured RBCs for blood typing and transfusion therapy, as well as the establishment of in vitro models of RBC-intrinsic disorders.

Disclosures

Palis: Rubies Therapeutics: Consultancy.
Long-Term Survival after Transplantation in Patients with Chronic Myeloid Leukemia from HLA-Compatible Unrelated Donors: May the Stem Cell Source Influence the Outcome?
Authors: Koldehoff Michael , Elmaagacli Ahmet H. , Ottinger Hellmut Dietmar , Dührsen Ulrich , Beelen Dietrich W.
Published: October 2019

Introduction

Allogeneic hematopoietic stem cell transplantation (alloSCT) is curative therapy for chronic myeloid leukemia (CML), but its long-term outcomes regarding graft sources are not well described.

Results

Here the outcomes of CML after alloSCT with bone marrow (BM, n=134) were compared to those of peripheral blood stem cell (PBSC, n=172) in the HLA-compatible unrelated donor setting. Patients were transplanted in 1st CP (Bone marrow transplantation (BMT), n=100, and Peripheral blood stem cell transplantation (PBSCT), n=103), in >1st CP (BMT, n=24, and PBSCT, n=52) and in blast crises (BMT, n=10, and PBSCT, n=17). Median follow-up were 54 months after BMT and 106 months after PBSCT. No significant differences were found in the incidence of acute and chronic graft-versus-host disease (GvHD) between both study-groups. The 5-year estimated probability of hematological relapse was 11% for patients in 1st CP CML and 49% for patients in advanced disease after BMT (p<0.001) and 18% for patients in 1st CP and 22% for patients in advanced disease after PBSCT (p= n.s.). The estimated probability for 10-year overall survival (OS) for patients in 1st CP and patients in advanced stages of CML were 58% and 20% after BMT and 59% and 52% after PBSCT, respectively (not significant for 1st CP and p<0.0001 for advanced stages). In the multivariate analysis patient age (age > 40 years), disease stage, acute GvHD, chronic GvHD and immunprophylaxis with use of ATG influenced OS significantly. For leukemia-free survival, the following risk factors were significantly in the multivariate analysis, graft source, patient age, gender constellation, disease stage, acute GvHD and chronic GvHD.

Conclusions

This large trial show significant difference between transplant recipients who received PBSC and those who received BM from unrelated donor. These finding may influence the selection of a graft source for alloSCT from unrelated donor.

Disclosures

Dührsen: Amgen: Research Funding; Gilead: Consultancy, Honoraria; Janssen: Honoraria; AbbVie: Consultancy, Honoraria; Roche: Honoraria, Research Funding; Celgene: Honoraria, Research Funding. Beelen:Medac: Consultancy, Other: Travel Support.
Prognostic Impact of Notch Signaling in Acute Myeloid Leukemia (AML)
Authors: Takam Kamga Paul , Resci Federica , Dal Collo Giada , Adamo Annalisa , Bazzoni Riccardo , Mercuri Angela , Bonifacio Massimiliano , Krampera Mauro
Published: October 2019

Background: Notch signaling is a developmental pathway involved in normal and malignant hematopoiesis. Mutations in Notch genes lead to T-ALL and are associated with poor prognosis in B-CLL. However its role in AML is still under investigation, with controversial results. In addition, although the pathway consists of 4 receptors and 5 ligands, many studies are mostly based on single receptor or ligand.

Aims: This study was conducted to determine if the expression level of each Notch receptor and ligand are associated with known prognostic factors and patient's survival.

Methods: AML primary cells were collected from 79 AML patients after informed consent. The follow-up was for 36 months. Flow cytometry analysis was used to study the expression of each Notch receptor and ligand. Prognostics or risk factors considered were sex, white blood counts, FAB and cytogenetics. The Mann-Whitney test was used for analyzing differences in expression levels between two groups. Spearman test was used for correlation between Notch expression levels and prognostics factors. Differences in overall (OS) and progression free (PFS) survival were established through Gehan-Breslow-Wilcoxon test.

Results: Consistently with previous findings, we found expression of Notch1, Notch2, Notch3, Notch4, Jagged1, Jagged2 and DLL3 in AML samples. There were no differences in expression levels between male and female. For each receptor or ligand, higher expression levels were found in more immature samples (M0, M1, and M2). Notch3, Notch4 and Jagged2 were enriched in adverse cytogenenetics risk groups compared to favorable cytogenetics risk patients. Then, for each receptor or ligand, patients were divided into two groups; patients with higher expression levels and patients with lower expression levels. Analyzing OS and PFS, we found that patients with lower expression levels of Notch4, Jagged2 and DLL3 displayed a longer survival compared to patients with higher expression levels.

Conclusion: Given its oncogenic role in T-ALL and other malignancies Notch1 has been the more studied receptors in hematological malignancies. However, we have previously demonstrated that Notch4, DLL3 and Jagged2 are all involved in survival of AML and B-ALL cells. Studying the prognostic value of all Notch receptors and ligands, the current research clearly shows that higher levels of Notch4 and Jagged2 are found in poor cytogenetics risk groups and are associated with a shorter patient's survival. These demonstrations suggest that expression levels of Notch receptors and ligands in AML patient's samples at diagnostic could stand as prognostic marker for clinical care.

Disclosures

No relevant conflicts of interest to declare.
Single Cell Transcriptomics Maps the Embryonic Emergence of HSC and Identifies Intercellular Interactions Regulating HSC Genesis
Authors: Hadland Brandon , Varnum-Finney Barbara , Dozono Stacey , Jackson Dana , Rafii Shahin , Bernstein Irwin D. , Trapnell Cole
Published: October 2019

Hematopoietic stem cells (HSC) are generated during a transient window of embryonic development from endothelial-like hemogenic precursors within specific arterial vessels such as the aorta of the AGM (aorta-gonad-mesonephros region). During HSC emergence, hemogenic precursors must acquire and maintain HSC-defining properties such as the ability to self-renew, home, and provide multilineage hematopoiesis, properties which distinguish rare HSC from a multitude of other hematopoietic progenitors arising simultaneously in the developing embryo. However, the precise niche-derived signals necessary and sufficient to support the acquisition and maintenance of these properties remains poorly defined. Toward identification of these signals, we generated a platform consisting of endothelial cells from the embryonic AGM (AGM-EC) which supports the specification and self-renewal of engrafting HSC from clonal embryo-derived hemogenic precursors in vitro. Using this platform to assay functional HSC potential at the single cell level, we determined a phenotype (VE-Caderin+CD61+EPCRhigh) that encompasses the population of hemogenic precursors during their asynchronous transition to HSC between E9.5 and E11.5 in murine embryonic development. To elucidate the transcriptional changes associated with the emergence of HSC from hemogenic precursors, we analyzed the global transcriptional profiles of FACS-purified VE-Caderin+CD61+EPCRhigh cells at various stages of embryonic development by single cell RNA-sequencing and reconstructed their developmental trajectory in "pseudotime" based on incremental changes in their transcriptional profiles. Complementary analysis of AGM-EC by bulk and single cell RNA-sequencing revealed a unique transcriptional profile of niche endothelial cells supporting HSC development enriched for immune/inflammatory signals. Combining the transcriptional profiles of emerging HSC with niche AGM-EC, we have identified candidate ligand-receptor pairs regulating intercellular interactions during HSC specification and self-renewal and have begun to validate the functional importance of these interactions in supporting HSC generation from hemogenic precursors in vitro. We expect these studies will enhance our understanding of the unique signal pathways necessary for the development of functional HSC, a critical step toward engineering HSC in vitro for clinical applications in disease modeling, drug discovery, and gene modification to identify novel therapies for hematologic and immunologic disorders.

Disclosures

Rafii: Angiocrine Bioscience: Equity Ownership.
Molecular Analysis of Factor V, Prothrombin and Methylenetetrahydrofolate Redutase in Thrombolic Patients
Authors: Paiva Aldair Sousa , Paiva Hugo Diogenes De Oliveira , Cavalcanti Geraldo Barroso , Leão Gioconda DR , Leão Marcos Dias , Bahia Frank , Vasconcelos Roberto C , Fernandes Andrea LAC , Gil Erica Aires , Soares Victor lima , Januario Geisa , Santos Karla Cristina , Lemos Telma Araujo , Jardim Alessandra Suelen
Published: October 2019

Background: The study of thrombotic events requires knowledge of changes in the hemostatic system associated with multiple acquired and hereditary risk factors that suggest predisposition to thrombosis. The factor V or Leiden (G1621A), Prothrombin (G21210A) and Methylenetetrahydrofolate redutase-MTHFR (C677T) mutations are the major genetic risk factors for venous thrombosis. This study assessed the frequency of mutations of the Factor V (Leiden), Prothrombin and MTHFR in patients with thrombophilia from the DNA Center Laboratory, Natal - RN. Methods: The detection of mutations was made by PCR-RFLP followed by enzymatic restriction with HindIII (Leiden and Prothrombin) and HinfI (MTHFR). Results: From 69 selected patients, 52 (75.36%) were females and 35 (24.64%) were males. The frequency of genotypes for the Factor V were: 3 mutated homozygous (4.35%), 4 heterozygous (5.80%) and 62 normal homozygous (89.85%). Regarding the mutation in the Prothrombin gene it was observed in 65 normal homozygous patients (94.2%) and 4 (5.8%) heterozygous. The analysis of the mutation in the gene MTHFR showed 35 (50.7%) normal homozygous patients, 5 (7.2%) mutated homozygous patients and 29 heterozygotes patients (42.1%). Conclusions: Approximately 50% of patients tested had at least one type of genetic alteration combined. Based on data obtained it is indicated the investigation of three markers (Factor V, Prothrombin and MTHFR) thrombophilia-related, targeting the real impact of the molecular mutations in thrombosis and the conduct of treatment.

Disclosures

No relevant conflicts of interest to declare.