bluuebunny/crossref_metadata_2025 huggingface dataset:
A Novel Role for Malate Dehydrogenase 1 in Acute Myeloid Leukemia Progression
Authors: Jin Jie , Huang Shujuan , Pan Jiajia
Published: November 2019

The tumor suppressor role of the metabolic genes is rarely reported. Here, we report that malate dehydrogenase 1 (MDH1) acts as an anti-oncogene in acute myeloid leukemia (AML) progression. In a screen of various leukemia cell lines, as well as peripheral blood mononuclear cells from patients with AML and healthy individuals, we observed that patients with AML had lower mRNA expression of MDH1 than healthy individuals. In addition, we found that overexpression of MDH1 in the AML cell lines MV4-11 and THP-1 in vitro promoted their differentiation, reduced their growth, induced apoptosis, and increased the proportion of cells in S phase, which were more sensitive to the chemotherapeutic drugs cytarabine and daunorubicin. Using an in vivo xenograft mouse model of AML, we found that disruption of MDH1 expression influenced tumor burden and mouse survival. These findings were consistent with an inverse relationship between MDH1 expression and overall survival in two independent cohorts of AML patients. We also identified changes in the expression of other genes, concomitant with reduced MDH1 expression in patients with AML, that largely explain the anti-oncogenic role of MDH1 in AML progression. Notably, downregulation of P53 and upregulation of the MAPK pathway in low MDH1-expressing MV4-11 cells were reversed upon overexpression of MDH1. In conclusion, we demonstrated that MDH1 plays a tumor-suppressing role in AML and can be used as a novel mimic drug therapy in the future.

Disclosures

No relevant conflicts of interest to declare.
Immunosenescent Characteristics of T Cells in Young Patients Following Haploidentical Stem Cell Transplantation from Parental Donors
Authors: Lee Ga Hye , Hong Kyung Taek , Choi Jung Yoon , Shin Hee Young , Lee Won-Woo , Kang Hyoung Jin
Published: November 2019

Introduction: Pediatric and adolescent patients in need of allogeneic hematopoietic stem cell transplantation generally receive stem cells from older, unrelated or parental donors when a sibling donor is not available. Despite encouraging clinical outcomes, it has been suggested that immune reconstitution accompanied by increased replicative stress and a large difference between donor and recipient age may worsen immunosenescence in pediatric recipients. Therefore, in this study paired samples were collected at the same time from donors and recipients of haploidentical hematopoietic stem cell transplantation (HaploSCT).

Methods: We conducted flow cytometry-based phenotypic and functional analyses and telomere length measurements of 21 paired T-cell sets from parental donors and children who received T cell-replete HaploSCT with post-transplant cyclophosphamide (PTCy) at Seoul National University Children's Hospital between February 2014 and January 2017. The conditioning regimen was comprised of targeted busulfan (total target area under the curve, 75,000 mg•h/L) with intensive pharmacokinetic monitoring, fludarabine and cyclophosphamide.

Results: Fourteen pediatric, adolescent, and young adult patients with malignant disease and seven with nonmalignant disease were included with a median post-transplantation period of 16.9 months (range, 12.4-38.8). Senescent T cells, CD28- or CD57+ subsets of both CD4+ and CD8+ T cells, were significantly expanded in patients compared with parental donors. Further, not only CD4+CD28- T cells, but also CD4+CD28+ T cells showed reduced cytokine production capacity and impaired polyfunctionality compared with parental donors, whereas their TCR mediated proliferation capacity was comparable. Of note, the telomere length in patient T cells was preserved, or even slightly longer, in senescent T cells compared with donor cells. We also found that the patients had a higher level of γ-H2AX-expressing CD28- senescent T cells compared with the donors, which is used as a DNA damage marker. Regression analysis showed that senescent features of CD4+ and CD8+ T cells in patients were influenced by donor age and the frequency of CD28- cells, respectively.

Conclusions: Our data suggest that T cells undergo premature immunosenescent changes and exhibit functional defects in pediatric HaploSCT recipients. Further, there is an increased level of DNA damage in patient CD4+ T cells compared to those of parental donors. Therefore, long-term, comprehensive immune monitoring of these patients is necessary.

Disclosures

No relevant conflicts of interest to declare.
Cytomegalovirus Infection in Bone Marrow-Derived Mesenchymal Stem Cells: Implications for Functions and Exploration of Relevant Mechanisms
Authors: Lv Weiran , Fan Zhiping , Huang Fen , Xu Na , Xuan Li , Lin Ren , Zhao Ke , Liu Qifa
Published: November 2019

Human cytomegalovirus (HCMV) infection could lead to life-threatening complications in immunocompromised and immunosuppressed populations, especially after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Reactivated CMV infections can contribute to poor graft function (PGF) characterized by multilineage hematopoietic failure after allo-HSCT. Bone marrow derived mesenchymal stem cells (BM-MSCs) play an important role in supporting hematopoiesis and modulating differentiation of hematopoietic primitive stem cells. Nevertheless, little is known about the dysfunctions and underlying mechanism closely associated with hematopoiesis of BM-MSCs after HCMV infection and no previous studies have concentrated on the dysfunctions of primary BM-MSCs from patients with CMV viremia following allo-HSCT. In the current study, we explored whether BM-MSCs could be infected by HCMV and evaluated the function of BM-MSCs with HCMV infection and the relationship between the dysfunction and mitogen activated protein kinases (MAPKs). Moreover, we retrospectively investigated the relation between CMV infection and PGF among 801 patients who received allo-HSCT in Nanfang Hospital. Our results showed that HCMV infection was detected both in BM-MSCs with HCMV AD169 infection in vitro and in BM-MSCs derived from patients with CMV viremia following allo-HSCT. Dysfunctional BM-MSCs with HCMV infection, which were characterized by impaired proliferation, osteogenesis potential, mRNA level of stem cell growth factor (SCF), colony-forming unit (CFU) plating efficiency and higher levels of apoptosis, adipogenesis potential, mRNA level of interleukin 6 (IL-6) and transforming growth factor β1 (TGF-β1) were revealed. Activation of p38 was detected in BM-MSCs with HCMV infection and function of infected BM-MSCs were enhanced by SB203580 treatment in vitro. Retrospective clinical data analysis showed that haploidentical donor (HRD) transplant and CMV infection (in 30 days) were risk factors of PGF. In summary, dysfunctional BM-MSCs were observed after HCMV infection which might induce hematopoiesis suppression and SB203580 treatment in vitro functionally improved BM-MSCs through downregulation of the p38MAPK pathway and our clinical analysis showed that CMV infection is a risk factor of PGF which further demonstrated hematopoietic inhibition of CMV.

Disclosures

No relevant conflicts of interest to declare.
Targeted-Capture Sequencing Is a Useful Method for MRD Markers Screening in KMT2A (MLL) Rearranged Leukemia
Authors: Sanada Masashi , Yamada Tomomi , Iijima-Yamashita Yuka , Nishijima Dai , Kanamori Takashi , Tomizawa Daisuke , Hori Toshinori , Horibe Keizo
Published: November 2019

Immunoglobulin (Ig)/T-cell receptor (TCR) gene rearrangement-based molecular minimum residual disease (MRD) monitoring has become one of the most powerful prognostic indicators for patients with acute lymphoblastic leukemia (ALL). Infant ALL has a poor prognosis and requires therapeutic development based on accurate MRD assessment. However, there are some difficulties associated with the highly sensitive detection of MRD based on Ig/TCR gene rearrangements in infant ALL cases, compared to other B-ALL cases, because of the immature Ig/TCR gene rearrangement patterns and their oligoclonality. Since KMT2A(MLL) gene rearrangements are identified in infant ALL at a high frequency, the use of KMT2A rearranged sequences is an alternative method for MRD detection in KMT2A rearranged ALL cases. However, identifying the genomic breakpoints in KMT2A rearrangements by conventional methods is laborious and difficult in routine practice. To detect molecular MRD markers in infant ALL cases, we designed capture probes covering the coding and recognition signal sequences of the V, D, and J genes of the Ig/TCR loci and common genomic breakpoints in KMT2A, AFF1 (AF4), MLLT3 (AF9), MLLT1 (ENL), MLLT10 (AF10), and ELL. 88 infant ALL cases from the JPLSG MLL-10 study, including 73 cases with KMT2A rearrangements, were analyzed by high-throughput target-capture sequencing on the Illumina platform. The sequenced data were analyzed with the Ig/TCR recombination analysis tool Vidjil and the Genomon pipeline for detection of KMT2A rearrangements.

Although 40% of cases in this study were marker-negative in the conventional marker screening analysis, clonal Ig/TCR targets for MRD measurements were identified in more than 80% of the cases by target-capture sequencing. Ig / TCR rearrangements detected in infant ALL were unique and related to age at diagnosis and type of KMT2A rearragement. KMT2A-rearranged cases showed a significantly higher frequency of incomplete rearrangements of IGH and TCRD compared to KMT2A-negative infant ALL cases.　The target-capture method also showed an increased sensitivity of Ig/TCR-based MRD measurements even in cases where MRD measurement was possible.　Genomic breakpoints of KMT2A rearrangements were detected in 80% of KMT2A-rearranged cases, in which KMT2A was most frequently rearranged with AFF1, followed by MLLT1, MLLT3, and MLLT10. Other rare fusion partners of the KMT2A gene were also captured by probes for KMT2A sequences. The determined patient-specific genomic breakpoint sequences could be used for monitoring the MRD of KMT2A-rearranged cases. Thus, target-capture sequencing is an easy and useful method for screening MRD markers in infant leukemia cases.

Disclosures

No relevant conflicts of interest to declare.
Role of Thrombopoietin in Stimulating Megakaryopoiesis of ITP Patients Via Rescuing Vascular Niche
Authors: Jiao Yingying , Zeng Xiaoyuan , Li Zongpeng , Chong Beng H , Zhang Yujiao , Ye Jieyu
Published: November 2019

Background: Previous studies have shown that the vascular niche was damaged in patients with immune thrombocytopenia (ITP). It is mainly manifested that the migration and tube-formation ability of bone marrow endothelial progenitor cells (BM-EPCs) were significantly reduced. Endothelial cells as an important part of the vascular niche play an important role in regulation of megakaryocytes and platelet hematopoiesis. Our previous studies suggested that thrombopoietin (TPO) has a protective effect on human endothelial cells. This study aim to investigate whether TPO has a protective effect in EPCs of ITP patients and to explore its mechanism.

Methods: BM-EPCs were isolated from ITP patients, and divided into two groups: TPO-treated group and TPO-free (control group). CCK8 was used to explore whether TPO has proliferative effect on BM-EPCs. BM-EPCs was identified by flow cytometry with co-expression of mouse anti-humanCD133, mouse anti-human CD34 and mouse anti-human vascular endothelial growth factor receptor (CD309) monoclonal antibodies. Moreover, the number of EPCs was evaluated with DiL-Ac-LDL uptake and FITC-UEA -I binding assay. Furthermore, the function of BM-EPCs was studied by using tube-formation and migration experiments.

Results: The ITP patient-derived EPCs in TPO-treated group exhibited longer fusiform and elliptical shape after 5 days and cobblestone-like morphology after 7-10 days. The result of CCK-8 showed TPO could promote the proliferation of EPCs and the optional drug concentration is 100 ng/ml. Much higher expression of CD34/CD133/CD309 in TPO-treated group was detected by flow cytometry. The number of cells with immunofluorescence double staining in TPO group was significantly higher than of control group (n=4, p<0.05). The tube formation (n=5, p<0.05) and migration capacity (n=5, p<0.05) were significantly enhanced in TPO-treated group.

Conclusion: Our studies indicates that TPO may exert megakaryopoiesis effect in ITP patients via protection of BM-EPCs. This may provide a supplemental explanation of how recombinant TPO and TPO receptor antagonist acts on ITP treatment.

Keywords: thrombopoietin; vascular niche; endothelial progenitor cell, megakaryopoiesis; endothelial cell

Disclosures

No relevant conflicts of interest to declare.
Clinico-Pathological Profiles of Obese Patients with Chronic Myeloid Leukemia (CML) and Their Response to Therapies (TKIs)
Authors: Yassin Mohamed A , Abdulla Mohammad Abdul-Jaber , Abbas Feryal , Chandra Prem , Nashwan Abdulqadir Jeprel , Ghasoub Rola S , Kassem Nancy , Al Sabbagh Ahmad , Nawaz Zafar , Akiki Susanna Jane
Published: November 2019

Background and Objectives:

Cancer patients in Qatar are managed in a standardized manner through a single center guided by the National Cancer Framework. Most cancer medication-dosing is based on weight and height (body surface area), however in CML, tyrosine kinase inhibitors (TKIs) dosing is fixed. Obesity, defined by a BMI of 30 or more, presents a risk to health yet to the best of our knowledge, there have been no large studies on obesity and response to TKI therapy in CML. The aim of this study was to explore the clinical characteristics of obese and non-obese CML patients, and compare their response to standard dose TKI therapy.

Methods:

A retrospective analysis was performed of 123 patients treated for CML in chronic phase at NCCCR over a 23-year period, 69 obese (BMI > 30) vs. 54 with normal BMI = 18.5 - 24.99 (controls). Comparisons were made of characteristics at diagnosis including: gender, age, ethnicity, CBC parameters, spleen size, Sokal score, bone marrow findings, TKI prescribed, together with response assessment using ELN guidelines.

Results:

Mean age at diagnosis was lower in obese patients compared with control group (39.0 vs 42.2). Obese patients had lower WBC count and higher Hgb at diagnosis (mean WBC 153.9 vs. 193.0; mean Hgb 11.4 vs. 10.3). A higher proportion of obesity group was females (31/69) compared to control group (14/54). Spleen size and Sokal scores were lower in obesity group (17.1 vs. 18.3 and 0.81 vs. 0.91 respectively). More obese patients received Imatinib (40 vs. 12 in control group) compared with other TKIs (21 vs. 37 in control group). Few patients in obesity group had to switch to an alternative TKI (13 vs. 20 in control group). Compared to control group more obese patients achieved a major molecular response (MMR defined as BCR-ABL/ABL (IS) value below 0.1%) (47 of 60 vs. 22 of 42; p = 0.011).

Conclusions:

The data suggests differences in clinical characteristics and response to treatment between obese and non obese patients that warrants further investigation. Obese patients with CML were younger at diagnosis, had better response to treatment and were less likely to need switch TKI due to failure of treatment.

Table

Disclosures

No relevant conflicts of interest to declare.
Microbial-Derived Metabolites Drive Protective Type-I Interferon Responses in Models of Gut Epithelial Damage and Limit Graft-Versus-Host Disease
Authors: Thiele Orberg Erik , Göttert Sascha , Poeck Hendrik
Published: November 2019

Background: Graft-versus-host disease (GVHD) is a dreaded complication after stem-cell transplantation (SCT). Standard treatment relies on immunosuppressants but is associated with an increased risk of infection and relapse. Up 50% of patients develop steroid-refractory GVHD, with dismal impact on SCT outcomes.

New studies in mice suggest that induction of type-I interferon (IFN-I) signalling or activation of IFN-I inducing pathways such as cGAS/STING or RIG-I/MAVS can promote gut barrier integrity and limit GVHD. However, the endogenous ligands that drive this "protective" IFN-I response are poorly defined. Recent data in mice and humans suggest that microbial-derived metabolites such as small-chain fatty acids or indoles can decrease GHVD mortality. Here, we describe IFN-I inducing metabolites that improve outcomes in mouse models of gut epithelial damage and acute GVHD via a mechanism dependent on STING signalling.

Methods: To investigate which cell types mediate protection and how, we generated intestinal organoids and antigen presenting cells (APC) from bone marrow of WT or genetically deficient mice (MAVS-/-, STING-/-, IFNAR-/-) under steady state conditions versus chemotherapy, total body irradiation and after allogeneic SCT in the presence or absence of microbial metabolites. Analysis was performed by microscopy, immunoblotting, qPCR, ELISA and flow cytometry. Outcomes of gut-injured mice were assessed by clinical scoring, histopathology, flow cytometry and organoid recovery.

Results: Metabolite treatment promoted regeneration of intestinal organoids as assayed by organoid numbers as well as proliferation. These effects were dependent on IFN-I and STING signalling. In addition, we found activated pro-inflammatory NFkB signalling and decreased apoptosis as evidenced by reduced caspase-3 cleavage. In APCs, IFN-I responses were enhanced in the presence of metabolites including increased IFN-β production and upregulation of IFN stimulated genes. Metabolite-treated mice showed improved recovery of intestinal stem cells following gut injury by chemotherapy and irradiation and increased survival in acute GVHD.

Conclusions: Our findings uncover a mechanism by which microbial metabolites amplify IFN-I signals, promote tissue regeneration and thereby prevent allo-activation and GVHD. We show for the first time that these IFN-I inducing metabolites are protective in diverse models of gut damage -- from chemotherapy and radiation to immune-mediated damage -- via a STING-dependent mechanism. Perhaps the poor prognosis of GVHD patients exhibiting a loss of microbiota diversity can be explained in part by an absence of "protective" metabolites able to amplify critical IFN signals. We are currently studying whether metabolite levels correlate with severity and outcome of GVHD in humans.

Figure

Disclosures

No relevant conflicts of interest to declare.
Changes in Platelet Utilization Via Implementation of Transfusion Guidelines in a Large Health System Hospital
Authors: Bowers Reid Christian , Thompson Michael A. , Van Dreser Julie , Sanchez Federico Augusto
Published: November 2019

Background: As part of system wide implementation of Patient Blood Management (PBM), transfusion guidelines were implemented at St. Luke's Medical Center, Advocate Aurora Health in Milwaukee, WI.

Methods: Beginning in 2015 Aurora Health Care made a platelet (PLT) count of ≤10K the standard threshold below which PLT transfusion is considered appropriate. Additionally, transfusing PLTs as single donor units with a PLT count between each unit was made standard. Education was provided to clinicians on the new standard including best practice advice in the transfusion ordering workflow. Data was gathered on institutional PLT use and individual clinician PLT ordering practices. Further education was provided based on the extent to which data showed conformity to the new standard.

Results: During PBM implementation 2015-2018 despite an initial decline there was a modest overall increase in institutional PLT usage from 2515 to 2933 units. The percentage of transfusions ordered with PLT ≤10K rose from 21.6% to 31.9%. The percentage of transfusions ordered as single donor units increased from 82.5% to 91.5%. See Graph/Table 1 & 2 for further analysis of individual practitioner PLT ordering. See Table 3 for reporting period volume data.

Conclusions: Implementation of PLT transfusion guidelines was associated with an initial annual reduction in PLT usage, but annual PLT utilization did go up overall during the reporting period. Individual clinician transfusion practices overall conformed more with guidelines at the end of the reporting period than at the beginning. Changes in treatment guidelines for acute promyelocytic leukemia, i.e. target PLT >50K, during the reporting period may account for a rebound in PLT usage and a decrease in the percentage of transfusions ordered with PLT ≤10K in the later vs. the earlier years of the reporting period. By contrast transfusions continued a positive trend toward being given as single donor units even in the later years of the reporting period.

Disclosures

Thompson: Doximity: Equity Ownership, Membership on an entity's Board of Directors or advisory committees; Celgene: Membership on an entity's Board of Directors or advisory committees; Lynx Bio: Research Funding; BMS: Research Funding; VIA Oncology: Membership on an entity's Board of Directors or advisory committees; Takeda: Membership on an entity's Board of Directors or advisory committees, Research Funding; GSK: Membership on an entity's Board of Directors or advisory committees; Adaptive: Membership on an entity's Board of Directors or advisory committees; UpToDate: Patents & Royalties: Myeloma reviewer; AbbVie: Research Funding.
Is It Time to Take MRD More Seriously? Relationship between MRD, Functional Cure and Life Expectancy Among Patients with Ph- B Cell Precursor ALL (BCP-ALL)
Authors: Shao Qiujun , Tiwana Simran , Despiegel Nicholas , Dirnberger Franziska
Published: November 2019

Background: Although BCP-ALL patients can achieve complete remission (CR) after induction chemotherapy, about 40% of them have residual leukemia cells in the bone marrow which is called minimal residual disease (MRD). Complete MRD response is defined as lack of MRD with an assay sensitivity of >= 10-4, which, if sustained for more than five years, may be regarded as a functional cure for ALL. On the contrary, presence of MRD is a signal of high risk of relapse leading to poor health outcomes. Blinatumomab is the only drug approved for BCP-ALL patients with MRD based on the results from BLAST, a single-arm phase 2 study conducted to assess the efficacy and safety of blinatumomab in adult BCP-ALL patients in CR with MRD. Currently, there is limited evidence on the relationship between MRD response and functional cure in ALL and life years gained.

Objectives: The objective of this study was to assess the relationship between complete MRD response after blinatumomab treatment and 1) functional cure; 2) life years and quality adjusted life years (QALYs) gained among BCP-ALL patients with MRD.

Methods: The relationship between MRD response, and clinical outcomes was analyzed using an existing cost-effectiveness analysis (CEA) based on combined decision tree and Markov cohort model over a lifetime horizon (50 years). The model includes survival conditional on treatment and MRD status for time to stem transplant, relapse and death. In the CEA, blinatumomab was compared to standard of care (SOC, defined as conventional maintenance chemotherapy). Clinical probabilities were derived primarily from blinatumomab clinical trial data and a non-interventional cohort study (used as a historical control). Life years gained were adjusted for quality based on utility values from trial EQ-5D scores. An annual discount rate of 3.5% was applied to estimate future outcomes.

Results: In the base case, the complete MRD response rate was 83.6%, and blinatumomab yielded more life years gained (8.26 vs. 5.68) and more QALYs (6.56 vs. 4.44) compared with SOC. The proportion of patients achieving a functional cure was 43.59% in blinatumomab group vs. 26.78% in the SOC group. Clinical response varied with variations in MRD response rate at 75%, 78%, 80%, 85%. Specifically, when the complete MRD response rate increased from 75% to 85%, the total life years gained in the blinatumomab group increased by 0.51, QALYS increased by 0.42, and the proportion of patients cured increased by 3.05%.

Conclusion: Achieving complete MRD response is associated with longer life expectancy, higher probability of being cured, and more QALYs among patients with BCP-ALL and MRD.

Disclosures

Shao: Amgen: Employment. Tiwana:Amgen: Employment, Equity Ownership. Despiegel:Amgen: Employment, Equity Ownership. Dirnberger:Amgen: Employment, Equity Ownership.
Safety Analysis of Five Randomized Controlled Studies of Daratumumab in Patients with Multiple Myeloma
Authors: Al Hadidi Samer , Kamble Rammurti , Miller-Chism Courtney Nicole , Mims Martha P.
Published: November 2019

ABSTRACT

BACKGROUND

Daratumumab is an anti-CD38 monoclonal antibody that is used more frequently in patients with Multiple Myeloma (MM). Multiple studies have demonstrated the efficacy and safety of daratumumab for relapsed/refractory MM and primary treatment for transplant eligible and ineligible patients.

METHODS

We conducted an integrated safety analysis to characterize the frequency, severity, natural history, and outcomes of adverse events (AEs) with daratumumab versus comparators. Data were pooled from 5 completed phase III randomized controlled studies that had included 1798 daratumumab-treated and 1797 comparator-treated patients with MM as a first line in both transplant eligible and transplant ineligible and for relapsed/refractory disease(Table.1). Safety analyses included reporting of AEs using crude and exposure-adjusted incidence rates.

RESULTS

The median follow up duration is 16.84 months (range: 7.4-28 months) for both daratumumab-treated and comparator-treated patients. Discontinuation for any reason occurred less often with daratumumab (22% vs. 33.9%) though discontinuation because of AEs occurred at similar rates (25% vs.26%) as well as deaths due to AEs occurred at similar rates (2.25% vs. 1.84%). When adjusted to exposure, neutropenia, lymphopenia, diarrhea, fatigue, dyspnea, pneumonia and hypertension were the only common grade 3 AEs more often reported with daratumumab than with the comparators(Table.2). The prevalence of common grade 3/4 AEs with daratumumab were <7% apart from neutropenia, lymphopenia and pneumonia (45.9 vs. 32.3, 13 vs. 7.5, and 10.6 vs.7.2)(Table 3). Grade 3/4 daratumumab infusion related reactions happened in 3.8%

CONCLUSIONS

These results from an integrated analysis support a favorable benefit/risk profile of daratumumab in patients with MM.

Disclosures

No relevant conflicts of interest to declare.
Hepatosplenic T Cell Lymphoma Case Report Presented with Hemphagocytosis Lymphohistiocytosis
Authors: Abu Haleeqa Mohamed , Alshehi Fatemah , Alghamrawi Gannat , Richard Lois
Published: November 2019

We are reporting a challenging case 37-year-old Pakistani male patient with Hepatosplenic T-Cell lymphoma (HSTCL) which is a rare aggressive form of Peripheral T-Cell lymphoma that is characterized by primary extra nodal disease with malignant T-cell proliferation in the liver, spleen, and bone marrow. He presented initially with nonspecific symptoms of generalized fatigue & vague abdominal pain . He was found to have hepatosplenomegaly (with liver & splenic lesions), derranged liver enzymes. He refused investigation and lost follow up.

He came back 3 months after in a bad condition with fever, abdominal pain & he deteriorated very rapidly. The diagnosis was challenging as he required an extensive investigations that ultimately showed the characteristic clinical, histopathologic, and cytogenetic features of hepatosplenic T-cell lymphoma with HLH presentation.

Patient had multiorgan failure,coagulopathy, shock(hepatic/renal/septic) and started on vasopressors, Ferritin was high >16000, Secondary Hemophagocytic Lymphohistiocytosis (HLH) was made at this point & patient was started on dexamethasone as per HLH protocol 2004 to reduce the tumor burden, and the plan was to start CHOP after 2 days, if patient condition allowed.

Bone marrow biopsy showed: hypercellular Bone marrow massively infiltrated by neoplastic mature T-lineage population with overall morphologic & phenotypic characteristics consistent with T cell lymphoma.

Flow cytometry was done from peripheral blood as bone marrow aspiration revealed dry tap.it showed presence of approximately 30-40% circulating lymphoma cells with aberrant mature T-cell phenotype, CD3+, TCRgd+, CD56+, CD8+/CD4-, and CD5-. The overall morphology and immunophenotypic features were consistent with hepatosplenic T-cell lymphoma (WHO classification updated 4th edition,2016).

patient received full CHOP. All through patient was steadily worsening and unfortunately passed away.

HSTCL is a rare and challenging diagnosis and presentation can be dramatic like this case with HLH. T cell lymphoma can present with HLH and treatment

Figure

Disclosures

No relevant conflicts of interest to declare.
Platelet Function Disorder Is More Common Than Von Willebrand Disease in Women with Inherited Bleeding Disorders: Report from a Public Sector University in Pakistan
Authors: Hossain Nazli , Shamsi Tahir S. , Naz Arshi , Abbas Sidra , Khan Nazeer
Published: November 2019

Abstract

Introduction: Inherited bleeding disorders are an important cause of menorrhagia.

Aim: To determine the prevalence of inherited bleeding disorders in women with menorrhagia in the absence of systemic and pelvic pathologies.

Methods: The study period was from March 2013 to September 2018. It was carried out at the Department of Obstetrics & Gynecology, Unit II, Dow Medical College and Ruth Pfau KM Civil Hospital in Karachi. The laboratory investigations were carried out partly at Dow Diagnostic Research Laboratory and mainly at the National Institute of Blood Disease & Bone Marrow Transplant. Included in the study were young girls and women who presented with menorrhagia at menarche or in the later years of reproduction, with normal bimanual pelvic examination and pelvic ultrasound results.

Results: Approximately 200 women with a mean menstrual cycle duration of 9.3+ 3.5 days were included. The mean age of the included patients was 32 years. Married women's partners were cousins in 50% of the women, while unmarried girls' parents were cousins in all cases. Platelet aggregation defect was presented in 33 (16.6%) women, von Willebrand disease in 26 (13.1%) women, factor IX deficiency in 20 (10%) women, combined defects in 29 (14.6%) women, and factor VII and VIII deficiencies in 4 and 11 (2%, 5.5%) women, respectively. Isolated defect with ADP, epinephrine, and collagen were decreased in 35 (17.6%), 41 (20%), and 33 (16%) women, respectively.

Conclusion: Inherited bleeding disorders were a common cause of menorrhagia in our population. Consanguinity was frequently seen in these subjects. Except for some forms of VWD, all other disorders have an autosomal recessive inheritance pattern. After excluding gynecological and medical causes, a work up for inherited bleeding disorders should be advised.

Keywords: menorrhagia, platelet function disorder, inherited bleeding disorders, Pakistan.

Disclosures

No relevant conflicts of interest to declare.
A GUT Microbiota Score Predicting for ACUTE GRAFT-Versus-Host Disease
Authors: Han Lijie , Peng Jie , Zhao Ke , Fan Zhiping , Jin Hua , Zhang Haiyan , Sun Hui , Liu Qifa , Jiang Zhongxing
Published: November 2019

Although some studies have reported that intestinal microbiota is associated with aGVHD, they inadequately lacked a satisfactory method for precisely predicting aGVHD occurrence. We collected stool and blood samples at day 15 post-transplantation from 150 patients, who underwent allogeneic hematopoietic stem cell transplantation (allo-HSCT) from two independent centers. Stool microbiota was detected by 16S rRNA gene sequencing, inflammatory factors in blood were detected by multiplex immunoassays, and T lymphocyte subsets were examined by flow cytometry. A gut microbiota score (GMS) from a LASSO (least absolute shrinkage and selection operator) model using the gut microbiota was developed and validated to predict aGVHD. The discovery cohort included most of the identified bacterial families. As determined by area under the curve (AUC) analysis of ROC plots, the GMS could predict aGVHD in grades II-IV (AUC = 0.904, P < 0.0001). Furthermore, the GMS model in the validation set also demonstrated a high AUC that was consistent with the discovery set (AUC = 0.887, P < 0.0001). The results also indicated that the ratio of Treg/Th17 cells in the low GMS subgroup was higher compared with the high GMS subgroup (P = 0.012), and the validation set was consistent with the discovery set (P = 0.003). In addition, high cytokine levels (including IL-1β, IL-6, IL-17, and TNF-α) were associated with high GMS. In conclusions, the GMS at neutrophil engraftment could predict the subsequent aGVHD, and it is a potential and novel method. The GMS was associated with the inflammatory factor levels and Treg/Th17 balance.

Disclosures

No relevant conflicts of interest to declare.
Comprehensive Genomic Profiling of Adult T-Cell Lymphoblastic Lymphoma in Chinese Patients
Authors: Chen Feili , Guo Hanguo , Huang Ling , Jiang Xinmiao , Wei Xiaojuan , Li Wenyu
Published: November 2019

Background:

T-cell lymphoblastic lymphoma (T-LBL) is a rare, aggressive neoplasm of precursor T cells that occurs mostly in adolescents and young adults. Extensive genomic analyses of T-LBL have discovered genes encoding adhesion molecules and extracellular matrix proteins in samples from T-LBL patients, with most studies being performed on patients in the Western hemisphere. We sought to define the mutational profile of T-LBL in Chinese patients.

Methods:

We collected 15 adult T-LBL biopsies from one center in China with confirmed pathologic diagnosis and treatment information. Median cohort age at diagnosis was 26.0 years. The male/female ratio was 13/2. DNA was extracted from formalin-fixed paraffin embedded samples, and targeted deep next generation sequencing (NGS) analyses were performed.

Results:

Mutation features were revealed in this Chinese T-LBL analysis. NOTCH1 mutations were found in 67% (10/15) of T-LBL patients (pts) with the highest mutation frequency. PHF6 mutations were seen in 53% (8/15) of cases. TP53mutations were seen in 53% (5/15) of T-LBL cases. JAK1 and JAK3 mutations were seen in 27% (4/15)of T-LBL cases respectively. Mutations at histone modification genesincluded SUZ12 (20%, 3/15), EZH2(13%, 2/15), and EP300 (7%,1/15). Other mutations included PTEN(27% 4/15), DNM2 (27% 4/15).

Discussion:

The aim of our study was to reveal the genomic landscape of Chinese T-LBL. The cohort collection was unbiased, the patients' features were in concordance with previous reports that the disease of T-LBL is mostly found in the young patients and shows male predominance. The most predominant somatic mutation wasNOTCH1, which is also usually seen in T-ALL. Mutations at histone modification genes were also seen in our study. NGS studies have identified epigenetic modifiers mutations as a hallmark of T-LBL, highlighting an attractive therapeutic target in this disease. In summary, this effort has identified novel mutation features in Chinese T-LBLs that can potentially lead to more effective treatments in Chinese population.

Disclosures

Li: Guangdong Province Hospital: Employment.
A Phase II Study of Lenalidomide Plus Rituximab in Patients with Newly Diagnosed Follicular Lymphoma: An Interim Analysis
Authors: Zheng Zhong , Wang Li , Cheng Shu , Xu Pengpeng , Zhao Weili
Published: November 2019

A Phase II Study of Lenalidomide Plus Rituximab in Patients with Newly Diagnosed Follicular Lymphoma: An Interim Analysis

Zhong Zheng1, Li Wang1,2, Shu Cheng1, Peng-Peng Xu1, Wei-Li Zhao1,2

1State Key Laboratory of Medical Genomics, Shanghai Institute of Hematology, Shanghai Rui Jin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China

2Pôle de Recherches Sino-Français en Science du Vivant et Génomique, Laboratory of Molecular Pathology, Shanghai, China

Abstract

Background: Follicular lymphoma (FL) is the most common subtype of indolent non-Hodgkin lymphoma (iNHL). Rituximab plus chemotherapy for FL significantly improves the outcome of the patients, nevertheless, most patients ultimately relapse. Therefore, novel agents along with Rituximab have been applied to increase treatment efficacy in this subset of iNHL patients. Tumor immune eascape plays a crucial role in lymphoma progression. Through modulating tumor microenvironment, lenalidomide, are emerging as effective therapeutic approaches to affect tumor immunity and inhibit lymphoma cells proliferation. However, its anti-tumor activity activity has not yet been assessed in de-novo chinese FL patients. This prospective phase II study is to evaluate the efficacy and safety of lenalidomide in combination with Rituximab (R2) in newly diagnosed FL patients (NCT 03715309).

Methods: Patients with newly diagnosed FL (grade 1 to 3a), aged 16 to 75 years, Eastern Cooperative Oncology Group performance status of 0 to 2 are enrolled. The doses and administration schedule are as follows: rituximab 375 mg/m2 on day 0, lenalidomide 25mg from day 1 to day 10 every 3 weeks for 6 cycles. The primary endpoint is complete response (CR) rate assessed by PET-CT, and secondary endpoints include progression-free survival (PFS), overall survival (OS), overall response rate (ORR) and main adverse events (AEs).

Results: To date, eighty-six patients have been enrolled, with median age of 48 years (range, 22-73). At diagnosis, Seventy-seven patients (89.5%) presented advanced Ann Arbor stage and 19 cases (22.0%) showed elevated serum LDH level. Twenty-nine patients (33.7%) had multiple extra-nodal sites involving bone marrow, bone, gastrointestinal, and spleen. Twenty-one patients (24.4%) had IPI scores ≥ 3. For Sixty-four patients available for response evaluation, the CR rate was 81.2% (52/64) and the ORR was 90.1% (58/64). Grade 3-4 neutropenia was found in 18 cases (20.9%). No grade 3-4 thrombocytopenia and grade 3-4 anemia were observed. For non-hematological AEs, cutaneous reactions and tumor flare reaction were observed in 12 cases (13.9%) and in 1 case (1.16%), respectively, while no grade 4 non-hematological AEs were presented.

Conclusion: Lenalidomide Plus Rituximab as first-line therapy for de novo patients with FL showed encouraging response and good tolerability.

Disclosures

No relevant conflicts of interest to declare.
AAV8-Specific Immune Adsorption Column: A Treatment Option for Patients with Pre-Existing Anti-AAV8 Neutralizing Antibodies
Authors: Kruzik Anita , Raim Roland , Voelkel Dirk , Weiller Markus , Hoellriegl Werner , Scheiflinger Friedrich , Rottensteiner Hanspeter , Reipert Birgit M , de la Rosa Maurus
Published: November 2019

Background: Pre-existing neutralizing antibodies (NAbs) are a major hurdle in adeno-associated virus 8 (AAV8) gene therapy and result in patients being excluded from clinical trials. Prevalence data published by Kruzik et al. suggest that up to 50% of patients have biologically relevant NAbs against AAV8, excluding them from current clinical trials. We developed a strategy to circumvent pre-existing anti-AAV8 immunity and specifically remove anti-AAV8 antibodies.

Aim: Develop an AAV8-specific immune adsorption column (IAC) to specifically remove anti-AAV8 antibodies.

Methods: An AAV8-specific IAC was developed. Protocols were optimized to couple AAV8 capsids to activated sepharose. All procedures complied with regulatory requirements to accelerate clinical development. Accompanying mouse studies tested potential concomitant immune suppressive regimens in Bl/6 mice using AAV8-FIX vectors. NAbs were assessed using validated NAb assays of confirmed biological relevance. Titers of AAV8 binding antibodies were assessed by ELISA and T-cell responses by IFN-g ELISpot.

Results: Optimized coupling protocols revealed that AAV8 empty capsids can be linked in a stable way to resin. IAC was tested in vitro and in macaques: An IAC could be developed fitting into approved and marketed platforms for immune apheresis. By applying standard experimental procedures for apheresis column development, we showed an IAC can deplete AAV8 binding antibodies and NAbs with greater efficacy than a pan-Ig adsorber (Therasorb-IgOmni). We mimicked treatment of patients in vitro by applying different treatment cycles to plasma reservoirs and showed anti-AAV8 titers of 1:5260 could be depleted to undetectable levels in 4 to 5 treatment cycles. These data are supported by in vivo studies. The IAC is part of a strategy that uses concomitant immunosuppressants to block potential anti-AAV8 T-cell responses.

Conclusion: IAC is an enabler for treatment of patients with pre-existing immunity against AAV8 and would also facilitate re-administration. IAC is intended to be applied in combination with Takeda's AAV8 based hemophilia programs.

Disclosures

Kruzik: Baxalta Innovations GmbH, a Takeda company: Employment. Raim:Baxalta Innovations GmbH, a Takeda company: Employment. Voelkel:Baxalta Innovations GmbH, a Takeda company: Employment. Weiller:Baxalta Innovations GmbH, a Takeda company: Employment. Hoellriegl:Baxalta Innovations GmbH, a Takeda company: Employment, Equity Ownership. Scheiflinger:Baxalta Innovations GmbH, a Takeda company: Employment, Equity Ownership. Rottensteiner:Baxalta Innovations GmbH, a Takeda company: Employment, Equity Ownership. Reipert:Baxalta Innovations GmbH, a Takeda company: Employment, Equity Ownership. de la Rosa:Baxalta Innovations GmbH, a Takeda company: Employment.
Epigenetic Plasticity of Type II Innate Lymphoid Cells in the Lower Gastrointestinal Tract
Authors: Laurie Sonia J. , Bruce Danny W. , Bommiasamy Hemamalini , Noel Melodie P , Foster Joseph P. , Davis Ian J. , Pattenden Samantha G. , Serody Jonathan S
Published: November 2019

Though hematopoietic stem cell transplantation (HSCT) is the preferred treatment for a variety of blood malignancies, its use is limited by the development of acute graft-versus-host disease (aGvHD). Type II innate lymphoid cells (ILC2s) are immune cells that play an important role in maintaining mucosal homeostasis, and our lab has previously shown that ILC2s in the gastrointestinal tract (GI) are sensitive to conditioning therapy prior to HSCT. Strikingly, we have demonstrated that the infusion of activated donor ILC2s markedly reduces aGvHD-associated mortality. We therefore wanted to investigate the mechanism of the loss of protective ILC2s from the GI tract. We hypothesized that ILC2s fail to repopulate the gut after HSCT due to inflammatory environmental cues that convert ILC2 precursors to an alternate, ILC1- or ILC3-like fate. Thus, we evaluated the impact of cytokines associated with commitment on murine ILC2s by exposing them to cytokines that may promote differentiation to an ILC1 or ILC3 fate (IL-1b/IL-12/IFN-γ and TGF-b/IL-6/IL-23, respectively). We found ILC2 cells acquired the ability to secrete TNF and IL-17 after in vitro skewing (with these lineage-defining cytokines. To test the ability of these "ex-ILC2" cells to home to other tissues in vivo, GFP-ILC2s were infused into recipients at the time of transplantation. We tracked GFP-ILC2s to the liver and spleen, where they made IFN-g and IL-17 and expressed transcription factors associated with the ILC1 and ILC3 lineages (Figure 1). Next we assessed the ability of cytokines alter ILC2 fate via epigenetic reprogramming by using ChIP-sequencing to evaluate the presence of histone marks that may underlie cellular plasticity. We show that these changes are associated with alterations in epigenetic marks around pioneer, lineage-determining factors. We therefore chose to test a screen of compounds known to modulate a variety of epigenetic targets to ask if they can maintain or convert ILC2s to alternate fates and identified a number of compounds that target bromodomains, methyltransferases, and histonedeacetylases, respectively, that alter the viability and differentiation of ILC2s into an "ex-ILC2"-like phenotype. Preliminary work suggests that maintenance ofG9a expression is able to rescue the loss of ILC2s, which is being tested in vivo. Taken together, these data provide new insights into mechanisms by which innate lymphoid cell precursors are epigenetically regulated, providing novel approaches to treating aGvHD following HSCT.

Figure 1

Disclosures

Davis: Triangle Biotechnology: Equity Ownership, Membership on an entity's Board of Directors or advisory committees. Pattenden:Triangle Biotechnology, Inc.: Equity Ownership, Other: Inventor on intellectual property. Serody:Merck: Research Funding; GlaxoSmithKline: Research Funding.
This Is a Title in Title Case: Prediction of CML Evolution By Telomere Length Analysis at the Single-Chromosome Level
Authors: Lu Jingyuan , Zhou Yingxing , Yan Xiaomei , Lu Quanyi
Published: November 2019

Telomeresplay a vital role in DNA repair activities and protecting chromosomes from degradation[1]. Telomeres are shortened during each cell division because of the end replication problem. After several cell division, telomeres are shortened to a critical length (< 3 kb), and eventually lead to overall genomic instability and triggering the DNA damage response[2-3], which is related to the cancerization of numerous cancers. The commonly used method to estimate telomere length isterminal restriction fragment (TRF) basedon southern blot, which requires thousands of cells and provides only a crude estimate of the average telomere length of all cells analyzed. However, it is believed that the frequency of critically short telomeres, rather than the mean telomere length, is a crucial factor for telomere dysfunction. Therefore, analysis of telomere length at the single-chromosome level is necessary to determine the frequency of critically short telomeres.

Here, we describe the development of a high-throughput method for telomere length analysis at the single-chromosome level by using a laboratory-built high-sensitivity flow cytometer (HSFCM)[4] combined with targeted fluorescent peptide nucleic acid (PNA) probes. The unambiguous detection of the telomere signalsfrom a single chromosome was achieved via HSFCM analysis. The fluorescence intensity of single chromosome was converted to the probe number by a calibration curve, and was further transformed to the base pair number of the telomere. Five representative cell lines were analyzed to compare their telomere length and the ratio of critically short telomeres at the single-chromosome level. The potential of using frequency of short telomere for disease treatment monitoring is examined by analyzing the telomere length in lymphocyte of leukemia patients. The abundance of short telomeres was compared between healthy donors and patients with chronic myeloid leukemia (CML) to see whether it can be used to predict the efficacy of therapeutics. Moreover, the quantity of short telomeres was compared among patients with acute leukemia, patients in different phases of CML and healthy donors to see whether it can be used as a marker for disease progression prediction in CML.

Reference

[1] Blackburn E. H., Epel E. S., Lin J., Science,2015, 350, 1193-1198.

[2]Collado M., Blasco M. A., Serrano M., Cell, 2007, 130, 223-233.

[3] Deng Y., Chan S. S., Chang S., Nat. Rev. Cancer, 2008, 8, 450-458.

[4] Yang LL, Zhu SB, Hang W, Wu LN, Yan XM, Anal. Chem., 2009, 81, 2555-2563.

Figure 1

Disclosures

No relevant conflicts of interest to declare.