bluuebunny/crossref_metadata_2025_split huggingface dataset:
Bone Marrow Memory CD8+ T Cells Play a Supporting Role in Hematopoiesis
Authors: Geerman Sulima , Pascutti Fernanda M , Bhushal Sudeep , Nolte Martijn A. 
Published: October 2019

Abstract
The bone marrow (BM) has an important function as primary lymphoid organ through the process of hematopoiesis. This process is sustained by hematopoietic stem cells (HSCs) and their life-long production of new blood cells, which is made possible by their unique ability to self-renew. Next to this, BM also functions as a secondary lymphoid organ, as it can mediate primary T cell responses against invading pathogens. We and others have shown that activated T cells can influence the hematopoietic process through the production of pro-inflammatory cytokines. This indicates that adaptive immune responses and hematopoiesis are intertwined in the BM, though most of the cellular and molecular interactions that occur during this crosstalk are yet unknown.
Based on previous observations that T cell deficient mice have altered hematopoiesis and that depletion of T cells from allogeneic BM grafts compromises HSC engraftment, we questioned to what extent BM T cells can directly affect the function of HSCs. To test this, we sorted and co-cultured murine BM T cells with HSCs (Lin-Sca-1+c-Kit+CD48-CD150+). We found that particularly BM CD8+ central memory (CD44+CD62L+) T cells (Tcm) enhance the capacity of HSCs to self-renew. Furthermore, we found that TCR-transgenic mice, which do not have memory T cells, have lower numbers of HSCs, which could subsequently be increased by transferring BM CD8+ Tcm. Remarkably, an increase in HSC numbers was also observed when HSCs were cultured with only supernatant derived from BM CD8+ Tcm. Moreover, HSCs cultured with supernatant from BM CD8+ Tcm and later transplanted in myeloablated hosts displayed a strongly enhanced ability to restore hematopoiesis. Importantly, the strong impact of BM T cells on HSCs was not only apparent in the steady state situation, but also following a viral infection with either acute or chronic lymphocytic choriomeningitis virus (LCMV). We could establish that both acute and chronic LCMV-specific CD8+ T cells or supernatant from these cells could increase the HSC self-renewal capacity.
In conclusion, our findings demonstrate that BM memory CD8+ T cells can positively influence the function of HSC through soluble mediators. We postulate that this process is particularly relevant after immune activation, in order to protect and/or restore the HSC pool and the subsequent hematopoietic recovery. We are currently using a proteomics approach to identify these soluble mediator produced by CD8+ T cells.

Disclosures
No relevant conflicts of interest to declare.
Diagnostic Efficacy of Whole-Exome Sequencing in 250 Patients with Congenital Bone Marrow Failure
Authors: Muramatsu Hideki , Okuno Yusuke , Yoshida Kenichi , Doisaki Sayoko , Hama Asahito , Wang Xinan , Narita Atsushi , Kawashima Nozomu , Xu Yinyan , Sakaguchi Hirotoshi , Takahashi Yoshiyuki , Sanada Masashi , Shiraishi Yuichi , Chiba Kenichi , Tanaka Hiroko , Miyano Satoru , Ogawa Seishi , Kojima Seiji 
Published: October 2019

Abstract
Introduction: Congenital bone marrow failure syndromes (CBMFSs) are a heterogeneous class of diseases with overlapping phenotypes. Therefore, a precise and comprehensive genetic diagnostic system is strongly warranted to arrive at appropriate clinical decisions to avoid ineffective therapies and/or lethal complications of allogeneic hematopoietic stem cell transplantation. However, a large panel of newly identified causative genes of CBMFSs have been identified in recent years; therefore, it is virtually impossible to establish a routine genetic diagnostic test using conventional Sanger sequencing. Whole-exome sequencing (WES) is a promising solution for the diagnosis of inherited diseases because it tests virtually all genes simultaneously. For the introduction of WES into clinical practice, it is necessary to clarify whether this technique has superior diagnostic efficacy to conventional clinical genetic tests.
Methods: We performed WES in 250 patients with CBMFSs lacking genetic diagnoses. Exome capture was performed using the SureSelect® Human All Exon V3–5 kit (Agilent Technologies, Santa Clara, CA, USA), which covers all known coding exons, followed by massively parallel sequencing using the HiSeq 2000 Sequencing System (Illumina, San Diego, CA, USA). Our established pipeline for WES (genomon: http://genomon.hgc.jp/exome/) detected >20,000 candidate variants per patient. Diagnoses were based on variants of 130 genes with pathogenicities confirmed by published studies.
Results: Genetic diagnoses were possible in 68 patients (27%). The best efficacy was achieved in patients with Fanconi anemia [35/73, 48%; FANCG (n = 17), FANCA (n = 14), FANCB (n = 1), FANCF (n = 1), SLX4 (n = 1), and BRCA2 (n = 1)], although Sanger sequencing was not applied because of the large sizes of its causative genes. Encouraging results were obtained in patients with Diamond–Blackfan anemia [11/ 61, 18%; RPS26 (n = 3), RPS7 (n = 2), RPS19 (n = 2), RPL5 (n = 2), RPL35A (n = 1), and RPL11 (n = 1)] and dyskeratosis congenita [7/29, 24%; TERT (n = 3), TINF2 (n = 2), and DKC1 (n = 2)]. Five genetic diagnoses (7%) were inconsistent with clinical diagnoses, possibly because of overlapping disease phenotypes.
Conclusion: Relative to conventional genetic testing, WES was found to be effective for the diagnoses of CBMFSs. Furthermore, the efficacy of WES will increase as our knowledge of gene mutations expands. In conclusion, the use of WES in clinical practice is warranted.

Disclosures
No relevant conflicts of interest to declare.
Expression of c-MYC and MMP9 in Alternatively Activated Macrophages (M2) of Classical Hodgkin Lymphoma
Authors: Yang Long , Harris Jonathan A. , Ibrahim Sherif , Liu Cynthia 
Published: October 2019

Abstract
Classical Hodgkin Lymphoma (CHL) is a monoclonal lymphoid neoplasm that accounts for approximately 30% of all lymphomas in the United States. Although multiagent chemotherapy and radiation therapy have made CHL a highly curable disease, approximately 15-20% of patients are refractory to the treatment or relapsed after remission. The rich inflammatory cellular microenvironment plays an important role in the pathogenesis of CHL. Gene-expression profiling has identified a gene signature of tumor-associated macrophages (TAMs) that was significantly associated with primary treatment failure. The potential of macrophage-specific markers, CD68 and CD163, as prognostic markers for resistance to therapy in CHL patients has been investigated; however, these studies generated inconsistent results. Roughly TAMs could be characterized into two distinct polarization states; the classically activated type 1 macrophages (M1) associated with acute inflammation and T-cell immunity and the alternatively activated type 2 macrophages (M2) associated with the promotion of the tumor growth. In the present study, we hypothesize that the specific type of TAMs, M2, may predict poor prognosis in CHL. To test this hypothesis, we examined the correlation between expressions of two M2-specific markers, c-MYC and matrix metalloproteinase 9 (MMP-9), with disease recurrence in a total of 42 cases of CHL, mostly nodular sclerosis subtype. Both c-MYC and MMP-9 are good candidate markers because c-MYC has been shown to be a specific M2 marker and plays a central role in M2 differentiation. In addition, matrix metalloproteinase 9 (MMP-9) is a M2 specific marker that directly related to its protumoral function. Immunohistochemical staining showed that both c-MYC and MMP9 are expressed in a subset of macrophages, but not in Hodgkin Reed-Sternberg (HRS) cells. By comparing the percentage of positive stains of these two markers within HRS rich areas in the primary tumor samples, we found that there was no significant difference in either c-MYC or MMP9 staining between patients who later recurred (n=6) or did not recur (n=36). The role of TAMs or an unidentified subtype of TAMs in predicting resistance to therapy in CHL patients remains to be determined.

Disclosures
No relevant conflicts of interest to declare.
Clinical Presentation and Outcome of ABVD-Treated Elderly Versus Younger Patients with Classical Hodgkin´s Lymphoma
Authors: Acosta-Maldonado Brenda Lizeth , Ramirez-Ibarguen Ana , Grimaldo-Gomez Flavio A , Rivas-Vera Silvia 
Published: October 2019

Abstract
Background
Approximately 20% of classical Hodgkin’s Lymphoma (cHL) patients are over 60 years of age. There is no standard of care for this age group. Although most patients receive ABVD (Doxorubicin, bleomycin, vinblastine and dacarbazine), a standard regimen in younger patients, little is known regarding the clinical presentation of cHL, and the long term efficacy of ABVD in young and elderly Latin American Mestizo patients.. Defining whether individuals in all age groups and social circumstances benefit from a common treatment approach requires investigation..
Thus we sought to evaluate and compare the clinical presentation and the efficacy of ABVD in the elderly (60 years of age and above) versus the younger population of individuals with cHD.
Methods
We retrospectively analyzed 44 consecutive cHL patients treated with ABVD at the National Cancer Institute in Mexico between 2006 and 2013, as compared to 218 patients under 60 years of age.
Results
Median age for the elderly was 65 (60-89y), without a difference in gender (21m/2 f). According to Ann Arbor staging 6-8-13-14 were stage I, II, III, or IV , respectively. The International Prognostic Score (IPS) was > 3 in all patients. B symptoms were present in 34 patients (77.3%). Mixed cellularity subtype was present in 52.3% of cases and nodular sclerosis subtype (34.1%). The great majority of patients (93%) received at least 1 ABVD (1-6), 27 patients received chemotherapy alone, 14 patients received combined modality treatment and 3 patients only received radiotherapy. Twenty four patients achieved a CR (54.5%), 5 partial responses, 5 had refractory /relapsed disease and 10 couldn’t be evaluated. Unlike younger patients a better response was not demonstrated with combined treatment modality (Table 1). Overall survival for the whole group at 7.2 years was estimated at 75.4% (95% CI 57.1-79.4) The OS (7 y) in elderly group was 75% versus 92% in younger group, however patients with age >75 had a lower survival, 50%. The median follow-up was of 8 years.
Summary
Our study showed no clinical differences between elderly vs younger population and confirms the efficacy of ABVD in elderly patients. The lower OS compared to other series may be related to the advanced stage of disease and poor IPS in our series Table 1.Clinical characteristics of elderly versus younger patientsFeature< 60 years (%)>60 years (%)P valueGenderMale/female61.8/38.247.5/52.30.10B Symptoms77.077.30.86Ann Arbor stage I II III IV7.424.931.835.913.618.224.538.60.45IPS >348.865.90.07Bulky disease48.838.60.26Histology Mixed cellularity Nodular Sclerosis43.848.852.334.10.60Treatment ABVD ABVD + RT RT Other42.455.32.32.361.431.86.84.50.37Response CR PR Relapse/progression NV60.89.718.910.654.511.411.422.70.37Overall survival (8 years)92%75%0.000

Disclosures
No relevant conflicts of interest to declare.
Peripheral Absolute Neutrophil, Mmonocyte and Lymphocyte Counts and Clinical Ooutcome in Diffuse Large B-Cell Lymphoma
Authors: Chen Yiming , Rodriguez Maria Alma , Feng Lei , Bi Weiqi , Zhang Liang , Wang Michael 
Published: October 2019

Abstract
Background
Baseline levels of peripheral absolute neutrophil (ANC), lymphocyte (ALC) and monocyte (AMC) counts have prognostic value in solid tumors. There are limited data on their impact on outcomes in patients with diffuse large B-cell lymphoma (DLBCL). The present study is to confirm whether the baseline levels of peripheral ANC, ALC and AMC can predict outcomes in patients with newly diagnosed DLBCL.
Patients and Methods
Data from lymphoma database at the University of Texas MD Anderson Cancer Center (UTMDACC) were used. The correlation of baseline ANC, AMC and ALC with progression-free survival (PFS) and overall survival (OS) were analyzed in 817 newly diagnosed previously untreated patients with DLBCL who received frontline treatments at UTMDACC between 2000-2011. Kaplan-Meier method was used for time-to event analysis including PFS and OS. Multi Cox proportional hazards model were fitted to evaluate important patient characteristics on the time-to event endpoints.
Results
Median ages at diagnoses were 58 years (range, 18 to 91 years). Seventy-nine percent of patients received conventional immunochemotherapy(CHOP-R) while 21 % of patients had intensive immunochemotherapy (HyperCVAD-R or R-EPOCH) or intensive chemotherapy(n=5; HyperCVAD or R-EPOCH) due to CD 20 negative; two patients received radiation therapy only. IPI low, intermediate and high were 38%, 46% and 16%, respectively. Overall response rate was 98% with a complete remission rate of 73%. Patients with abnormal ANC (> 7.3 X109/L) had an inferior PFS (P<0.001) and OS (P<0.001) compared to patients with ANC ≤ 7.3 X109/L; patients with abnormal ALC (< 1.0 X109/L) had significant short PFS (P<0.001) and OS (P<0.001) compared to patients with ALC ≥ 1.0 X109/L; similarly, patients with abnormal AMC (> 0.7X109/L) had inferior PFS (P< 0.001) and OS (P< 0.001) compared to patients with AMC≤ 0.7 X109/L. Multivariate analysis identified abnormal ALC and AMC as an independent adverse prognostic factors for PFS (hazard ratio [HR] 1.39 and 1.49, respectively; P<0.016 and P=0.004, respectively) and abnormal ANC and AMC as an independent adverse prognostic factors for OS (P=0.017 and P=0.024, respectively; HR 1.56 and 1.42, respectively).
Conclusions
An abnormal Neutrophil, lymphocyte and monocyte counts in peripheral blood are independent, poor prognostic factor for PFS and OS in patients with DLBCL. Incorporation of these immunological markers into IPI prognostic model may improve outcome prediction.

Disclosures
No relevant conflicts of interest to declare.
Phase 2 Trial of Entospletinib (GS-9973), a Selective SYK Inhibitor, in Follicular Lymphoma (FL)
Authors: Sharman Jeff P. , Klein Leonard M. , Boxer Michael , Kolibaba Kathryn S. , Hawkins Michael J. , Abella Steve , Wu Meihua , Yasenchak Christopher A. 
Published: October 2019

Abstract
Background: Spleen tyrosine kinase (Syk) is a mediator of B-cell receptor signaling in normal and transformed B-cells. Entospletinib is an orally bioavailable, selective inhibitor of Syk (Kd 7.6 nM, no other kinase < 100 nM).
Methods: This Phase 2 trial is evaluating Entospletinib 800 mg BID in a 41 subject cohort with previously treated FL in a study of 165 subjects with lymphoid malignancies. Tumor imaging was planned at weeks 8, 16, 24 and then every 12. Tumor response was assessed per Cheson 2007 criteria.
Results: For the subjects with FL included in this analysis, median age was 67 years (range 41 – 89), 49% were male. The median number of prior Rx regimens was 2 (range 1-8). Prior treatments (Rxs) included anti-CD20 antibodies (rituximab 100%, ofatumumab 5%), alkylating agents (95%; bendamustine 51%) and anthracyclines (51%). Baseline risk factors: Ann Arbor Stg III-IV (66%), Gr 3a FL (27%), FLIPI ≥3 (34%). At the time of this analysis, 41 subjects with follicular lymphoma were enrolled and 38 subjects have been treated through first response assessment (1 subject ongoing prior to first response assessment, 1subject discontinued due to AE and 1 subject withdrew consent). Entospletinib was generally well tolerated. The most common TEAEs (any Grade/≥Gr 3, independent of causality) were fatigue (46%/12%), nausea (42%/2%), diarrhea(27%/0%), decreased appetite (22%/0%), vomiting (22%/0%) and common laboratory abnormalities were increased AST (37%/17%), increased ALT (34%/20%), increased total bilirubin (27%/5%), anemia (34%/10%) and neutropenia (22%/10%). Reversible Grade 3 or 4 ALT/AST elevations occurred in 8 (19.5%) FL subjects. 3 subjects died while on study: 2 from progressive disease and 1 from acute renal failure investigator reported as unrelated to study drug. Investigator response assessments are available for 29 subjects, 16/29 (55%) subjects experienced reduced tumor bulk measured by SPD; 3 (10%) achieved a decrease of ≥ 50%. CR has not been observed at this early evaluation. 14/41 subjects continue on Rx. Median duration of Rx for all patients was 15 weeks. Among all patients who experienced reduction in tumor volume, median duration of Rx was 25 weeks (range 4-51).
Conclusions: Entospletinib monotherapy given with this dose and schedule was generally well tolerated and demonstrated moderate activity in subjects with advanced relapsed FL, including those with poor prognostic features. Updated data with longer follow-up duration will be presented at the meeting.

Disclosures
Sharman: Gilead Sciences: Research Funding. Klein:Gilead Sciences: Research Funding. Boxer:Gilead Sciences: Research Funding. Kolibaba:Gilead Sciences: Research Funding. Hawkins:Gilead Sciences: Research Funding. Abella:Gilead Sciences: Employment, Equity Ownership. Wu:Gilead Sciences: Employment, Equity Ownership. Yasenchak:Gilead Sciences: Research Funding.
GADD45a Is a Tumor Suppressor in BCR-ABL-Driven Leukemogenesis
Authors: Mukherjee Kaushiki , Sha Xiaojin , Skorski Tomasz , Bhatia Ravi , Hoffman Barbara , Liebermann Dan 
Published: October 2019

Abstract
The bcr-abl oncogene causes chronic myelogenous leukemia (CML). Growth arrest DNA damage 45a (Gadd45a) gene, a member in the gadd45 family of genes including Gadd45b & Gadd45g, is upregulated during myeloid lineage terminal differentiation. It is involved in G2/M cell cycle arrest and apoptosis in response to multiple stressors, including genotoxic and oncogenic stress . To investigate the effect of GADD45A in the development of CML, syngeneic wild type lethally irradiated mice were reconstituted with either wild type or gadd45a null myeloid progenitors transduced with a retrovirally expressed 210-kD BCR-ABL fusion oncoprotein. It was observed that loss of gadd45a accelerates BCR-ABL driven CML resulting in the development of a more aggressive AML like disease. Loss of Gadd45a resulted in increase of BCR-ABL expressing leukemic Stem/progenitor cells (GFP+Lin- cKit+Sca+). BCR-ABL transformed GADD45A deficient progenitors exhibit increased proliferation and decreased apoptosis, associated with enhanced PI3K-AKT-mTOR-4E-BP1 signaling and upregulation of p30C/EBPα and MCL-1 expression. Moreover, loss of Gadd45a was observed to facilitate transformation of BCR-ABL expressing BM cells to factor independence. These results provide novel evidence that gadd45a functions as a suppressor of BCR/ABL driven myeloid leukemogenesis. Since Gadd45a functions as a tumor suppressor in murine BCR-Abl driven leukemia, the status of Gadd45a mRNA expression levels was also investigated in human CML samples by utilizing real time PCR analysis. It was found that Gadd45a transcript levels were significantly up-regulated in chronic phase I CML samples. However in chronic phase-II, accelerated phase and blast crisis phase samples, expression was significantly down-regulated relative to normal controls. Thus, Gadd45a expression was observed to be altered in human CML samples correlating with disease progression. These results provide novel evidence that gadd45a functions as a suppressor of BCR/ABL driven myeloid leukemogenesis. These data also provide the impetus to further elucidate the role Gadd45a plays in suppressing the development of CML, and explore how its loss contributes to the progression of CML to a more aggressive leukemic phenotype.

Disclosures
No relevant conflicts of interest to declare.
Index to Predict MMR at 12 Months in Chronic Phase Chronic Myeloid Leukemia Patients Based on the Area Under the Lymphocyte Curve
Authors: Murai Kazunori , Ito Shigeki , Kowata Shugo , Oyake Tatsuo , Ishida Yoji 
Published: October 2019

Abstract
BACKGROUND: Dasatinib is a second-generation BCR-ABL inhibitor that has a 325-fold higher potency than imatinib and a 16-fold higher potency than nilotinib in vitro. Recent reports suggested that absolute lymphocytosis in response to dasatinib for chronic phase (CP) –chronic myeloid leukemia (CML) might be associated with favorable response. However, as the fluctuation of lymphocytosis was often observed, it is sometimes difficult to define lymphocytosis. To evaluate how much increase and how long continuation of lymphocytosis affects the better clinical efficacy, we developed an index (DL index; Dasatinib induced Lymphocytosis index) that combines duration and increase of lymphocytosis. The aim of this study was to test the DL index as a predictor of Major Molecular Response (MMR) at 12 month.
PATIENTS and METHODS: The DL area was based on a graph plotting the absolute lymphocyte counts and was the area under the lymphocyte curve using trapezoidal method. DL index was DL area divided by weeks. For example, DL index12W was defined as (DL area from 4 to 12 weeks) divided by (12-4) weeks. We tested the DL index12W in 30 patients who developed MMR at 12 months (MMR group) and 9 patients without MMR (non-MMR group).
RESULTS and DISCUSSION: DL score12w in MMR group and non-MMR group were 2714.0 (95% CI; 2286.0-3142.2, n=30) and 1559.9 (95% CI; 1006.9- 2113.0, n=9) respectively. There was a significant difference between MMR group and non-MMR group in DL index12W by t-test(Figure 1, p=0.0016). A significant linear relationship was obtained between DL index12w and the peaked lymphocyte counts within 12 weeks by Spearman's correlation coefficient rank test (Figure 2, p=1.97E-07, correlation coefficient=0.91).
The DL index12w, 2,113, which was the upper limit of non-MMR group 95% CI, is equivalent to 2,909/µl in lymphocyte count, and the DL index12w, 2,286, which is the lower limit of MMR group 95% CI, equivalent to 3,123/µl in lymphocyte count. Therefore, we divided all patients (n=39) into two groups, equal or more than 3,123/µl and less than 3,123/µl in the peaked lymphocyte counts. The patients with the peaked lymphocyte counts ≥ 3,123/µl reached MMR at 12 months (19/20=95.0%) to compare those less than 3,123/µl (11/19=57.9%) (p=0.0075).
CONCLUSION: The peaked lymphocyte number, induced by the DL index12w, which calculation was extremely easy, was associated with molecular response. These data suggests that this tool can be useful in interpreting the results of molecular response at 12 months.
Figure 1 Figure 1.
Figure 2 Figure 2.

Disclosures
No relevant conflicts of interest to declare.