bluuebunny/crossref_metadata_2025 huggingface dataset:
Malignant Lymphoma in Primary Sjögren Syndrome: Incidence and Risk Factors.
Authors: López Andrés , Solans-Laque Roser , Bosch-Gil Josep A , Gironella Mercedes , Purroy Noelia , Marta Campillo , Vallespi Teresa
Published: October 2019

Abstract 4991

Background

Primary Sjögren Symdrome (PSS) is the second common systemic autoimmune rheumatic disease whose main histological feature is a focal infiltration of the exocrine glands by activated B and T lymphocytes. PSS is considered a benign autoimmune condition with a chronic indolent course. However, several reports have noted an increased incidence of malignant lymphoma with an estimated risk of up to 44 times greater than in the normal population. The aims of the present study are to assess the cumulative incidence of lymphoma in patients diagnosed of PSS, as well as to identify risk factors for developing this condition.

Methods

All patients diagnosed since 1988 and followed-up for longer than 2 years at our institution, were included. The pathological, immunological, and clinical features of each case of PSS were retrospectively examined. These data were cross-examined with the cumulative incidence of lymphoma in this group of patients, in order to determine common findings in patients later developing lymphoid malignancies.

Results

272 patients had been diagnosed as having a PSS in this time period, of which 244 [9 males, 235 females; median age 58 years (range 17 to 88)] had been followed for more than 2 years. After a median follow up of 9.6 years, 11 (4.5%) patients developed a lymphoma, after 6 to 171 months from diagnosis. The cumulative risk for developing lymphoma at 5, 10 and 15 years was 3.4, 4.1, and 9.3%, respectively. Risk factors associated with the development of lymphoma were skin purpura/vasculitis, lymphopenia and low levels of C3 and C4 complement fractions. Low levels of C3 and C4 were also found to be associated with an earlier development of lymphoma. Conversely, skin purpura/vasculitis and lymphopenia were both associated with the development of lymphoma at a later stage. The hystological analysis showed mucosal-associated lymphoid tissue lymphoma in 5 patients, diffuse large B cell in 3, and follicular lymphoma in 3 cases. Clinically, lymphomas had extranodal presentation in 8 out of the 11 cases, involving the salivary glands in 5, lacrimal glands in 1, and soft tissues and lung in 1 case, respectively. Only 3 patients had received immunosuppressive therapy prior to the diagnosis of lymphoma, and EBV was not found in the histological examination of any of these cases.

Conclusion

Patients with PSS are at a higher risk for developing lymphoma. Our study suggests low levels of C3 or C4, as well as the presence of purpura or lymphopenia delimitates a group of patients at a higher risk, so that such patients may benefit from a closer follow up.

Disclosures

No relevant conflicts of interest to declare.
Primary Mediastinal Large B-Cell Lymphoma: A Single Center Experience in Korea.
Authors: Ahn Heui June , Cho Yoo Jin , Kang Myoung Joo , Choi Dae Ro , Kim Shin , Lee Dae Ho , Kim Sang We , Lee Jung Shin , Huh Jooryung , Suh Cheolwon
Published: October 2019

Abstract 5013

Introduction

Primary mediastinal large B-cell lymphoma (PMBCL) was formally established as a distinct subtype of diffuse large B-cell lymphoma (DLBCL). Some studies indicated that patients with PMBCL have an aggressive clinical course with short median survival but more recent studies reported a relatively good response rate and survival. Therefore, controversies still exist regarding the response to therapy and prognosis of patients with PMBCL.

Patients and methods

Between July 1993 and July 2008, a total of 26 patients with PMBCL were identified at Asan Medical Center, Seoul Korea. We retrospectively reviewed the clinic-pathologic features and clinical outcomes of them in comparison with 597 patients diagnosed with non-mediastinal DLBCL during the same period.

Result

Out of the 26 patients, 17 (65.4%) were females and 9 (34.6%) males, while out of the 597 patients, 257 (43.0%) were females and 340 (57.0%) males (p=0.025). The median age of the PMBCL patients was 31.5 years old (range 15-78 years old), while that of the DLBCL patients was 56.0 years old (range 15-85 years old). Out of the 26 patients, 14 (53.8%) had a Ann Arbor stage III or IV disease, 7 (26.9%) had B symptoms, and by the IPI, 11 were in low, 9 in low intermediate, 2 in high-intermediate and 4 in high risk group. Out of 24 patients treated with front-line therapy CHOP or R-CHOP, 17 (70.8%) reached a CR, while 1 PR patient reached a CR after being treated with high-dose chemotherapy followed by autologous stem cell transplantation. Five refractory patients were treated with high-dose chemotherapy followed by autologous stem cell transplantation, but among them only one reached a CR and 4 died of disease progression. With a median follow-up of 41.5 months (range 1-92 months), 5-year survival rates of PMBCL and non-mediastinal DLBCL patients were 69% and 65.7%, respectively (p=0.982, Log Rank).

Conclusion

There was no difference between PMLBL and non-mediastinal DLBCL in terms of clinical features and outcomes.

Disclosures

No relevant conflicts of interest to declare.
Is Skull-Base to Proximal Thigh PET/CT Enough? Do We Need Vertex to Toe PET/CT Imaging for Cutaneous T-Cell Lymphoma (CTCL) Patients?.
Authors: Woodward Catherine S , King Nassoma , Prince Kenneth , Parker Sareeta , Halkar Raghuveer , Lechowicz Mary Jo
Published: October 2019

Abstract 5020

INTRODUCTION

Positron emission tomography (PET) imaging using [F-18] fluorodeoxyglucose (FDG) provides metabolic information valuable to staging and restaging lymphoma patients. The Society of Nuclear Medicine recommends skull-base to proximal thigh imaging to survey the body and recommends vertex to toe PET/CT when the tumor may be in the extremities, e.g. melanoma (Delbeke 2006). As CTCL may develop in any region of the body, we aimed to assess the usefulness of vertex to toe PET/CT images in evaluating CTCL patients.

METHODS

We retrospectively analyzed 79 PET/CT scans from 33 CTCL patients (15 men, 18 women, age range: 7-83, median: 56) who were imaged between 2004 and 2008. All studies were performed on GE Discovery LS 8 slice MDCT PET/CT scanner or GE Discovery DST 16 slice MDCT PET/CT scanner (Milwaukee, WI) and the images were interpreted on a GE AW or Xeleris workstation by four board certified nuclear medicine physicians.

RESULTS

The length distribution of the 79 scans is detailed in Figure 1. The 79 scans showed an average maximum SUV of 8.6 +/- 5.2 (range: 2.3 – 21.7). Of the 79 scans, 15 scans showed only cutaneous involvement; 25 scans showed lymph node involvement (as defined by radiologic criteria of lymph node > 1.5 cm, and SUV > 2.5); 10 scans showed likely benign, non-specific uptake in the lymph nodes; 8 scans showed hypermetabolic lymphadenopathy (as defined by SUV> 2.5, lymph node measurement <1.5 cm); 4 scans suggested organ involvement; and 18 scans showed no FDG uptake suggestive of malignancy, usually representing post-treatment patients. 71% of patients with elongated scans (17 of 24 patients) had FDG uptake suggestive of malignancy in areas of the body that would not have been imaged following the standard of skull-base to mid-thigh procedure. For 14 of these 17 patients, the elongated scans demonstrated systemic involvement which did not affect disease management, due to widespread uptake. However, in 3 patients, focal cutaneous uptake noted in the upper or lower extremities played a critical role in staging as there was no FDG uptake suggestive of malignancy in the skull-base to mid-thigh region. These results are demonstrated in Figure 2.

CONCLUSION

3 patients (13% of patients with at least one elongated scan) had FDG uptake only in the extremities. If these patients had been imaged from skull-base to proximal thigh, no suspicious FDG uptake would have been noted, and treatment plan may have been altered. Further studies prospective PET studies are necessary to determine its role in the staging and management of CTCL.

Disclosures

No relevant conflicts of interest to declare.
Prognostic Factors in Aggressive, Non-Primary Cutaneous Peripheral T-Cell Lymphoma in Peru: A Study of 227 Cases.
Authors: Beltran Brady , Morales Domingo , Quinones Pilar , Desposorio Carlos , Sokol Lubomir , Castillo Jorge
Published: October 2019

Abstract 5021

Background

Peripheral T-cell lymphoma (PTCL) is a heterogeneous family of entities with a worse prognosis, stage by stage, than their B-cell counterparts. We have previously reported that Peruvian population has a higher incidence of PTCL, akin to the Asian population. The goal of this study is to therefore investigate the clinical characteristics and prognostic factors in patients with PTCL in a reference center in Peru.

Methods

A total of 227 cases of aggressive, non-primary cutaneous PTCL diagnosed between January 1997 and December 2008 were reviewed, reappraised according to their morphological, immunological and clinical characteristics, and reclassified according to the new WHO classification of lymphoid neoplasms. Kaplan-Meier method was used to estimate survival curves, which were compared using the log-rank test. The multivariate analysis was performed using the Cox proportional-hazard regression test.

Results

The mean age at diagnosis was 57 years (range 14 – 92 years) with a male-to-female ratio of 1:1. According to the new WHO classification of lymphoid neoplasms, 97 cases (43%) were classified as adult T-cell leukemia/lymphoma (ATLL), 84 cases (37%) as PTCL, unspecified (PTCLU), 28 cases (12%) as anaplastic large cell lymphoma (ALCL), 10 cases (4%) as extranodal NK/T lymphoma, nasal type, 4 cases (2%) as angioimmunoblastic lymphoma and 3 cases (1%) were diagnosed with more rare PTCL subtypes. Fifteen percent were stage I, 12% stage II, 16% stage III and 56% stage IV. Distribution based on the International Prognostic Index (IPI) score was: low 24%, low-intermediate 24.5%, intermediate-high 26% and high 25.5%. B symptoms were present in 64% of patients. The median overall survival (OS) for the whole group was 8.4 months, the 2-year OS was 34% and the 5-year OS was 27%. ATLL patients showed a median overall survival of 6.1 months, PTCL of 11.4 months, ALCL of 12.4 months and extranodal NK/T cell lymphoma of 7.8 months. The IPI score, the Prognostic Index for PTCLU (PIT) score and the presence of B symptoms showed statistical significance in both the univariate and the multivariate survival analysis (p=0.001, p=0.005 and p=0.005, respectively).

Conclusions

The distribution of PTCL subtypes in Peru is comparable to the series of patients reported in Asian countries. ATLL and PTCL-U are the most frequent aggressive, non-primary cutaneous PTCL subtypes found. PTCL tends to present with advanced stages and has a poor prognosis, with a 5-year OS of less than 30%. In the multivariate analysis, the IPI score, the PIT score and presence of B symptoms were independent prognostic factors for survival in PTCL.

Disclosures

No relevant conflicts of interest to declare.
Central Nervous System Involvement in Patients with Diffuse Large-B Cell Lymphoma: A Retrospective Analysis of a Single Center in Brazil.
Authors: Rocha Talita , Pinto Maeva , Fortier Sergio Costa , Campos Igor , Paes Roberto , Chiattone Carlos
Published: October 2019

Abstract 5029

Background

Relapse in the central nervous system (CNS) after initial treatment of diffuse large-B cell lymphoma (DLBCL) is an uncommon but serious complication. This retrospective study of a single institution in a Latin American country investigated the incidence of CNS involvement in patients with DLBCL and compared it with results in developed countries.

Patients and methods

A total of 134 patients treated for DLBCL from January 2001 to April 2008 were retrospectively analyzed for incidence of secondary CNS involvement of lymphoma.

Results

Twenty six (19,4 %) patients of this cohort had used rituximab as first line treatment and nine (6,7%) had done IT chemoprophylaxis. Seventy (52%) were stage III and IV. Forty seven (35%) had high intermediate or high international prognostic index. Nine of 134 (6,7%) developed CNS disease after a median observation of 36 months. The median time for the CNS relapse or progression was 7,7 months after diagnostic and all but one patient died despite the treatment proposed. Among the 9 patients that relapsed, seven (77,7 %) had parenquimal CNS involvement. Seven (77,7%) had stage III or IV disease. One (11,1 %) had bone marrow involvement. Two (22,2%) received IT chemoprophylaxis and 3 (33,3%) had used rituximab.

Discussion and Conclusion

We describe here our cases and emphasize that as we know, this is the only Brazilian study investigating this kind of involvement. Different than expected, we found a similar CNS infiltration compared to developed countries cohorts. This suggests that CNS disease is probably related to biologic features than circumstantial issues normally present in developing countries such as late diagnoses and treatment. The evaluation of the risk factors for CNS involvement had not been done due to the small cohort.

Disclosures

No relevant conflicts of interest to declare.
The Prohibitin 3′ Untranslated Region Polymorphisms Are Associated with AML Susceptibility.
Authors: Kim Hye-Ran , Shin Myung-Geun , Kweon Sun-Seog , Kim Hee-Nam , Kim Hyeoung-Joon , Lee Il-Kwon , Shin Jong-Hee , Suh Soon-Pal , Ryang Dong-Wook
Published: October 2019

Abstract 5038

The prohibitin gene, which is located on human chromosome 17q21 encodes a certain regulatory RNA molecule responsible for inhibitory effects and the respective tumor suppressor function. Common C/T and A/G polymorphisms occur at nucleotide 10701 and 10730 respectively in the 3' untranslated region (UTR) of the prohibitin gene. Recent findings indicate that presence of at least one mutant allele within the 3' UTR prohibitin gene polymorphism causes inactivation of bioactive RNA, resulting in loss of its pro-apoptotic function and subsequent risk for malignant growth. These observations led to the hypothesis that individual carrying the prohibitin T allele has increased susceptibility to acute myeloid leukemia (AML). To assess this, we carried out a case-control study of the prohibitin genotype in 381 patients with AML and 372 healthy controls. The distribution of genotypes for the prohibitin 3' UTR polymorphism in patients with AML (P=0.79) and controls (P = 0.10) were in Hardy–Weinberg equilibrium. The prohibitin 3'UTR 10701TT genotype was associated with an increased risk for AML (odds ratios (OR)=2.6; 95 percent confidence interval (CI)=1.90 to 6.19, P = 0.031). However, the prohibitin 3'UTR 10730 polymorphism was not related to AML susceptibility. Haplotype (10701T-10730A) increased risk (OR=2.11; CI=1.28-2.94) for AML. There was a significant difference in the frequency of T-allele carriers between the patients and controls (OR=1.34; CI=1.01-1.77; P=0.046). Interestingly, AML patients having 10730GG genotype had shorter overall survival rate than those with AA and AG genotypes. In conclusion, the prohibitin 3'UTR region genotyping is valuable in assessing risk of AML and estimating prognosis.

Disclosures

No relevant conflicts of interest to declare.
Effects of SHIP Over Expression On the Invasion and Migration of K562 Cells and Its Mechanism.
Authors: Yang Lin , Luo Jianmin , Liu Xiaojun , Wen Shupeng
Published: October 2019

Abstract 5051

Introduction

The phosphatidylinositol-3 kinase (PI3K) pathway plays a central role in regulating numerous biologic processes, including survival, adhesion, migration, metabolic activity, proliferation, differentiation, and end cell activation through the generation of the potent second messenger PI-3,4,5-trisphosphate (PI-3,4,5-P(3)). The SH2 domain-containing inositol polyphosphate 5-phosphatase (SHIP) is known to play an important role in the negative regulation by PI3K-dependent signaling cascades activated by several tyrosine-kinase coupled cytokine receptors. SHIP knockout mice display myeloproliferation and hyper-responsiveness to growth factor stimulation and have short life span with myeloid cell infiltration into vital organs. We have proved SHIP inhibits growth of leukemia cells, and it may have an important role in apoptosis of leukemia cells. However, to date the role of anti-migrated effect of SHIP and its mechanism is unclear.

Methods

Based on the results of our previous studies, with different human leukemia cell lines K562 as study samples, including K562 transfected with wtSHIP (Group A), K562 transfected with muSHIPC1076T (Group B) and K562 transfected with empty vector (Group C); Expression of SHIP at mRNA and protein level was detected by real-time PCR and Western blot, respectively; By Transwell chamber assay was used to count the numbers of Group A and Group B cells that penetrated the matrigel to the back of PVPF membrane after transfection; Primary migration associated factor FAK, p-FAK and NFκB were screened by Western blot. The expression of MMP-2 and MMP-9 was examined by zymography.

Results

Compared with Group C and Group B, the migration of Group A cells was decreased (32±6 vs 78±13 and 83±16)(P<0.01); Over expression of wild-type SHIP does not affect the MMP2 secretion in K562, but site-directed mutant in C terminus of SHIP can significantly block MMP-9 secretion in K562 cells, By Western blot analysis, the expression of p-FAK and NFκB protein in Group A cells were down-regulated to 44% and 63% of the Group C, respectively.

Conclusions

The results confirmed SHIP as a negative regulator for cell migration and invasion in bcr/abl transformed cells through decreasing the MMP-9 expression, which may be induced by reduced phopha-FAK and NFαB expression, and implied that it may function through its PXXP domain.

Disclosures

No relevant conflicts of interest to declare.
Casein Kinase CK2 Inhibition Results in Tumor Suppression.
Authors: Mohammad Hanan , Lindner Daniel j , Kalafatis Michael
Published: October 2019

Abstract 5052

Protein Kinase II (CK2) is a pleiotropic, and ubiquitous serine/threonine kinase taht utilizes both ATP and GTP as phosphate donors. Protein kinase CK2 mostly exists as a tetramer composed of two catalytic subunits α and α‘, which exists in heterogeneous or homogenous nature, and two regulatory β subunits. CK2 is a key regulator of signaling pathways involved in cell cycle, proliferation and apoptosis. It is consistently overexpressed in cancer tissue and capable of shuttling between cellular compartments but mainly localized in the nuclear matrix of cancer cells. CK2 is highly involved in apoptosis suppression, oncogene activation and tumorigenesis. It is also considered a bad prognostic marker in cancer tissue and is suggested to be a promising target for cancer therapy. In this study, we examined the effect, of a specific protein kinase inhibitor, and ATP competitor, 2-dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole (DMAT), on the growth of various cancer types. We have reported DMAT as a potent cancer therapy. In vitro analysis of the viability of hematologic neoplasms and solid tumors, including human lymphoma U937, hormone dependent breast cancer MCF7, hormone independent breast cancer MDA-MB231 and MDA-MB468, and human cervical cancer HeLa cell lines, revealed marked reduction of cellular viability upon treatment with different DMAT concentration at varying time periods. Lymphoma cell line U937 showed an IC50 between 6 -12 μM. A sharp decrease in cancer cells growth was specifically observed following DMAT treatment of cervical carcinoma HeLa cell line with IC50 between 0.2-0.3 μM. Each of the breast cancer cell lines showed IC50 of 6, 10, and 20 μM DMAT for MDA-MB468, MCF7 and MDA-MB231 respectively. The more cancer cell lines we screen, the more evidence we have to suggest DMAT as a potential anti-cancer therapy. However, the specific mechanism of action of DMAT-inhibited-CK2 pathway in cancer ablation is not clear yet. Using Propidium-Iodide staining in conjunction with flow cytometry techniques, we analyzed and compared cell cycles of treated U937, MCF-7, MDA-MB231, MDA-MB468, and HeLa cell lines. Our data indicate that DMAT induces cell cycle arrest in HeLa cell line at G0/G1 phase. No effect on cell cycle was observed for all other cell lines tested. However, all cell lines underwent apoptosis following treatment with DMAT. Thus far our results suggest that DMAT can induce cell cycle arrest, apoptosis and maybe necrosis or multiple processes at the same time. We suggest that the mechanism of DMAT in cancer inhibition could be of multiple actions which further validate this molecule as a potent cancer therapy that could be suitable for clinical investigation.

Disclosures

No relevant conflicts of interest to declare.
ADAMTS13 ACTIVITY and VON Willebrand FACTOR LEVELS IN PATIENTS with Livedoid VASCULOPATHY.
Authors: da Costa França Andrea Fernandes Eloy , de Moraes Mazetto Bruna , Orsi Fernanda A. , de Paula Erich Vinicius , Machado Tania F.G.S , Souza Elemir M , Annichino-Bizzacchi Joyce Maria
Published: October 2019

Abstract 5064

Introduction

Livedoid vasculopathy is a rare skin disease caused by the extensive formation of microthrombi in dermis vessels, which leads to infarction and ulceration of the epidermis. The pathogenesis of livedoid vasculopathy is unknown, and hypercoagulation states have been implicated. Moreover, success using anticoagulation was reported in some patients. We hypothesized that decreased levels of ADAMTS13 activity and Von Willebrand factor (vWF) levels could contribute to the pathogenesis of local thrombus formation in livedoid vasculopathy.

Objective

Here we evaluated the levels of ADAMTS13 activity and vWF in patients with livedoid vasculopathy.

Patients and methods

Fourteen patients (93% female) with livedoid vasculopathy, median age of 41.5 years (22 - 48) were included in the study. All patients were evaluated for the presence of classical acquired and hereditary thrombophilia markers. In addition, vWF antigen levels (Elisa) and activity (collagen binding assays), as well as ADAMTS13 activity (evaluated by residual collagen binding) were evaluated.

Results

nine patients (64%) were carriers of the MTHFR C677T mutation (6 heterozygous and 3 homozygous); 1 patient was a carrier of factor V Leiden mutation, and 1 patient was a carrier of the FII G20210A mutation. None of the patients presented deficiency of antithrombin, protein C or S, and antiphospholipid antibodies were not identified in our study population. ADAMTS13 and VWF activities were normal in all patients. Median vWF antigen was 120.5U/dl (reference value of 40.0 - 232.0%), and vWF activity was 102.3% (reference value of 45.5 - 203.7%). ADAMTS13 activity was 138.9% (reference value of 56.0 - 227.2 %). Two patients had a history of thromboembolic diseases: one patient with 2 strokes and one patient with central retinal artery occlusion.

Conclusion

We show here that ADAMTS13 activity and vWF levels are not altered in patients with livedoid vasculopathy. The MTHFR C677T mutation is frequent in patients with livedoid vasculopathy but quantification of homocysteine levels is necessary to consider any relationship with this disease.

Disclosures

No relevant conflicts of interest to declare.
Hereditary Spherocytosis (HS) Associated with a Heterozygous MTHFR C677T Mutation and with Elevated Anticardiolipin Antibodies.
Authors: Noble Luis S. , Velazquez Maria , Gupta Anuradha , Landeros Mark M. , Herrada Juan
Published: October 2019

Abstract 5069

Background

Hereditary spherocytosis (HS) is a spectrum of inherited erythrocyte membrane defects that result in hemolysis and varying degrees of anemia, usually not associated with increased thrombotic risk. However, there is some concern about post-splenectomy thrombocytosis, since both arterial and venous thromboembolic events have been reported in patients splenectomized for HS.

A review of the medical literature revealed only a few cases of thrombosis associated with pregnancy and hereditary spherocytosis, and none of them had methyl tetrahydrofolate reductase (MTHFR) mutations or the presence of elevated anticardiolipin antibodies (both considered risk factors for thromboembolic disease).

Methods

Case report

Results

A 34-year-old Caucasian female with history of hereditary spherocytosis and splenectomy at age five, underwent evaluation for infertility. The patient was found to have a high titer of anticardiolipin antibodies and the presence of heterozygosity for the MTHFR C677T mutation. She denied any personal or family history of deep venous thrombosis. Subsequently the patient became pregnant. Because the combination of thrombophilic states appears to increase the risk of thromboembolic events, it was decided to start the patient prophyllactically with oral aspirin 81 mg daily, Foltx® (Folic acid 2.5 mg, cyanocobalamin 2000 mcg, and pyridoxine hydrochloride 25 mg), and heparin 5000 mg subcutaneously twice a day. On her most recent evaluation, at 26 weeks of pregnancy, no medical or obstetric problems have been observed.

Conclusion

Our case illustrates an unusual combination of thrombophilic states. Further studies are needed to evaluate the clinical significance of this association.

Disclosures

No relevant conflicts of interest to declare.
Thrombophilia in Retinal Vessel Thrombosis.
Authors: Herrada Juan , Gupta Anuradha , Landeros Mark M. , Vazquez Carlos
Published: October 2019

Abstract 5072

Background

Retinal vessel thrombosis (RVT) is the second most common retinal vascular disease after diabetic retinopathy, and is a common cause of visual morbidity. The role of thrombophilia in the pathogenesis of RVT remains under investigation.

Methods

Retrospective analysis. Between 2001 and 2009, the medical records and laboratory reports performed on patients (pts) with retinal vessel thrombosis who underwent thrombophilic evaluation were reviewed. The thrombophilic laboratory evaluation consisted of: Factor V mutation analysis, Factor II mutation analysis, Protein C activity, Protein S activity, antithrombin-III activity, Factor VIII activity, homocystein level, Methyl tetrahydrofolate reductase (MTHFR) C677T mutation analysis, and anticardiolipin antibodies.

Results

9 pts were identified. All but 2 pts were females. Median age at diagnosis was 50 years (31-78). Factor V Leiden was found in 1 pt. MTHFR C677T heterozygous mutation was present in 4 pts, and in 2 of these, hyperhomocystinemia was present as well. Anticardiolipin antibodies were elevated in 6 pts, and in 2 pts it was the only laboratory abnormality. One pt with hyperlipidemia had a negative thrombophilic laboratory testing.

Conclusions

Although newer laboratory methods are able to detect coagulation defects, their clinical relevance in retinal vessel thrombosis requires additional research.

Disclosures

No relevant conflicts of interest to declare.
Clinical Outcome of Patients with Myelophthisic Anemia Arising From Advanced Gastric Cancer.
Authors: Kwon Ji Yeon , Kim Han Jo , Kim Kyoung Ha , Kim Se-Hyung , Lee Sang-Cheol , Kim Hyun Jung , Bae Sang Byung , Kim Chan Kyu , Lee Nam Su , Lee Kyu-Taek , Park Sung Kyu , Won Jong-Ho , Hong Dae-Sik , Park Hee-Sook
Published: October 2019

Abstract 5095

Background

Although it is not common to encounter patients with myelophthisic anemia arising from advanced gastric cancer, clinical features and optimal treatment are not yet to be elucidated. Prognosis for gastric cancer patients with bone marrow metastases is extremely poor. The current study was performed to evaluate clinical outcome of patients with myelophthisic anemia arising from advanced gastric cancer.

Methods

We retrospectively reviewed the medical records of 26 advanced gastric cancer patients with bone marrow metastases between September 1986 and February 2009 at Soonchunhyang University Hospital.

Results

The median age was 46 years (range 24-61 yeras). All patients had poorly differentiated adenocarcinoma, including 17 signet ring cell carcinomas. The majority of the patients showed thrombocytopenia, anemia, and elevation of lactate dehydrogenase. Sixteen patients (61.5%) were received palliative chemotherapy with a median of 4 cycles. (range, 1-13 cycles). Median survival durations after bone marrow metastases for entire patients were 37 days (95% CI, 12.5-61.5 days). The median survival times from bone marrow involvement were 11 days in the best supportive care group (range 9.5-12.5 days) and 121 days (range 94.7-147.3 days) in the palliative chemotherapy group (p <0.001). Patients died of tumor progression (11 patients, 45%), brain hemorrhage (6 patients, 25%), infection (5 patients, 21%), and DIC (1 patient, 4%). There were no chemotherapy related deaths.

Conclusion

It is difficult to decide whether to proceed with aggressive treatment for gastric cancer patients with bone marrow metastases because of the hematologic findings, e.g. anemia, thrombocytopenia, and DIC. However, this study suggests that palliative chemotherapy should be actively considered in patients with myelophthisic anemia arising from advanced gastric cancer.

Disclosures

No relevant conflicts of interest to declare.
Low Incidence of Cardiac Siderosis in Heavily Iron Overloaded Egyptian Thalassemics.
Authors: El-Beshlawy Amal M. , Hamdy Mona Mohamed , Elgamrawy Mona , Azim Khaled Abdel , Salem Doria , Samir Ahmed , Eltagui Mona
Published: October 2019

Abstract 5110

Introduction

Myocardial siderosis in thalassemia major remains the leading cause of death in developing countries. Once heart failure develops, the outlook is usually poor with precipitous deterioration and death. Cardiovascular magnetic resonance (CMR) technology can measure cardiac iron deposition directly using the magnetic relaxation time T2*. This allows earlier diagnosis and treatment and help to reduce mortality from this cardiac affection.

Patients and Methods

89 β thalassemia patients (10 to 43 years, mean age of 20.78±6.36) were recruited in this study. All patients were receiving chelation therapy of subcutaneous desferroxamine. Evaluation of hemosiderosis was based on CMR, liver magnetic resonance R2 and serum ferritin.

Results

T2* values ranged between 4.3 to 53.8 ms with a mean of 28.50±11.74 ms among our study group. The left ventricular ejection fraction (LVEF) as measured by CMR ranged between 55 and 78%; mean=67.66±4.69%.and liver iron concentration (LIC) ranged between 1.5 to 56 mg/g dry weight with a mean of 26.13±13.37 mg/g. Serum ferritin varied among our study group from 533 to 22363ng/ml; mean=4514.27±2847.58ng/ml with 83.15% above 2500ng/ml. The prevalence of myocardial siderosis (T2*<20ms) among our patients was 22/89 patients (24.7%) aged 20.86±7.54 years with a mean T2* value of 12.718±4.36ms and LVEF of 68.59 ±5.84%. LIC and serum ferritin results were 30.85±13.48 mg/g and 6122.86±4185.67ng/ml respectively. There was no correlation between T2* results and the age, LVEF, LIC and serum ferritin of this group (P=0.651, P=0.085, P=0.999 and P=0.627 respectively). Those patients with severe cardiac siderosis (T2*<10ms) constituted 7/89 (7.9%) with a mean age of 18.43±4.35 years. Although these patients had a mean T2* of 7.8±1.73 ms, the LVEF value was 65.14±6.20 % and only one patient had clinical cardiac disease (T2*=4.3 ms and LVEF =55%). LIC and serum ferritin results were 29.84±16.99mg/g and 7202.14±6953.79ng/ml respectively. In this group of severe cardiac siderosis, T2* was not correlated to age (P=0.5), LVEF (P=0.144), LIC (P=0.969) and serum ferritin (P=0.818).

Conclusion

Low prevalence of myocardial siderosis in the Egyptian thalassemic patients in spite of the very high serum ferritin. In severely iron overloaded patients the cardiac function was not affected. T2* is the best test that can identify at risk patients who can be treated with optimization of their chelation protocols. The possibility of a genetic component for the susceptibility of cardiac iron loading in our population should be considered.

Disclosures

No relevant conflicts of interest to declare.
Regulation of Complement Activation by Thrombomodulin.
Authors: Delvaeye Mieke , DeVriese Astrid , Moons Michael , Esmon Naomi , Esmon Charles , Conway Edward M.
Published: October 2019

Abstract 5127

Thrombomodulin (TM) is an integral membrane glycoprotein, ubiquitously expressed by vascular endothelial cells. Its epidermal growth factor (EGF)-like repeats amplify thrombin-mediated generation of activated protein C and activated thrombin activatable fibrinolysis inhibitor, thereby suppressing coagulation, inflammation and fibrinolysis, and inactivating the anaphylatoxins, C3a and C5a. TM also has direct anti-inflammatory properties, interfering with leukocyte adhesion, and preventing complement activation via its lectin-like domain. We recently established that TM is a physiologically important negative regulator of the alternative pathway (AP) of complement activation, defects of which increase the risk of the thrombotic microangiopathy, atypical hemolytic uremic syndrome. In this report, we assessed the role of TM in the other complement activation pathways. In serum, TM interferes with classical pathway (CP) mediated erythrocyte cell lysis, and protects against CP-induced cell death. TM co-precipitates with C4b, and enhances its inactivation and cleavage to C4c and C4d by complement factor I in the presence of the cofactor C4b binding protein. TM also interferes with the coagulation complement pathway by interfering with thrombin cleavage and activation of C5 to C5a. Overall, TM negatively regulates complement via the major recognized pathways. The findings implicate defects in TM in a wide range of diseases in which complement plays a role, and underlines the potential of TM as a therapeutic target.

Disclosures

No relevant conflicts of interest to declare.
Endothelial Progenitor Cells(EPC) in Peripheral blood in Diabetic Foot with and without PAD.
Authors: Krutikov Alexander , Anisimov Sergey , Gabaidulina Malika , Meln Irina , Grineva Elena , Vavilov Valery , Semernin Eugene , Zaritskey Andrey
Published: October 2019

Abstract 5130

EPC is not very well defined population. These cells have either angiopoietic or angiostimulating function, the latter seems to be more relevant. It is well known that

Mobilization of EPC to peripheral blood is decreased in diabetic rats. From the opposite, ischemia proved to stimulate mobilization. That is why, we decided to study EPC in diabetic patients with and without PAD

Patients and methods

40 pts with diabetes(type I- 4, type II-36) were studied. Duration of diabetes -1-46 years. The duration of more than 10 years was in 57% patients. Age- 49-73 years(mean age- 62 years). The patients were divided into two groups- diabetic foot without PAD(25 pts) – Group A, and diabetic foot with PAD- group B. PAD was diagnosed with sonography, duplex sonography and Rx arteriography. Normal volunteers were use as controls(mean age 51.6+2.0)

CD34+ and CD133+ cells were numerated with CytoFlow(BD).

5 Day CFU-Hill Colony Assay- non-adherent mononuclear cells (MNCs) was used to study EPC

In this method, peripheral blood MNCs are plated on fibronectin-coated dishes (6-well). After a 48 h pre-plating step to deplete the sample of adherent macrophages and mature endothelial cells, the non-adherent cells are removed and re-plated on fibronectin-coated dishes (24-well). Unique colonies that are formed in the 5 Day CFU-Hill Colony Assay are referred to as colony-forming unit-Hill colonies (CFU-Hill colonies) or colony-forming unit-ECs (CFU-ECs). Count the number of colonies per well for each sample. CFU-Hill colonies are defined as a central core of round cells with radiating elongated spindle-like cells at the periphery. Colonies without the CFU-Hill morphology may also be present but are not scored as CFU-Hill colonies. CFU-Hill Colonies are fixed with methanol and stained with a Giemsa solution. Plasma concentration of VEGF was studied by ELISA

Results

EPC colony forming ability in group A was 4.0+ 0.7, in group B-27.4+3.9. The differences between these two groups is highly significant(p 0.001). Low EPC level in diabetic neuropathy is probably related to decreased mobilization(G.Fadini e.a.2006). Rather unusual was higher EPC level in group B. It was shown that age-related decrease in EPC is due to decline in HIF-1 signaling(M. Hoenig e.a 2008). Probably, diabetic defect of mobilization is partly overcome with ischemia induced up-regulation of HIF-1 and VEGF. EPC in group B patients does not differ from controls(26.1+6.5).

There were now correlations between EPC number and the number of either CD34+, CD133+, or KDR+ cells. We also failed to find any correlations between VEGF and EPC

Conlusion

EPC number in peripheral blood in diabetic patients without PAD is severely decreased. Diabetic patients with PAD have near normal EPC number. Thus, ischemia in diabetic patients is unable adequately mobilize EPC

Disclosures

No relevant conflicts of interest to declare.
Autologous Hematopoietic Stem Cell Transplantation (HCT) is a Safe and Effective Treatment for Primary Plasma Cell Leukemia: The CIBMTR Experience.
Authors: Mahindra Anuj , Vesole David , Kalaycio Matt E. , Vela-Ojeda Jorge , Shrestha Smriti , Zhang Mei-Jie , Dispenzieri Angela , Milone Gustavo A. , Lonial Sagar , Hari Parameswaran N.
Published: October 2019

Abstract 532

Primary Plasma cell leukemia (PCL) is a rare plasma cell disorder characterized by an aggressive clinical course. The efficacy of autologous and allogeneic HCT in PCL is uncertain due to the small number of patients that have been reported in the literature. To determine the potential for long-term disease control using HCT in PCL, we performed a retrospective review of 160 patients with primary PCL who received autologous (n=107), allogeneic (n=52) or syngeneic (n=1) HCT and were reported to the CIBMTR between 1995 and 2006. Kaplan–Meier analysis was used for overall survival (OS) and progression free survival (PFS). Cumulative incidence estimates of non-relapse mortality (NRM), relapse, acute and chronic GVHD were calculated.

Median age at the time of autologous HCT was 57 years (range, 32-74 years) and allogeneic HCT was 48 years (range, 24-62 years). Median time from initial treatment to autologous HCT was 7 months and allogeneic HCT was 6 months. At the time of transplant, in the autologous HCT group, 19% patients were in complete remission, 56% in partial remission and 1% had relapsed/progressive disease; in the allogeneic HCT group, 17% were in complete remission, 46% in partial remission and 10% had relapsed/progressive disease. In the allogeneic HCT group, 83% of the patients received myeloablative conditioning regimens and 17% received nonmyeloablative/reduced-intensity conditioning. The rate of acute GVHD was 35% (95% confidence interval [CI], 22%-49%) and the rate of chronic GVHD at 3 years was 27% (95% CI, 13%-40%). After a median follow-up of 38 months, 20 of the 52 patients (38%) are alive in the allogeneic HCT group and 68 of the 107 pts (64%) are alive in the autologous HCT group (Figure 1). Progressive disease accounted for 38% of the deaths in the allogeneic HCT group and 86% of the deaths in the autologous HCT group. The outcomes are summarized in table 1. The remarkable overall survival of patients after autologous HCT in this analysis is noteworthy and indicates that autologous HCT is a safe and acceptable treatment option for patients with primary PCL. Although relapse rates are lower, allogeneic HCT carries a much higher risk of NRM and should be considered primarily in the context of a clinical trial.

Disclosures:

Lonial: Celgene: Consultancy; Millennium: Consultancy, Research Funding; BMS: Consultancy; Novartis: Consultancy; Gloucester: Research Funding.
HS1 (HCLS1) Protein, a Binding Partner of HAX1, Induces G-CSF-Triggered Granulopoiesis Via LEF-1 Transcription Factor.
Authors: Skokowa Julia , Lan Dan , Uenalan Murat , Gupta Kshama , Welte Karl
Published: October 2019

Abstract 548

Mutations in HS1 (HCLS1) associated protein × 1 (HAX1) have been described in patients with severe congenital neutropenia (CN). HAX1 is a ubiquitously expressed and until now it was unclear how HAX1 mutations cause isolated neutropenia in CN patients. So we were interested in investigating G-CSF-dependent HS1 (HCLS1) protein expression and phosphorylation in CN patients compared to healthy individuals, since HS1 (HCLS1) is a hematopoietic-specific protein and may link HAX1 to the G-CSFR signaling via the tyrosine kinases lyn and syk. Indeed, we found that treatment of CD34+ hematopoietic cells with G-CSF leads to phosphorylation of HS1 (HCLS1) protein and recruitment of receptor thyrosine kinases lyn and syk. In CN patients harboring HAX1 mutations, G-CSF failed to upregulate expression and phosphorylation of HS1 (HCLS1), suggesting that for HS1 phosphorylation functional HAX1 protein is required. Indeed, inhibition of HAX1 in CD34+ hematopoietic cells from healthy individuals significantly abrogated G-CSF-triggered HS1 (HCLS1) phosphorylation. Previously, we found that LEF-1 is a master regulator of granulopoiesis and is severely diminished in myeloid cells of CN patients with HAX1 mutations. Moreover, these cells also exhibited severe defects in F-actin assembly. Using immunoprecipitation assay of endogenous proteins, we found that upon G-CSF stimulation of CD34+ hematopoietic cells, HS1 (HCLS1) protein additional to HAX1, also binds to LEF-1 protein modulating its expression, transcriptional activity and subsequently granulopoietic differentiation. Using CFU, qRT-PCR and reporter gene assays we demonstrated that inhibition of HS1 (HCLS1) severely disturbed G-CSF-triggered granulocytic differentiation in line with downregulation of LEF-1 expression and functions, as well as with pathological redistribution of F-actin.

Taken together, in this study we demonstrated for the first time the connection between G-CSF receptor, HAX1 mutations, downregulation of HS1 (HCLS1) and LEF-1, abnormal F-actin assembly and the defective granulocytic differentiation in CN patients. Moreover, we demonstrated for the first time an ultimate direct connection between Wnt signaling and G-CSF cytokine cascade in granulopoiesis.

Disclosures:

No relevant conflicts of interest to declare.
Lyl1 and Scl Regulate Bcl2 Expression and Control Apoptosis at Different Stages of Hematopoietic Development.
Authors: Souroullas George , Goodell Margaret
Published: October 2019

Abstract 569

The Lymphoblastic Leukemia gene 1 (Lyl1) encodes a basic-helix-loop-helix (bHLH) transcription factor that is involved in T cell acute lymphoblastic leukemia via chromosomal translocations. Lyl1 has an almost identical bHLH motif to Scl, a known master regulator of hematopoiesis. In our previous work, we used a double knock-out mouse model to demonstrate functional redundancy between Lyl1 and Scl in adult hematopoietic stem cells (HSCs), where in the absence of both genes in adult mice, HSCs and progenitors undergo rapid apoptosis. Despite their common and overlapping functions, the two genes appear to also function at different aspects of hematopoietic development. Lyl1 knock-out mice exhibit a unique defect in B-cell development. This phenotype is recapitulated in vitro, where Lyl1 progenitors grown on a layer of OP9 cells and allowed to differentiate with the addition of cytokines fail to differentiate to B cells as well as wild type progenitors. Rescue assays, where Lyl1 was transduced using a retrovirus into Lyl1 knock-out progenitors and transplanted into lethally irradiated recipients, showed complete rescue of the B cell lineage. Forced expression of Scl into Lyl1 knock-out progenitors failed to rescue, giving rise to only about 5% of peripheral blood B cells. This demonstrates the distinct functions of the two genes in differentiated cells types, despite their functional redundancy in the HSCs. We further sought to dissect the functions and differences between Lyl1 and Scl based on their target genes. Using bioinformatics analysis and a literature search, we identified a number of genes that were candidates for Lyl1 transcriptional regulation; these interactions with Lyl1 and Scl were confirmed by chromatin immunoprecipitation. We found that Bcl2, and genes belonging to this family, are direct target genes of both Lyl1 and Scl in progenitor cells, and that they are downregulated in Lyl1 and Scl single knock-out mouse models. Since Bcl2 is critical for B cell survival, we hypothesized that the B cell deficiency in Lyl1 knock-out mice could be partly due to a loss of Bcl2. We performed rescue assays where Bcl2 was transduced using a retrovirus into Lyl1 knock-out progenitors which were transplanted into lethally irradiated recipient mice. Bcl2 overexpression rescued the B-cell defect in Lyl1 knock-out bone marrow. Since Bcl2 was identified as a target gene for Scl as well, we tested whether Bcl2 could also rescue the rapid apoptosis phenotype of the Lyl1/Scl double knock-out progenitors. Forced expression of Bcl2 in these cells, followed by deletion of the floxed Scl allele, resulted in a delayed onset of apoptosis, but incomplete rescue of the progenitor cell death. These findings distinguish the activities of Lyl1 and Scl in B cells and demonstrate a possible mechanism for their functions, through in part by their activation of the antiapoptotic gene Bcl2.

Disclosures:

No relevant conflicts of interest to declare.
Gabp Transcription Factor Is Required for Myeloid Differentiation, In Part, through Its Control of Gfi-1 Expression
Authors: Yang Zhong-Fa , Drumea Karen , Cormier James , Wang Junling , Rosmarin Alan G.
Published: October 2019

Abstract 740

GABP is an ets transcription factor which regulates genes that are required for myeloid differentiation. The tetrameric GABP complex includes GABPα, which binds DNA via its ets domain, and GABPβ, which contains the transcription activation domain. In order to examine the role of Gabp in myeloid differentiation, we created mice in which exons that encode the ets DNA-binding domain are flanked by LoxP recombination sites (Gabpa fl/fl or floxed mice), and bred them to mice that bear Cre recombinase under the control of the pIC-inducible promoter, Mx1. Injection of these mice with pIC induces conditional deletion of Gabpa in hematopoietic tissues, and these mice are referred to as Gabpa knock-out mice (KO). Control mice that carry floxed Gabpa but lack Mx1-Cre were also treated with pIC. KO mice rapidly lost myeloid cells in peripheral blood, bone marrow, and spleen. Bone marrow cells from control and KO mice were sorted for CD11b and Gr1 to distinguish mature granulocytes from immature granulocytes and monocytes. KO mice had few mature myeloid cells, and the residual cells exhibited a dysplastic phenotypic appearance that did not resemble either normal maturing monocytes or granulocytes; instead they exhibited morphologic characteristics of both lineages. These KO myeloid cells simultaneously expressed differentiation-associated monocyte-restricted and granulocyte-restricted genes and markers of immaturity (Ly6C and c-kit). Thus, the patterns of gene and antigen expression by these dysplastic KO myeloid cells did not correspond to either normal monocytes or granulocytes. We recognized that these KO myeloid cells resemble those seen in mice with disruption of the transcriptional repressor, Gfi-1. We isolated granulocyte-monocyte progenitor (GMP) cells from control and KO bone mice, and found that expression of Gfi-1 was significantly reduced in KO GMPs, compared to controls. Chromatin immunoprecipitation demonstrated that Gabp bound the Gfi-1 promoter in vivo and transfection analysis showed that Gabp activated the Gfi-1 promoter. To test the hypothesis that Gabp controls myeloid differentiation through its regulation of Gfi-1, we grew bone marrow cells from both control and KO mice in liquid culture with SCF, Il-3 and Il-6, or GM-CSF. We transduced these cells with retrovirus bearing Gfi-1, and analyzed myeloid colony formation, and expression of CD11b and Gr1. Transduction of KO bone marrow cells with Gfi-1 partially restored myeloid colony formation in semi-solid media, and the percentage of Gr1+CD11b+ cells in liquid culture. We conclude that GABP plays a non-redundant role in myeloid development, and it controls differentiation of myeloid cells, at least in part, through its regulation of the transcriptional repressor, Gfi-1.

Disclosures:

No relevant conflicts of interest to declare.