bluuebunny/crossref_metadata_2025_split huggingface dataset:
Microrna (miR)-17-92 Contributes to Therapy Resistance in Burkitt Lymphoma Cells
Authors: Miles Rodney R. , Rodic Vladimir , Barth Matthew J. , Cairo Mitchell S. , Hermiston Michelle L. 
Published: October 2019

Abstract
Background
Burkitt lymphoma (BL) is curable, but patients with relapse or refractory disease have very poor outcomes. The underlying biological differences leading to relapse/refractory disease are unknown. We previously identified a potential association between microRNA (miR)-17-92 and relapse in patient samples. MiRs encoded by this gene have been implicated in therapy resistance, so we are seeking relationships between miRNA expression and therapy resistance in BL cells.
Methods
The miR-17-92 locus was targeted for deletion in Raji BL cells using custom CRISPR-Cas9 lentiviral vectors. Single cell-derived clones were established and locus deletion was determined by genomic DNA PCR. Inducible miR-17-92 expression in Raji cells was established by transduction of a cumate switch inducible lentivector system containing the miR-17-92 gene along with a CymR repressor expression vector (Raji/CymR/miR-17-92). MiR expression was assessed by TaqMan assay, and protein expression and caspase-3 cleavage were assessed by western blotting. Cells were treated with 4HC, the active metabolite of cyclophosphamide, or doxorubicin, and viability was assessed by AlamarBlue assay.
Results
Genomic DNA PCR verified hemizygous deletion of the miR-17-92 gene in several Raji BL clones; no homozygous deleted clones were identified. In two hemizygous clones R5 and R24, miR-17 expression was decreased to 0.24-fold and 0.37-fold, respectively, relative to the parental Raji line. The miR-17-92 target proteins PTEN and Bim were increased in the R5 and R24 clones by western blotting. Compared to Raji, R5 and R24 showed increased basal caspase-3 cleavage and much greater induction of caspase-3 cleavage with cytotoxic chemotherapy (10 µM 4HC). In addition, 0.5 and 1 µM doxorubicin showed a dose dependent induction of cleaved caspase in R5 and R24 cells that was much greater than in parental Raji cells. The IC50 for doxorubicin in R5 and R24 miR-17-92 hemizygous cells was markedly decreased (0.00025 and 0.00032 µM, respectively) compared to parental Raji cells (0.064 µM). Finally, miR-17 expression was inducible by treating cumate repressor vector transduced Raji/CymR/miR-17-92 cells with 30 µg/mL cumate (+2.8-fold at 3 days, +7.5-fold at 5 days).
Conclusion
We established miR-17-92 hemizygous-deleted cell lines that show increased PTEN and pro-apoptotic Bim, increased basal caspase-3 cleavage, and increased chemosensitivity. We have established Raji BL cell lines with inducible miR-17-92 expression to extend these experiments by determining the effect of increased miR-17-92 on chemosensitivity. Our findings show that high miR-17-92 expression in BL cells can contribute to therapy resistance, which may be important for understanding refractory BL in patients and could lead to more effective targeted therapies.

Disclosures
Cairo: Celgene: Research Funding.
Limited Role of Workup Including PET-CT, Esophagogastroduodenoscopy and Bone Marrow Biopsy in Staging of Ocular Extranodal Marginal Zone Lymphoma of Mucosa-Associated Lymphoid Tissue (MALT) Type
Authors: Chae Heejung , Suh Cheolwon , Yoon Dok Hyun , Hong Jung Yong , Park Chan-Sik , Huh Jooryung , Sa Ho-Seok , Kim Kyoung Won 
Published: October 2019

Abstract
Background
Lymphoma of the ocular adnexa accounts for approximately 1 to 2% of non-Hodgkin lymphomas and the most common subtype is extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue (MALT) type (80-90%). Ocular MALT lymphoma is mostly localized disease with indolent clinical course. The workup for non-gastric MALToma is usually done similar to the workup for other non-Hodgkin lymphoma. However, as there are paucity of data in the role of PET, esophagogastroduodenoscopy and bone marrow biopsy,weaimed toinvestigate these issues in patients with ocular MALT lymphoma.
Patients and Method
One hundred thirty patients diagnosed as ocular MALT lymphoma between Jan 2001 and Dec 2015 at a single tertiary hospital were included in this retrospective analysis. We retrospectively reviewed clinical characteristics, diagnosis, natural history of disease, treatment and outcomes.
Result
The median age of all patients was 48 years (range, 20-70 years). Fifty-nine (45%) were male patients. The majority of patients (n=87, 67%) presented with stage I disease, followed by bilateral involvement, defined as stage I+I (n=21, 16%), stage IV (n=16, 12%), and stage II (n=6, 5%). Nodal involvement was present in 8% (n=10), and 12% (n=16) of patents accompanied extra-nodal involvement other than orbits at initial diagnosis. Positron emission tomography (PET) had low sensitivity in detecting primary orbit lesions (n=44/120, 37%). Moreover, only in 3% of cases (n=4), PET imaging after CT scans on neck, chest, abdomen and pelvis could provide additional clinically meaningful information that led to upstaging. Accompanying gastric MALToma was found in 2% (n=2/82) of patients by EGD. Those two patients all survived until last follow-up. Bone marrow (BM) involvement was found in 6% (n=7/120). None of them had clinically significant cytopenia. With a median follow-up period of 74 months, 5-year overall survival (OS) rate was 99% in all patients. There was no significant difference in 5-year OS rates according to BM involvement (100% with and 99% without BM involvement).
Conclusion
PET-CT, EGD and BM biopsy has limited role in staging workup of primary ocular MALT lymphoma. Those workups should be performedonly in selected cases.

Disclosures
No relevant conflicts of interest to declare.
Chimeric Antigen Receptor-Modified T Cells for the Treatment of Acute Myeloid Leukemia Expressing CD33
Authors: Song Degang , Swartz Michael H. , Biesecker Steve G. , Borda Fernando , Shah Rutul R. , Emtage Peter , Wierda William G. , Cooper Laurence J.N. , Chan Tim 
Published: October 2019

Abstract
Relapsed acute myeloid leukemia (AML) is an aggressive disease with very poor outcomes. Redirection of T-cell specificity via chimeric antigen receptor (CAR) has shown promising anti-tumor activity in clinical trials, particularly for B cell linage malignancies. CD33 is a transmembrane protein expressed on normal and malignant myeloid-derived cells as well (as on subsets of activated T cells and NK cells). Since this protein is commonly expressed on AML cells, we sought to evaluate the efficacy of targeting AML with CD33-specific CAR-T cells. We generated a lentiviral construct to co-express CD33-specific CAR and a kill switch based on a tag derived from the epidermal growth factor receptor. The latter allows for the conditional elimination of CAR-T cells in vivo. Following transduction of primary T cells, we confirmed CAR and kill switch co-expression by flow cytometry and western blot analyses. Elimination of genetically modified T cells was demonstrated using the clinically-available antibody, cetuximab. CD33 CAR-T cells demonstrated specific cytotoxicity to CD33+ target cell lines. CD33 CAR-T cells were also activated to produce IFNg, TNF, and IL-2 cytokines in response to CD33+ target cells. Furthermore, adoptive transfer of CD33 CAR-T in immunocompromised (NSG) mice bearing established CD33+(CD19neg) AML (MOLM-13) tumor resulted in reduction of tumor burden and improvement of overall survival, compared to control mice receiving CD19 CAR-T cells or no immunotherapy (Figure). Sampling of blood demonstrated the persistence of the CD33 CAR-T cells with no detection of AML (MOLM-13) tumor cells. These pre-clinical data demonstrate the effectiveness of CD33 CAR-T cells in targeting CD33+ AML tumor cells and provide a rationale for future clinical evaluation in AML patients with unmet medical need.

Disclosures
Song: Intrexon Corporation: Employment, Equity Ownership. Swartz:Intrexon Corporation: Employment, Equity Ownership. Biesecker:Intrexon Corporation: Employment, Equity Ownership. Borda:Intrexon Corporation: Employment. Shah:Intrexon Corporation: Employment, Equity Ownership. Wierda:Genentech: Research Funding; Gilead: Research Funding; Abbvie: Research Funding; Novartis: Research Funding; Acerta: Research Funding. Cooper:MD Anderson Cancer Center: Employment; Intrexon: Equity Ownership; Sangamo BioSciences: Patents & Royalties; Targazyme,Inc.,: Equity Ownership; City of Hope: Patents & Royalties; ZIOPHARM Oncology: Employment, Equity Ownership, Patents & Royalties; Miltenyi Biotec: Honoraria; Immatics: Equity Ownership. Chan:Intrexon Corporation: Employment, Equity Ownership.
Foxp3+ regulatory T Cells Maintain Bone Marrow Microenvironment for B Cell Differentiation from Hematopoietic Stem Cells
Authors: Nishikii Hidekazu , Pierini Antonio , Yokoyama Yasuhisa , Kimura Takaharu , Kwon Hye-Sook , Alvarez Maite , Baker Jeanette , Shizuru Judith , Wu Joy , Chiba Shigeru , Negrin Robert S. 
Published: October 2019

Abstract
Background: Foxp3+regulatory T cells (Treg) are a subpopulation of T cells, which regulate the immune system, maintain self-tolerance and enhance immune tolerance after transplantation. It was also reported that recipient derived Treg could provide immune privilege niche to allogeneic hematopoietic stem cells (HSC) after transplantation. However, the precise role of Treg in hematopoiesis has not been fully elucidated.
Methods: We used Foxp3-DTR mice (B6, CD45.2) for in vivo depletion of Treg through diphtheria toxin (DT) injection and investigated whether Treg depletion would affect hematopoiesis derived from HSC. To investigate whether Treg depletion affects the function of the bone marrow microenvironment, we transplanted wild type bone marrow cells into lethally irradiated Foxp3-DTR mice after Treg depletion.
Results: We found 1) a significant defect on B cell progenitors including mature B cells (IgM+B220+, P<0.001), pre-B cells (IgM-B220+CD19+cKit-, P<0.001) and pro-B cells (IgM-B220+CD19+cKit+, P<0.05), 2) LT-HSC population (CD34-/lowFlit3-cKit+Sca1+Lin-) was significantly expanded (p<0.01) and entered into cell cycle, 3) the residual Foxp3-CD4+ or CD8+ T cells in the bone marrow had an activated immune phenotype and clustered at sinusoids when bone marrow cells from Treg depleted mice were analyzed. Expanded LT-HSC from Treg depleted mice had reduced long-term reconstitution capacity when we performed competitive repopulation experiments using purified LT-HSC from Foxp3-DTR mice with or without Treg depletion (100 cells/mice, CD45.2), total bone marrow cells (2x10e5/mice, B6-F1, CD45.1/CD45.2) and congenic recipient mice (lethally irradiated B6, CD45.1). B cell reconstitution was also severely abrogated following transplantation using Treg depleted mice as recipients (p<0.01). In those mice, we observed a significant reduction of IL-7 production (p<0.01). Interestingly, we found that a subpopulation of CD45-TER119-CD31- ICAM1+ perivascular stromal cells are a major source of IL-7 in the bone marrow. ICAM1+ perivascular stromal cells also secrete SCF and CXCL12, which is crucial for the maintenance of LT-HSC. In Treg depleted BM cells, a significant reduction in IL-7 secretion from ICAM1+ perivascular stromal cells was observed, suggesting that this population is the target of activated T cells after Treg depletion.
Conclusions: These data demonstrate that Treg play a key role in B cell differentiation from HSCs by maintaining the immunological homeostasis in the bone marrow microenvironment. These data provide new insights into Treg biology and function in normal and stress hematopoiesis.

Disclosures
Negrin: Stanford University: Patents & Royalties.
Hepatitis in Association with Prolonged 6-Mercaptopurine Therapy
Authors: McILVANIE SAMUEL K. , MacCARTHY JOHN D. 
Published: October 2019

Abstract
Four patients are presented who developed toxic hepatitis while being treated for leukemia with 6-mercaptopurine and steroids. The onset was rapid and suggested an acute hepatitis with intrahepatic blockage. Pathologic examination of the liver in two patients who expired with hepatitis is presented and the pathology described. In the third autopsied case the hepatitis had resolved while receiving massive steroid therapy, even though the leukemia progressed rapidly when 6-mercaptopurine was withdrawn. Early recognition of the toxic hepatitis becomes especially important whero 6-mercaptopurine is the first drug to be used in the management of the patient with acute leukemia.
Abnormal Human Hemoglobins. I. Biosynthesis in Vitro of Fe59-Labeled Human Hemoglobin A and Hemoglobin S
Authors: KRAUS LORRAINE M. , MORRISON DEMPSIE B. , KRAUS ALFRED P. 
Published: October 2019

Abstract
The synthesis of Fe59 labeled-hemoglobin during incubation of bone marrow from normal and sickle cell anemia (crisis and non-crisis) patients has been investigated. The in vitro synthesis of hemoglobin S in the steady state (noncrisis) sickle cell anemia individual was found to proceed at a much faster rate than the synthesis of hemoglobin A from normal individuals. In the crisis of sickle cell anemia, the uptake of Fe59 and synthesis of hemogobin S was approximately at the same level as synthesis of hemoglobin A from the normal individual. The results of this study indicate that one of the mechanisms involved in sickle cell anemia crisis is a decrease in synthesis of hemoglobin S as compared to the synthesis of hemoglobin S during the steady state of noncrisis.
The Collection and Storage of Viable Human Fetal Hematopoietic Tissue for Intravenous Use
Authors: FERREBEE JOSEPH W. , LOCHTE HARRY L. , SWANBERG HERBERT , THOMAS E. DONNALL 
Published: October 2019

Abstract
Livers and spleens from human fetuses in the 12 to 20 weeks’ gestation period were preserved in a sterile condition in glycerol and serum at -70 C. for periods of several weeks. Studies of DNA synthesis rates with C14-formate, and radioautographs after incubation with tritiated thymidine, revealed that hematopoietic elements concentrated in these tissues at this stage of gestation maintained their viability during these periods of storage. Intravenous infusion of cellular suspensions prepared from stored frozen tissue, or from fresh tissue in this same age group, produced no observed untoward clinical event in the three patients studied.
Some of the arguments favoring the use of fetal hematopoietic tissue, in contrast to adult marrow, in the attempt to restore marrow function after radiation injury are briefly reviewed.
Correlation of Radioactive Hemin Formation with Morphologic Alterations in Cultures of Human Bone Marrow
Authors: POWSNER EDWARD R. , BERMAN LAWRENCE 
Published: October 2019

Abstract
A method for measurement of radioactive hemin formation has been applied to the in vitro system for culture of human erythroblasts previously described. The results of these measurements have been shown to correlate with a morphologic parameter of maturation, the increase of orthochromic cells, in the same in vitro cell culture system.
The significance of these findings as well as the differences in the times of formation of heme and the increase of orthochromic cells has been discussed.
It is proposed that both the formation of radioactive hemin and the increase of orthochromic cells may be useful in the measurement of different aspects of the effects of various plasmas on the maturation of normal and abnormal human erythroblasts.
The Relationship of Tumor Necrosis to Red Blood Cell Changes in the Hamster
Authors: SHERMAN JOSEPH D. 
Published: October 2019

Abstract
1. Sterile, cell-free saline extracts of the necrotic portion of methylcholanthrene-induced hamster sarcoma injected into non-tumor bearing hamsters produced a statistically significant decrease in hemoglobin, hematocrit and red blood cell levels.
2. These changes in hemoglobin, hematocrit and red blood cells are comparable to those observed during sarcoma growth.
3. Sterile, cell-free, extracts of viable sarcoma injected into hamsters failed to produce alterations in the hemoglobin, hematocrit or red blood cell values.
4. The necrosis produced during sarcoma growth in the hamster cheek pouch is concluded to be responsible for alterations in the red blood cell counts, hemoglobin and hematocrit values.
Studies on Erythropoiesis. XII. The Effect of Transfusion-Induced Polycythemia in the Mother on the Fetus
Authors: JACOBSON LEON O. , MARKS EDNA K. , GASTON EVELYN O. 
Published: October 2019

Abstract
Erythropoiesis is reduced to zero in the mouse by repeated homologous red cell transfusion.
Mating of transfusion-induced polycythemic mice was allowed, and observations were made on maternal and fetal erythropoiesis. It was observed that erythropoiesis in the pregnant, polycythemic mouse was suppressed throughout pregnancy, this observation suggesting that erythropoietin production was concomitantly suppressed.
Erythropoiesis in the fetuses of polycythemic mothers was initiated by 10 days and maintained in an active state throughout fetal life. Hematologic values of these fetuses at birth were higher than those of the newborn normal pregnant mice of the same strain.
It is possible that the fetus initiates and maintains erythropoietin independently of erythropoietin from the maternal circulation. The higher hematologic values of the newborn of polycythemic mothers are probably the result of the greater availability of iron from the breakdown of transfused cells, since it was found that comparable hematologic values were achieved in the newborn when iron-dextran (Imferon) was injected into the maternal tissues during pregnancy.
Long-Term Mixed Cultures of Human Hemic Cells, with Granulocytic, Lymphocytic, Plasmocytic and Erythrocytic Series Represented
Authors: BROOKE JOHN H. , OSGOOD EDWIN E. 
Published: October 2019

Abstract
New culture strains containing humman hemic cells of monocytic, lymphocytic, granulocytic, plasmocytic and erythrocytic series are described with suggestions as to maintenance of the desired cell series in culture.
The addition of cells from lymphocytic, plasmocytic and erythrocytic series to a previously well established culture has made possible long-term culture of these cell types, since this procedure aids in maintaining the necessary high gradient factor for their growth and multiplication.
Although the life span of these cells has been considerably shortened in culture and immature forms predominate, the cells show recognizable characteristics of the cell series of origin. A high population density must be maintained in cultivation of the mixed cultures or the desired cell type may be lost.
Notes on the Mechanism of Leukocytosis
Authors: POLAK H. , NEMEC J. , Zita Z. , Blazková J. , Kucerová M. 
Published: October 2019

Abstract
The authors studied the influence of a six hour infusion of 25 I.U. of ACTH on the contractility and motility of leukocytes and on the absolute number of neutrophils in 10 convalescent patients.
The findings were as follows:
1. A significant increase occurred in leukocyte motility after the ACTH infusion, together with an increase in contractility.
2. The authors assume that leukocytosis following ACTH is due to an increase in the contractility of neutrophil leukocytes.
3. In view of the differences in the bone marrow reserve of mature neutrophils, it is impossible to demonstrate a direct correlation between the increase in motility, contractility and the absolute number of neutrophils.
4. An indirect estimate of the bone marrow granulocyte reserve can be made from the absolute number of neutrophils and the degree of leukocyte motility.
Studies of the Rate of Production and Life Span of Erythrocytes in Acute Leukemia
Authors: NATHAN DAVID G. , BERLIN NATHANIEL I. 
Published: October 2019

Abstract
The rate of red cell formation and the red cell life span were determined in six adult patients with acute myelogenous leukemia by the use of Fe59, Cr51 and glycine-2-C14. One patient with acute monocytic leukemia was studied with Fe59 alone.
Erythropoiesis was normal or increased in two patients, deficient in four patients and absent in one. The data indicate that in the cases of deficient erythropoiesis a decreased number of red cells were produced in the marrow, but that they were released at a rapid rate.
The red cell life span appeared to be normal in one patient, short but finite in four patients, and short owing to random destruction in one patient. One study did not include the measurement of red cell life span.
The liver removed an abnormally large proportion of the plasma iron when marrow erythropoiesis was deficient.
The bone marrow examination may be an unreliable indicator of the rate of erythropoiesis.
The Measurement of Folic Acid Activity in Serum: A Diagnostic Aid in the Differentiation of the Megaloblastic Anemias
Authors: HERBERT VICTOR , BAKER HERMAN , FRANK OSCAR , PASHER INEZ , SOBOTKA HARRY , WASSERMAN LOUIS R. , Weill Hazel , Bandel Lee 
Published: October 2019

Abstract
A modified L. casei microbiologic assay for the "folic acid" content of fasting serum appears to reflect the presence or absence of folic acid deficiency in patients with megaloblastic anemia of various causes.
Advantages of this assay method for fasting serum "folic acid" level over other procedures presently in use for evaluating possible folic acid deficiency are discussed.
Autocannibalism: The Etiology of Nerve-Resection Anemia
Authors: BEUTLER ERNEST , HOFSTRA DIANA 
Published: October 2019

Abstract
1. The finding of Hollán that severe anemia follows nerve resection in rats has been confirmed.
2. By the use of total-body labeling with radioiron, it has been demonstrated that this anemia is due to blood loss.
3. The blood lost by the nerve-sectioned rat is found in the stools, but autopsy reveals no source of gastrointestinal bleeding.
4. Anemia occurs only in animals with excoriation of the denervated foot. Anemia may be precipitated by withdrawing food for 48 hours. Hollán has found that amputation of the denervated limb prevents onset of the anemia.
5. It is concluded that nerve-resection anemia in the rat is due to autocannibalism of the denervated foot.
The Cellular Composition of Inflammatory Exudates in Human Leukemias
Authors: BOGGS DANE R. 
Published: October 2019

Abstract
An induced inflammation in patients with the leukemias was studied with reference to the cellular character of the exudate. Patients with the chronic forms of leukemia generally exhibited a normal cellular response. The granulocytic character of the exudate was proportional to the number of circulating mature neutrophils and metamyelocytes. Immature leukemic cells were not found in the exudates. The patients with acute leukemia with markedly reduced circulating mature granulocytes responded with acellular exudates or with marked qualitative and quantitative alteration in cellular composition. Bacterial infection was most frequent in the patients with few or no cells in the exudate.
The nature of the defect or defects in leukemia relating to increased susceptibility to bacterial infection are reviewed.