bluuebunny/crossref_metadata_2025 huggingface dataset:
Nongastric Marginal Zone B-Cell Lymphoma: A Prognostic Model from a Retrospective Multicenter Study.
Authors: Oh Sung Yong , Kwon Hyuk-Chan , Kim Won Seog , Park Yeon Hee , Kim Kihyun , Kim Sung-Hyun , Kim Hyun Jung , Kwon Jung Mi , Lee Jeeyun , Ko Young Hye , Ahn Yong Chan , Oh Suk Joong , Lee Soon Il , Bang Soo Mee , Kim Jung Han , Park Jinny , Ryoo Baek-Yeol , Lee Seung-Sook , Kim Ho Young , Kim Hyo-Jin
Published: October 2019

Purpose: The International Prognostic Index (IPI) and Follicular Lymphoma Prognostic Index (FLIPI) are used as prognostic indices for NHL and indolent lymphoma. However, marginal zone B-cell Lymphoma (MZL) evidences a distinctive clinical presentation and a natural course; thus, in this study, we attempted to devise an adequate prognostic index for MZL.

Patients and method: From 1990 to 2005, 205 patients diagnosed with MZL were retrospectively reviewed. After analysis of the prognostic factors, progression free survival (PFS) and overall survival (OS), we constructed a prognostic index of MZL (MZLPI) via the summation of each factor. We then compared PFS and OS with IPI, FLIPI, and MZLPI.

Results: According to our multivariate analysis of PFS and OS of MZL, nodal MZL, ECOG performance ≥ 2 and advanced stage were composed of MZLPI. MZLPI was grouped as follows: score 0 as a low risk group, score 1 as an intermediate risk group, and score ≥2 as a high risk group. The PFS curve, according to MZLPI results, evidenced a more discriminated pattern than IPI and FLIPI, and this was especially true in the intermediate risk group. In OS, MZLPI (P=0.0007) evidenced a more discriminated pattern than IPI (P=NS) or FLIPI (P=0.0044).

Conclusion: MZLPI, which is constructed of relatively simple factors, may represent a useful prognostic index for the prediction of PFS and OS in MZL, and may also be used as a substitute for IPI or FLIPI.
Phase I/II Study of Rituxan (R) in Combination with Doxil (D) in Patients (pts) with Relapsing or Refractory B-Cell Lymphoma: Clinical Efficacy without Cardiac Toxicity.
Authors: Czuczman Myron S. , Skipper Marion , Mohr Alice , Bernstein Zale P. , Hernandez-Ilizaliturri Francisco J. , Chanan-Khan Asher A.
Published: October 2019

In the past, the most common initial combination chemotherapy regimen used by most oncologists to treat B-cell lymphoma was the CHOP regimen. Although doxorubicin (DOX) is a major component of CHOP, it is associated with myelosuppression, alopecia, and potential cardiotoxicity. In vitro data has demonstrated synergistic activity between rituximab and certain drugs, including DOX. Because of D’s unique liposomal encapsulation, delivery, and toxicity profile, it may prove to be a more effective, less toxic anthracycline to combine with R. A formal Phase I/II trial to evaluate the safety and efficacy of R+D in pts with relapsing B-cell lymphoma was undertaken. R (375 mg/m2/dose) was given on day 1 and D (30 mg/m2/dose) on day 3 on q 21 day cycles for 6 cycles. Thirty-six of 42 pts have been enrolled to date. Four pts (n=3 with aggressive histologies) progressed while on treatment and did not complete the planned 6 cycles of therapy; 1 pt has not completed therapy, but is a PR mid-treatment. Demographics: 19M:17F; Median age = 63 (range 35–83); Follicular lymphoma (FL) grade 1 or 2 (n=14); FL, grade 3 (n=7); SLL (n=5); de novo DLBCL (n=3); Transformed NHL (n=6); Mantle cell lymphoma (n=1); Median number of prior treatments = 2 (range 1–8); prior anthracycline exposure (n=24; 67%). Overall, R + D was very well-tolerated. Reversible grade 3 (n=8) or grade 4 (n=4) neutropenia was seen and responded to growth factor support. Reversible grade 3 thrombocytopenia was seen in 2 pts. Of note, grade 3 palmar/plantar erythrodysesthesia occurred only in 3 pts whom were non-compliant with instructions given to avoid this side-effect associated with D therapy. Overall response rate (ITT) = 64% (42% CR, 22% PR); responses were equally distributed between both indolent and aggressive histologies. Median time-to-progression (TTP) = 12 months (range 1M to 57+M); mean TTP = 17 months. Median TTP in CRs = 14 months (range 4M to 57+M); Median TTP in PRs = 5 months (range 1M to 17M). Fifteen of 24 (63%) pts with prior anthracycline exposure demonstrated objective responses (9 CR; 6 PR). No clinical cardiac toxicity has been seen and comparison of pre-Rx to post-Rx LVEF remained >50% in all pts completing therapy. D + R immunochemotherapy is a unique, well-tolerated, non-cardiotoxic and effective salvage therapy for pts with either indolent or aggressive relapsed B-cell lymphoma. An update of the completed database will be presented at the annual ASH meeting.
RNA Based Cancer Vaccines - Clinical Trials in Patients with Prostate Cancer and Malignant Melanoma.
Authors: Kvalheim Gunnar , Aamdal Steinar , Gaudernack Gustav
Published: October 2019

Anti-tumour vaccines targeting the entire tumour antigen repertoire represent an attractive immunotherapeutic approach. This repertoire is present in the total mRNA isolated from the tumour. The mRNA from each cancer patient may be amplified and thus overcome the problem of limitation of material that has hampered the development of individualized vaccines. We have developed immuno-gene-therapy for malignant melanoma and prostate cancer. Monocyte-derived dendritic dells are transfected with autologous melanoma-mRNA or mRNA from three prostate cancer cell lines (DU-145, LN-CaP and PC-3). The vaccines may generate T cell responses against a broad repertoire of tumor-associated epitopes, and the melanoma approach moreover target patient-specific tumor antigens. Effective protocols were established for mRNA-transfection by square wave electroporation and for generation of clinical grade dendritic cells. A full scale preclinical evaluation demonstrated in vitro T cell responses in 6/6 advanced melanoma patients. The responses were specific to antigens encoded by the transfected tumor-mRNA. Recently, we have conducted two phase I/II trials, in advanced malignant melanoma and androgen-resistant prostate cancer. Successful vaccine preparations were obtained for all 41 patients elected. No serious adverse effects were observed. Specific T cell responses (T cell proliferation and/or IFNγ ELISPOT) were demonstrated in 9/19 evaluable melanoma patients and in 12/19 prostate cancer patients. The response rates were higher for patients receiving intradermal vaccination, compared to intranodal injection. Thirteen prostate cancer patients developed a decrease in log-slope PSA. The PSA-response was significantly related to the T cell response (p=0,002). We conclude that the tumour mRNA based DC-vaccine is feasible and safe, and that T cell responses are elicited in about 50% of patients. In the next generation clinical protocols, patients will undergo Treg depletion by chemotherapy (Temozolomide) prior to DC vaccination. Subsequently, T cells will be expanded ex vivo using the Dynabeads Clin Ex Vivo system before re-infusion into the patients.
The Clinical Outcomes of Autologous Stem Cell Transplantation in Peripheral T Cell Lymphoma: from Retrospective Multicenter Study in Korea.
Authors: Yang Deok-Hwan , Kim Won Seog , Kim Seok Jin , Bae Sung Hwa , Kim Sung Hyun , Kim In Ho , Mun Yeung-Chul , Shin Ho-Jin , Chae Yee Soo , Kwak Jae-Yong , Kim Hawk , Kim Min Kyoung , Kim Jin Seok , Baek Jin Ho , Suh Cheol Won , Park16 Hee Sook
Published: October 2019

Peripheral T cell lymphoma (PTCL) is a heterogenous group of aggressive lymphoma, which T-cell phenotype itself associated with unfavorable prognostic factors when compare to patients with B-cell phenotype lymphoma. High-dose chemotherapy (HDT) followed by autologous stem cell transplantation (ASCT) has represented a rescue option from poor prognosis. We retrospectively investigated the clinical outcomes of ASCT in 139 Korean patients with PTCL between December 1994 and June 2007. The overall survival (OS) and progression-free survival (PFS) were measured from the date of transplantation and were estimated using the Kaplan-Meier method. The median age at transplantation was 41 years. The histological subtypes had 45.3% PTCL-unspecified, 13.7% anaplastic large cell (ALK +/−: 15.7/57.8%), 10.8% angioimmunoblastic T cell, 21.6% extranodal NK/T cell, and 8.6% others. Disease status at transplantation consisted of 54 patients (38.8%) with first complete response (CR)/partial response (PR), 58 patients (41.7%) with chemosensitive-relapsed CR/PR, 15 patients (10.8%) with refractory CR/PR and 12 patients (8.6%) with progressive or refractory. At a median follow-up of 29.3 months, the 5-year probability of OS and PFS was 42.3±4.9% and 42.2±5.9%, respectively. The 5-year OS and PFS rates in patients with 1st CR/PR were 64.9±7.2% and 61.1±7.6%, compared to those in patients with chemosensitive-relapsed and refractory CR/PR were 33.5±6.8% and 33.2±8.3%, respectively. Early transplant-related mortality was 10.1%. Histological subtypes, bulky (≥10cm), prognostic index for PTCL (PIT) at diagnosis (0–2 vs 3–4), age-adjusted IPI at diagnosis, disease status at transplantation, the transplant timing (front-line vs second-line) and conditioning regimen were significant differences of survivals in univariate analysis. PIT, the transplant timing and the achievement of CR before transplantation were the variable prognostic for OS and PIT, bulky, and the achievement of CR provided significant independent factors for PFS in multivariate analysis. Our data showed that patients with PTCL who had low PIT and achieved CR before ASCT improved the OS and PFS regardless of the transplant timing and front-line ASCT of primary chemosensitive patients was effective in the OS.

Figure Figure
Effect of Thrombotic Events on Overall Survival in Patients with Newly Diagnosed Myeloma: Analysis from a Randomized Phase III Trial of Thalidomide Plus Dexamethasone Versus Dexamethasone in Newly Diagnosed Multiple Myeloma (E1A00).
Authors: Kumar Shaji , Zhang Lijun , Greipp Philip R. , Rajkumar S. Vincent
Published: October 2019

Background: Patients with newly diagnosed multiple myeloma is at increased risk of thrombotic events, primarily venous thrombosis. The risk is significantly increased in patients receiving certain therapeutic regimens such as the combination of thalidomide and dexamethasone. Deep vein thrombosis, especially if complicated by pulmonary embolism, can result in death. It is not clear if venous thrombotic events can result in compromised survival in this group of patients.

Methods: We studied patients enrolled in the Eastern Co-operative Group trial (E1A00) that randomized patients with newly diagnosed myeloma to single agent dexamethasone or thalidomide and dexamethasone. Patients with grade 3 or greater thrombotic events (DVT, or myocardial ischemia, or stroke) were identified. Wilcoxon rank sum test (for continuous variables) or Fisher’s exact test (for categorical variables) was used to compare age, baseline M-protein, hemoglobin, WBC, platelets count, creatinine, beta2-microglobulin, or ECOG PS between patients with and without thrombotic events.

Results: Among the 207 patients enrolled, 36 patients (17.4%) had grade 3 or greater thrombotic events with any treatment attribution, of whom, 27 patients had DVT, 3 patients had an MI and 6 pts had a stroke. However, in only 31 of these patients were the event considered treatment-related (possible, probable, or definite) and among these 24 had a DVT. The baseline characteristics did not differ between the patients with thrombosis and those without, irrespective of its relationship to treatment. Similarly, the baseline characteristics did not diff for those with venous or arterial thrombosis. The overall survival was similar among the patients developing a DVT compared to those without a thrombotic event, when all DVT events were considered irrespective of the treatment attribution (Figure). However, majority of the patients with an arterial event (3 patients with MI and 6 with stroke) had a short overall survival.

Conclusion: Venous thrombosis remains a common complication in patients with myeloma, especially when treated with thalidomide and dexamethasone. While occurrence of thrombosis clearly contributes to morbidity and often leads to dose reductions and missed doses, it does not appear to have an adverse impact on the overall survival of these patients.

Figure Figure
Anti-CD3 Activated T Cells from Cord Blood Can Be Expanded Ex Vivo and Armed with Bispecific Antibodies To Lyse Her2/Neu + and CD20+ Targets: A Strategy for Providing Anti-Tumor/Lymphoma Effects after Cord Blood Transplant.
Authors: Sorenson Carly B. , Frandsen Thomas D. , Dorr Suanne , Ratanatharathorn Voravit , Uberti Joseph P. , Lum Lawrence G.
Published: October 2019

Our previous studies showed that anti-CD3 activated T cells (ATC) from peripheral blood mononuclear cells could be expanded in interleukin-2 (IL-2) for 14 days and armed with anti-CD3 x anti-Her2/neu (Her2Bi)[J Hemat & Stem Cell Res10:247,2001], anti-CD3 x anti-CD20 (CD20Bi)[Exp Hemat33:452,2005], or anti-CD3 x anti-EGFR [Clin Cancer Res12:183,2006] bispecific antibody (BiAb)and can kill Her2/neu, CD20, and EGFR+ tumor targets, respectively. In this study, we asked whether anti-CD3 activated cord blood T cells (CBATC) could be expanded and targeted with Her2Bi and CD20Bi to tumors or hematologic malignancies for infusions after cord blood stem cell transplant (CBSCT). CB mononuclear cells were activated with anti-CD3 (20 ng/ml) and expanded for 14 days in IL-2 (100 IU/ml). CBATC were armed with Her2Bi or CD20Bi and tested for specific cytotoxicity directed SK-BR-3, Raji, or B9C targets and cytokine secretion or IFNγ EliSpots after binding to tumor cells. Our results show the mean expansion of CBATC to be 43-fold (n=8) after 14 days of culture. By the end of culture, the proportions of CD8+ and CD4+ were 82% and 18%, respectively. The proportion of cells expressing CD19 or CD20 did not exceed 6.3%, CD56+ cells were <3.6% and CD3-CD16+CD56+ cells was <0.7%. Cells positive for CD4+CD25+ or CD8+ CD25+ were 4.2% or 7.1%, respectively (n=2). Specific cytotoxicity was optimized when CBATC were armed with 50 ng/106 cells of Her2Bi or CD20Bi (arming dose ranged from 5, 50, and 500 ng/106 cells); arming significantly increased cytotoxicity of the armed CBATC over that seen for unarmed CBATC. Cytotoxicity peaked between days 12 and 14 for both BiAbs. The ability of CD20Bi armed ATC to produce Elispots for IFNγ were tested by arming ATC the CD20Bi after 0, 8, 11, and 13 days of culture peaked on day 8. Only CD20+ targets induced Elispots and day 8 armed ATC exhibited peak numbers (2,200 Elispots(ranged from 1,700 to 1,800 on day 8)/106 armed ATC plated). At an effector/target ratio (E:T) of 25:1, the mean cytotoxicity of CBATC armed with Her2Bi or CD20Bi was 60% (n=4) and 35% (n=1), respectively. In an extended culture to day 47, mean cytotoxicity for Her2Bi-armed CBATC was 36% at an E/T of 25:1 compared to 4.35% for unarmed CBATC. Unarmed CBATC did not kill Daudi targets. Armed CBATC mediated both specific cytotoxicity and secreted IFN-γ as measured by ELISA or EliSpots. Both fresh and frozen CB could be used in the assays. In a clinical application, specific cytotoxicity of armed CBATC could be used to augment anti-tumor and anti-lymphoma effects after CBSCT.
Onset of Cardiac Iron Loading in Pediatric Patients.
Authors: Wood John C. , Origa Raffaella , Agus Annalisa , Matta Gildo , Coates Thomas D. , Galanello Renzo
Published: October 2019

Patients with thalassemia major develop life threatening cardiac complications in their teens and twenties from iron overload. Cardiac MRI allows us to diagnosis preclinical cardiac iron deposition, but it is not known at what age T2* screening should be initiated. Historical data and pilot MRI studies suggest that patients must reach a critical transfusional exposure prior to cardiac iron uptake. This study was a two-institution characterization of the prevalence of cardiac iron overload in 77 pediatric patients.

Methods: Study was performed at the Ospedale Regionale Microcitemie in Cagliari and the Childrens Hospital Los Angeles (CHLA). Retrospective review of medical records was authorized by the IRB for pediatric patients undergoing cardiac MRI prior to July of 2007. Subjects at both institutions had assessment of cardiac T2* and cardiac function using validated techniques on a 1.5 T General Electric CVi scanner; patients at CHLA also underwent MRI-based liver iron measurements. Complete transfusional iron burden was measured in the Italian patients. Serial MRI data was recorded in 30 patients, but associations between cardiac T2* and age were determined using linear and logistic regression using only results from the first patient MRI scan.

Results: Patient ages ranged from 8.0 to 18 years of age at Cagliari (n=36) and 2.5 to 17.9 years of age at CHLA (n=41), reflecting the use of MRI to monitor liver iron in patients < 8 years of age. Patients were moderately iron loaded with hepatic iron concentrations of 12.7 ± 9.8 (CHLA) and ferritin values of 2329 ± 1162 (Cagliari). Median cardiac T2* was 30.2 and ranged from 3.4 to 72.8. Cardiac T2* and its reciprocal were uncorrelated with liver iron (CHLA) and ferritin levels (Cagliari). Figure 1 demonstrates the decrease in cardiac T2* with increasing chronologic age. Serial data are connected by lines. Although linear regression was weakly positive (r2 = 0.06, p = 0.03), the relationship was fundamentally nonlinear. No patient below the age of 9.5 years of age demonstrated an abnormal cardiac T2* (< 20 ms) while 36% of patients between the ages of 15–18 years had detectable cardiac iron. Figure 2 demonstrates the logistic regression curve modeling the prevalence of detectable cardiac iron as a function of age (r2= 0.13, p< 0.002). Chronologic age was highly correlated (r2 = 0.88) with both transfusional iron burden and with duration of transfusion therapy, making it impossible to separate the relative importance of these three variables.

Conclusion: Thalassemia major patients did not accumulate cardiac iron until early in their second decade of life. Consequently, cardiac iron monitoring can be safely deferred until children are able undergo MRI examination without sedation.

Figure Figure Figure Figure
Improved Outcome of Adult Acute Lymphoblastic Leukemia Treated with a Pediatric Protocol: Results of a Pilot Study.
Authors: Haiat Stephanie , Vekhoff Anne , Marzac Christophe , Calendini OrsAnton , Lapusan Simona , Marjanovic Zora , Rio Bernard , Marie Jean-Pierre , Legrand Ollivier
Published: October 2019

Adult ALL still have poor outcome compared with childhood ALL, with an expected OS of less than 40%. Recent retrospective studies have shown that adolescents and young adults (<30 yo) treated with pediatric protocol have a better prognosis than those treated with adult protocols. The aim of this pilot study was to assess the feasibility and efficacy of the French pediatric protocol FRALLE 2000 to treat 28 oldest adult ALL aged from 16 to 57 years.

Methods: 28 consecutive adults Philadelphia negative ALL aged from 16 to 57 years, with a median follow-up of alive patients of 35 months, received treatment courses according to FRALLE 2000 from 2001 to 2007. After a prednisone prephase and a four-drugs induction (prednisone, daunorubicin, vincristine and 9 infusions of L-asparaginase), patients in CR received a consolidation course, two delayed intensifications with L-asparaginase separated by an interphase, and a maintenance chemotherapy during two years. Results were compared with the outcome from 20 consecutive patients treated in our institution with the historic EORTC ALL-4 adults protocol from 1998 to 2001.

Results: All the clinical (age, WHO performance status, gender) and biological (WBC, phenotype, cytogenetic) parameters of patients treated in FRALLE protocol were statistically similar with those of patients treated in ALL-4 adults protocol. CR rate was achieved in 82% of patients after FRALLE induction, and 100% after a salvage therapy with high doses cytarabine. The good early response was evaluated by cortico-sensitivity and chemo-sensitivity (after 14 days of chemotherapy). All patients who achieved cortico and chemo-sensitivity were alive in persistant CR, with a better survival than other patients (p=0.008) (fig1). When we compared patients treated by FRALLE or ALL-4 protocol, the 4-years DFS and OS are 90% +/−6% vs 47%+/−12%, (p= 0,01) and 83%+/−9% vs 35%+/−16%, (p=0,05) respectively (figures 2 and 3). This better outcome is not explained by significant differences in patients characteristics nor by a better CR rate but rather by a lower relapse rate in the pediatric treatment group. This indicates a major role of the dose intensity, especially for L-asparaginase, corticosteroid, methotrexate, and purinethol. No treatment related mortality and no severe side effect, except one pulmonary embolism, were observed during treatment with supportive cares including parenteral nutrition, granular growth factor, infectious prophylaxis, and antithrombine III infusions. This pilot study shows that adults up to 57 years with Ph negative-ALL have a dramatically better outcome when there are treated with childhood ALL protocol without any major side effect. This therapeutic strategy has to be confirmed by the current prospective study performed by the EORTC/HOVON group.

Figure Figure
Novel Synthetic Histone Deacetylase Inhibitor, SK-7041, Has Potent Anti-Proliferative Activity in Acute Myeloid Leukemia Cell Lines.
Authors: Yoon Sung-Soo , Bae Eunkyung , Park Juwon , Cha Yongjun , Lee Young Y. , Kim Byoung K.
Published: October 2019

Histone acetyltransferase (HAT) and histone deacetylase (HDAC) activities determine the acetylation status of histones, which regulates gene expression through chromatin remodeling. Aberrant histone acetylation is known to play a key role in leukemogenesis. Thus, histone deacetylase inhibitors (HDACIs) are emerging as a new class of anti-cancer agents for leukemia. In this study, we examined anti-proliferative effects of novel hybrid synthetic HDACI, SK-7041, in various acute myelogenous leukemia (AML) cell lines (KG-1, HL-60, HEL, U937, ML-1). SK-7041 induced the time-dependent hyperacetylation of histones H3 and H4 in AML cell lines. It preferentially inhibited the enzymatic activities of HDAC1 and HDAC2, as compared with the other HDAC isotypes, indicating that class I HDAC is the major target of SK-7041. All the cell lines tested showed similar patterns of growth inhibition by SK-7041. Their IC50 values were approximately 0.5 mM at 72 hours incubation. SK-7041 effectively induced the apoptosis via activation of caspase-3, -7, -9, and PARP, but not caspase-8. SK-7041 enhanced G1 arrest via decreasing Cyclin D1 expression and increasing p21 expression. Changes in gene expression profiles after treating KG-1 cells with various concentrations of SK-7041 were assessed using a cDNA microarray consisting of distinct cDNAs of cancer-related genes. 7 genes, namely RGL1, FYN, CARD9, ABCA7, TNFRSF6B, CASP9, and ENPP2 were induced substantially (Global M >2.0) and 8 genes, namely PTPN7, CD34, INSIG1, IL16, LHX6, TRIB3, BID, and PDCD4 were reduced substantially (Global M < 2.0). In conclusion, this study showed SK-7041 inhibited cancer cell growth and caused characteristic gene expression profile changes in AML cells. The alteration in levels of acetylated histones was closely associated with expression of specific cancer-related genes in AML cells.
Detection of Nucleophosmin Gene Mutations in Plasma from Patients with Acute Myeloid Leukemia: Clinical Significance and Implications.
Authors: Ma Wanlong , Zhang Xi , Jilani Iman , Ravandi Farhad , Estey Elihu , Kantarjian Hagop , Giles Francis , Sheikholeslami Mohammad , Keating Michael , Albitar Maher
Published: October 2019

Nucleotides insertion in the nucleophosphamin (NPM1) gene has been reported in about one third of patients with acute myeloid leukemia (AML). Multiple studies showed that the presence of NPM1 mutations associated with better outcome in patients with AML. Studies reported to date have analyzed leukemic cells obtained from bone marrow or peripheral blood. We tested for mutations in the NPM1 gene using peripheral blood plasma and compared results with clinical outcome from a single institution. Analyzing plasma from 98 newly diagnosed patient with AML showed NPM1 mutation in 24 (23%) of patient while only one (4%) of 28 previously untreated patients with myelodysplastic syndrome (MDS) showed NPM1 mutation. Compared with peripheral blood cells, 2 (8%) of the 24 positive patients were negative by cells; none were positive by cells and negative by plasma. Most of the mutations detected (45%) were in patients with FAB classification M2, M4 and M5. In addition to the reported 4 bp insertion, we also detected 4 bp deletion in one patient in cells and plasma. Patients with NPM1 mutation had a significantly higher white blood cell count (P = 0.0009) and a higher blast count in peripheral blood (P = 0.002) and in bone marrow (P = 0.002). Blasts in patients with NPM1 mutant expressed lower levels of HLA-DR (P = 0.005), CD13 (P = 0.02) and CD34 (P < 0.0001), but higher CD33 levels (P = 0.0004). Patients with NPM1 mutation appear to have better chance of responding to standard therapy (P = 0.06). Event free survival of patients with NPM1 mutation was longer (P = 0.056) than in patients with intermediate cytogenetic abnormalities. The most striking difference in survival was in patients who required >35 days to respond to therapy (Figure). Survival was significantly longer in patients with NPM1 mutation requiring >35 days to respond (P = 0.027). This data not only support that NPM1 plays a significant role in the biology and clinical behavior of AML, but also show that plasma DNA is enriched with leukemia-specific DNA and is a reliable source for testing.

Figure Figure
Survival Is Poorer in Patients with Secondary Core Binding Factor Acute Myelogenous Leukemia Compared with De Novo Core Binding Factor Leukemia.
Authors: Borthakur Gautam , Jain Nitin , Kantarjian Hagop , Lin E. , Ravandi Farhad , Pierce Sherry , Estey Elihu
Published: October 2019

Translocation (8;21)(q22;q22) and inversion of chromosome 16 [inv(16) (p13q22)] are considered good-risk cytogenetic abnormalities in acute myeloid leukemia (AML) accounting for 15% of primary AML cases (Blood. 2002 Dec 15;100(13):4325–36) and are characterized at the molecular level by disruption of genes encoding subunits of core binding factor (CBF). Increasing age, increasing peripheral blast percentage and complex cytogenetics are associated with poor overall survival in these patients (Br J Haematol. 2006 Oct;135(2):165–73). Here we assess differences in patient characteristics and outcomes between the primary and secondary core binding factor AML. One hundred eighty nine CBF AML patients treated at our institution were studied; 18 (9.5%) had secondary AML. Patients with secondary CBF AML were older (p= 0.02, median ages 57 vs. 44 years) and had lower WBC counts (p=0.03) (Table1). Overall survival (OS) was worse in the secondary AML patients (94 weeks versus 621 weeks, p=0.0016) (Figure 1). Age, hemoglobin, platelet count, and bilirubin were significantly associated with OS in the univariate analysis. In the multivariate analysis, after adjusting for age, hemoglobin, WBC, and bilirubin, secondary CBF AML was marginally significantly associated with worse OS (hazard ratio 1.884, CI95% 0.979–3.625, p=0.058) but not worse PFS (p= 0.15) (Table 2). These data suggest that the secondary CBF AML has much poorer prognosis than the primary CBF AML, further indicating that “CBF AML” is not a homogeneous entity with a uniformly good prognosis.

Table 1: Summary statistics of patients’ continuous characteristics by abnormality status Patient characteristics Total patients Primary AML (n=171, male=96) Secondary AML (n=18, male=8) p-value (primary vs secondary AML) Median (range) Median (range) Median (range) Age (years) 44 (16–88) 44 (16–88) 57 (31–75) 0.02 WBC (103/mm3) 14.7 (0.6–387) 16.4 (0.6–387) 6.6 (0.8–103.8) 0.03 Hemoglobin (g/dl) 8.1 (2.5–14.3) 8.2 (2.5–14.3) 7.8 (4.8–10.8) 0.33 Platelet count (103/mm3) 38 (5–350) 38 (5–350) 39 (9–139) 0.88 Bilirubin (mg/dl) 0.6 (0.0–15.3) 0.6 (0.0–5.0) 0.6 (0.1–15.3) 0.94 Creatinine (mg/dl) 0.9 (0.5–2.9) 0.9 (0.5–2.9) 0.9 (0.5–1.8) 0.66 Table 2: Multivariate Cox proportional hazards model in estimating the association between covariates and patients’ survival Variable Hazard ratio (95% CI) p-value Abnormality (secondary vs primary) 1.884 (0.979–3.625) 0.058 Age (1 year increase) 1.028 (1.012–1.044) 0.0004 Hemoglobin (1gm/dl increase) 0.864 (0.769–0.970) 0.014 WBC (103/mm3 increase) 1.003 (1.000–1.007) 0.061 Bilirubin (1 mg/dl increase) 1.179 (1.044–1.332) 0.008

Figure Figure
Prognostic Value of Serum Lactate Dehydrogenase (LDH) in Patients with Primary Systemic Amyloidosis Undergoing Stem Cell Transplantation.
Authors: Kumar Shaji , Lacy Martha Q. , Dispenzieri Angela , Hayman Suzanne , Buadi Francis K. , Dingli David , Litzow Mark R. , Gertz Morie A.
Published: October 2019

Background: Primary amyloidosis (AL) is characterized by multi-organ deposition of light chain derived amyloid protein, resulting in organ dysfunction of varying severity. High dose chemotherapy and autologous stem cell transplantation (ASCT) is an effective treatment for AL, but is limited in applicability due to high transplant related mortality in patients with advanced organ dysfunction. Serum LDH, an enzyme that catalyzes the conversion of lactate to pyruvate, is present in large amounts in heart, kidney, liver, and muscle and is elevated with abnormalities in one or more of the organs. It is commonly elevated in a variety of hematologic malignancies. Given the multi-organ involvement in AL, we examined the prognostic value of LDH in AL.

Methods: We examined our transplant database to assess the prognostic value of LDH. Laboratory values from pre-ASCT evaluation were used for the analysis.

Results: Three hundred and thirteen pts who underwent ASCT between March 1997 and March 2007 were included in this analysis. The median age at ASCT was 57.5 years (range, 33–73); and 183 pts (58.3%) were male. The median duration from diagnosis to transplant was 4.1 months (range, 1–75) and the median follow up post transplant was 28.3 months (range, 1 m to 10 years) for the 229 pts (73%) alive at the time of analysis. The median LDH was 194 IU (range, 92-441) and 138 pts (44%) had an LDH > 200. The median overall survival post-ASCT for patients with an LDH > 200 was 59.4 mos (95% CI; 37.9, 80.8) compared to not reached for the rest (Figure). The prognostic value of LDH for post transplant OS was independent of cardiac troponin (cTnT). The median OS post ASCT for patients with cTnT > 0.035 (n=208) was 37.4 months (95% CI; 17, 57.9) compared to 97.5 months (95% CI; 64.8, 130.2) for the rest. In a Cox proportional hazards model, cTnT > 0.035 was associated with a 2.5 fold risk of death (p = 0.0013) and an LDH > 200 was associated with 1.7 fold risk (p = 0.04). We then examined if LDH added to the cTnT in a prognostic model for survival post ASCT. Pts with both cTnT < 0.035 and LDH < 200 (n=133) had a median OS that was not reached compared to 71 mos (95% CI; 48.3, 94) if one factor was abnormal (n=89) and 37.4 mos (95% CI; 16.1, 58.8) if both cTnT and LDH were abnormal as above (n=45); p < 0.001 (Figure 2).

Conclusions: In this study we have demonstrated the prognostic value of serum LDH in AL pts undergoing ASCT. Serum LDH appears to add to the value of troponin in prognostic staging of these patients. These results need to be examined in a larger cohort of patients undergoing different treatments. It is possible that LDH is able to globally assess abnormalities of multiple organs including heart, liver and lung. Prospective studies looking at the different LDH fractions are ongoing.

Figure Figure Figure Figure
Hepatic Response after High-Dose Melphalan and Stem Cell Transplantation for AL Amyloidosis Associated Liver Disease.
Authors: Girnius Saulius , Seldin David C. , Skinner Martha , Finn Kathleen T. , Sanchorawala Vaishali
Published: October 2019

Immunoglobulin light chain (AL) amyloidosis is a plasma cell dyscrasia with extracellular protein deposition in various organ systems, including the liver. The natural history of AL amyloidosis with liver involvement has a poor prognosis, with median survival of only 8.5 to 9 months. High-dose melphalan chemotherapy and autologous peripheral blood stem cell transplantation (HDM/SCT) has been shown to result in durable hematologic response and prolonged overall survival in systemic AL amyloidosis. Patients were included who had hepatic involvement and received HDM/SCT from 1998 to 2006. Data was collected with the approval of the Institutional Review Board. Patients receiving HDM/SCT had to meet eligibility criteria to qualify for this aggressive treatment. Stem cells were mobilized using G-CSF alone. Intravenous melphalan was administered at 100–200 mg/m2 over 2 days in divided doses and stem cells were re-infused 24–72 hours after HDM. Liver response was determined by criteria set up by the Consensus Opinion from the 10th International Symposium on Amyloid and Amyloidosis. Sixty-nine patients qualified for this study, including 47 males and 22 females. The median age was 56 years (range, 37–75), median bilirubin was 0.5 mg/dL (range, 0.1 to 5.7), median alkaline phosphatase was 193 U/L (range, 59 to 1243) and the median liver size was 3 cm (range, 0–20cm) below the costal margin. Nine patients (13%) died from treatment-related mortality. The hematologic CR one year after treatment was achieved by 53% (31/58) of evaluable patients. The overall survival was 84% at 1 year and 49% at 5 years, by Kaplan-Meier estimates. Hepatic response occurred in 21% (10/47) at 1 year after HDM/SCT and 40% (12/30) at 2 years after HDM/SCT. In summary, hepatic involvement with AL amyloidosis does not increase treatment-related mortality from HDM/SCT. Hepatic response follows a hematological response and can occur as far as 2 years from HDM/SCT. Forty % of surviving evaluable patients showed remission of their hepatic disease from AL amyloidosis by 2 years.
Reduction of Prion Infectivity in Packed Red Cells by Separation of Whole Blood and Washing of the Red Cell Fraction.
Authors: Morales Rodrigo , Buytaert-Hoefen Kimberley A. , Hansen Eric T. , Hlavinka Dennis , Goodrich Raymond , Soto Claudio
Published: October 2019

Although prion diseases are rare in humans, the established link between a new variant form of CJD (vCJD) and the consumption of cattle meat contaminated by BSE have raised concerns about a possible outbreak of a large epidemic in the human population. Over the past few years, BSE has become a significant health concern in several countries, and it now seems apparent that vCJD can also be iatrogenically transmitted from human to human by blood transfusion. Exacerbating this state of affairs is the lack of a reliable test to identify individuals incubating the disease during the long and silent period from the onset of infection to the appearance of clinical symptoms. The purpose of this research study was to evaluate the effectiveness of separation of whole blood and washing of the red cell fraction for the removal of infectious scrapie prion protein (PrPSc) from red blood cell (RBC) suspensions. Samples of human, whole blood were spiked with 5 × 106 LD50 263K PrPSc. Analysis of the treated sample supernatants by Western blot revealed that approximately >88% of the PrPSc was removed with the initial plasma expression and the equivalent of 6% was detected in a saline wash (300 mL; 0.9% saline). The final sample of RBCs revealed no detectable levels of PrPSc by Western blots. Further analysis of the treated RBCs using the PMCA assay indicated detectable amounts of PrPSc only after 2 consecutive amplification rounds. Semi-quantitative analysis of PMCA amplification enabled us to estimate that the treated RBCs contained less than 1 × 104 LD50 PrPSc. This corresponded to removal levels exceeding ≥99% of spiked material in whole blood. These in vitro estimations were confirmed by in vivo infectivity studies in a hamster model of disease transmission.

Results from in vivo studies displayed significant differences in the incubation periods of the spiked blood inoculated hamsters (100.1 ± 1.7) versus washed RBCs (135.8 ± 6.7). Moreover, a substantial difference in the attack rate (6/15: 40% in washed RBC, versus 13/13: 100% in spiked blood) further indicated a substantial removal of infectious prions. Comparison of this data with results of animals inoculated with different dilutions of infectious material, indicated a >99.94% reduction of infectivity. Washed, packed human red cells produced by this procedure were able to be stored in standard additive solutions (AS-3) for 42 days while still meeting all in vitro blood bank standards for acceptable red cell quality.

Conclusion This data suggests that separation of plasma coupled with a simple, low volume wash of red cells may represent an efficient method to remove prions from red blood cell fractions, thus reducing possible infectivity of these products.
Recovery of Donor Peripheral Blood Platelet Count Following Platelet Apheresis.
Authors: Dumont Larry J. , Lassahn L. , Tomasulo Peter A. , Harpool Dennis , Pinkard S. , Rugg Neeta , Cancelas Jose A. , Landry S. , Ambruso Daniel R. , Clementi Chris , Schuyler Robert
Published: October 2019

BACKGROUND: Apheresis platelet collection from healthy normal blood donors can reduce the donor peripheral blood platelet concentration by 50% or more. The kinetics of peripheral blood platelet count (PLT) recovery in the apheresis donors over the first 24 hours has not been described. The objective of this study was to determine the recovery kinetics of the donor peripheral blood platelet count following apheresis platelet donation.

METHODS: Healthy apheresis platelet donors were enrolled following informed consent. The apheresis platelet collection was performed using the Gambro Trima system (Gambro BCT, Lakewood, CO) following local SOP and manufacturer’s directions for use. The minimum predicted post-count was configured in the Trima to no less than 78K plt/μL. PLT was determined pre-procedure (Pre), immediately post procedure (Post), 4–11 h (FU1) and 11–41 h (FU2) post-donation using standard methods. The PLT recovery was evaluated as the increase in PLT following the donation (Delta). The effects of study site, time of sample, and the fraction of platelets collected (Fpc) at donation on Delta were evaluated using a random effects generalized linear regression model. A full regression model of Delta as a function of study site, follow-up period and Fpc with all main and interaction effects was used to test hypotheses.

RESULTS: 548 subjects were entered into the study at 3 study sites; Pre-PLT 276±59 × 103 plt/μL, Post-PLT 205±47 × 103 plt/μL, Fpc 25±10%. No adverse events were reported by any subjects. Recovery of platelet count following apheresis platelet donation is variable between subjects; and the independent variables of study site, follow-period and Fpc accounted for 25% of the total variation in Delta. PLT increased 12.4±0.9 × 103 plt/μL by the time of follow-up sampling (p=0.01), although there was no difference between PLT at FU1 (214±49 × 103 plt/μL) and FU2 (212±47 × 103 plt/μL; p=0.15). None of the donors reached their pre-donation platelet count during the follow-up period. There was no difference in Delta between centers (p=0.23). Fpc had a significant affect on Delta (p<0.0001); with estimated Delta of 5.2±0.9 × 103 plt/μL at Fpc=15% and 16.0±.8 × 103 plt/μL at Fpc=30%.

CONCLUSION: Platelet recovery following apheresis platelet donation was observed to be dependent on the fraction of platelets donated. Surprisingly, the recovery observed within the first 11 h was equivalent to that observed between 11–42 h, averaging 17.5% of the drop observed during apheresis. Recovery was not complete when observed for up to 41 h following donation in this study. Additional investigation of PLT recovery following apheresis donation is indicated to describe and differentiate the potential roles of de novo production, early pro-platelet release and platelet release from peripheral pools over the early post-donation period.
High Levels of Circulating HSP70 Levels in Patients with Chronic Myeloid Leukemia and Correlation with Resistance to Therapy with Imatinib.
Authors: Tseng Richard , Yeh Chen-Hsiung , Jilani Iman , Zhang Zhong , Kantarjian Hagop , Cortes Jorge , O’Brien Susan , Giles Francis , Hannah Alison , Estrov Zeev , Keating Michael , Albitar Maher
Published: October 2019

Cellular increase in the heat shock protein 70 (HSP70) inhibits death receptor and mitochondria-initiated signaling for apoptosis. In Vitro and in vivo data from patients with chronic myeloid leukemia (CML) indicates that overexpression of HSP70 in leukemic cells is associated with resistance to imatinib. The HSP70 protein is detectable in the plasma and has been reported to represent a marker for cardiovascular stress as well as prostate cancer. We measured HSP70 in the plasma of 139 patients with CML using a sandwich assay based on meso scale technology and correlated levels of HSP70 protein in plasma with clinical behavior. All samples were collected prior to initiating imatinib therapy. The levels of HSP70 in CML patients in chronic phase (N=93) were not significantly (P=0.08) different from those in patients with CML in accelerated/blast crisis (N=46) (median 33.24 ng/ml, range: 3.892–128.172 ng/ml Vs median 26.57 ng/ml, range: 4.498–114.746 ng/ml, respectively). However, CML patients had significantly (P<0.0001) higher levels of HSP70 than normal control (N=95, median=4.17 ng/ml, range:1.746–24.684 ng/ml). There was significant correlation between plasma levels of HSP70 and platelets (r=0.54), WBC (r=0.32), and basophils (r=0.31) in patients in chronic phase and with platelets (r=0.72), blasts (r=0.39), and basophils (r=0.50) in patients in Acc/Bl phase. Patients in chronic phase and high levels above the median had significantly (P=0.02) higher rate of progression to ACC/Bl phase while on therapy with imatinib (figure). In addition these patients appear to have a tendency toward shorter survival (P=0.07). There was no significant correlation between plasma HSP70 levels and survival in patients with ACC/Bl phase. This data support the reported role of HSP70 in the resistance to imatinib in patients with CML and potentially plasma HSP70 levels may represent a marker for resistance in patients with chronic phase CML.

Figure Figure
Gfi-1 Is Overexpressed on Chronic Myeloid Leukemic Cells.
Authors: Cheng Hui , Chang Wei , Huang Ming , Sun Ying , Zhang Yicheng
Published: October 2019

Gfi-1 oncogene was first discovered as an integration site for Moloney murine leukemia virus(MoMulv) in virally infected cells that were selected for interleukin-2 (IL-2) independence; In synergy with pim-1 and L-Myc genes, Gfi-1 was involved in the pathogenesis of lymphoid malignancies. Recent studies also indicated that Gfi-1 regulates self-renewal and engraftment of hematopoietic stem cells and the development of neutrophils. Normally, Gfi-1 zinc finger protein is expressed almost exclusively on thymocytes or stem cells. Chronic myeloid leukemia (CML) is a typical hematopoietic stem cell disease, we ask whether Gfi-1 is expressed on CML cells and the possible role in the leukemogenesis. By RT-PCR and flow cytometry, we analysed the expression of Gfi-1 in the transcription and the protein levels respectively on bone marrow mononuclear cells(MNC) from 53 CML patients. Control samples were taken from the ribs of 5 patients undergoing thoracic surgery for lung cancer. Results Gfi-1 mRNA was detected in K562 cell line and all the MNCs from CML patients but not or very weak in control MNCs; Flow cytometry study show that both the positive percentage (87.2±7.5% Vs 12.4±6.6%, p<0.01) and the relative fluorescence intensity (represented by mean fluorescence intensity, 58.94 ±22.09 Vs 18.67±7.88, p<0.01) were significantly increased in CML MNCs. Considering the complexity of the composition of bone marrow MNCs, we also sorted CD34+ cells from CML or control samples and examined by Rt-PCR and flow cytometry; The Gfi-1 was found to be overexpressed in CD34+ cells of CML; On confocal microscopy, Gfi-1 was mainly sublocalized in nucleus. Conclusion Gfi-1 was overexpressed in MNCs or CD34+ cells from CML patients; Gfi-1 may be implicated in the leukemogenesis of CML and potentially be targeted in the future for treatment of CML.
Monitoring Endothelial Cell Activation In Vivo during Thrombus Formation.
Authors: Atkinson Ben T. , Nandivada Prathima , Furie Bruce , Furie Barbara C.
Published: October 2019

The endothelium serves as a metabolically active interface between the blood and underlying tissues and offers response to changes in its microenvironment. The endothelium is rapidly activated in response to stimuli associated with fluctuations in shear stress, physical trauma, oxidative stress, and thrombotic and inflammatory mediators. To study endothelial cell activation we have monitored calcium mobilization in vitro in cultured endothelial cells and also in situ in the living mouse, following vessel wall injury. The addition of exogenous ADP (10 μM) or thrombin (1 U/mL) to Human Umbilical Vein Endothelial Cells (HUVECs) loaded with the calcium sensitive dye, Fluo-4 AM, is followed by rapid elevation of intracellular calcium, with a sustained peak observed within 10 sec. To further investigate the ability of the endothelium to activate in response to mediators potentially localized in the microenvironment of the endothelium during thrombus formation, we investigated the ability of IL-6 (0.1 ng/mL), IL-8 (80 ng/mL), MCP-1 (15 ng/mL), NAP-2 (10 ng/mL) and SDF-α (60 ng/mL) to mobilize calcium in HUVECs. These potential agonists were selected based on their primary roles in inflammation and thrombosis, and proposed roles in atherosclerosis and tumor angiogenesis. SDF-α and NAP-2 rapidly mobilized calcium in HUVECs, with similar maximum responses, but NAP-2 initiated a more prolonged (4-fold longer) rise in intracellular calcium. IL-6, IL-8, and MCP-1 also mobilized calcium, but the rise in intracellular calcium was almost 4-fold lower than that observed with SDF-α and NAP-2. Comparable calcium mobilization occurs in HUVECs subjected to a single pulse of a nitrogen dye-tuned laser. In addition, targeting of single cells within a confluent culture of endothelial cells initiated calcium elevation in the targeted cell and was followed by a wave of calcium elevation in surrounding cells. To determine whether this endothelial cell activation, and more specifically the calcium elevation, occurs in vivo, we conducted experiments using the laser-induced model of thrombus formation to look for calcium elevation in the arteriolar endothelium in live mice. Calcium elevation was monitored by Fluo-4AM introduced intravenously into the mouse circulation via the jugular vein. Fluo-4AM is non-specific in its uptake among cell types, and endothelial cell uptake of the dye in the cremaster muscle microcirculation was observed. Upon laser-induced injury, calcium elevation in the endothelium could be monitored by intravital microscopy. The observed calcium elevation was rapid (within 30 s) and preceded detection of platelets in the developing thrombus. These results add to the evidence of a dynamic endothelium and demonstrate that the endothelium activates rapidly prior to thrombus formation in the laser induced thrombosis model.