M1 Cases Research Articles

Diabetic maculopathy screening in England; are we seeing too much?

PurposeThe England screening service classification of diabetic retinopathy has strict, quality assured criteria to identify potential diabetic maculopathy termed ‘M1’. All new M1 cases identified by the screening service are referred to a hospital service. We aimed to evaluate the effectiveness of the England National Diabetic Eye Screening R1M1 classification of diabetic maculopathy as a criteria for secondary care referral in Portsmouth, UK.MethodsRetrospective audit of all patients referred to Portsmouth Hospitals NHS Trust with R1M1 pathology from April 2013 to January 2014. The total number of referrals received for this period was noted as well as the number of patients followed up in subsequent care pathways. Follow‐up data on those who remained under hospital care is presented for three years.ResultsA total of 306 diabetic patients were referred to Portsmouth Hospitals NHS Trust for R1M1 pathology over a 10 month period. At the first hospital appointment 135 (44%) had no fluid present on macular SD OCT and were either referred back to screening if the M1 features had resolved (65) or followed up with retinal images (70). 115 (38%) patients were considered to require further follow‐up in secondary care. Of those patients remaining in secondary care 70 remained in active follow up 3 years later.ConclusionsThese results would suggest that 44% of those with M1 features have no evidence of diabetic maculopathy on OCT at the initial hospital appointment and were discharged to community screening. Follow‐up for over three years in a secondary care setting is required by 23%. Secondary service could be better utilised by streamlining referrals either by refining the R1M1 classification or developing community based OCT service.

Variation in treatment and outcome in the early stage oral cavity squamous cell carcinoma.

This study aims to determine the survival impact of patient characteristics and treatment options associated with the early stage oral cavity squamous cell carcinoma, OCSCC. The methods are analysis of Irish cancer database examining T1/2, N0, and M0 cases of OCSCC from 1997 to 2007 inclusive. In total, 397 cases were identified. Anterolateral tongue accounted for 52.9% of cases. Increased age at diagnosis and smoking are independent prognostic survival indicators associated with poorer outcomes. Surgery as the initial intervention was associated with significantly better survival outcomes, while surgery and adjuvant radiotherapy significantly worse outcomes. Surgical intervention is recommended as the first-line treatment in the early stage OCSCC in combination with elective neck dissection.

Contemporary operative management of T4 breast cancer

Guidelines advise modified radical mastectomy following neoadjuvant systemic therapy for T4 breast cancer. We studied the influence of current systemic therapy and tumor subtype on pathologic stage and practice patterns to identify patients for whom less aggressive operative treatment might be considered. We identified 98 clinical T4 M0 cases operated on at our institution from October 2008-July 2015. Patient, tumor, and treatment variables were analyzed. Clinical T4 substage was 7% T4a, 32% T4b, 3% T4c, and 58% T4d. Tumor biologic subtype was 41% ER+/HER2-, 36% HER2+, and 23% ER-/HER2-. A total of 86 patients (88%) had neoadjuvant systemic therapy; 87% of patients underwent total mastectomy, 9% skin-sparing mastectomy, and 4% breast conservation. Axillary dissection was performed in 74% of patients and sentinel node surgery with (14%) or without (11%) axillary dissection in the remainder; 41/98 (42%) were lymph node negative at operation. The pathologic complete response rate in the breast (31%) and axilla (39%, cN+cases) correlated with biologic subtype (P<.0001). Overall 5-year, disease-free, and breast cancer-specific survival were 68% and 86%. Alignment with guidelines was substantial for both breast and axillary operation. Favorable breast cancer-specific survival suggests current multidisciplinary treatment has improved outcomes. Careful assessment of pathology and treatment response may identify clinical T4 patients appropriate for breast or axillary conservation.

Effect of idarubicin combined with medium-dose arabinoside in intensive chemotherapy on the prognosis of acute myeloid leukemia

Objective To explore the efficacy and feasibility of idarubicin combined with medium-dose arabinoside in intensive chemotherapy for acute myeloid leukemia(AML). Methods Fifty remittent patients with induced chemotherapy who underwent the intensive consolidation therapy with medium-dose arabinoside from January 2010 to January 2015 in hematology department of General Hospital of Beijing Military Area were considered as the treatment group. The single arabinoside (2 g/m2, 1/12 h, d1-3) treatment for 50 cases in the same period were considered as the control group. Two groups were performed with 6 courses sequential. There were 28 males, 22 females in the treatment group, and average age was 27.6 years (18-52 years), with 3 cases of M1, 27 cases of M2, 8 cases of M4, 12 cases of M5. In the control group, there were 30 males, 20 females, and average age was 26.8 years (16-50 years), with 2 cases of M1, 30 cases of M2, 8 cases of M4, 10 cases of M5. Then the complications and disease-free survival of two groups were observed respectively. Results Follow-up was done until June 2015. In the treatment group, 1 case died of pulmonary infection, 2 cases died of septic shock and 3 cases died of pulmonary infection, 1 case died of septic shock in the control group. The treatment related mortality of both groups were 6% (3/50), 8% (4/50), the related relapse rates were 32% (16/50), 52% (26/50), the disease-free survival rates were 62% (31/50), 40% (20/50) respectively in two groups. Conclusion Compared with the single arabinoside, the intensive consolidation therapy with medium-dose arabinoside and idarubicin for the treatment of AML has gained lower relapse rate and the related mortality, and the DFS rate has been improved significantly, which need further clinical application. Key words: Leukemia, myeloid, acute; Idarubicin; Arabinoside; Intensive consolidation therapy

Characteristics and Prognosis of 24 Cases of Primary Acute Myeloid Leukemia with Trisomy 8

To explore the clinical features and prognosis of primary acute myeloid leukemia (AML) with trisomy 8. The clinical data of 24 cases diagnosed as primary AML with trisomy 8 were collected. The clinical characteristics such as sex, age, subtype of FAB, blood routine and bone marrow blast at the first visit were analyzed and the relationship of the characteristics with CR rate and the prognosis was explored. 12 out of 24 AML patients were diagnosed as M5 (50%), while M2, M3, M4 and M6 had 3 cases, respectively (12.5%); one case did not receive the chemotherapy. 23 cases received 1-2 cycles of standard induction chemotherapy. Among them 3 cases of M3 achieved complete response (CR) and survived until the last following up with 100% 5-year OS rate. Among 20 cases of non-M3, 12 cases achieved CR1 (60%), 4 cases achieved partial response (PR) (20%), 4 cases did not respond (NR); 5 cases relapsed in follow-up for 3 years after CR1 (41.7%), 3 cases achieved CR2 after re-induction chemotherapy, and 2 cases remained NR. Among 20 cases of non-M3, 1 case failed to be followed-up after diagnosis within 1 month. The mean follow-up time of 19 cases was 26.2 (1.5-84) months, 9 cases died (6 cases of M5, 1 case of M4 and 2 cases of M2), who achieved PR and NR, or relapsed after CR1; the 3-year DFS and OS were 21%, 31.5% respectively. 2 cases of non-M3 accepted allo-HSCT with HLA-matched sibling donor and kept disease-free survival until the last following up, and survived for 58 and 66 months respectively. Except for 3 cases of M3, 2 cases received allo-HSCT and the cases without chemotherapy, the other 18 cases with initial WBC count less than 10×10(9)/L had OS and DFS longer than those of 10 cases with initial WBC count no less than 10×10(9)/L (P<0.05, P<0.01). The OS of 10 cases with CR1 was longer than OS of those cases without CR1 (P<0.01). The incidence of trisomy 8 in M5 is higher than the other AML subtypes, and the prognosis of M5 is poor. The initial WBC count above 10×10(9)/L is a high-risk factor. M3 with trisomy 8 and RARA gene has a very good prognosis. Trisomy 8 may increase the risk of primary AML except for M3, so allo-HSCT with HLA-matched sibling donor should be carried out as much as possible after CR1. The gene mutation of FLT3, MLL, HOX11, C-kit, NPM1 may possess an important significance on prognosis.

MP21-02 NOVEL QUANTITATIVE IMAGING ALGORITHMS DISTINGUISH LOCALIZED AND METASTATIC HIGH-GRADE PRIMARY PROSTATE CANCERS

INTRODUCTION AND OBJECTIVES: Prostate cancers (PC) associated with overt metastases at diagnosis (stage M1) or that progress to this stage are lethal. Although pathologic features of most M1 cases are indistinguishable from high-grade, non-metastatic PC (stage M0), profound disparities in outcome are apparent. More than 70% of M1 patients progress to castration-resistant prostate cancer (CRPC) and death within 5 years compared to 100% survival of highgrade M0 cases. Improved tools that detect features of lethal high-grade disease in prostate needle biopsies (PNBXs) will enable early identification of patients at greatest risk of metastatic progression. We have applied novel quantitative imaging (QI) technology and validated machine learning software tools to extract histomorphometric features from PNBX specimens that distinguish M1 and M0 cancers. This 00computer vision00 approach can be applied to routine diagnostic biopsies with the goal of detecting lethal PC as early as possible and improving patient stratification for entry into clinical trials. METHODS: We created a diverse, annotated biorepository with detailed clinical information corresponding to PNBX tissue from 427 patients with high-grade M0 or M1 PC. Retrospective analysis was used to assemble cohorts matched for age, race, and Gleason grade that were either M0 (n1⁄48) or M1 (n1⁄410). Archival PNBX slides stained with hematoxylin and eosin were quality checked and digitally scanned at 40X magnification. Fifteen image tiles were collected from each slide and two classifiers were constructed to assign each image tile to the M0 or M1 groups. QI analysis was used to investigate a set of features, including Fractal Dimensions (FD), Lacunarity (LA) [1st classifier], and Nuclear Features (NF) [2nd classifier] in M0 and M1 PC. RESULTS: Approximately 332 image tiles from the M0-RRP (n1⁄4122) and M1-ADT (n1⁄4210) cases were converted to nuclear masks. Using LA and FD tests, a trained classifier achieved 70% accuracy in distinguishing images from M1 and M0 cases, in which LA features were significantly different. We also identified 63 NFs differentially expressed in the M0 and M1 cases after random selection of 30% of nuclei and calculation of 1,500 NFs using partial least squares method. With this method, a classifier trained with the selected NFs to characterize nuclear intensity, texture, size, and shape achieved an accuracy of 75.5% in M0-RRP cases and 74.8% in M1-ADT cases. CONCLUSIONS: This innovative cohort of patients at the extremes of high-grade PC provides the opportunity to generate and test the performance of novel QI algorithms.

391 How accurate is the PSA test? A prevalence study of disturbed PSA values in a tertiary referral hospital

INTRODUCTION AND OBJECTIVES: Prostate cancers (PC) associated with overt metastases at diagnosis (stage M1) or that progress to this stage are lethal. Although pathologic features of most M1 cases are indistinguishable from high-grade, non-metastatic PC (stage M0), profound disparities in outcome are apparent. More than 70% of M1 patients progress to castration-resistant prostate cancer (CRPC) and death within 5 years compared to 100% survival of highgrade M0 cases. Improved tools that detect features of lethal high-grade disease in prostate needle biopsies (PNBXs) will enable early identification of patients at greatest risk of metastatic progression. We have applied novel quantitative imaging (QI) technology and validated machine learning software tools to extract histomorphometric features from PNBX specimens that distinguish M1 and M0 cancers. This 00computer vision00 approach can be applied to routine diagnostic biopsies with the goal of detecting lethal PC as early as possible and improving patient stratification for entry into clinical trials. METHODS: We created a diverse, annotated biorepository with detailed clinical information corresponding to PNBX tissue from 427 patients with high-grade M0 or M1 PC. Retrospective analysis was used to assemble cohorts matched for age, race, and Gleason grade that were either M0 (n1⁄48) or M1 (n1⁄410). Archival PNBX slides stained with hematoxylin and eosin were quality checked and digitally scanned at 40X magnification. Fifteen image tiles were collected from each slide and two classifiers were constructed to assign each image tile to the M0 or M1 groups. QI analysis was used to investigate a set of features, including Fractal Dimensions (FD), Lacunarity (LA) [1st classifier], and Nuclear Features (NF) [2nd classifier] in M0 and M1 PC. RESULTS: Approximately 332 image tiles from the M0-RRP (n1⁄4122) and M1-ADT (n1⁄4210) cases were converted to nuclear masks. Using LA and FD tests, a trained classifier achieved 70% accuracy in distinguishing images from M1 and M0 cases, in which LA features were significantly different. We also identified 63 NFs differentially expressed in the M0 and M1 cases after random selection of 30% of nuclei and calculation of 1,500 NFs using partial least squares method. With this method, a classifier trained with the selected NFs to characterize nuclear intensity, texture, size, and shape achieved an accuracy of 75.5% in M0-RRP cases and 74.8% in M1-ADT cases. CONCLUSIONS: This innovative cohort of patients at the extremes of high-grade PC provides the opportunity to generate and test the performance of novel QI algorithms.

Evaluating PSA nadir drift in high-risk and metastatic prostate cancer.

310 Background: Prostate cancer (PC) is a heterogeneous, clinically disparate, and often unpredictable disease whereby nearly 3% of patients present with bone metastatic PC (BMPC). Virtually all patients diagnosed with stage M1 disease fail to achieve complete remission and rapidly progress to castration-resistant PC (CRPC) and death (28% 5-year survival). Androgen deprivation therapy (ADT), the current standard of care for M1 disease, does not induce durable remissions. Improved understanding of tumor biology associated with lethal progression is desperately needed to guide clinical trials. Methods: We assembled a diverse, clinically annotated biorepository with diagnostic biopsy tissue from 428 patients with high-risk M0 and M1 disease. Retrospective analysis was performed on M0 (n = 157) and M1 (n = 112) patients stratified by treatment sequence. Clinical and pathological variables analyzed included age at diagnosis, race, primary Gleason sum, tumor burden, initial treatment received, PSA at time of biopsy, PSA velocity and length of follow-up. Primary endpoints included PSA nadir per treatment round, time from biopsy to CRPC, and death. Results: Compared to M0 cases, M1 patients were older, displayed significantly higher PSAs at the time of biopsy, and were more likely to display primary Gleason pattern 5. Patients with stage M1 disease had significantly worse survival, shorter time to CRPC, and median PSA nadir following each treatment round never fell below 2ng/ml. Median PSA nadirs associated with each treatment round demonstrated marked upward drift in the M1 cohort when compared to the M0 cohort. Conclusions: The presence of distant metastases is a major obstacle in achieving durable remissions for very high-risk PC. Our data confirms that ADT has limited effect on controlling cancer progression in M1 patients. PSA nadir drift in M1 compared to M0 patients may indicate intrinsic tumor resistance to currently available therapies. Multimodal profiling of M1 primary tumors may reveal actionable targets for initiation of combination treatment regimens at diagnosis.

Histiocytic differentiation in acute monocytic leukemia

ABSTRACTMyeloid histocytes of dendritic cells (DCs), Langerhans cells (LCs), and macrophages in varied tissues, as leukemic blasts in acute monoblastic and monocytic leukemia (AML-M5a and M5b), are derived from monocyte progenitors in bone marrow. Based on DC induction from hematopoietic stem cells, myeloid progenitors, and monocytes, and occasional expressions of histocyte-related antigens (HRAs) in M5, we presume some M5 cases share histiocytic phenotypes originally. To clarify the conception, 93 M5 cases were tested with antibodies for HRAs, CD1a, CD163, S100, fascin, and langerin by immunostaining, and their morphologic characteristics were studied by light and transmission electron microscopy. The study revealed that 23 M5 cases were positive for two or more kinds of HRAs and shared a serial of histocytic immunophenotype and morphologic features, which were closely associated with M5b subtype and expression of CD14 in M5.

Novel immunophenotyping marker in acute myeloid leukemia: does it implicate prognosis?

Introduction AML is a heterogeneous group of neoplasms that affect hematopoietic cells responsible for the production of myeloid lineages in bone marrow (BM). Novel immunophenotyping marker in AML (CD30) is a 120 kDa cell membrane glycoprotein that shares sequence homology with tumor necrosis factor (TNF) receptors; it is expressed by myeloblasts (CD34+ and/or CD117+) in a substantial number of cases of non Monocytic AML, either de novo or arising from MDS. CD30 expression is more common in AML patients with unfavorable chromosomal abnormality. It is suggested that CD30 could be a target for therapy by using anti-CD30 antibodies in a subset of patients with non Monocytic AML; also it might provide useful information for patient prognosis and stage of disease. Aim of work Is to assess novel immunophenotyping marker in AML (CD30) and cytogenetic abnormality and its correlation with prognosis. Materials and methods This prospective cohort study included 120 patients with newly diagnosed acute myeloid leukemia. Their ages ranged from (2 to 75) years with a mean of (40.15±23.55) years. Chromosomal analysis by karyotyping was done in all AML patients and Multicolor flow-cytometry immunophenotypic analysis was performed on bone marrow aspirates of the AML patients using fluroisothiocyanate (FITC) conjugated antibodies to assess CD30 expression on BM myeloblasts. Results The previous study showed that CD30 was expressed in all AML cases rather than M4 and M5 cases that showed negative expression. In addition, this study showed that there is more CD30 expression in myeloblasts with unfavorable chromosomal abnormalities. A significant association between platelets counts and CD30 expression was also observed. There was a higher degree of thrombocytopenia and a greater tendency to have higher leucocytic counts in patients having +ve CD30 expression than those with –ve CD30 expression. Conclusion The analysis of CD30 expression has a potential role to be used as a prognostic marker in AML.

Immunophenotypic Analysis of Patients with CD56⁺ Acute Myeloid Leukemia

To explore the immunophenotype characteristics of newly diagnosed patients with CD56⁺ acute myeloid leukemia (AML). Combining with cytomorphology, four-color flow cytometry was used to analyze the immunophenotype of 342 AML patients with CD56⁺ or CD56⁻. In 342 AML patients, the CD56⁺ expression was found in 83 AML patients who accounted for 24.27% and included 10 cases of M1, 45 cases of M2, 5 cases of M3, 6 cases of M6 and 17 cases of M5. The statistical analysis showed that there was statistical difference between CD56⁺ and CD11b⁺ patients (P < 0.05), but there was no statistical difference between CD56⁺ and HLA-DR, CD34, CD38, CD13, CD33, CD15, CD117, CD14, CD64, CD2, CD7, CD5, CD3, CD4, CD10, CD19, CD20, CD22 (P > 0.05). AML with only CD56 positive always has poor prognosis, thus the prognosis of patients with CD56⁺ AML accompanied by other antigens still needs more research.

CD117 expression on blast cells in acute myeloid leukemia.

The aim of the present work was to analyze the frequency of CD117 (c-KIT) antigen expression on the blast cells in acute myeloid leukemia (AML), evaluation of the presence of the relationship between the expression of the c-KIT and leukemia according to the FAB classification and definition of co-expression of the antigen CD117, antigens CD33 and CD34. The data of 47 patients with AML were diagnosed. M0 AML variant was established in 3 (6%) patients, M1 – in 2 (4%), M2 – in 9 (20%), M4 – in 22 (47%) and M5 – in 11 (23%). For immunophenotypic stu­dies monoclonal antibodies (mAb) that detect antigens of anti-CD34, anti-CD33 and anti-CD117 (Becton Dickinson, USA) were used. The presence of the antigen CD117 was detected in 39 people, accounting for 83% of all surveyed. Antigen c-KIT was present in 48.1±17.0% cells on average: in all 3 cases – AML M0, in2 cases of AML M1, in 6 cases – AML M2, 20 of 22 cases – AML M4 and in 8 of 11 AML M5 cases. Average levels of CD117 in investigated leukemia cases statistically differed significantly (p=0.0067). Among 39 CD117- positive patients in 25 (53%) co-expression of CD117+/CD34+ was revealed. Expression of CD117+/CD34- was observed in 14 cases (30%), CD117-/CD34+ – in 4 cases (8,5%), CD117-/CD34- – in 4 cases (8.5%). CD34 had of 64% of cells of myeloid origin. A high positive cor­relation between expression of CD117 and CD34 (r=+0,5169) was determined, being statistically significant (p 0,0067).

Abstract S6-05: Characterization of male breast cancer: First results of the EORTC10085/TBCRC/BIG/NABCG International Male BC Program

Abstract Intro: Male BC is a rare disease (&amp;lt;1% male tumors); knowledge is limited and management extrapolated from female BC. An international consortium, coordinated by EORTC and TBCRC, was created to better characterize and manage this disease, with 3 parts: 1) retrospective joint analysis 2) prospective registry 3) clinical trial(s). We report 1st results of part 1. Methods: Joint analysis of male BC pts with available FU &amp; FFPE samples, treated in 1990-2010, in 23 centers from 9 countries. Clinical data, long term outcomes, local pathology were centrally analyzed at EORTC. FFPE samples were analyzed at 3 central labs (UK, NL, US) for histology, grade, ER, PR, AR, HER-2, Ki67. Cut-off for positivity: Allred score ≥ 3 for ER/ AR/PR; 20% for Ki67; ASCO-CAP for HER2. Outcome data (OS &amp; RFS) analyzed per Kaplan Meier, p-values correspond to logrank test, CI’s by Brookmeyer &amp; Crowley. Reported % use number of non-missing values as denominator. Results: 1822 pts enrolled; 349 (19%) ineligible (no valid central lab assessment); of 1473 eligible pts (1384 from EU, 89 from US) 63% diagnosed in 2001-2010. Median age at diagnosis (Dx) was 68.5 ys. Of pts with known M status at Dx, 56 (5.1%) were M1; of 1046 M0 cases, 60% were N0 and 51% had T1 tumors at Dx; 4% of pts had BCS and 18% had SLNB; half received adjuvant RT. (Neo)adjuvant CT was used in 30% of M0 pts, most (44%) anthracycline-only; 77% of M0 pts received adjuvant ET, most tamoxifen (88.4%). Central pathology: 697 cases with central histology &amp; grade (full series ongoing): 87% ductal, 53% grade 2. In 1473 pts, at least 1 biomarker centrally assessed; 92% ER highly+ (Allred 7-8), PR had wider variation (35% highly+); 87% AR highly+; 25% Ki67 high; 5% HER2pos. Using IHC surrogates: 58% Luminal A-like, 35% LuminalB-like/HER2neg, 6% LuminalB-like/HER2pos; 0.1% HER2pos/non-Luminal; 1% TNBC (16% not classified). Outcome: For 1046 M0 pts, median FU was 5.7 yrs (0-19.2); 63% alive at analysis; in 88% cause of death was reported, mainly 33% non-cancer and 28% progression(PD)/toxicity. Significant OS improvement over time is seen. 75% (42/56) M1 pts died, mainly due PD. In M0 pts, median OS (yrs) was significantly correlated with ER+ p=0.001 [Allred 0-2: 3.9 (0.1, 6.0); 3-6: 7.1 (4.7, 11.7); 7-8: 8.8 (8.3, 10.0)] &amp; with PR+ p=0.022 [Allred 0-2: 7.3 (6.1, 10.3), 3-6: 8.0 (7.0, 9.3), 7-8: 9.5 (8.4, 12.8)]; less correlation for AR; no major differences for HER2 or Ki67. Median OS &amp; grade (394 cases; full series ongoing): Grade1: 12.8 yrs (6.1-15.6); Grade2: 7.9 (6.5-10.7); Grade3: 9.3 (4.9-21.1). Median OS for IHC surrogates: 8.7 (7.8-9.7) for Luminal A; 8.3 (6.9-9.4) for Luminal B/HER-2neg; 9.3 years (5.9-21.1) for Luminal B/HER-2+. Similar results were seen for RFS. ER/PR/AR using histoscores will be presented. Conclusions: a) 56% pts had T1 tumors at Dx but only 4% had BCS; b) ER was highly + in &amp;gt;90% but adjuvant ET given in only 77% pts; c) Male BC is usually ER+, PR+ &amp; AR+ and of Luminal A-like subtype (5% HER2pos &amp; 1% TNBC); d) Significant improvement in OS over time; e) ER and PR (Allred) are prognostic (high expression/better prognosis), less for AR, not for Ki67 nor IHC surrogates; f) In-depth characterization of samples is ongoing. Funding: BCRF, EBCC Council, Pink Ribbon NL, BRO. Citation Format: Fatima Cardoso, John Bartlett, Leen Slaets, Carolien van Deurzen, Elise van Leewen-Stok, Peggy Porter, Barbro Linderholm, Ingrid Hedenfalk, Carolien Schroder, John Martens, Jane Bayani, Christi van Asperen, Melissa Murray, Clifford Hudis, Lavinia Middleton, Joanna Vermeij, Stephanie Peeters, Judith Fraser, Monica Nowaczyk, Isabel Rubio, Stefan Aebi, Catherine Kelly, Kathryn Ruddy, Eric Winer, Cecilia Nilsson, Lissandra Dal Lago, Larissa Korde, Kim Benstead, Danielle Van Den Weyngaert, Oliver Bogler, Theodora Goulioti, Nicolas Dif, Carlo Messina, Konstantinos Tryfonidis, Jan Bogaerts, Sharon Giordano. Characterization of male breast cancer: First results of the EORTC10085/TBCRC/BIG/NABCG International Male BC Program [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr S6-05.

Abstract T MP8: Faster Angiographic Reperfusion is Associated with Improved Outcomes Across the Full Rankin Distribution: An Ordinal Analysis of the IMS III Trial

INTRODUCTION: In the NIH-funded IMS III trial, faster angiographic reperfusion led to a greater likelihood of good clinical outcome based on the dichotomized modified Rankin Score (mRS) score of 0-2 (minimal or no disability) vs. 3-6. From 240 to 390 minutes, among 1000 patients, every 15 minute acceleration in reperfusion was associated with 32 more patients with mRS 0-2. We hypothesized that faster reperfusion would lead to better clinical outcomes across the full mRS distribution (0, 1, 2, 3, 4, 5-6). METHODS: IMS III randomized subjects to IV/endovascular vs IV tPA alone. We analyzed the endovascular cohort with M1, M2 and ICAT occlusions reperfused (TICI 2-3) during the procedure (up to 7 hours). Time to reperfusion was defined as time from stroke onset to procedure termination. The ordinal 90-day mRS as a function of time to reperfusion was assessed using a proportional odds model with adjustment for pre-morbid mRS&gt;0, ASPECTS 0-4, and NIHSS strata (10-19 vs ≥20). “Typical patient” covariate values were used to obtain mRS distributions at prespecified time points. An algorithmic joint table analysis was applied to generate number-needed-to-treat values. RESULTS: Among 177 M1, M2, and ICAT occlusion cases with angiographic reperfusion (after excluding 2 cases with missing 90-day mRS and 3 with missing baseline ASPECTS), the common OR for improvement across all levels of the mRS was 0.78 (95% 0.66-0.91) adjusted and 0.77 (95% 0.66-0.89) unadjusted for each 30 minute increase in time. See Figure for adjusted probabilities based on the fitted model. From 240 to 390 minutes, among 1000 patients, every 15 minute acceleration in reperfusion was associated with 57 more patients with improvement of ≥ 1 mRS disability level. CONCLUSION: Faster time to reperfusion improves clinical outcomes for substantially more patients when considering the potential effects across all mRS levels, as opposed to considering a single transition between binary endpoints 0-2 vs 3-6.

A Short and Readable Proof of Cut Elimination for Two First-Order Modal Logics

A well established technique toward developing the proof theory of a Hilbert-style modal logic is to introduce a Gentzen-style equivalent (a Gentzenisation), then develop the proof theory of the latter, and finally transfer the metatheoretical results to the original logic (e.g., [1, 6, 8, 18, 10, 12]). In the first-order modal case, on one hand we know that the Gentzenisation of the straightforward first-order extension of GL, the logic QGL, admits no cut elimination (if the rule is included as primitive; or, if not included, then the rule is not admissible [1]). On the other hand the (cut-free) Gentzenisations of the first-order modal logics M3 and ML3 of [10, 12] do have cut as an admissible rule. The syntactic cut admissibility proof given in [18] for the Gentzenisation of the propositional provability logic GL is extremely complex, and it was the basis of the proofs of cut admissibility of the Gentzenisations of M3 and ML3, where the presence of quantifiers and quantifier rules added to the complexity and length of the proof. A recent proof of cut admissibility in a cut-free Gentzenisation of GL is given in [5] and is quite short and easy to read. We adapt it here to revisit the proofs for the cases of M3 and ML3, resulting to similarly short and easy to read proofs, only slightly complicated by the presence of quantification and its relevant rules.

Value of CD11a and CD18 in flow cytometric immunophenotypic diagnosis of acute promyelocytic leukemia

As acute promyelocytic leukemia (APL) has the highest curability among different acute amyeloid leukemia (AML) subtypes and requires a unique form of treatment, it is important to quickly establish or exclude the diagnosis of this subtype of leukemia. Furthermore, accurate and rapid diagnosis of APL is critical due to its association with disseminated intravascular coagulation. Patients and methods: This study was conducted on 100 newly diagnosed AML M0-M3 patients and a comprehensive flow cytometric immunophenotypic (FCM IPT) analysis of their peripheral blood (PB) and/or bone marrow samples with the inclusion of CD18 and CD11a monoclonal antibodies was done. Results: Cases of AML-M3 did not express human leukocyte antigen (HLA)-DR, CD11a and CD18, a highly significant different finding compared to non M3 cases (P Conclusion: Testing for CD18 and CD11a in conjunction with routine diagnostic FCM IPT panel will append extra diagnostic power to FCM IPT in the differentiation of difficult cases of AML-M3 from AML-(M0-M2).

Relationship of the urine cortisol level with the performance status of patients with lung cancer: a retrospective study.

Cortisol plays an important role in the physical status of patients with end-stage lung cancer, but the association of urine cortisol levels with TNM stage/performance status (PS) is unclear in patients with advanced lung cancer receiving chemotherapy. The objective of this study was to examine this association. In this single-center, retrospective, observational study, cortisol concentrations in 24-h pooled urine from 22 patients with advanced lung cancer were measured over 2days. The mean concentration in each patient was compared with PS, TNM stage, and serum sodium and potassium ion levels. The 24-h urine cortisol levels were higher in PS2 or PS3 cases compared to PS1 (p < 0.05) and increased proportionally with PS. Urine cortisol also increased in N2 or N3 cases compared to N1 (p < 0.01) and also increased in M1 cases (p < 0.05). Urine cortisol levels were negatively correlated with serum sodium (R = -0.49, p < 0.05) and had a tendency for a positive correlation with serum potassium (R = 0.40, p = 0.06). The 24-h urine cortisol level increased in patients with advanced lung cancer undergoing chemotherapy. Low serum levels of potassium and high levels of sodium may indicate relative adrenal insufficiency.

Flt3/ITD Enhances Hematopoietic Cell Migration Towards Chemokine CCL2 Independent of Blocking the Negative Feedback Mechanism on Rho-Associated Kinase

Internal tandem duplication mutations in the Flt3 gene (Flt3/ITD) found in patients with AML is associated with extremely poor prognosis. Our previous report demonstrating Flt3/ITD-mediated enhancement of hematopoietic cell migration towards chemokine CXCL12 (SDF1) suggests that Flt3/ITD likely facilitates dissemination of AML cells in the patients. Following studies showed that CXCL12 transiently up-regulated the expression of Rho-associated kinase-1 (Rock1) but subsequently down-regulated Rock1 expression in the control cells, whereas this effect was abolished by Flt3/ITD. The results demonstrated that CXCL12 generates negative regulatory feedback on Rock1 expression to prevent excessive migration to CXCL12 in the control cells, whereas this mechanism is abrogated by Flt3/ITD, thereby inducing deregulated cell migration (Onishi et al. ASH 2012). However, it is not known if Flt3/ITD augments migration to other chemokines and whether Flt3/ITD-induced blockage of the negative feedback loop on Rock1 expression reflects a specific effect on CXCL12/CXCR4 signaling pathway or more global change by ITD-Flt3. We found that mRNA for CCR2, a receptor for chemokine CCL2, is expressed in human AML cells. Herein, we investigated the effect of Flt3/ITD on migration to CCL2 in hematopoietic cells.Expression of CCR2 was significantly higher in the M4 and M5 cases with AML compared to other FAB subtypes that are deposited in the public gene expression database. However, there was no difference in the mRNA level for CCR2 between Flt3/ITD+ and Flt3/ITDneg cases. Consistent with AML samples, expression of surface CCR2 protein in Ba/F3 cells transfected with Flt3/ITD is equivalent to the control cells lacking Flt3/ITD. While the control Flt3/ITDneg Ba/F3 cells failed to migrate towards 1, 5, 10 and 50ng/mL of CCL2, Flt3/ITD marginally but significantly enhanced the cell migration towards 5ng/mL of CCL2 in Ba/F3 cells within 4 hours compared to control. In the Flt3/ITDneg cells exposed to 5ng/mL of CCL2, the mRNA expression of Rock1 continued to increase without being down-regulated to the basal level within 4 hours, and did not show any biphasic changes. In the Flt3/ITD+ Ba/F3 cells, however, Rock1 expression was significantly elevated compared to Flt3/ITDneg cells prior to incubation with CCL2, but down-regulated to 50% of the original level by 5ng/mL of CCL2 within 30 minutes. In contrast, Rock1 expression was barely affected and remained elevated by CXCL12 in Flt3/ITD+ Ba/F3 cells.Elevated expression of CCR2 in the M4 and M5 AML suggests that CCR2 signaling pathways can regulate migration of AML cells with monocytic lineage. Similar to CXCL12, migration towards CCL2 was also enhanced by Flt3/ITD without up-regulating CCR2 expression, suggesting that the enhanced chemotaxis by Flt3/ITD is not specific to CXCL12 and likely attributed to qualitative changes in the CCL2/CCR2 signaling pathway rather than their quantitative increase. Down-regulation of Rock1 expression by CCL2 in Flt3/ITD+ Ba/F3 cells may represent one of the qualitative changes in the CCL2 signaling pathway. However, blocking the CXCL12-induced negative regulatory mechanism on Rock1 expression existing in the Flt3/ITD+ cells was not identified in the CCL2 signaling. These data indicate that while enhancement in cell migration to chemokines by Flt3/ITD is not specific to CXCL12, blocking the negative feedback mechanism on Rock1 expression is not necessarily used in other chemokine signaling pathways. Our data suggests that Flt3/ITD mutations regulate trafficking of AML cells by modulating various chemokine signaling, but divergent molecular mechanism is involved in regulating cell migration towards different chmeokines. DisclosuresNo relevant conflicts of interest to declare.

Flow Cytometric Detection of Asparagine Synthetase Protein in Leukemia Cells; Indication for L-Asparaginase Therapy

Modern clinical treatments of childhood acute lymphoblastic leukemia (ALL) employ enzyme-based methods for depletion of blood asparagine in combination with standard chemotherapeutic agents. L-asparaginase (L-asp) therapy causes depletion of plasma asparagine followed by the loss of intracellular asparagine. Due to the lack of a rapid up-regulation of asparagine synthetase (ASNS) protein content in ALL cells, they are preferentially killed by L-asp. Elevated expression of ASNS within the leukemia cells causes decreased sensitivity to L-asp. The proof of ASNS deficiency in leukemia cells is considered to predictive for effectiveness of L-asp even in acute myeloid leukemia (AML) other than ALL patients. The establishment of quantitative estimation of ASNS protein content would be useful for the L-asp treatment in leukemia therapy.Objective: Our aim was to set up a flow cytometry system to check ASNS deficiency in leukemia cells and to investigate the sensitivity to L-Asp and the ASNS expression in AML leukemia cells.Methods: AML (KG-1, HL-60, U937) and ALL (MOLT-4, RS4;11) and CML (K562) cell lines were grown in RPMI1640 medium with 10% FCS. Primary leukemic cells from the peripheral blood or bone marrow of 20 AML patients were harvested on EDTA and isolated by Ficoll density gradient within 72h. ASNS expression was evaluated by cytosolic flow cytometry with Z5808 McAb (Hybridoma 31: 325-332.2012) and expressed as a ΔMFI(Difference of Mean Fluorescence Intensity(MFI) between by Z5808 and isotypic control) or MFI ratio(MFI by Z5808/MFI by isotypic control). When a sufficient amount of leukemic cells was available, sensitivity to L-asp (expressed as an IC50 - concentration inhibiting 50% of cell viability) was evaluated in vitro by incubating various concentrations of E. coliL-asp with the cells and by measuring the cell viability with a counting kit (WST1 viability assay) at day 3.Results: Determination of IC50 for the HL-60 (⊿MFI 48 ± 8.01, MFI ratio 1.77 ± 0.03) and U937 (⊿MFI 16.7 ± 0.47, MFI ratio 1.19 ± 0.02) demonstrated that these cells were equally sensitive to L-asp than the ALL cell line MOLT-4 in vitro (0.37 and 0.02IU/mL versus 0.15 IU/mL, respectively). K562 and KG-1 (⊿MFI 135.7 ± 5.66, MFI ratio 2.48 ± 0.09) cells with the highest ASNS expression exhibited resistance to L-asp (>10 IU/ml). Both of ASNS Expression by ⊿MFI and MFI ratio was inversely correlated with L-asp sensitivity judging from cell line studies. Judging from cell line study, the threshold for ASNS protein expression effective for L-asp treatment was considered to be <25 for ⊿MFI and <1.8 for MFI ratio respectively. Fresh leukemia cases contained three ALLs, Ph1ALL, and 13 AML; M0, 1; M1, 3; M2, 2; M4, 1; M5, 3; M7, 3; and Acute Mixed lineage leukemia, 2 cases. IC50 determination was possible on 11/20 patients. Four displayed a high sensitivity to L-asp (IC50 < 0.01 IU/mL) whereas two displayed resistant to L-asp (IC50 >10 IU/mL) Remaining 5 patients were a moderate sensitivity (IC50 < 0.5 IU/mL). ASNS Expression in ALL was almost near zero. ASNS expression in fresh AML was low in all cases except for one M2 and one M4 cases. Indeed, at high ASNS expression, these cases were resistant in vitro to L-asp. The patients with blasts sensitive to L-asp had mainly M1, M5 or M7 AML. A good inverse correlation between ASNS expression and sensitivity to L-asp was observed also in primary leukemic cells from AML patients.Conclusions: We demonstrated here that some of AML cell lines with low ASNS expression are more sensitive to L-asp than leukemia cells with high ASNS expression. Also, fresh AML cells with low ASNS expression are more sensitive to L-asp than with high ASNS expression. 11/13AML or two AMLL cases were supposed to be effective for L-asp. Plasma asparagine depletion by L-asp in selected patients having low ASNS may be a promising therapeutic approach even for AML.This work was supported by Japan Society for the Promotion of Science Grant-in-Aid for Scientific Research (C) KAKENHI Grant Number 24590713. DisclosuresNo relevant conflicts of interest to declare.

Aberrant phenotype in Iranian patients with acute myeloid leukemia.

The aim of this study was to evaluate the incidence of aberrant phenotypes and possible prognostic value in peripheral and bone marrow blood mononuclear cells of Iranian patients with AML. 56 cases of de novo AML (2010-2012) diagnosed by using an acute panel of monoclonal antibodies by multiparametric flowcytometry. Immunophenotyping was done on fresh bone marrow aspirate and/or peripheral blood samples using the acute panel of MoAbs is stained with Phycoerythrin (PE) /fluorescein isothiocyanate (FITC), Allophycocyanin (APC) and Peridinin-chlorophyll protein complex (perCP). We investigated Co-expression of lymphoid-associated markers CD2, CD3, CD7, CD 10, CD19, CD20 and CD22 in myeloblasts. Out of the 56 cases, 32 (57.1%) showed AP. CD7 was positive in 72.7% of cases in M1 and 28.5% in M2 but M3 and M4 cases lacked this marker. We detected CD2 in 58.35 of M1cases, 21.40% of M2 cases, 33.3 of M3 and 20% of M5; but M4 patients lacked this marker. The CBC analysis demonstrated a wide range of haemoglobin concentration, Platelet and WBC count which varied from normal to anaemia, thrombocytopenia to thrombocytosis and leukopenia to hyper leukocytosis. Our findings showed that CD7 and CD2 were the most common aberrant marker in Iranian patients with AML. However, we are not find any significant correlation between aberrant phenotype changing and MRD in our population. Taken together, this findings help to provide new insights in to the investigation of other aberrant phenotypes that may play roles in diagnosis and therapeutic of AML.