Cytological Subtypes Research Articles

Standardized Ultrasound Evaluation for Active Surveillance of Low-Risk Thyroid Microcarcinoma in Adults: 2024 Korean Society of Thyroid Radiology Consensus Statement.

Active surveillance (AS) has been widely adopted as an alternative to immediate surgery owing to the indolent nature and favorable outcomes of papillary thyroid microcarcinoma (PTMC). AS is generally recommended for tumors measuring ≤1 cm without aggressive cytological subtypes, risk of gross extrathyroidal extension (ETE), lymph node metastasis (LNM), or distant metastasis. AS requires careful patient selection based on various patient and tumor characteristics, and ultrasound (US) findings. Moreover, during AS, regular US is performed to monitor any signs of tumor progression, including tumor growth, new US features of potential gross ETE, and LNM. Therefore, appropriate imaging-based assessment plays a crucial role in determining whether AS or surgery should be pursued. However, detailed recommendations concerning US evaluation are currently insufficient, necessitating the formulation of this guideline. The Korean Society of Thyroid Radiology has developed a consensus statement for low-risk PTMC, covering US assessment methods when considering AS as a management option and conducting follow-up imaging tests during AS. This guideline aims to provide optimal scientific evidence and expert opinion consensus regarding a standardized US-based assessment protocol for low-risk PTMC.

Immunocytochemical detection by a fully automated immunostainer for assisting subclassification of pulmonary tumors on ThinPrep slides in real-life clinical samples: A single-institution experience.

This study aims to investigate the feasibility and reliability of ThinPrep slides in detecting the subclassification of lung cancer and develop a process for immunocytochemistry (ICC) with optimized staining steps of an automated immunostainer. Cytomorphology and ancillary ICC by automated immunostainer on ThinPrep slides were performed to subclassify 271 cytology cases of pulmonary tumor, which were stained with 2 or more of the following antibodies: p40, p63, thyroid transcription factor-1 (TTF-1), Napsin A, synaptophysin (Syn), and CD56. The accuracy of cytological subtyping was improved from 67.2% to 92.7% (p < .0001) after ICC. The accuracy of cytomorphology combined with ICC results for lung squamous-cell carcinoma (LUSC), lung adenocarcinomas (LUAD), and small cell carcinoma (SCLC) was 89.5% (51 of 57), 97.8% (90 of 92), and 98.8% (85 of 86), respectively. The sensitivity and specificity of 6 antibodies were as follows: p63 (91.2%, 90.4%) and p40 (84.2%, 95.1%) for LUSC, TTF-1(95.6%, 64.6%) and Napsin A (89.7%, 96.7%) for LUAD and Syn (90.7%, 60.0%) and CD56 (97.7%, 50.0%) for SCLC, respectively. P40 expression on ThinPrep slides had the highest agreement (κ=0.881) with immunohistochemistry (IHC) results, followed by p63 (κ=0.873), Napsin A (κ=0.795), TTF-1 (κ=0.713), CD56 (κ=0.576), and Syn (κ=0.491). The result of ancillary ICC on ThinPrep slides by fully automated immunostainer was in good agreement with the gold standard in pulmonary tumors subtype and immunoreactivity, objectively achieving accurate subtyping in cytology.

Transcriptome analysis of tissue microbiota diversity in tumor and non-tumor lymph nodes

Metagenomic studies in recent years have demonstrated that all tissues of the human body studied by genomic and transcriptomic sequencing methods, both in pathological processes and in normality, contain fragments of DNA and RNA from a variety of microorganisms. The composition of tissue microbiota and its relationship with development of pathological changes are still poorly understood, despite increasing number of studies in this area every year. In this study, gene expression of the lymph node microbiome in reactive follicular hyperplasia and follicular lymphoma was investigated. To study expression of lymph node microbiome genes in reactive follicular hyperplasia and follicular lymphoma. The work included 38 biopsy samples of lymph nodes with follicular lymphoma of different cytological subtypes and 10 biopsy samples of lymph nodes with reactive follicular hyperplasia. Verification of diagnosis was carried out using standard histological, histochemical and immunohistochemical methods. Using sequencing method, the transcriptome was examined. Statistical analysis and data visualization were performed using the R programming language (version 4.2.1). Tumor lymph nodes are characterized by large Simpson and Shannon alpha diversity values (p-value = 0.026465 and p-value = 0.007122, respectively). Two clusters were discovered, characterized by different levels of relative abundance of microorganisms. It has been proven that diversity of microorganisms present in tumor tissue and their number are statistically significantly higher than corresponding indicators in the lymph nodes with follicular hyperplasia.

Cell-mediated immunity and expression of MHC class I and class II molecules in dogs naturally infected by canine transmissible venereal tumor: Is there complete spontaneous regression outside the experimental CTVT?

The canine transmissible venereal tumor (CTVT) is a transplantable cancer with the ability evade the immune system, despite strict immune surveillance of the host; in this context, the relationship between inflammatory infiltrate and CTVT prognosis is not entirely understood. Natural canine transmissible venereal tumors of 22 dogs were evaluated for tumor/host interaction through clinical and epidemiological data, cyto-histopathological and cytogenetic findings and, mainly, cell-mediated immune response. We performed analysis on dogs with naturally acquired disease to provide information from the study of CTVT biology in its natural course, as the clinical evolution of the natural tumor in the host is not yet as well known as in the laboratory. Populations for T cell labeling (CD3+ CD4+ CD8+), B cells, NK cells, and macrophages were analyzed by flow cytometry in blood and tumor samples and expressions of MHC class I and class II molecules were quantified by immunohistochemistry and compared mainly between the phases of progression and regression in the natural CTVT. Dogs were also treated with vincristine sulfate and evaluated for chemotherapeutic response. Chemotherapy was effective in 88% of cases and there was no recurrence of the disease 12 months after the cure. Tumor cells displayed a numerical chromosomal variation between 54 and 72, not correlating with the host genotype. Although a greater expression of MHC molecules [18.6 ± 5.8% class I (P < 0.004) and 38.5 ± 6.5% class II (P < 0.003)] was observed in the regression phase, no significant effect was observed between the clinical phase of the tumor and cellular immune response in the analysis by flow cytometry (P > 0.05). We also found no correlation between cytological subtype of the tumor (plasmacytoid, lymphocytoid and mixed) and cellular immune response, suggesting that there is no difference in tumor immunogenicity. Here, we found no immunological evidence to support the theory of the immune-induced complete spontaneous regression in CTVT.

Different methylation signatures at diagnosis in patients with high-risk myelodysplastic syndromes and secondary acute myeloid leukemia predict azacitidine response and longer survival

BackgroundEpigenetic therapy, using hypomethylating agents (HMA), is known to be effective in the treatment of high-risk myelodysplastic syndromes (MDS) and acute myeloid leukemia (AML) patients who are not suitable for intensive chemotherapy and/or allogeneic stem cell transplantation. However, response rates to HMA are low and there is an unmet need in finding prognostic and predictive biomarkers of treatment response and overall survival. We performed global methylation analysis of 75 patients with high-risk MDS and secondary AML who were included in CETLAM SMD-09 protocol, in which patients received HMA or intensive treatment according to age, comorbidities and cytogenetic.ResultsUnsupervised analysis of global methylation pattern at diagnosis did not allow patients to be differentiated according to the cytological subtype, cytogenetic groups, treatment response or patient outcome. However, after a supervised analysis we found a methylation signature defined by 200 probes, which allowed differentiating between patients responding and non-responding to azacitidine (AZA) treatment and a different methylation pattern also defined by 200 probes that allowed to differentiate patients according to their survival. On studying follow-up samples, we confirmed that AZA decreases global DNA methylation, but in our cohort the degree of methylation decrease did not correlate with the type of response. The methylation signature detected at diagnosis was not useful in treated samples to distinguish patients who were going to relapse or progress.ConclusionsOur findings suggest that in a subset of specific CpGs, altered DNA methylation patterns at diagnosis may be useful as a biomarker for predicting AZA response and survival.

The metabolic reprogramming in acute myeloid leukemia patients depends on their genotype and is a prognostic marker.

Leukemic cells display some alterations in metabolic pathways, which play a role in leukemogenesis and in patients' prognosis. To evaluate the characteristics and the impact of this metabolic reprogramming, we explore the bone marrow samples from 54 de novo acute myeloid leukemia (AML) patients, using an untargeted metabolomics approach based on proton high-resolution magic angle spinning-nuclear magnetic resonance. The spectra obtained were subjected to multivariate statistical analysis to find specific metabolome alterations and biomarkers correlated to clinical features. We found that patients display a large diversity of metabolic profiles, according to the different AML cytologic subtypes and molecular statuses. The link between metabolism and molecular status was particularly strong for the oncometabolite 2-hydroxyglutarate (2-HG), whose intracellular production is directly linked to the presence of isocitrate dehydrogenase mutations. Moreover, patients' prognosis was strongly impacted by several metabolites, such as 2-HG that appeared as a good prognostic biomarker in our cohort. Conversely, deregulations in phospholipid metabolism had a negative impact on prognosis through 2 main metabolites (phosphocholine and phosphoethanolamine), which could be potential aggressiveness biomarkers. Finally, we highlighted an overexpression of glutathione and alanine in chemoresistant patients. Overall, our results demonstrate that different metabolic pathways could be activated in leukemic cells according to their phenotype and maturation levels. This confirms that metabolic reprogramming strongly influences prognosis of patients and underscores a particular role of certain metabolites and associated pathways in AML prognosis, suggesting common mechanisms developed by leukemic cells to maintain their aggressiveness even after well-conducted induction chemotherapy.

Expression of Glioma-associated oncogene homolog 1 as biomarker with sonidegib in advanced basal cell carcinoma.

The pivotal BOLT (Basal cell carcinoma Outcomes with LDE225 [sonidegib] Treatment) study established the durable efficacy and manageable toxicity of sonidegib 200 mg once daily (QD) through 42 months in patients with advanced basal cell carcinoma (BCC). This secondary analysis used expression of Glioma-associated oncogene homolog 1 (GLI1) as a biomarker to assess the extent of Hedgehog pathway inhibition by sonidegib in patients with locally advanced BCC (laBCC) and metastatic BCC (mBCC). The study enrolled 230 patients, 79 and 151 receiving sonidegib 200 and 800 mg QD, respectively. At week 17, GLI1 expression was reduced from baseline by a median percentage (95% confidence interval) of 88.7% (54.6%–93.0%) and 97.0% (77.5%–98.9%) for aggressive laBCC, 97.5% (80.3%–98.8%) and 95.0% (80.7%–97.5%) for nonaggressive laBCC, and 99.1% (96.4%–99.6%) and 99.3% (95.9%–99.9%) for mBCC in the 200 and 800 mg groups, respectively. Substantial repression of GLI1 was observed in patient subgroups stratified by age, sex, BCC cytological subtype, Eastern Cooperative Oncology Group performance status, lesion site, baseline number of BCCs, and prior radiotherapy. Results support further studies on the inhibition of Hedgehog pathway genes by sonidegib in patients with laBCC and mBCC.

The role of cytology in endobronchial ultrasound-guided transbronchial needle aspiration: A study of 813 cases focusing on diagnostic yield, an analysis of misdiagnosed cases and diagnostic accordance rate of cytological subtyping.

BackgroundEndobronchial ultrasound‐guided transbronchial needle aspiration (EBUS‐TBNA) is a minimally invasive technique for cytological and histological diagnosis. The objective of this study was to explore the role of cytological diagnosis in EBUS‐TBNAs.MethodsEight hundred and thirteen consecutive cases performed EBUS‐TBNA with both cytological and histological diagnoses were retrospectively reviewed. All patients were followed up for clinical data.ResultsBefore immunohistochemical examination, the cytological sensitivity, specificity, and diagnostic accuracy of EBUS‐TBNAs were 92.9% (421/453), 98.9% (348/352), 95.5% (769/805), respectively. After immunohistochemical examination, the sensitivity, specificity, and diagnostic accuracy were 93.0% (423/455), 99.4% (348/350), 95.8% (771/805), respectively. The majority of false‐negative were cases whose cytological diagnosis was “atypical” or the cytological diagnosis suggested “inadequate.” “Neoplastic” were also prone to false‐negative cytology. The diagnostic accordance rate of cytological subtyping was 90.3% for squamous‐cell carcinoma, 99.2% for adenocarcinoma, and 98.1% for small‐cell carcinoma before immunohistochemical examination, and became 85.9%, 98.5%, and 98.2% after immunohistochemical examination, respectively.ConclusionCytological diagnosis in EBUS‐TBNAs had a good sensitivity and high specificity. The sensitivity and specificity of cytological diagnosis were proved to be higher after the immunohistochemical examination. At the same time, cytology had high accordance rate in subtype diagnosis. False‐negative results occurred more commonly in cases whose cytological diagnosis was “atypical” or the cytological diagnosis suggested “inadequate” or the corresponding histological diagnosis was “Neoplastic.”

Subtyping and EGFR mutation testing from blocks of cytological materials, based on liquid‐based cytology for lung cancer at bronchoscopic examinations

Liquid-based cytology (LBC) allows immunohistochemistry (IHC), fluorescence in situ hybridization, and molecular testing to be performed in fixed cell materials. We examined the feasibility of subtyping and EGFR mutation testing of bronchoscopic samples from patients with lung cancer using cell blocks (CB) based on LBC fixation (LBC-CB). We included 35 consecutive patients with peripheral lung nodules who underwent endobronchial ultrasonography with a guide sheath in our hospital. Thirty of these patients were diagnosed with lung cancer by obtaining cytological samples. Cytological subtyping was performed with IHC using LBC-CB, and the Cobas EGFR Mutation Test ver. 2 was performed using extracted genomic DNA from the LBC-CB, formalin-fixed paraffin-embedded (FFPE) tissue, and matched plasma. Of the 30 cases, 25 were classified cytomorphologically as adenocarcinoma (ADC, n = 17) and squamous-cell carcinoma (SQCC, n = 8). The remaining five cases were classified by IHC as favor ADC (n = 3) and favor SQCC (n = 2) according to the WHO criteria. In the final ADC group (n = 20), EGFR mutations on the LBC-CB were identified in eight cases (40%; 1 exon 19 deletion, 6 L858R, and 1 L861Q). Mutations in FFPE samples were identified in seven cases (35%) at the same site in each case. Plasma EGFR mutations were identified in four cases (20%) at the same site. The CB detection rate was higher than for FFPE and plasma. LBC-CB is suitable for subtyping and EGFR mutation testing in lung cancers.

Reproducibility of histologic prognostic parameters for mantle cell lymphoma: cytology, Ki67, p53 and SOX11.

Mantle cell lymphoma (MCL) shows a clinical aggressiveness that varies from patient to patient. Despite major advances in outcomes with current immunochemotherapy, the future development of therapies requires risk stratification to tailor therapy intensity. Within the group of reference pathologists for the ongoing trials of the European MCL Network, we performed a round robin test on a tissue microarray to evaluate the reproducibility in assessing the biomarkers of outcome in MCL. Cytological subtype, Ki67-index and expression of p53 and SOX11 were evaluated on 20 diagnostic tumour samples by eight participating labs independently. We demonstrate that the assessment of the proliferation index by counting the Ki67 positive cells as well as assessment of SOX11 and p53 expression status is reproducible between labs. For the most established prognostic biomarker, Ki67, the intra-class correlation coefficient was very good when assessed as a continuous parameter (0.87). The agreement was lower when the values were analysed in a dichotomized way applying the commonly used cutoff of 30% (kappa = 0.65, complete concordance of all labs in 13/20 (65%)). Cases with discrepant results between labs in the dichotomized analysis showed mean values close to the cutoff of 30%. Centralised scoring and digital image analysis revealed results in line with the scores from individual labs. All cases in our cohort were additionally assessed for gene expression signatures and of TP53 gene alterations. Given the good reproducibility when guidelines of assessment are applied, the biomarker studied in this inter-laboratory test presents potential candidates to be enhanced for risk-stratification in the future clinical trials.

Malignancy rate of Bethesda category III thyroid nodules according to ultrasound risk stratification system and cytological subtype.

The risk of malignancy is considered to be 10% to 30% for cases of thyroid nodules with atypia or follicular lesion of undetermined significance (AUS/FLUS). However, only a minority of patients with AUS/FLUS undergo surgery; therefore, the risk of malignancy might be overestimated due to selection bias. To overcome this problem, we categorized cases of thyroid nodules with AUS/FLUS using the ultrasound risk stratification system (US-RSS) to calculate the malignancy rate and identify the patients most suitable for surgical treatment.In this retrospective observational study, we subcategorized 382 pathologically confirmed thyroid nodules with AUS/FLUS using current US-RSSs (American Thyroid Association, Korean-Thyroid Imaging Report and Data System, American College of Radiology-Thyroid Imaging, Reporting and Data System, European Thyroid Imaging Report and Data System) and calculated the malignancy rate. Additionally, cases of nodules with AUS/FLUS were categorized according to their cytological subtypes, and the malignancy rate was calculated.Current US-RSSs showed good or moderate agreement among them. The overall malignancy rate for thyroid nodules with AUS/FLUS was 38.7%. On categorization of the nodules with AUS/FLUS, the malignancy rates were found to be 60% to 67.5% for the high suspicion category, 32.2–36.6% for the intermediate suspicion category, and 12.4% to 16.3% for the low suspicion category. The malignancy rate for nodules with cytologic atypia was significantly higher than that for nodules with architectural atypia, especially in the intermediate suspicion category.Categorization of thyroid nodules with AUS/FLUS using current US-RSSs helps to determine the optimal course of management of patients, especially when combined with cytological subtype characterization.

CMTTdb: the cancer molecular targeted therapy database.

The cancer molecular targeted therapy has achieved unprecedented progress in the past decade and is thought to be the most promising direction for cancer treatment in future. As the fast growing of the clinical trials of targeted anticancer agents for different cancer types, it is critical to collect and integrate such information to guide clinical practice. We constructed the Cancer Molecular Targeted Therapy database (CMTTdb) to store and retrieve molecular targeted therapy data about randomized clinical trials (RCTs) of targeted agents and also accompanied targets, biomarkers, targeted cancer subtypes, etc. Different with some existing resources, CMTTdb particularly focuses on clinical application of the trails. Design of the trails, such as treatment modalities (monotherapy or combination with other therapies), as well as results on clinical efficacy parameters, adverse events are also collected. In this current version, CMTTdb contains data for 1,088 clinical trials which cover 165 agents, 80 targets, 15 cancer types (95 molecular subtypes and 56 histological or cytological subtypes) from public literatures. This database is freely available at http://www.biosino.org/CMTTdb. A user-friendly web interface was designed so that these data can be easily retrieved. CMTTdb will be a valuable source for providing access to information of clinical trials on the rapidly growing number of novel targeted agent and be useful in guiding oncologists for the optimization of the therapy strategy for cancer treatment.

Bone Marrow Morphology of Multiple Myeloma with Non-bone-related Extramedullary Disease

To explore the morphological characteristics of bone marrow cells of multiple myeloma with non-bone-related extramedullary disease and thier clinical significance. Bone marrow smears, peripheral blood smears and bone marrow biopsy sections as well as thier examination results of 20 cases of multiple myloma with non-bone-related extramedullary disease were collected at initial diagnosis in First Affiliated Hospital of Xi'an Jiaotong University from March 2013 to March 2016, and morphological characterisistes of bone marrow cells were analysed in combination with clinical data. The morphology of plasma cells in 20 patients showed 2 cytologic subtypes: primitive cell type (16 cases) and pleomorphic type (4 cases). Immature plasma cells were found in the 5 patients' peripheral blood smear, accounting for about 1%-4% of the number of peripheral blood cells. In bone marrow tissue, plasma cells hyperproliferated with nodular and packed type, and secondary myelofibrosis counted for 12 cases (60%). 13 MM patients whose non-bone-related extramedullary disease occurred during therapy were divided into 2 groups according to the marrow fibrosis density. The median time from diagnosis of MM to extramedullary lesions resulting from fibrosis 0-1 grade and 2-3 grade was 23.7±3.7 months and 10.5±3.2 months ahead of the former(P=0.025). Bone marrow plasma cell morphology of multiple myeloma with non-bone-related extramedullary disease at the initial diagnosis is mostly immature type, and plasma cells proliferate with nodular and packed type and accompanied by different degree of fibrosis. The degree of myelofibrosis indirectly reflects the degree of proliferation and malignancy of the bone marrow plasma cells, which maybe possess some value in predicting extramedullary disease in the early stages of the MM.

3:45 PM Abstract No. 376 Clear cell hepatocellular carcinoma is associated with non-viral cirrhosis and arterial-phase hypo-enhancement but does not predict outcome after locoregional therapy

Clear cell hepatocellular carcinoma (HCC) is a less common HCC cytologic variant associated with improved outcomes in studies prior to the advent of locoregional therapy (LRT). This study was undertaken to determine whether clear cell HCC predicts better outcomes after LRT. We retrospectively identified 124 consecutive patients (92 men, 32 women; median age 59 y) with 132 HCC (mean diameter 4.8 ± 3.9 cm) diagnosed by percutaneous biopsy between 2008-2017 prior to transarterial chemoembolization (TACE) (n = 51, 41%), yttrium-90 radioembolization (n = 17, 13%), percutaneous thermal ablation (n = 41, 33%), and combination TACE/ablation (n = 15, 12%). Baseline Barcelona Clinic Liver Cancer (BCLC) stage was 0/A (n = 48, 38%), B (n = 33, 26%), C (n = 27, 22%), and D (n = 16, 13%). Cytological subtype was determined from percutaneous core needle biopsy as 100% clear cells, focal clear cells, or no clear cells. Baseline clinical and imaging features, and radiologic response by mRECIST were correlated with cytological subtype using multinomial regression analysis. Time to progression (TTP) and transplant free survival (TFS) were analyzed by Cox proportional hazard models. Percutaneous core biopsies comprised entirely of clear cell subtype were associated with non-viral etiologies of HCC (OR 5.3 95% CI 1.64-17.16; p = 0.005), solid histological arrangement (OR 4.32 95% CI 1.14-16.37; p = 0.031), and hypo-enhancement on arterial phase CT or MRI images (OR 3.08 95% CI 1.02-9.3; p = 0.046). The clear cell subtype was not associated with improved objective response by mRECIST (OR 0.93 95% CI 0.37-2.33; p = 0.87), delayed TTP (HR 1.17 95% CI 0.86-1.6; p = 0.32), or better TFS (HR 1.01 95% CI 0.69-1.47; p = 0.96) in comparison to specimens with focal or no clear cells. There remained no significant association between the clear cell subtype and TTP or TFS after stratifying by LRT modalities. Clear cell HCC is more common in non-viral etiologies of cirrhosis and is associated with atypical enhancement on radiologic images. It does not, however, impart independent prognostic information prior to LRT.

The potential significance of alpha-enolase (ENO1) in lung adenocarcinomas - A utility of the immunohistochemical expression in pathologic diagnosis.

We herein analyzed the relationships among the immunohistochemical expression of alpha-enolase (ENO1) and clinicopathological factors in order to define the significance of ENO1 in lung adenocarcinomas (ADCs). ENO1 expression was detected in most of the ADCs examined (95.8%), but not in bronchial and alveolar epithelia. ENO1 expression was typically observed in the cytoplasm among most ADCs (95.8%), but was also detected in the nucleus (56.3%). The levels were significantly higher in terminal respiratory unit (TRU) cytological subtype ADCs. Neither cytoplasmic nor nuclear expression was associated with any other clinicopathological factors including post-operative survival and growth activity. These results suggest that ENO1 is a crucial factor promoting neoplastic transformation exclusively in TRU subtype ADCs. We also investigated the potential utility of the immunohistochemical expression of ENO1 to differentiate TRU-type ADC cells from the reactive hyperplasia of pneumocytes and bronchiolar epithelial cells because difficulties are associated with discriminating these lesions in small biopsy specimens. The sensitivity and specificity of ENO1 (cytoplasmic/nuclear) were 87.5%/37.5% and 88.9%/100%, respectively, which are superior to those of p53 (18.8% and 100%). ENO1 has potential as a biomarker to assist in the histopathological detection of TRU subtype ADC cells.

Prospective Evaluation of the Effect of Deferasirox on Hematologic Response in Transfusion-Dependent Patients with Low-Risk MDS and Iron Overload: The Rythmex Study

▪Background:Long-term blood transfusions remain the mainstay of supportive care in patients with low-risk myelodysplastic syndromes (MDS). Blood transfusions can result in iron overload, which can lead to impaired organ function, significant morbidity and mortality. Although debate exists around its use in patients with MDS, iron chelation therapy is recommended in patients with low-risk MDS to prevent iron-overload toxicities and to improve survival. In addition, hematologic improvement has been reported during chelation therapy (Jensen PD 1996, Gattermann N 2012). Here we report results from a prospective, observational study that evaluated a hematologic response to deferasirox in a large population of regularly transfused patients with low- or intermediate-1-risk MDS.Methods:inclusion criteria: Transfusion-dependent (at least 4 packed red blood cells (PRBC)/8 weeks) adult patients with IPSS low- or intermediate-1-risk MDS who had initiated or had been receiving chelation therapy for less than 3 months. Primary endpoint was reduction in transfusion requirements at 3 month, (i.e. the number of PRBC received during the 8 weeks before deferasirox initiation compared with the 8 weeks before month 3). Secondary endpoints included: hematologic improvement (HI; IWG 2006 criteria for erythroid [HI-E], platelet [HI-P] and neutrophil [HI-N]) at 3, 6 and 12 months, duration of response, serum ferritin levels and safety.Results: 70 patients were included, 57 were evaluable. Most exclusions (19%) were related to a low level of transfusion before inclusion. Median age was 78.3 years. The cytologic subtypes were: sideroblastic anemia, 31.6%; refractory anemia, 19.3%; refractory anemia with excess blasts-1, 17.5%; refractory cytopenia with multilineage dysplasia, 17.5%; 5q syndrome, 10.5%; unclassified SMD, 3.6%. Median time since diagnosis was 2.79 years (33 months). Before inclusion, 87% patients had received a treatment for MDS (erythropoiesis-stimulating agents, 72%; lenalidomide, 17%;G-CSF, 16%; azacitidine, 7%; thalidomide: 2%). Patients received a median of 5.8 ± 2.8 PRBC during the 8 weeks before inclusion. The mean number of PRBC received per month during the 6 months before inclusion was 2.11. The median serum ferritin level at inclusion was 1 543 ng/ml.The mean number of PRBC received in the whole population post inclusion (eight weeks before month 3) was 5.9, which was not significantly different from the number of PRBC given during the 8 weeks before inclusion (5.8). However at 3 months, 4 patients had a positive effect on transfusion requirement. At 6 and 12 months, positive effect occurred in 3 and 4 additional patients, respectively. 10 patients achieved HI-E during the study and according to Kaplan-meier analysis, the probability of HI-E was 12.7% at 6 months and 24.2% at 1 year. During the study, 17 patients (32%) achieved an erythroid response, i.e positive effect on transfusion requirement and/or HI-E; 19% had an erythroid response at month 12. Median duration of erythroid response was 123 days. 3.5%, 10%, 21% of patients had received a concomitant treatment with deferasirox at 3, 6, and 12 months, respectively. However,deferasirox was the sole specific treatment received in 12/17 patients with erythroid response. 27 patients experienced adverse events (AEs) related to deferasirox, among them 4 patients with grade > 2 . 32 patients underwent deferasirox dosing modifications, and 20 patients had discontinued treatment by 3 months ( median duration of treatment 11.3 months). In patients receiving deferasirox for more than 9 months, the median serum ferritin level at 12 months was 1 438 ng/ml . In contrast, the median serum ferritin level at 12 months was 2247 ng/ml in the whole group. There were no predictive factors of erythroid response (cytologic classification, time since diagnosis, level of ferritin at inclusion, the number of PRBC transfused before inclusion), but the number of responders was low.Conclusions: After 3 months of treatment, deferasirox had no impact on transfusion requirement in regularly transfused low risk MDS patients. However, deferasirox could induce 19% of specific erythroid response at 12 months. These results suggest that iron chelation therapy with deferasirox may induce an effect on hematopoiesis in a small subset of patients with MDS and iron overload. DisclosuresRose:Genzyme, Novartis, Celgene: Honoraria, Research Funding. Beyne-Rauzy:Celgene, Novartis: Honoraria. Dreyfus:Novartis: Honoraria.

Molecular Genetic Profiling in Chronic Myelomonocytic Leukemia with Low Risk Cytogenetic Features

Background: Chronic myelomonocytic leukemia (CMML) is a heterogeneous disease that can vary from a myelodysplastic (MD) predominant profile to a myeloproliferative (MP) one. CMML has a variable course, with a median overall survival of 20 months and 15-30% of progression to acute myeloid leukemia (AML). Cytogenetic abnormalities are present in 20-40% of cases and recurrent gene mutations have been reported in several genes. Patients with low risk cytogenetic features (normal karyotype and isolated -Y) account for approximately 80% of CMML patients and often fall into the low risk categories of CMML prognostic scores. Our hypothesis is that mutational study can contribute to diagnosis and prognostic stratification in this subset of patients.Methods: A retrospective study was performed on 57 patients with CMML. Cases with normal karyotype (n=53), isolated -Y (n=2) and no metaphases (n=2) were selected. DNA was extracted from BM (n=52) and PB (n=5) samples at diagnosis. Targeted deep-sequencing was performed in a panel of 83 myeloid-related genes. Libraries were prepared with 1μg of genomic DNA using the KAPA Library Preparation Kit (Kapa Biosystems) and then enriched using the SeqCap EZ capture chemistry (Nimblegen, Roche). Libraries were sequenced with 150 bp paired-end reads on an Illumina MiSeq. Herein we present the results of 43 cases which were preliminarily analyzed using the Illumina MiSeq Reporter and Variant Studio softwares. High-probability oncogenic mutations were called by eliminating sequencing and mapping errors and known SNPs based on the available databases. An in-house bioinformatics pipeline will be designed to analyze the whole series of patients. A preliminar statistical analysis was performed with SPSS. Fisher's exact test was used to compare variables between patient subsets. Complete study, including the correlation of the sequencing findings with the clinical data, will be presented in the meeting.Results: Median age at diagnosis was 70 years (range 48-87) and there was a 2:1 male predominance. Median follow up of patients was 23 months (range 1-116) during which 23% (11/43) of cases progressed to AML. Morphological WHO subtypes were CMML-1 in 36 (84%) cases and CMML-2 in 7 (16%). According to the FAB criteria 34 (79%) cases were classified as CMML-MD and 9 (21%) as CMML-MP. According to the CMML-specific scoring system (CPSS) 28/43 (65%) patients belonged to the low-risk category, 10/43 (23%) to the intermediate-1 and 4/43 (12%) to the intermediate-2. The mean depth of the targeted resequencing per base per sample was 810-fold. After excluding sequencing and mapping errors a mean of 293 single nucleotide variants (snv) and insertions/deletions (indels) were called per sample. After filtering non-silent variants and excluding known polymorphisms a mean of 6 variants per sample were called as high-probability somatic changes. Distribution of detected variants across the patients can be seen in Figure 1. Most frequently affected genes were TET2 (70%), ASXL1 (47%) and SRSF2 (35%); followed by RUNX1 (23%), NRAS (16%), CBL (12%), EZH2 (12%), SETBP1 (12%) and ZRSR2 (12%). Variants detected in 5-10% of patients included IDH2, CRBBP, SH2B3, UMODL1, DNMT3A, JAK2, PTPN11, SF3B1 and U2AF1 genes. Statistical analysis revealed that some variants correlated with CMML subtypes: SH2B3 (P=0.010) and STEBP1 (P=0.024) associated with CMML-2; JAK2 (0.007), NRAS (P=0.026) and EZH2 (P=0.05) associated with CMML-MP. Variants in NRAS also correlated with progression to AML (P=0.04) and patients in intermediate groups of CPSS associated with JAK2 (P=0.008) and EZH2 (P=0.011) variants.Conclusions: Genetic profiling using targeted deep-sequencing is a highly promising approach for CMML diagnosis and varies according to the cytological subtypes. With the correlation of the results with the clinical data of patients, we expect to determine if targeted molecular profiling can contribute to prognostic stratification of patients with CMML and low risk cytogenetic features. For the moment, we have already found a correlation with progression to AML.Acknowledgments: Instituto de Salud Carlos III, Ministerio de Sanidad y Consumo, Spain (PI 11/02519; PI 11/02010); RTICC, FEDER (RD12/0036/0044); 2014 SGR225 (GRE) Generalitat de Catalunya; Fundació Josep Carreras, Obra Social "La Caixa" and Celgene Spain. [Display omitted] DisclosuresSole:Celgene: Honoraria, Membership on an entity's Board of Directors or advisory committees.

The animal-dependent risk factors in canine T-cell lymphomas.

Malignant lymphomas are one of the most common malignancies occurring in dogs; among them T-cell tumours are less commonly recognized. Recently, many authors have recommended cytology as a sufficient diagnostic method for canine lymphomas, especially if supported by immunocytochemistry or flow cytometry. The aim of the study was to characterize animal-dependent risk factors in canine T-cell lymphomas (TCLs) in Poland, including specific cytological subtypes. Determination of the type and subtype of the tumour was made based on the updated Kiel cytological classification adopted for dogs as previously described. Two breeds turned out predisposed to TCL (dog de Bordeaux and Boxer) while no predisposition to B-cell lymphoma could be evidenced. Dogs with low-grade lymphoma were significantly older than those with high-grade lymphoma.

Cytological variations and typical diagnostic features of endocervical adenocarcinoma in situ: A retrospective study of 74 cases.

Background:The sensitivity of Papanicolaou smears for detecting endocervical adenocarcinoma in situ (AIS) is very low. A comprehensive cytological analysis of endocervical AIS is necessary to increase diagnostic accuracy.Methods:The subjects were 74 patients with pathologically-diagnosed AIS. A total of 140 Papanicolaou smears were reviewed to calculate the sensitivity of the Papanicolaou smears for detecting AIS and the incidence of sampling/screening/diagnostic errors. The cytological review was performed by 6 cytotechnologists, and the final cytological diagnosis was obtained at the consensus meeting. We classified the cases into three differentiation types; typical type (well-differentiated AIS), polymorphic type (poorly differentiated AIS), and mixed typical and polymorphic type. Three cytological subtypes (endocervical, endometrioid and intestinal subtypes) of AIS were also analyzed.Results:The sensitivity of the original Papanicolaou smears for the detection of AIS was 44.6%, while that for the detection of AIS and adenocarcinoma was 63.5%. The diagnostic accuracy of AIS increased to 78.5% in the final diagnosis. The common characteristic features were microbiopsies/hyperchromatic crowded groups (HCG) (82.0%) and mitotic figures (72.2%). The appearance of single cells (2.8%) was rare, and all the cervical cytology smears showed no evidence of necrotic tumor diathesis. The most common AIS was the typical type (41 cases, 67.2%) among all cytologically-diagnosed AIS or adenocarcinoma cases (61 cases). Although mixed typical and polymorphic AIS existed in 17 cases (27.9%), pure polymorphic AIS was very rare (3 cases, 4.9%). The endocervical subtype was the most predominant subtype (67.2%), followed by a few mixed subtypes. The important diagnostic keys for AIS cytology are as follows: (1) The appearance of microbiopsies/HCG (single-cell pattern is rare), (2) mitotic figures in the microbiopsies/HCG, (3) a lack of necrotic tumor diathesis in cases with polymorphic AIS, and (4) recognition of typical cytological subtypes.Conclusions:The relatively low diagnostic accuracy AIS was caused by the underestimation of microbiopsies/HCG and the overestimation of polymorphic components. The typical cytological features of AIS are the presence of microbiopsies/HCG with mitotic figures in the absence of necrotic tumor diathesis in specimens containing endocervical samples. The recognition of infrequent AIS subtypes (endometrioid and intestinal subtypes) is also important.

Bronchial Brushing Increases the Diagnostic Yield of Fiberoptic Bronchoscopy in Bronchogenic Carcinoma

Background : The importance of rapid and accurate cytopathological diagnosis in bronchial cancers is increasing due to advances in treatment modalities. Aims : We evaluated the diagnostic methods and cytologic subtypes of bronchial cancers to determine the diagnostic reliability of different bronchoscopic techniques. Material and methods: Retrospective data were obtained from the hospital files and pathological specimens of the patients with diagnosis of primary lung cancer from a period of 36 months. Cytological tumor typing was determined using histopathology of bronchoscopic forceps biopsy (FB), bronchial-bronchoalveolar lavage (BL), bronchial brushing (BB), transbronchial fine-needle biopsy. Computed tomography or ultrasonography guided transthoracic biopsy and surgical biopsies were used where the other interventional methods were inadequate for diagnosis. Results : A total of 124 patients were diagnosed during study period. 119 (96%) of them were male. The median age was 68, ranging between 36 and 88 years. Histopathologic subtypes were determined as non-small cell carcinoma (NSCC) in 104 (83.9%), squamous cell carcinoma in 64 (51.6%), adenocarcinoma in 16 (12.9%), NSCC not otherwise specified in 24 (19.3%) and small cell carcinoma in 20 (16.1%) patients. The combination of FB, BL and BB established the diagnosis of bronchogenic carcinoma in most of the cases (92.6%). Conclusions : Lung cancer is seen commonly in elderly male patients with smoking history and squamous cell carcinoma is the most common cytologic type. High diagnostic accuracy can be achieved by a combination of bronchoscopic FB, BB and BL procedures. Keywords: Bronchoscopy; Lung cancer; Bronchial brushing