Presence Of Malignant Cells Research Articles

Role of Alpha-Smooth Muscle Actin and Fibroblast Activation Protein Alpha in Ovarian Neoplasms

Background/Aims: Studies show that tumor growth is not just determined by the presence of malignant cells, since interactions between cancer cells and stromal microenvironment have important impacts on the cancer growth and progression. Cancer-associated fibroblasts play a prominent role in this process. The aims of the study were to investigate 2 cancer-associated fibroblasts markers, alpha-smooth muscle actin (α-SMA), and fibroblast activation protein alpha (FAP) in the stromal microenvironment of benign and malignant ovarian epithelial neoplasms, and to relate their tissue expression with prognostic factors in ovarian cancer. Methods: α-SMA and FAP were evaluated by immunohistochemistry in malignant (n = 28) and benign (n = 28) ovarian neoplasms. Fisher’s exact test was used with a significance level lower than 0.05. Results: FAP immunostaining was stronger in ovarian cancer when compared to benign neoplasms (p = 0.0366). There was no significant difference in relation to α-SMA expression between malignant and benign ovarian neoplasms as well as prognostic factors. In ovarian cancer, FAP stainings 2/3 was significantly related to histological grades 2 and 3 (p = 0.0183). Conclusion: FAP immunostaining is more intense in malignant neoplasms than in benign ovarian neoplasms, as well as in moderately differentiated and undifferentiated ovarian carcinomas compared to well-differentiated neoplasms, thus indicating that it can be used as a marker of worse prognosis.

Harnessing the power of the immune system in non-Hodgkin lymphoma: immunomodulators, checkpoint inhibitors, and beyond.

Non-Hodgkin lymphoma is a malignancy of B lymphocytes that typically infiltrate sites of disease, including the lymph nodes, spleen, and bone marrow. Beyond the presence of malignant cells, many immune cells are also present within the tumor microenvironment. Although these immune cells have the potential to regulate the growth of malignant B cells, intratumoral immune cells are unable to eradicate lymphoma cells and most patients with lymphoma have clinical evidence of disease progression. Recent data have identified some of the mechanisms that account for the suppressed antitumor immune response and have created opportunities for treatment to overcome the deficiencies. Two general categories of immunological therapies are available. The first approach is to use agents that prevent inhibitory signals via immune checkpoint receptors that downregulate immune cell function. Blockade of suppressive programmed cell death 1 (PD-1) or CTLA-4 signaling has resulted in significant clinical activity by allowing intratumoral T cells to remain activated and target malignant cells. A second approach is to additionally activate T cells that are suboptimally active or suppressed, by providing signals through costimulatory molecules including CD27 or CD40 or by adding immunostimulatory cytokines. There has been significant heterogeneity in the responses to these treatment approaches. Clinical responses are seen in many diseases, but the most promising responses have been with PD-1 blockade in Hodgkin lymphoma. In other lymphomas, responses are seen but only in a subset of patients. Further research is needed to identify the mechanisms that account for response and to identify patients most likely to benefit from immune modulation.

Malignant Pseudo-Meigs Syndrome Presenting as Recurrent Eosinophilic Pleural Effusion

SESSION TITLE: Pulmonary Manifestations of Systemic Disease 3 SESSION TYPE: Affiliate Case Report Poster PRESENTED ON: Tuesday, October 31, 2017 at 01:30 PM - 02:30 PM INTRODUCTION: Pseudo-Meigs’ syndrome is a clinical syndrome where pleural effusion (PEf) and/or ascites are associated with ovarian tumor other than fibroma. We report a challenging case of pseudo-Meigs’ syndrome associated with ovarian mucinous carcinoma in a patient with recurrent right-sided exudative, eosinophilic PEf without cytologic evidence of malignancy in effusion. CASE PRESENTATION: A 51-year-old woman with a history of chronic lymphedema and venous stasis was admitted to the hospital for acute hypoxemic respiratory failure presenting with shortness of breath for 24 hours. On examination, she was in respiratory distress. Her temperature was 38.1°C; pulse 114 beats/minute; respiratory rate 22 breaths/minute; and oxygen saturation 93% breathing 3L/minute of supplemental oxygen through nasal cannula. Auscultation of the chest revealed crackles in her lower lung fields, pronounced on the right. Laboratory findings include white blood cells 19,500/mm3, erythrocyte sedimentation rate 130 mm/hour, and creatinine kinase 18 U/L. Computed tomography (CT) of the chest showed a large right-sided PEf. 900 mL of exudative fluid was drained by thoracentesis with improvement of symptoms. PEf was remarkable for eosinophilia (17%) with no malignant cells. On day 2 of hospitalization, her shortness of breath recurred. Chest radiograph showed recurrent right sided PEf. Another thoracentesis drained 1000 ml of eosinophilic effusion with no malignant cells. Repeat CT showed no underlying lung pathology. 48 hours later, her symptoms worsened and chest radiograph showed right sided PEf for the third time. Given the recurrent effusion with persistent negative cytology, CT of the abdomen was performed for a possible underlying malignancy. This revealed a large multiloculated cystic pelvic mass extending into the abdomen with omental caking and/or peritoneal carcinomatosis and lymph node involvement. Her labs then revealed CA 125 of 78 U/mL and CEA of 4.8 ng/mL. A debulking surgery was performed, revealing a 27x25x20cm ovarian mucinous carcinoma, histologic type G2: moderately differentiated without other organ involvement. Peritoneal ascetic fluid and lymph nodes were negative for malignancy. Three days later, 1000mL was drained via thoracentesis with cytology negative for malignant cells. 17 days later, follow up chest radiograph showed persistent effusion, however the fourth thoracentesis drained only 150 cc, and she remained asymptomatic. DISCUSSION: Recurrent right-sided exudative eosinophilic PEf, even without the presence of malignant cells, may be a marker for underlying malignancy when no obvious lung pathology is found. CONCLUSIONS: Ruling out an underlying malignant tumor, is essential as this could be from pseudo-Meig’s syndrome as seen in our case. Reference #1: Pleural Effusion in Meigs' Syndrome-Transudate or Exudate?: Systematic Review of the Literature. Krenke R, Maskey-Warzechowska M, Korczynski P, Zielinska-Krawczyk M, Klimiuk J, Chazan R, Light RW. Medicine. 12/2015 DISCLOSURE: The following authors have nothing to disclose: Reema Qureshi, Anna Hilton, Humnah Khudayar, Somwail Rasla, Taro Minami No Product/Research Disclosure Information

Swirling pattern in patients with exudative pleural effusion

ObjectiveThoracic ultrasound is the gold-standard method for studying pleural effusions. It is more sensitive than chest radiography or computed tomography.AimThe aim of the paper was to determine whether the echogenic swirling pattern identifiable on real-time chest ultrasonographic images is a predictor of malignant pleural effusions.DesignMedical records of patients undergoing chest ultrasonography in the Chest Department in Kasr Alaini Cairo University Hospital in the period from July 2013 to December 2014 were reviewed retrospectively. Patients with an echogenic swirling pattern in the pleural effusion, or with malignant diseases associated with pleural effusions, whose pleural fluids had been examined cytologically or whose pleural tissues had been examined pathologically, were enrolled in this study. A total of 45 patients were included. Malignant pleural effusions were diagnosed by the presence of malignant cells in the pleural fluid identified by pleural biopsy. The echogenic swirling pattern was defined as numerous echogenic floating particles within the pleural effusion, which swirled in response to respiratory movement or heartbeat.ResultsThere was a statistically significant relation between the swirling sign and the diameter, type, and amount of pleural fluid, with P value of 0.001. There was no statistical significance between the swirling sign and each of the diagnosis, whether malignant or nonmalignant, malignant cases either primary or secondary malignancy, and side of effusion.ConclusionThe presence of the swirling sign is related to the nature and amount of fluid and has no predilection to the diagnosis of the cause of pleural effusion.

Cytopathology of pericardial effusions : Experience from a tertiary center of cardiology.

Pericardial effusion (PE) is a common clinical condition that can develop as a result of systemic or cardiac diseases. Here, we report the results of cytology for patients who underwent pericardiocentesis for PE. The study comprised 283 patients who underwent primary percutaneous pericardiocentesis between 2007 and 2016. The mean age of the patients was 60.0± 16.6years; 162 (57.2%) were male and 121 (42.8%) were female. The presence of reactive mesothelial cells, acute and chronic inflammatory cells, and/or blood without evidence of malignant cells was considered as benign. The presence of malignant cells with/without reactive mesothelial cells, inflammatory cells, and/or blood was considered as malignant. The vast majority of PE specimens (219 cases; 77.4%) were classified as benign. Only 20 cases (7.1%) were classified as atypical, and malignant cells were present in the PE specimens of 44 cases (15.5%). The most common diagnosis was benign PE. The most commonly encountered malignancy was lung cancer. The rate of malignancy was 1.9% in the serous group and 24% in the hemorrhagic group, which was statistically significant. Benign PE was the most frequent cytological diagnosis in our study. Chronic nonspecific pericarditis was the most frequent type of pericarditis in the benign PE group, while lung adenocarcinoma was the most frequent malignancy in the malignant PE group. The rate of malignancy was significantly higher in the hemorrhagic group than in the serous group.

Morphological and phenotypical features of ovarian metastases in breast cancer patients

BackgroundAutotransplantation of frozen-thawed ovarian tissue is a method to preserve ovarian function and fertility in patients undergoing gonadotoxic therapy. In oncology patients, the safety cannot yet be guaranteed, since current tumor detection methods can only exclude the presence of malignant cells in ovarian fragments that are not transplanted. We determined the need for a novel detection method by studying the distribution of tumor cells in ovaries from patients with breast cancer. Furthermore, we examined which cell-surface proteins are suitable as a target for non-invasive tumor-specific imaging of ovarian metastases from invasive breast cancer.MethodsUsing the nationwide database of the Dutch Pathology Registry (PALGA), we identified a cohort of 46 women with primary invasive breast cancer and ovarian metastases. The localization and morphology of ovarian metastases were determined on hematoxylin-and-eosin-stained sections. The following cell-surface markers were immunohistochemically analyzed: E-cadherin, epithelial membrane antigen (EMA), human epidermal growth receptor type 2 (Her2/neu), carcinoembryonic antigen (CEA), αvβ6 integrin and epithelial cell adhesion molecule (EpCAM).ResultsThe majority of ovarian metastases (71%) consisted of a solitary metastasis or multiple distinct nodules separated by uninvolved ovarian tissue, suggesting that ovarian metastases might be overlooked by the current detection approach. Combining the targets E-cadherin, EMA and Her2/neu resulted in nearly 100% detection of ductal ovarian metastases, whereas the combination of EMA, Her2/neu and EpCAM was most suitable to detect lobular ovarian metastases.ConclusionsExamination of the actual ovarian transplants is recommended. A combination of targets is most appropriate to detect ovarian metastases by tumor-specific imaging.

Fibrin in Reproductive Tissue Engineering: A Review on Its Application as a Biomaterial for Fertility Preservation.

In recent years, reproductive medicine has made good use of tissue engineering and regenerative medicine techniques to develop alternatives to restore fertility in cancer patients. For young female cancer patients who cannot undergo any of the currently applied strategies due to the possible presence of malignant cells in their ovaries, the challenge is creating an in vitro or in vivo artificial ovary using carefully selected biomaterials. Thanks to its numerous qualities, fibrin has been widely used as a scaffold material for fertility preservation applications. The goal of this review is to examine and discuss the applications and advantages of this biopolymer for fertility restoration in cancer patients, and consider the main results achieved so far.

High-throughput screening of rare metabolically active tumor cells in pleural effusion and peripheral blood of lung cancer patients

Malignant pleural effusion (MPE), the presence of malignant cells in pleural fluid, is often the first sign of many cancers and occurs in patients with metastatic malignancies. Accurate detection of tumor cells in pleural fluid is crucial because the presence of MPE denotes an advanced stage of disease and directs a switch in clinical managements. Cytology, as a traditional diagnostic tool, has limited sensitivity especially when tumor cells are not abundant, and may be confounded by reactive mesothelial cells in the pleural fluid. We describe a highly sensitive approach for rapid detection of metabolically active tumor cells in MPE via exploiting the altered glucose metabolism of tumor cells relative to benign cells. Metabolically active tumor cells with high glucose uptake, as evaluated by a fluorescent glucose analog (2-NBDG), are identified by high-throughput fluorescence screening within a chip containing 200,000 addressable microwells and collected for malignancy confirmation via single-cell sequencing. We demonstrate the utility of this approach through analyzing MPE from a cohort of lung cancer patients. Most candidate tumor cells identified are confirmed to harbor the same driver oncogenes as their primary lesions. In some patients, emergence of secondary mutations that mediate acquired resistance to ongoing targeted therapies is also detected before resistance is manifested in the clinical imaging. The detection scheme can be extended to analyze peripheral blood samples. Our approach may serve as a valuable complement to cytology in MPE diagnosis, helping identify the driver oncogenes and resistance-leading mutations for targeted therapies.

Clinical Presentation, Diagnosis, and Radiological Findings of Neoplastic Meningitis

Neoplastic meningitis is a complication of solid and hematological malignancies. It consists of the spread of malignant cells to the leptomeninges and subarachnoid space and their dissemination within the cerebrospinal fluid. A literature review was conducted to summarize the clinical presentation, differential diagnosis, laboratory values, and imaging findings of neoplastic meningitis. Neoplastic meningitis is an event in the course of cancer with a variable clinical presentation and a wide differential diagnosis. In general, characteristic findings on gadolinium-enhanced magnetic resonance imaging and the presence of malignant cells in the cerebrospinal fluid remain the cornerstones of diagnosis. However, both modalities do not always confirm the diagnosis of neoplastic meningitis despite a typical clinical picture. Clinicians treating patients with cancer should be aware of the possibility of neoplastic meningitis, especially when multilevel neurological symptoms are present. Neoplastic meningitis can be an elusive diagnosis, so clinician awareness is important so that this malignant manifestation is recognized in a timely manner.

P2.04-040 Pleural Effusion Characteristics and Relationship with Outcomes in Cancer Patients: Topic: Esophageal Cancer and Other Malignancies

Pleural effusion is a common occurrence in cancer patients, with an estimated annual incidence of 150,000 in the United States. It generally represents advanced malignant disease with median survival of 3-12 months. It can lead to debilitating symptoms such as dyspnea and pain, adversely affecting the quality of life. We performed a retrospective analysis of a dataset of 62 cancer patients with pleural effusion after first thoracentesis procedure. The recorded data included: age, gender, type of cancer, number of thoracentesis, pleural fluid volume characteristics and volume. We studied 62 patients, with ages between 20-77 years, median 57 years; 64% were females and 36% were males. The number of patients deceased by the end of follow up was 43 (69.4%), alive at the end of follow up was 8 (12.9%), lost to follow up 11 (17.7%). Median overall survival was 3 months. The cancer type was divided into breast, 32.3%, non-small cell lung, 25.8% and 41.9% other cancers. Thirty-three (53.2%) patients had malignant pleural effusion with a median of 3 months of survival. Twenty-nine (46.8%) patients had non-malignant effusion with a similar median survival of 3 months. We analyzed the correlation of the radiologically estimated effusion volume, thoracentesis volume and presence of blood and malignant cells with outcomes (overall survival, OS). There is a negative correlation between the radiologically estimated effusion volume and OS (Pearson rank correlation of -0.43). This negative correlation is maintained in subgroup analysis (breast -0.56; lung -0.33). The thoracentesis volume was also negatively correlated with OS (Pearson rank correlation of -0.45), finding also maintained in subgroup analysis (breast -0.39, lung -0.66). We found no statistically significant difference in OS between malignant effusions (average OS 6.1 months, median 3 months) and non-malignant effusions (average OS 4.9 months, median 3 months). There was no statistically significant difference in survival between bloody effusions (average survival 3.5 months, median survival 1.25 months) and non-bloody effusions (average survival 6.2 months, median 3.5 months). We found an inverse correlation between the radiologically estimated pleural fluid volumes and OS as well as between thoracentesis volume and OS among cancer patients. This correlation is maintained in subset analysis of the two most common types of cancer in our sample, breast and NSCLC. Survival was not influenced by the presence of malignant cells or blood in the pleural fluid. A prospective study to better characterize the prognostic value of first thoracentesis may be warranted.

Evaluation of minimal disseminated disease in cryopreserved ovarian tissue from bone and soft tissue sarcoma patients.

What is the risk of finding malignant cells in cryopreserved ovarian tissue from sarcoma patients? Minimal disseminated disease (MDD) was not detected in frozen-thawed ovarian tissue from 26 patients by any of the sensitive methods applied. In case of leukemia, the risk of malignant cell transmission through the graft is well known and widely documented. However, for bone cancer, like Ewing sarcoma or osteosarcoma, only a small number of case reports, have been published. These cancers often affect prepubertal girls, in whom ovarian tissue cryopreservation and transplantation is the only option to preserve fertility. The presence of malignant cells in cryopreserved ovarian tissue from patients with bone/soft tissue sarcoma was investigated with disease-specific markers for each patient, using immunohistochemistry (IHC), FISH and real-time quantitative RT-PCR (qPCR), with the original tumor serving as a positive control. Forty-eight sarcoma patients were enrolled in the study, 12 of whom subsequently died. In each case, tissue from the primary tumor was investigated in order to identify markers (immunohistochemical and/or molecular) to analyze the ovarian tissue case by case. Ovarian tissue from osteosarcoma (n=15), liposarcoma (n=1) and undifferentiated sarcoma (n=5) patients could not be evaluated, as no specific markers were detected by FISH or sensitive IHC in any of their primary tumoral tissue. One patient with Li-Fraumeni syndrome was also excluded from the study. IHC analyses were therefore performed on ovarian tissue from 26 patients and qPCR on 19. The primary tumors involved were Ewing sarcoma family of tumors (n=14), rhabdomyosarcoma (n=7), synovial sarcoma (n=2), clear cell sarcoma (n=2) and a malignant peripheral nerve sheath tumor (n=1). MDD was not detected in any of the 26 analyzed samples using sensitive techniques in this largest reported series, even from patients who subsequently died and/or those who presented with metastasis (11/26), hence the most aggressive forms of bone cancer. Indeed, anti-CD99 IHC and PCR performed on patients presenting with Ewing sarcoma family of tumors (n=14) was negative in all cases. In patients with soft tissue sarcoma (n=12) primitive tumor markers were detected by IHC and were negative in ovarian tissue. PCR could only be performed in 6/12 of these patients, again proving negative. Cryopreserved ovarian fragments to be transplanted cannot be tested, so this analysis of malignant cells cannot guarantee that all cryopreserved fragments will not contain any disseminated disease. Moreover, molecular markers are not readily available for all types of tumors. These results are reassuring regarding the risk of malignant cells in the ovary for transplantation, as the study involves a large series including different types of sarcomas. We believe this will help clinicians in their patient counseling for fertility preservation and restoration. This work was supported by the Fonds National de la Recherche Scientifique de Belgique-FNRS under Grants Nos 7.4578.14 (Télévie to MS) and 5/4/150/5 to MMD. The authors declare no competing financial interests.

OR48 Resolution of conflicting hla assignment due to loss of heterozygosity in the hla region by NGS typing

57 year old female patient diagnosed with acute myeloid leukemia (AML) was referred to bone marrow transplant. Transplant workup and search for compatible unrelated donor was started immediately, along with induction therapy for AML. HLA typing performed from peripheral blood using high-resolution SSO method showed homozygous typing for all loci, except HLA C. However, the quality of the typing data, along with the unusual B and C allele association prompted confirmatory typing by next-generation sequencing (NGS), which revealed the presence of a previously undetected HLA B∗40:01:02 allele at 14%, and confirmed the presence of HLA C∗03:03:01 allele at 16%, consistent with a dramatic allelic imbalance. Research on the patient’s clinical history revealed that at the time of diagnosis and sampling, a significant population of atypical monocytic cells with promonocytes/monoblasts was detected by flow cytometry, accounting for at least 55% of the total leukocytes in the peripheral blood. Genomic microarray testing showed a 35.3 Mb terminal long contiguous stretch of homozygosity 6p25.3p21.31, which includes the HLA locus, resulting in copy-neutral loss of heterozygosity (LOH) acquired likely due to the malignancy. The LOH explains why one HLA haplotype was underrepresented in the initial blood sample acquired before the treatment. Buccal DNA sample was requested and tested with SSO method, which confirmed the presence of the alleles detected by NGS. In addition, confirmatory HLA typing performed 3 months later from peripheral blood post-treatment gave identical results. This supports the finding that the initial typing discrepancy was due to the overwhelming presence of malignant cells with LOH involving the HLA locus in the patient’s peripheral blood, collected before initiation of treatment. NGS typing using the Omixon Holotype platform was able to provide accurate typing of a sample with allelic disequilibrium, and allowed not only for detection but also for quantification of the minor alleles. This case highlights the importance of confirmatory typing pre-transplant for patients with hematological malignancies, since the presence of malignant cells in the peripheral blood may bias typing data if LOH involving the 6p region is present, which is a recurrent finding in patients with aplastic anemia or AML.

Concurrent Presence of Cryptococcal Meningitis and Neoplastic Meningitis in a Recipient of Hematopoietic Stem Cell Transplantation: A Case Report

Introduction: Cryptococcal meningitis is a rare disorder that is caused by Cryptococcus neoformans in immunocompromised host. Case Presentation: The case was a 64-year-oldmanwith a history of stem cell transplant for acute leukemia, whopresented chronic meningitis caused by Cryptococcus neoformans and concurrent neoplastic meningitis with presence of malignant cells only in his cerebrospinal fluid (CSF). Conclusions: Chronic meningitis in immunocompromised hosts can be due to more than one pathologic etiology and we should do all diagnostic evaluation to rule out all possible differential diagnosis

Evaluation of the accuracy of the CellaVision™ DM96 in a high HIV-prevalence population in South Africa.

IntroductionThe CellaVision™ DM96 (DM96) is a digital microscopy system which performs well in developed countries. However, to date it has not been evaluated in Africa, where the pathology spectrum encountered is very different. In particular, its utility in a setting with high HIV prevalence has not been assessed, which is of interest because of the morphological aberrations often seen in HIV-positive patients.ObjectivesThis study aimed to evaluate the accuracy of the DM96 in a South African laboratory, with emphasis on its performance in samples collected from HIV-positive patients.MethodsA total of 149 samples submitted for a routine differential white cell count in 2012 and 2013 at the Chris Hani Baragwanath Academic Hospital in Johannesburg, South Africa were included, of which 79 (53.0%) were collected from HIV-positive patients. Results of DM96 analysis pre- and post-classification were compared with a manual differential white cell count and the impact of HIV infection and other variables of interest were assessed.ResultsPre- and post-classification accuracies were similar to those reported in developed countries. Reclassification was required in 16% of cells, with particularly high misclassification rates for eosinophils (31.7%), blasts (33.7%) and basophils (93.5%). Multivariate analysis revealed a significant relationship between the number of misclassified cells and both the white cell count (p = 0.035) and the presence of malignant cells in the blood (p = 0.049), but not with any other variables analysed, including HIV status.ConclusionThe DM96 exhibited acceptable accuracy in this South African laboratory, which was not impacted by HIV infection. However, as it does not eliminate the need for experienced morphologists, its cost may be unjustifiable in a resource-constrained setting.

CYTOLOGICAL ANALYSIS OF THYROGLOSSAL DUCT CYSTS.

Thyroglossal duct cysts are regarded as congenital anomalies. They arise from the residual segments of thyroglossal duct. A cancer with the incidence rate from 1% to 1.5% may develop in thyroglossal duct cysts. Approximately 30% of patients asked to be examined due to inflammatory cysts. This Study was aimed at determining the cytological characteristics of thyroglossal duct cyst smear, Such as cell specificity, cellularity and the content of the extracellular matrix. Thyroglossal duct cyst smears were analyzed in 28 adult patients who had undergone the fine-needle aspiration cytology in the period from 2004 to 2014. Three patients underwent the Surgery following the Sistrunk procedure. The rest of the patients are in the process of monitoring. The monitoring process lasts from I to 10 years. As many as 27 Out of 28 smears contained macrophages, 9 contained stratified squamious.epithelial cells and only 4 smears contained follicular cells. Granulocytes were present in 4 smears. Two smears had scarce cellularity, 21 had moderate and 5 high cellularity. Malignant cells were not observed. In 19 smears, extracellular matrix consisted of cholesterol crystals. Thyroglossal duct cysts have no specific cytological featres. Macrophages are the dominant cell population. Cholesterol crystals and stratified squamous epithelial cells enable differentiation of thyroglossal duct cysts from thyroid gland Cystic nodules. Fine-needle aspiration cytology is necessary when assessing the cellular composition of thyroglossal duct cysts in order to promptly detect the possible presence of malignant cells and to conduct a surgical treatment.

Adequacy Criteria of Rapid On-Site Evaluation for Endobronchial Ultrasound-Guided Transbronchial Needle Aspiration: A Simple Algorithm to Assess the Adequacy of ROSE

Rapid on-site evaluation (ROSE) for endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) has been advocated to qualitatively diagnose biopsy samples. However, adequate ROSE criteria during EBUS-TBNA are unclear. The aim of this study was to determine adequacy criteria of ROSE in EBUS-TBNA samples and suggest an appropriate algorithm. Patients who underwent EBUS-TBNA for nodal evaluation between March and July 2013 at Seoul National University Hospital were included prospectively. The ROSE slides were reviewed independently by two pathologists, and the results were compared to the final pathologic results. Diagnostic yields, sensitivity, specificity, and accuracy were calculated in order to make nodal evaluations. EBUS-TBNA was performed on 300 lymph nodes in 133 patients. Samples were nondiagnostic in 7.7%, 6.3%, and 1.7% of the cytologic, histologic, and overall pathologic results, respectively. On the ROSE slides, a large tissue core size (≥2 cm), microscopic anthracotic pigment (MAP), and increased lymphocyte density (LD; ≥40 cells/field [40×, mean of 10 fields]) were significantly associated with adequate final cytologic or histologic results. Malignant cells were not statistically associated with adequacy but were considered a parameter indicating an adequate diagnosis. Using four sequential criteria, tissue core size, the presence of malignant cell, MAP, and LD ≥40 cells/field, the sensitivity and accuracy rates of ROSE increased from 64.4% to 98.6% and from 64.7% to 97.3%, respectively. A high adequacy rate of ROSE in EBUS-TBNA can be achieved by sequentially applying four criteria: tissue core size, malignant cells, MAP, and increased LD.

Abstract P2-03-08: Mutational landscape of metaplastic breast carcinomas

Abstract Introduction: Metaplastic breast carcinoma (MBC) is an aggressive histologic type of breast cancer, which preferentially displays a triple-negative phenotype. These tumors are characterized by the presence of malignant cells exhibiting differentiation towards squamous epithelium or mesenchymal elements, including spindle, chondroid, osseous and rhabdoid differentiation. Unlike other rare histologic types of breast cancer such as adenoid cystic and secretory carcinomas, which are underpinned by the MYB-NFIB and ETV6-NTRK3 fusion genes respectively, pathognomonic genetic alterations have not been identified in MBC. It has been suggested, however, that the frequency of PIK3CA somatic mutations would be significantly higher in MBCs than in other forms of triple-negative disease. Here we sought to characterize the mutational landscape of MBCs by means of high-depth whole exome sequencing analysis. Material and Methods: Twenty-one triple-negative MBCs were retrieved from the authors’ institutions. Representative sections from frozen blocks were microdissected to ensure tumor cell content greater than 50%. DNA samples extracted from microdissected tumor and matched peripheral blood leukocytes were subjected to high-depth (250x) whole exome sequencing on an Illumina GAIIx or HiSeq2000. Somatic point mutations were called using MuTect and somatic insertions and deletions (indels) were called using Strelka, Varscan2 and Haplotype Caller. Potentially pathogenic mutations were predicted using computational algorithms including PolyPhen-2, Mutation Taster, Mutation Assessor, CHASM and FATHMM. Significantly mutated genes were identified using MutSigCV. Pathway and network enrichment analysis of mutations was performed with Ingenuity Pathway Analysis and HOTNET. The genomic landscape of MBCs was compared with that of triple-negative breast cancers (TNBCs) analyzed as part of The Cancer Genome Atlas project. Results: A mean of 135 somatic non-synonymous point mutations and indels were identified per MBC. The most frequently mutated gene was TP53, found in 12/21 cases (57%), and the only significantly mutated gene as defined by MutSigCV (q<0.01). The repertoire of somatic mutations found in MBCs was qualitatively similar to that of TNBCs of no special type, and recurrently mutated genes were altered at similar frequencies in MBCs and TNBCs of no special type. When somatic mutations were annotated in pathways and networks, MBCs were found to have potentially pathogenic mutations affecting genes directly related to the PI3K pathway, including pathogenic non-synonymous mutations affecting PIK3CA, PIK3R1, PIK3R2, PIK3C2B, PIK3C2G and PTEN, significantly more frequently than TNBCs of no special type (10 out of 21 MBCs vs. 11 out of 62 TNBCs; Fisher's exact test p-value=0.0099). Conclusion: The majority (57%) of MBCs harbored non-synonymous mutations affecting TP53. While the frequencies of mutations affecting recurrently mutated genes in MBCs are similar to those found in other forms of TNBCs, MBCs significantly more frequently harbor mutations affecting PI3K pathway-related genes than TNBCs of no special type. Citation Format: Charlotte KY Ng, Britta Weigelt, Salvatore Piscuoglio, Y Hannah Wen, Maria R De Filippo, Luciano G Martelotto, Rachael Natrajan, Raymond Lim, Edi Brogi, Larry Norton, Anne Vincent-Salomon, Jorge S Reis-Filho. Mutational landscape of metaplastic breast carcinomas [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 P2-03-08.

In Vitro Efficient Expansion of Tumor Cells Deriving from Different Types of Human Tumor Samples

Obtaining human tumor cell lines from fresh tumors is essential to advance our understanding of antitumor immune surveillance mechanisms and to develop new ex vivo strategies to generate an efficient anti-tumor response. The present study delineates a simple and rapid method for efficiently establishing primary cultures starting from tumor samples of different types, while maintaining the immuno-histochemical characteristics of the original tumor. We compared two different strategies to disaggregate tumor specimens. After short or long term in vitro expansion, cells analyzed for the presence of malignant cells demonstrated their neoplastic origin. Considering that tumor cells may be isolated in a closed system with high efficiency, we propose this methodology for the ex vivo expansion of tumor cells to be used to evaluate suitable new drugs or to generate tumor-specific cytotoxic T lymphocytes or vaccines.

The Importance of Wound Biopsy in the Accurate Diagnosis of Acral Malignant Melanoma Presenting as a Foot Ulcer

Neoplastic changes arising at the sites of chronic, nonhealing wounds are not uncommon; however, they often go undiagnosed. We report a case of rapidly progressing plantar melanoma presenting as a chronic, nonhealing ulcer. A 46-year-old patient presented at a specialized Wound Healing Center with an enlarging painful ulcer on the right heel of 3 months duration. The wound was biopsied and specimens were sent for examination at the Wound Pathology service at the Department of Dermatology and Cutaneous Surgery, University of Miami. Histology demonstrated features consistent with acral malignant melanoma. Immunohistochemistry using melanocytic markers MART-1, S-100, HMB-45 revealed positive staining indicating the presence of malignant cells, and D2-40 staining showed lymphatic invasion of the tumor in the wound biopsy specimen. The case presented here underscores the importance of wound biopsying in the diagnosis of malignancies associated with nonhealing wounds.

Prognostic significance of minimal pleural effusion in non-small cell lung cancer patients.

e19152 Background: Malignant pleural effusion (MPE) was defined by the presence of malignant cells in pleural space. Accurate characterization of pleural effusion (PE) is an essential part in staging the disease for adequate management. At diagnostic evaluation, minimal pleural effusion (mPE) that is too small amount PE to examine the presence of malignant cells through thoracentesis is present outside scope of current staging system. Therefore whether mPE has prognostic impact or not is an important clinical issue needed to be investigated. Methods: 1,382 consecutive stage IIIB, IV NSCLC patients were retrospectively enrolled from Inha Lung Cancer Cohort. Information regarding smoking habits, ECOG PS, Charlson comorbidity index score, weight loss in 6 months before diagnosis, levels of hemoglobin, albumin, alkaline phosphatase and calcium at the time of diagnosis, treatment, followed-up and survival status was analyzed as potential confounding factors. All patients were performed staging work-up and followed-up at Inha university hospital. The mPE was defined as less than 1 cm of maximal thickness on contrast enhanced chest CT scan at diagnosis and its presence was not considered in staging the disease. Results: According to status of PE at diagnosis, patients were classified into no PE, mPE and MPE. No PE was presented in 614 patients (44.4%); mPE in 339 patients (24.5%); malignant PE in 356 patients (25.8%). PEs of 73 (5.3%) patients was not evaluated. Overall survival of patients with mPE or MPE was shown to be significantly shorter than those of patients with no PE (MST, 9.67 for no PE, v 6.60 for mPE v 5.37 months for MPE, log-rank p<.0001), even after adjustment for age, smoking habits, ECOG PS, Charlson comorbidity index score, weight loss, hemoglobin, albumin, alkaline phosphatase, calcium, histology, T subset, N subset, M subset, and treatment status (hazard ratios, 95% CIs and Cox p: 1.18, 1.023 to 1.373 and .024 for mPE; 1.545, 1.314 to 1.816 and <.0001 for MPE). Conclusions: The mPE is commonly encountered clinical situation in advanced stage NSCLC patients. These results suggest that presence of mPE at diagnosis confers grave prognosis like MPE, in spite of its nature.