37 results on '"Jan Blancato"'
Search Results
2. Hypoxia-activated neuropeptide Y/Y5 receptor/RhoA pathway triggers chromosomal instability and bone metastasis in Ewing sarcoma
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Congyi Lu, Akanksha Mahajan, Sung-Hyeok Hong, Susana Galli, Shiya Zhu, Jason U. Tilan, Nouran Abualsaud, Mina Adnani, Stacey Chung, Nada Elmansy, Jasmine Rodgers, Olga Rodriguez, Christopher Albanese, Hongkun Wang, Maureen Regan, Valerie Zgonc, Jan Blancato, Ewa Krawczyk, G. Ian Gallicano, Michael Girgis, Amrita Cheema, Ewa Iżycka-Świeszewska, Luciane R. Cavalli, Svetlana D. Pack, and Joanna Kitlinska
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Multidisciplinary ,Chromosomal Instability ,Humans ,General Physics and Astronomy ,Bone Neoplasms ,Neuropeptide Y ,Sarcoma, Ewing ,General Chemistry ,Hypoxia ,rhoA GTP-Binding Protein ,General Biochemistry, Genetics and Molecular Biology ,Receptors, Neuropeptide Y - Abstract
Adverse prognosis in Ewing sarcoma (ES) is associated with the presence of metastases, particularly in bone, tumor hypoxia and chromosomal instability (CIN). Yet, a mechanistic link between these factors remains unknown. We demonstrate that in ES, tumor hypoxia selectively exacerbates bone metastasis. This process is triggered by hypoxia-induced stimulation of the neuropeptide Y (NPY)/Y5 receptor (Y5R) pathway, which leads to RhoA over-activation and cytokinesis failure. These mitotic defects result in the formation of polyploid ES cells, the progeny of which exhibit high CIN, an ability to invade and colonize bone, and a resistance to chemotherapy. Blocking Y5R in hypoxic ES tumors prevents polyploidization and bone metastasis. Our findings provide evidence for the role of the hypoxia-inducible NPY/Y5R/RhoA axis in promoting genomic changes and subsequent osseous dissemination in ES, and suggest that targeting this pathway may prevent CIN and disease progression in ES and other cancers rich in NPY and Y5R.
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- 2022
3. Circulating Cell-free miRNA Expression and its Association with Clinicopathologic Features in Inflammatory and Non-Inflammatory Breast Cancer
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Raja Marrakchi, A Gat, Irene Anne Jillson, F. Ben Ayed, Bilel Neili, A Benammar Elgaaied, David Goerlitz, Jan Blancato, Christopher A. Loffredo, Khouloud Hamdi, Islam, Ahmed Abidi, and S Chivi
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Adult ,0301 basic medicine ,CA15-3 ,Oncology ,Cancer Research ,medicine.medical_specialty ,Epidemiology ,Real-Time Polymerase Chain Reaction ,Inflammatory breast cancer ,Statistics, Nonparametric ,03 medical and health sciences ,0302 clinical medicine ,Breast cancer ,Internal medicine ,microRNA ,Biomarkers, Tumor ,medicine ,Humans ,skin and connective tissue diseases ,Aged ,Neoplasm Staging ,business.industry ,Public Health, Environmental and Occupational Health ,Case-control study ,Cancer ,Middle Aged ,Prognosis ,medicine.disease ,Gene Expression Regulation, Neoplastic ,MicroRNAs ,030104 developmental biology ,Real-time polymerase chain reaction ,Case-Control Studies ,030220 oncology & carcinogenesis ,Cancer cell ,Female ,Inflammatory Breast Neoplasms ,Neoplasm Grading ,business ,Follow-Up Studies - Abstract
Recent discovery showing the presence of microRNAs (miRNAs) in the circulation sparked interest in their use as potential biomarkers. Our previous studies showed the diagnostic potential of miR-451 as a serological marker for inflammatory breast cancer (IBC), miR-337- 5p and miR-30b for non-inflammatory breast cancer (non-IBC). The aim of this study is to investigate the prognostic values of circulating miRNAs by comparing the amounts of 12 circulating miRNAs in the serum of IBC and non-IBC from Tunisian breast cancer patients, and by determinating whether correlated pairs of miRNAs could provide useful information in the diagnosis of IBC and non-IBC patients. TaqMan qPCR was performed to detect circulating expression of miRNAs in serum of 20 IBC, 20 non-IBC and 20 healthy controls. Nonparametric rank Spearman rho correlation coefficient was used to examine the prognostic value of miRNAs and to assess the correlation profile between miRNAs expression. Further, a large number of miRNAs were highly correlated (rho>0.5) in both patients groups and controls. Also, the correlations profiles were different between IBC, non-IBC and healthy controls indicating important changes in molecular pathways in cancer cells. Our results showed that miR-335 was significantly overexpressed in premenopausal non-IBC patients; miR-24 was significantly overexpressed in non-IBC postmenopausal patients. Patients with previous parity had higher serum of miR-342-5p levels than those without. Furthermore, patients with HER2+ IBC present lower serum levels of miR-15a than patients with HER2- disease. Together, these results underline the potential of miRNAs to function as diagnostic and prognostic markers for IBC and non-IBC, with links to the menopausal state, Her2 status and parity.
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- 2016
4. Exome sequencing of oral squamous cell carcinoma in users of Arabian snuff reveals novel candidates for driver genes
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Maged El-Setouhy, Shiyong Li, Akram Thabet Nasher, Christopher A. Loffredo, Nezar Noor Al-hebshi, Rashad Mohammed Al-Sanosi, and Jan Blancato
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0301 basic medicine ,Genetics ,Mouth neoplasm ,Cancer Research ,Genetic heterogeneity ,Biology ,stomatognathic diseases ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,Oncology ,030220 oncology & carcinogenesis ,Genetic variation ,TP63 ,HRAS ,Exome ,Gene ,Exome sequencing - Abstract
The study sought to identify genetic aberrations driving oral squamous cell carcinoma (OSCC) development among users of shammah, an Arabian preparation of smokeless tobacco. Twenty archival OSCC samples, 15 of which with a history of shammah exposure, were whole-exome sequenced at an average depth of 127×. Somatic mutations were identified using a novel, matched controls-independent filtration algorithm. CODEX and Exomedepth coupled with a novel, Database of Genomic Variant-based filter were employed to call somatic gene-copy number variations. Significantly mutated genes were identified with Oncodrive FM and the Youn and Simon's method. Candidate driver genes were nominated based on Gene Set Enrichment Analysis. The observed mutational spectrum was similar to that reported by the TCGA project. In addition to confirming known genes of OSCC (TP53, CDKNA2, CASP8, PIK3CA, HRAS, FAT1, TP63, CCND1 and FADD) the analysis identified several candidate novel driver events including mutations of NOTCH3, CSMD3, CRB1, CLTCL1, OSMR and TRPM2, amplification of the proto-oncogenes FOSL1, RELA, TRAF6, MDM2, FRS2 and BAG1, and deletion of the recently described tumor suppressor SMARCC1. Analysis also revealed significantly altered pathways not previously implicated in OSCC including Oncostatin-M signalling pathway, AP-1 and C-MYB transcription networks and endocytosis. There was a trend for higher number of mutations, amplifications and driver events in samples with history of shammah exposure particularly those that tested EBV positive, suggesting an interaction between tobacco exposure and EBV. The work provides further evidence for the genetic heterogeneity of oral cancer and suggests shammah-associated OSCC is characterized by extensive amplification of oncogenes.
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- 2016
5. Somatic mutation signatures in primary liver tumors of workers exposed to ionizing radiation
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Jan Blancato, Garrett T. Graham, Islam, David S. Goerlitz, Valentina Revina, Archana Ramesh, Christopher A. Loffredo, Evgeniya Kirillova, Jay Zeck, and Bhaskar Kallakury
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Male ,Carcinoma, Hepatocellular ,Neoplasms, Radiation-Induced ,Angiogenesis ,DNA Mutational Analysis ,Hemangiosarcoma ,lcsh:Medicine ,Pilot Projects ,Biology ,Chromatin remodeling ,Article ,Russia ,Cholangiocarcinoma ,Cohort Studies ,03 medical and health sciences ,0302 clinical medicine ,Germline mutation ,Occupational Exposure ,Exome Sequencing ,medicine ,Cancer genomics ,Humans ,lcsh:Science ,Protein kinase B ,Gene ,Exome sequencing ,030304 developmental biology ,Aged ,0303 health sciences ,Multidisciplinary ,lcsh:R ,Liver Neoplasms ,Wnt signaling pathway ,Middle Aged ,medicine.disease ,3. Good health ,Occupational Diseases ,Waste Disposal Facilities ,Liver ,030220 oncology & carcinogenesis ,Radioactive Waste ,Mutation ,Cancer research ,lcsh:Q ,Female ,Liver cancer - Abstract
Liver cancer is associated with genetic mutations caused by environmental exposures, including occupational exposure to alpha radiation emitted by plutonium. We used whole exome sequencing (WES) to characterize somatic mutations in 3 histologically distinct primary liver tumors (angiosarcoma of the liver (ASL), cholangiocarcinoma (CCA) and hepatocellular carcinoma (HCC)) from Mayak worker subjects occupationally exposed to ionizing radiation (IR) to investigate the contribution of IR to the mutational landscape of liver cancer. DNA sequence analysis revealed these tumors harbor an excess of deletions, with a deletions:substitutions ratio similar to that previously reported in radiation-associated tumors. These tumors were also enriched for clustered mutations, a signature of radiation exposure. Multiple tumors displayed similarities in abrogated gene pathways including actin cytoskeletal signaling and DNA double-strand break (DSB) repair. WES identified novel candidate driver genes in ASL involved in angiogenesis and PIK3CA/AKT/mTOR signaling. We confirmed known driver genes of CCA, and identified candidate driver genes involved in chromatin remodeling. In HCC tumors we validated known driver genes, and identified novel putative driver genes involved in Wnt/β-catenin signaling, chromatin remodeling, PIK3CA/AKT/mTOR signaling, and angiogenesis. This pilot study identifies several novel candidate driver mutations that are likely to be caused by IR exposure, and provides the first data on the mutational landscape of liver cancer after IR exposure.
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- 2018
6. Knowledge and Practice of Colorectal Screening in a Suburban Group of Iraqi American Women
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Zainab Faeq, Irene Anne Jillson, Khaled W. Kabbara, William Mumford, Jan Blancato, and Carolyn Cousin
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Adult ,Gerontology ,Health Knowledge, Attitudes, Practice ,medicine.medical_specialty ,Referral ,Population ,Alternative medicine ,Disease ,Article ,Young Adult ,Ethnicity ,Humans ,Medicine ,Young adult ,education ,Health Education ,Early Detection of Cancer ,education.field_of_study ,business.industry ,Incidence (epidemiology) ,Public Health, Environmental and Occupational Health ,Middle Aged ,United States ,Suburban Population ,Cancer registry ,Oncology ,Iraq ,Female ,Perception ,Health education ,Colorectal Neoplasms ,business ,Follow-Up Studies - Abstract
Colorectal cancer (CRC) was the second most common cancer among women in 2008, accounting for 571,000 cases, and 9.4 % of all cancer cases afflicting women worldwide. According to the World Health Organization (WHO) and the Iraqi National Cancer Registry (INCR), Iraq has seen a steady rise in CRC rates among its general population over the past several decades. Despite Iraq’s increasing national incidence of CRC and the growth of the US’ Iraqi immigrant population over the last 10 years, little remains known about the prevalence of CRC among the latter population, their knowledge of CRC and associated risk factors, or their behavioral intent and practices regarding CRC screening. The aims of this study were to (1) examine the knowledge of and adherence to National Cancer Institute screening recommendations for CRC among a population of Iraqi women living in the Washington D.C. Metropolitan Area and (2) test the efficacy of a one-time educational intervention conducted using linguistically and culturally appropriate materials to raise awareness of, and promote future adherence to, CRC screening methods. This descriptive study used a pre/post design with a 12-month follow-up. Following extensive dissemination of information regarding the study in the Iraqi American community in the study location, 50 women were initially recruited, of whom 32 participated in the study. The study’s findings revealed that the participants generally had low baseline levels of CRC screening adherence and preventive knowledge that significantly improved after the intervention as demonstrated by pre- and post-assessments of knowledge and behavior. These findings could be used to raise awareness (1) among clinicians regarding the need for early detection and screening of and referral for CRC treatment among Iraqi American women and (2) among Iraqi American women about risk factors for this disease and the importance of early detection and screening. The study also highlights the need for a larger study of knowledge, attitudes, and perceptions among both this population and the clinicians who serve them.
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- 2015
7. Preimplantation genetics and other reproductive options in Huntington disease
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Jan Blancato, Preston C Sacks, and Erin M Wolfe
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0301 basic medicine ,Infertility ,Genetics ,Pregnancy ,In vitro fertilisation ,Offspring ,medicine.medical_treatment ,Prenatal diagnosis ,Embryo ,Disease ,Fertilization in Vitro ,030105 genetics & heredity ,Biology ,Preimplantation genetic diagnosis ,medicine.disease ,Polymerase Chain Reaction ,Article ,03 medical and health sciences ,Huntington Disease ,Prenatal Diagnosis ,medicine ,Humans ,Female ,Preimplantation Diagnosis - Abstract
Preimplantation genetic diagnosis (PGD) is a form of prenatal diagnosis applied to potential parents with known carrier status of a genetic disease, such as Huntington disease. It employs the use of polymerase chain reaction to amplify single cells from early embryos obtained with in vitro fertilization (IVF) techniques. PGD allows the couple the chance to have a pregnancy and livebirth child without Huntington disease, although there are some risks and expenses related to the procedures. Success of the procedure may be greater than standard IVF because the patients are not infertility patients, but are undergoing the procedure to avoid passing a highly deleterious disease gene to offspring. Recent advances in sequencing may allow for higher success rates as the chromosomally abnormal embryos will be identified more easily and the embryos with the highest chance of survival will be transferred.
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- 2017
8. Expanding primary cells from mucoepidermoid and other salivary gland neoplasms for genetic and chemosensitivity testing
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Weisheng Wang, Ahmad M. Alamri, Xiaogang Zhong, Jan Blancato, Bruce J. Davidson, Bassem R. Haddad, Bhaskar Kallakury, Ewa Krawczyk, Sujata Choudhary, Priscilla A. Furth, and Xuefeng Liu
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0301 basic medicine ,lcsh:Medicine ,Medicine (miscellaneous) ,Gene mutation ,Transcriptome ,0302 clinical medicine ,Immunology and Microbiology (miscellaneous) ,Akt Inhibitor MK2206 ,Tumor Cells, Cultured ,Medicine ,0303 health sciences ,Salivary gland ,High-Throughput Nucleotide Sequencing ,Cell Differentiation ,3. Good health ,Neoplasm Proteins ,Gene Expression Regulation, Neoplastic ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Mucoepidermoid carcinoma ,lcsh:RB1-214 ,Signal Transduction ,Research Article ,Drug sensitivity ,Cell Survival ,Neuroscience (miscellaneous) ,Antineoplastic Agents ,Biology ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,lcsh:Pathology ,Humans ,Primary cell culture ,Protein kinase B ,Protein Kinase Inhibitors ,030304 developmental biology ,Cell Proliferation ,Salivary gland neoplasms ,business.industry ,AKT ,lcsh:R ,Cancer ,medicine.disease ,Dd ,Xenograft Model Antitumor Assays ,030104 developmental biology ,Cell culture ,Culture Media, Conditioned ,Immunology ,Cancer research ,Next-generation sequencing ,Carcinoma, Mucoepidermoid ,Salivary gland neoplasm ,business ,Chemosensitivity assay - Abstract
Restricted availability of cell and animal models is a rate-limiting step for investigation of salivary gland neoplasm pathophysiology and therapeutic response. Conditionally reprogrammed cell (CRC) technology enables establishment of primary epithelial cell cultures from patient material. This study tested a translational workflow for acquisition, expansion and testing of CRC-derived primary cultures of salivary gland neoplasms from patients presenting to an academic surgical practice. Results showed that cultured cells were sufficient for epithelial cell-specific transcriptome characterization to detect candidate therapeutic pathways and fusion genes, and for screening for cancer risk-associated single nucleotide polymorphisms (SNPs) and driver gene mutations through exome sequencing. Focused study of primary cultures of a low-grade mucoepidermoid carcinoma demonstrated amphiregulin-mechanistic target of rapamycin-protein kinase B (AKT; AKT1) pathway activation, identified through bioinformatics and subsequently confirmed as present in primary tissue and preserved through different secondary 2D and 3D culture media and xenografts. Candidate therapeutic testing showed that the allosteric AKT inhibitor MK2206 reproducibly inhibited cell survival across different culture formats. By contrast, the cells appeared resistant to the adenosine triphosphate competitive AKT inhibitor GSK690693. Procedures employed here illustrate an approach for reproducibly obtaining material for pathophysiological studies of salivary gland neoplasms, and other less common epithelial cancer types, that can be executed without compromising pathological examination of patient specimens. The approach permits combined genetic and cell-based physiological and therapeutic investigations in addition to more traditional pathologic studies, and can be used to build sustainable bio-banks for future inquiries. This article has an associated First Person interview with the first author of the paper., Summary: Genetics and therapeutic responses of uncommon neoplasms, including salivary gland cancers, are investigatable through isolation of primary cells from patient material using conditional reprogramming cell technology without compromising pathological diagnosis.
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- 2017
9. HPV positive neuroendocrine cervical cancer cells are dependent on Myc but not E6/E7 viral oncogenes
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Nancy Palechor-Ceron, Xuefeng Liu, Ewa Krawczyk, Thomas Ried, Jan Blancato, Siddartha Paul, Seema Agarwal, Richard Schlegel, Bassem R. Haddad, Naidong Wang, Danny Wangsa, Tung Wei Hou, Faris Alkhilaiwi, Christopher Albanese, Hang Yuan, Shuang Fang, Dan Zhou, Yukari Usuda, Maura L. Gillison, Sujata Choudhary, Yun Ling Zheng, David E. Symer, Aleksandra Dakic, and Dan P. Hartmann
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0301 basic medicine ,Papillomavirus E7 Proteins ,Cell ,Viral Oncogene ,Uterine Cervical Neoplasms ,Biology ,Models, Biological ,Article ,Proto-Oncogene Proteins c-myc ,Fusion gene ,Mice ,03 medical and health sciences ,Tumor Cells, Cultured ,medicine ,Animals ,In Situ Hybridization, Fluorescence ,Cell Proliferation ,Recombination, Genetic ,Human papillomavirus 16 ,Gene knockdown ,Multidisciplinary ,Oncogene ,Cell growth ,Oncogene Proteins, Viral ,medicine.disease ,Primary tumor ,Repressor Proteins ,Cell Transformation, Neoplastic ,030104 developmental biology ,medicine.anatomical_structure ,Cell culture ,Karyotyping ,Cancer research ,Female ,Gene Fusion - Abstract
Using conditional cell reprogramming, we generated a stable cell culture of an extremely rare and aggressive neuroendocrine cervical cancer. The cultured cells contained HPV-16, formed colonies in soft agar and rapidly produced tumors in immunodeficient mice. The HPV-16 genome was integrated adjacent to the Myc gene, both of which were amplified 40-fold. Analysis of RNA transcripts detected fusion of the HPV/Myc genes, arising from apparent microhomologous recombination. Spectral karyotyping (SKY) and fluorescent-in-situ hybridization (FISH) demonstrated coordinate localization and translocation of the amplified Myc and HPV genes on chromosomes 8 and 21. Similar to the primary tumor, tumor cell cultures expressed very high levels of the Myc protein and, in contrast to all other HPV-positive cervical cancer cell lines, they harbored a gain-of-function mutation in p53 (R273C). Unexpectedly, viral oncogene knockdown had no effect on the growth of the cells, but it did inhibit the proliferation of a conventional HPV-16 positive cervical cancer cell line. Knockdown of Myc, but not the mutant p53, significantly inhibited tumor cell proliferation. On the basis of these data, we propose that the primary driver of transformation in this aggressive cervical cancer is not HPV oncogene expression but rather the overexpression of Myc.
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- 2017
10. Fluorescence In Situ Hybridization of Cells, Chromosomes, and Formalin-Fixed Paraffin-Embedded Tissues
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Ahmad M. Alamri, Jan Blancato, and Jun Yeb Nam
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0301 basic medicine ,Tissue Fixation ,DNA Copy Number Variations ,In situ hybridization ,Article ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Formaldehyde ,medicine ,Fluorescence microscope ,Humans ,Metaphase ,Cells, Cultured ,In Situ Hybridization, Fluorescence ,Chromosome Aberrations ,Paraffin Embedding ,medicine.diagnostic_test ,Hybridization probe ,Molecular biology ,030104 developmental biology ,Microscopy, Fluorescence ,chemistry ,Cell culture ,030220 oncology & carcinogenesis ,DNA Probes ,DNA ,Fluorescence in situ hybridization ,Comparative genomic hybridization - Abstract
Fluorescence in situ hybridization (FISH) with DNA probes allows the visualization of gene copy number and localization of specific DNA targets with fluorescence microscopy. Cells in culture, metaphase chromosomes, and tissue sections are fixed and prepared on glass slides. Both the DNA in the cells and fluorescently labeled probe are denatured, and the labeled probe is allowed to hybridize to the cellular DNA. The slides are washed, counterstained, and viewed via fluorescence microscopy. We describe the basic method for preparing slides and probes for studies involving DNA copy number changes and structural chromosome rearrangements in formalin-fixed paraffin-embedded (FFPE) tissue sections and cell culture preparations.
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- 2017
11. Molecular profiling (MP) for malignancies: Knowledge gaps and variable practice patterns among United States oncologists (Onc)
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Arden Buettner, Brynn Tiscione, Jan Blancato, Petra Prins, Joanne Willey, Maria L. Lankford, Chao Yin, Mitchell Scharf, John L. Marshall, Bhavana Pendurthi Singh, and Susan Lynne Britton
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03 medical and health sciences ,Cancer Research ,0302 clinical medicine ,Oncology ,Practice patterns ,business.industry ,030220 oncology & carcinogenesis ,Medicine ,Profiling (information science) ,Computational biology ,business ,030215 immunology - Abstract
10510 Background: Clinically impactful therapies for malignancies require the identification of specific molecular alterations. Onc must be aware of these targets and how to interpret them to provide optimum care. The use of MP has become the standard of care for many cancers, and is recently FDA approved. Using 2 data sets, we assessed the current awareness and incorporation of MP in the treatment of cancer; comparing data from community based Onc (C) to academic Onc (A). Methods: C consisted of 292 physicians polled using an audience response system during 6 case-based research events across the US. Questions focused on various aspects of molecular testing. Data for A was obtained from a chart review focused on timing and extent of MP in disease specific academic practices (lung, breast, GI) (N = 59). Results: Within C, 257 (88%) were Onc from community-based practices. The frequency at which Onc ordered MP significantly varied depending on tumor type; 33% in lung cancer (LC), 18% in colorectal cancer (CRC) and less commonly in breast cancer (BC) (8%). In A, MP was ordered more frequently; 74% in LC, 27% in CRC and 0% in BC. These results reflect a gap in practice among community versus academic Onc, as C had lower utilization of MP for both LC and CRC. In C, Onc were also asked to match the molecular alteration with the appropriate targeted therapy. Onc incorrectly matched the molecular alteration to the targeted therapy or marked unknown in up to 69%. This reflects a large knowledge gap among community Onc with regards to the correct application of MP to currently FDA approved targeted therapies. Conclusions: Given the significant knowledge and practice gap, we conclude there is an urgent need for focused educational activities that facilitate improved knowledge of MP and corresponding personalized therapeutic strategies for Onc in the US.
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- 2019
12. MicroRNAs in glioblastoma multiforme pathogenesis and therapeutics
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Zainab Afzal, Jan Blancato, Habib Kedir, Varsha Harish, Arpita Roy, Amanda Shea, Juliet Chijioke, Deepak Kumar, Shahnoza Dusmatova, Brent T. Harris, Malathi Ramalinga, and Mukesh Verma
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0301 basic medicine ,Cancer Research ,Angiogenesis ,Reviews ,Drug resistance ,Review ,Biology ,Bioinformatics ,Metastasis ,Pathogenesis ,03 medical and health sciences ,glioblastoma multiforme ,0302 clinical medicine ,RNA interference ,microRNA ,medicine ,Animals ,Humans ,Radiology, Nuclear Medicine and imaging ,Brain Neoplasms ,Cancer ,Clinical Cancer Research ,Genetic Therapy ,medicine.disease ,3. Good health ,Gene Expression Regulation, Neoplastic ,MicroRNAs ,030104 developmental biology ,Oncology ,030220 oncology & carcinogenesis ,cancer therapy ,RNA Interference ,Glioblastoma - Abstract
Glioblastoma multiforme (GBM) is the most common and lethal cancer of the adult brain, remaining incurable with a median survival time of only 15 months. In an effort to identify new targets for GBM diagnostics and therapeutics, recent studies have focused on molecular phenotyping of GBM subtypes. This has resulted in mounting interest in microRNAs (miRNAs) due to their regulatory capacities in both normal development and in pathological conditions such as cancer. miRNAs have a wide range of targets, allowing them to modulate many pathways critical to cancer progression, including proliferation, cell death, metastasis, angiogenesis, and drug resistance. This review explores our current understanding of miRNAs that are differentially modulated and pathologically involved in GBM as well as the current state of miRNA‐based therapeutics. As the role of miRNAs in GBM becomes more well understood and novel delivery methods are developed and optimized, miRNA‐based therapies could provide a critical step forward in cancer treatment.
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- 2016
13. High-throughput, Quantitative Analysis of Acrolein-derived DNA Adducts in Human Oral Cells by Immunohistochemistry
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Fung-Lung Chung, Marcin Dyba, Jan Blancato, Hanjoo Lee, Tierra Johnson, Kepher Mekambi, Deborah L. Berry, Emily Greenspan, Jishen Pan, and Susette C. Mueller
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musculoskeletal diseases ,Cell type ,Histology ,medicine.drug_class ,Bronchi ,Biology ,Monoclonal antibody ,Tandem mass spectrometry ,Cell Line ,Lipid peroxidation ,DNA Adducts ,Mice ,chemistry.chemical_compound ,medicine ,Animals ,Humans ,Acrolein ,Cells, Cultured ,Mice, Inbred BALB C ,Mouth ,Articles ,Immunohistochemistry ,Molecular biology ,High-Throughput Screening Assays ,Microscopy, Fluorescence ,chemistry ,Cell culture ,biology.protein ,Anatomy ,Antibody ,Quantitative analysis (chemistry) ,Software ,Mutagens - Abstract
Acrolein (Acr) is a ubiquitous environmental pollutant as well as an endogenous compound. Acrolein-derived 1,N2-propanodeoxyguanosines (Acr-dG) are exocyclic DNA adducts formed following exposure to cigarette smoke or from lipid peroxidation. Acr-dG is mutagenic and potentially carcinogenic and may represent a useful biomarker for the early detection of cancers related to smoking or other oxidative conditions, such as chronic inflammation. In this study, we have developed a high-throughput, automated method using a HistoRx PM-2000 imaging system combined with MetaMorph software for quantifying Acr-dG adducts in human oral cells by immunohistochemical detection using a monoclonal antibody recently developed by our laboratory. This method was validated in a cell culture system using BEAS-2B human bronchial epithelial cells treated with known concentrations of Acr. The results were further verified by quantitative analysis of Acr-dG in DNA of BEAS-2B cells using a liquid chromatography/tandem mass spectrometry/multiple-reaction monitoring method. The automated method is a quicker, more accurate method than manual evaluation of counting cells expressing Acr-dG and quantifying fluorescence intensity. It may be applied to other antibodies that are used for immunohistochemical detection in tissues as well as cell lines, primary cultures, and other cell types.
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- 2012
14. Multiple pregnancies, hepatitis C, and risk for hepatocellular carcinoma in Egyptian women
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Ghada R Nasr, Kirti Shetty, Sania Amr, Doa’a A. Saleh, Jan Blancato, Christopher A. Loffredo, and Emily A Iarocci
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Oncology ,Cancer Research ,medicine.medical_specialty ,Carcinoma, Hepatocellular ,Hepatocellular carcinoma ,Epidemiology ,Gravidity ,Women’s health ,03 medical and health sciences ,0302 clinical medicine ,Pregnancy ,Risk Factors ,Internal medicine ,Genetics ,medicine ,Humans ,Obstetrics ,business.industry ,Incidence ,Incidence (epidemiology) ,Liver Neoplasms ,Case-control study ,virus diseases ,Odds ratio ,Hepatitis C ,medicine.disease ,digestive system diseases ,3. Good health ,Case-Control Studies ,030220 oncology & carcinogenesis ,Women's Health ,Egypt ,Female ,030211 gastroenterology & hepatology ,Viral hepatitis ,business ,Research Article - Abstract
Background The reasons for the worldwide sex disparity in the incidence of hepatocellular carcinoma (HCC) remain elusive. We investigated the role of multiple pregnancies on the associations between viral hepatitis C (HCV) infection and HCC risk among Egyptian women. Methods We used data collected from blood specimens and questionnaires administered to female HCC cases and controls in Cairo, Egypt, from 1999 through 2009. HCV infection was defined as being sero-positive for either anti-HCV antibodies or HCV-RNA. Using logistic regression models we calculated odds ratios (OR) and 95% confidence intervals (CI) to estimate the associations between being HCV positive and HCC risk, and how it is modified by the number of pregnancies, after adjustment for other factors, including hepatitis B status. Results Among 132 confirmed female cases and 669 controls, the risk of HCV-related HCC increased with the number of pregnancies. Women infected with HCV had higher risk for HCC if they had more than five pregnancies, as compared to those who had five or fewer pregnancies (adjusted OR (95% CI): 2.33 (1.29-4.22)). The association of HCV infection with HCC risk was significantly greater among the former (21.42 (10.43-44.00)) than among the latter (6.57 (3.04-14.25)). Conclusion Having multiple pregnancies increases the risk of HCV-related HCC among Egyptian women, raising questions about the roles of estrogens and other pregnancy-related hormones in modulating HCV infection and its progression to HCC.
- Published
- 2014
15. Diagnostic FISH probes for del(17)(p11.2p11.2) associated with Smith-Magenis syndrome should contain theRAI1gene
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Ann C.M. Smith, Sarah H. Elsea, Jan Blancato, Meredith Wilson, and Christopher N. Vlangos
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Genetics ,Mutation ,medicine.diagnostic_test ,Retinoic acid induced 1 ,Chromosome ,Biology ,Smith–Magenis syndrome ,medicine.disease ,medicine.disease_cause ,Molecular biology ,Gene mapping ,medicine ,Haploinsufficiency ,Gene ,Genetics (clinical) ,Fluorescence in situ hybridization - Abstract
Smith-Magenis syndrome (SMS) is a mental retardation syndrome with distinctive behavioral characteristics, dysmorphic features, and congenital anomalies usually associated with an interstitial deletion of chromosome 17p11.2. While high quality G-banding will identify most SMS patients, fluorescent in situ hybridization (FISH) is the recommended test for confirmation of an SMS diagnosis. Recently, haploinsufficiency of the RAI1 gene due to deletion or mutation was determined to be the likely cause of SMS. All diagnostic FISH probes available commercially contain the FLII gene and are approximately 580 kb centromeric to RAI1. We present two patients with SMS who have interstitial deletions at 17p11.2 but are not deleted for currently available commercial FISH probes that include FLII; both patients have deletions that are demonstrated with probes containing the RAI1 gene. We recommend that for diagnostic accuracy, all future FISH tests for SMS be performed with probes containing the RAI1 gene, as some atypical deletions in the region critical to the SMS phenotype will otherwise be missed.
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- 2004
16. Clinical and molecular delineation of the Greig cephalopolysyndactyly contiguous gene deletion syndrome and its distinction from acrocallosal syndrome
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J. Edward Spence, Lynne M. Bird, Joyce T. Turner, R. Neil Schimke, Heidi A. Heilstedt, Lakshmi Mehta, Leslie G. Biesecker, Jan Blancato, Jennifer J. Johnston, Isabelle M. Olivos-Glander, and Kyrieckos A. Aleck
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Male ,Genotype ,DNA Mutational Analysis ,Kruppel-Like Transcription Factors ,Nerve Tissue Proteins ,Biology ,Contiguous gene syndrome ,Frameshift mutation ,Craniofacial Abnormalities ,Diagnosis, Differential ,Zinc Finger Protein Gli3 ,Intellectual Disability ,medicine ,Humans ,Abnormalities, Multiple ,Hypertelorism ,In Situ Hybridization, Fluorescence ,Genetics (clinical) ,Greig cephalopolysyndactyly syndrome ,Genetics ,Base Sequence ,Polydactyly ,Macrocephaly ,Syndrome ,medicine.disease ,Acrocallosal syndrome ,DNA-Binding Proteins ,Phenotype ,Karyotyping ,Female ,Agenesis of Corpus Callosum ,Chromosome Deletion ,medicine.symptom ,Cognition Disorders ,Haploinsufficiency ,Chromosomes, Human, Pair 7 ,Gene Deletion ,Transcription Factors - Abstract
Greig cephalopolysyndactyly syndrome (GCPS) is caused by haploinsufficiency of GLI3 on 7p13. Features of GCPS include polydactyly, macrocephaly, and hypertelorism, and may be associated with cognitive deficits and abnormalities of the corpus callosum. GLI3 mutations in GCPS patients include point, frameshift, translocation, and gross deletion mutations. FISH and STRP analyses were applied to 34 patients with characteristics of GCPS. Deletions were identified in 11 patients and the extent of their deletion was determined. Nine patients with deletions had mental retardation (MR) or developmental delay (DD) and were classified as severe GCPS. These severe GCPS patients have manifestations that overlap with the acrocallosal syndrome (ACLS). The deletion breakpoints were analyzed in six patients whose deletions ranged in size from 151 kb to 10.6 Mb. Junction fragments were found to be distinct with no common sequences flanking the breakpoints. We conclude that patients with GCPS caused by large deletions that include GLI3 are likely to have cognitive deficits, and we hypothesize that this severe GCPS phenotype is caused by deletion of contiguous genes.
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- 2003
17. Human T-Lymphotropic Virus Type 1 Oncoprotein Tax Promotes Unscheduled Degradation of Pds1p/Securin and Clb2p/Cyclin B1 and Causes Chromosomal Instability
- Author
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Chou-Zen Giam, Jan Blancato, Min-Hui Liang, Imre Boros, Wei Liao, Yu-Liang Kuo, Baoying Liu, and Tami Kleinberger
- Subjects
Time Factors ,Cyclin D ,Cyclin A ,Cyclin B ,Cell Cycle Proteins ,Cyclin B1 ,Cell Growth and Development ,biology ,Cell Cycle ,Temperature ,Nuclear Proteins ,Ubiquitin-Protein Ligase Complexes ,Gene Products, tax ,Cell cycle ,Flow Cytometry ,Cell biology ,Securin ,Plasmids ,G2 Phase ,Saccharomyces cerevisiae Proteins ,Cell Survival ,Green Fluorescent Proteins ,Immunoblotting ,Mitosis ,Saccharomyces cerevisiae ,CDC20 ,Apc8 Subunit, Anaphase-Promoting Complex-Cyclosome ,Anaphase-Promoting Complex-Cyclosome ,Chromosomes ,Adenoviridae ,Cell Line ,Animals ,Humans ,Molecular Biology ,Metaphase ,Cell Nucleus ,Cell Biology ,Fibroblasts ,Aneuploidy ,Precipitin Tests ,Molecular biology ,Luminescent Proteins ,Karyotyping ,Mutation ,biology.protein ,Anaphase-promoting complex ,Protein Kinases ,HeLa Cells ,Thymidine - Abstract
Human T-lymphotropic virus type 1 (HTLV-1) is the causative agent of adult T-cell leukemia. The HTLV-1 transactivator, Tax, is implicated as the viral oncoprotein. Naïve cells expressing Tax for the first time develop severe cell cycle abnormalities that include increased DNA synthesis, mitotic arrest, appearance of convoluted nuclei with decondensed DNA, and formation of multinucleated cells. Here we report that Tax causes a drastic reduction in Pds1p/securin and Clb2p/cyclin B levels in yeast, rodent, and human cells and a loss of cell viability. With a temperature-sensitive mutant of the CDC23 subunit of the anaphase-promoting complex (APC), cdc23(ts); a temperature-sensitive mutant of cdc20; and a cdh1-null mutant, we show that the diminution of Pds1p and Clb2p brought on by Tax is mediated via the Cdc20p-associated anaphase-promoting complex, APC(Cdc20p). This loss of Pds1p/securin and Clb2p/cyclin B1 occurred before cellular entry into mitosis, caused a G(2)/M cell cycle block, and was accompanied by severe chromosome aneuploidy in both Saccharomyces cerevisiae cells and human diploid fibroblasts. Our results support the notion that Tax aberrantly targets and activates APC(Cdc20p), leading to unscheduled degradation of Pds1p/securin and Clb2p/cyclin B1, a delay or failure in mitotic entry and progression, and faulty chromosome transmission. The chromosomal instability resulting from a Tax-induced deficiency in securin and cyclin B1 provides an explanation for the highly aneuploid nature of adult T-cell leukemia cells.
- Published
- 2003
18. Absence of donor-derived keratinocyte stem cells in skin tissues cultured from patients after mobilized peripheral blood hematopoietic stem cell transplantation
- Author
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Cristian Carvallo, Carole Yee, Michael R. Albert, Peiman Hematti, Richard W. Childs, Cynthia E. Dunbar, J. Barrett, Jonathan C. Vogel, Monika M Fuehrer, Jan Blancato, Elaine M. Sloand, and W.G. Kearns
- Subjects
Adult ,Keratinocytes ,Male ,Cancer Research ,Pathology ,medicine.medical_specialty ,X Chromosome ,Biopsy ,medicine.medical_treatment ,Hematopoietic stem cell transplantation ,Biology ,Dental Enamel Proteins ,Y Chromosome ,Granulocyte Colony-Stimulating Factor ,Genetics ,medicine ,Humans ,Transplantation, Homologous ,Cell Lineage ,Molecular Biology ,Cells, Cultured ,In Situ Hybridization, Fluorescence ,Hematopoietic Stem Cell Mobilization ,Stem cell transplantation for articular cartilage repair ,Transplantation Chimera ,Amelogenin ,integumentary system ,Stem Cells ,Graft Survival ,Hematopoietic Stem Cell Transplantation ,Hematopoietic stem cell ,Cell Differentiation ,Amniotic stem cells ,Cell Biology ,Hematology ,Middle Aged ,Tissue Donors ,Endothelial stem cell ,medicine.anatomical_structure ,Epidermal Cells ,Microscopy, Fluorescence ,Organ Specificity ,Leukocyte Common Antigens ,Female ,Stem cell ,Follow-Up Studies ,Adult stem cell - Abstract
Objective Recent studies suggest that primitive bone marrow-derived cells contribute to regeneration of many tissues, including muscle, endothelium, myocardium, neural tissues, liver, and skin. Conversely, primitive cells resident in muscle and other tissues have been reported to reconstitute hematopoiesis. We investigated the contribution of cells with a primitive hematopoietic phenotype to human epidermal skin formation in recipients of allogeneic mobilized peripheral blood hematopoietic stem cell (HSC) transplantation. Patients and Methods Our study population included female patients who had received granulocyte colony-stimulating factor mobilized peripheral blood HSC transplants from male donors for a variety of benign and malignant hematologic disorders at least 6 months before study entry, with a history of skin graft-vs-host disease. Epidermal skin cells (keratinocytes) obtained from punch biopsies of the skin were cultured under conditions specific for growth and expansion of homogenous populations of keratinocytes from keratinocyte stem cells. After multiple passages, DNA was extracted from cultured cells and evaluated by two different polymerase chain reaction (PCR) method for detection of Y chromosome specific sequences. Results Neither sensitive PCR-based technique revealed the presence of male donor-derived keratinocyte stem cells in keratinocytes cultured from skin biopsies of female allogeneic transplantation recipients. Conclusions We could not confirm the contribution of donor mobilized peripheral blood hematopoietic stem cells to keratinocyte stem cell populations after HSC transplantation. These results cannot explain the presence of donor-derived cells with keratinocyte phenotypic markers in tissue sections of HSC transplant recipients.
- Published
- 2002
19. Hypercholesterolemia in children with Smith-Magenis syndrome: del (17)(p11.2p11.2)
- Author
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Lorraine Potocki, Andrea L. Gropman, Joan E. Bailey-Wilson, Ann C.M. Smith, Ozlem Goker-Alpan, James R. Lupski, Jan Blancato, and Sarah H. Elsea
- Subjects
Male ,medicine.medical_specialty ,Percentile ,Adolescent ,Hypercholesterolemia ,Biology ,Contiguous gene syndrome ,Body Mass Index ,chemistry.chemical_compound ,High-density lipoprotein ,Internal medicine ,medicine ,Humans ,Abnormalities, Multiple ,Child ,Triglycerides ,Genetics (clinical) ,Cholesterol ,Cholesterol, HDL ,Infant ,Cholesterol, LDL ,Syndrome ,Smith–Magenis syndrome ,medicine.disease ,DNA-Binding Proteins ,Chromosome 17 (human) ,Endocrinology ,chemistry ,Child, Preschool ,Low-density lipoprotein ,CCAAT-Enhancer-Binding Proteins ,Female ,lipids (amino acids, peptides, and proteins) ,Sterol Regulatory Element Binding Protein 1 ,Body mass index ,Gene Deletion ,Chromosomes, Human, Pair 17 ,Transcription Factors - Abstract
Purpose: Smith-Magenis syndrome (SMS), a probable contiguous gene syndrome due to an interstitial deletion of chromosome 17 band p11.2, is associated with a distinct and complex phenotype, including physical, developmental, and neurobehavioral features. The majority of SMS patients are deleted for a common ∼ 4 Mb interval that includes the gene SREBF1, a transmembrane transcription factor that regulates the low density lipoprotein (LDL) receptor and plays a crucial role in cholesterol homeostasis. A systematic study of fasting lipid profiles of patients with SMS was conducted to determine the frequency of cholesterol abnormalities. Methods: Fasting lipid profiles were examined in 49 children (27F/22M) between the ages of 0.6 years to 17.6 years (mean, 6.9 years) with a cytogenetically confirmed diagnosis of SMS. Observed values for serum total cholesterol (TC), triglycerides (TG), LDL cholesterol, and high density lipoprotein cholesterol were compared with published norms. The body mass index (BMI) was used as a measure of nutritional status. Results: Mean TC was significantly higher than published NHANES III pediatric norms (P < 0.0008). Overall 28 of 49 (57%) SMS subjects had lipid values greater than the 95th percentile for age and gender for at least one or more of the following: TC, TG, and/or LDL. Only 16 SMS subjects (32%) were within normal limits for all three of these variables. BMI values showed minimal positive correlation to SMS lipid values; however, no consistent effect was found. Thus BMI values alone do not explain the marked trend in increased TC, TG, and/or LDL observed in the SMS group. Based on the American Academy of Pediatrics recommended lipid levels for children and adolescents, only one third of SMS subjects fall within normal range for TC and LDL; an additional one third each measure “borderline” or “high” for these values. Conclusion: Hypercholesterolemia is common in SMS and may serve as a useful early clinical biochemical marker of the syndrome.
- Published
- 2002
20. miRNAs in Sera of Tunisian patients discriminate between inflammatory breast cancer and non-inflammatory breast cancer
- Author
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Khouloud Hamdi, Bilel Neili, Mohammed Islam, Raja Marrakchi, Farhat Benayed, Christopher A. Loffredo, Amel Benammar Elgaaied, Neila Stambouli, Simon Chivi, Olfa Baroudi, Irene Anne Jillson, Jan Blancato, and David Goerlitz
- Subjects
Oncology ,medicine.medical_specialty ,Multidisciplinary ,Inflammatory breast cancer ,microRNA ,business.industry ,Research ,RT-qPCR ,Early detection ,medicine.disease ,Circulating MicroRNA ,Circulating microRNA ,Breast cancer ,Prognostic biomarkers ,Internal medicine ,Potential biomarkers ,Immunology ,medicine ,Mirna profiling ,business ,skin and connective tissue diseases ,Pathological - Abstract
In recent years, circulating miRNAs have attracted interest as stable, non-invasive biomarkers for various pathological conditions. Here, we investigated their potential to serve as minimally invasive, early detection markers for inflammatory breast cancer (IBC) and non-inflammatory breast cancer (non-IBC) in serum. miRNA profiling was performed on serum from 20 patients with non-IBC, 20 with IBC, and 20 normal control subjects. Real-time reverse transcription-polymerase chain reaction (qRT-PCR) was applied to measure the level of 12 candidate miRNAs previously identified in other research(miR-342-5p, miR-342--3p, miR-320, miR-30b, miR-29a, miR-24, miR-15a, miR-548d-5p, miR-486-3p, miR-451, miR-337-5p, miR-335). We found that 4 miRNAs (miR-24, miR-342-3p, miR-337-5p and miR-451) were differentially expressed in serum of IBC patients compared to non-IBC, and 3 miRNAs (miR-337-5p ,miR-451and miR-30b) were differentially expressed in IBC and non-IBC patients combined compared to healthy controls. miR-24, miR-342-3p, miR-337-5p and miR-451 were found to be significantly down-regulated in IBC patients compared to non-IBC. Likewise, the expression level of mir-451 showed significant down-regulation in IBC serum, while mir-30b and miR-337-5p were up-regulated in non-IBC serum comparatively to normal controls. Using receiver operational curve (ROC) analysis, we show that dysregulated miRNAs can discriminate patients with IBC and non-IBC from healthy controls with sensitivity ranging from 76 to 81% and specificity from 66 to 80%, for three separate miRNAs. In conclusion, our data suggest that circulating miRNAs are potential biomarkers for classifying IBC and non-IBC, and may also be candidates for early detection of breast cancer.
- Published
- 2014
21. A syndrome of overgrowth and acromegaloidism with normal growth hormone secretion is associated with chromosome 11 pericentric inversion
- Author
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Antony R Lafferty, Constantine A. Stratakis, Jorge R. Toro, Jeanne M. Meck, Jan Blancato, Suvimol Hill, and Maria L. Turner
- Subjects
Proband ,medicine.medical_specialty ,Pediatrics ,Cutis ,Autosomal dominant trait ,Biology ,medicine.disease ,Insulin resistance ,Endocrinology ,Internal medicine ,Acromegaly ,Genetics ,medicine ,Differential diagnosis ,Abnormality ,Letters to the Editor ,Genetics (clinical) ,Chromosomal inversion - Abstract
An acromegalic phenotype in late childhood or early adulthood is shared by a variety of clinical conditions, including growth hormone (GH) excess.1 Exclusion of an abnormality of the somatotrophic axis in a young patient with acromegaloid features should lead the differential diagnosis towards diagnoses such as pachydermoperiostosis (MIM 1671002)3-5 or insulin mediated pseudoacromegaly, a disorder associated with severe insulin resistance.6 In the absence of insulin resistance and findings characteristic of pachydermoperiostosis, such as thickening of the periosteum (visible mostly in skull x rays) or the skin, acrolysis, or alopecia,4 5 7 another genetic syndrome associated with acromegaloid features may be considered.8-13 These are rare conditions, having each been described in individual kindreds, and their causes remain unknown. Inheritance, when present, appears to be as an autosomal dominant trait. They are almost always associated with abnormalities of the skin, the mucosa, and its appendages, such as keratitis,9thickened mucosa,10 hypertrichosis,12 and cutis verticis gyrata.8 13 In this report, we identify a chromosomal anomaly that was confirmed by fluorescence in situ hybridisation (FISH) in a patient with acromegaloid features and his family. The patient, his mother, and sibs participated in protocol 97-CH-0076 (National Institute of Child Health and Human Development, National Institutes of Health (NIH)) and consented to cytogenetic and DNA studies, and the use of the proband's photographs for the purposes of medical education and publication. The proband was a 14 year 3 month old male (fig 1) who was referred to our clinic with the diagnosis of possible acromegaly. He was born at term after an uncomplicated pregnancy. His birth weight was 5018 g (over the 95th centile for a newborn and on the 50th centile for a 2½ month old boy), and his length was 60 cm (over …
- Published
- 2001
22. Characterization of a supernumerary marker derived from chromosome 17 by microdissection in an adult with MR/MCA
- Author
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Chahira Kozma, Jeanne M. Meck, Jan Blancato, and Yuan Jiang
- Subjects
Adult ,Genetic Markers ,Male ,Pathology ,medicine.medical_specialty ,Marker chromosome ,Ring chromosome ,Aneuploidy ,Trisomy ,Biology ,Intellectual Disability ,medicine ,Humans ,Supernumerary ,In Situ Hybridization, Fluorescence ,Genetics (clinical) ,Microdissection ,medicine.diagnostic_test ,Syndrome ,medicine.disease ,Molecular biology ,Chromosome 17 (human) ,Scoliosis ,Karyotyping ,Microcephaly ,Chromosomes, Human, Pair 17 ,Fluorescence in situ hybridization - Abstract
We describe a 38-year-old adult who has a supernumerary marker chromosome in 40% of metaphase cells which was identified by reverse in situ hybridization with a DNA probe made by microdissection to be derived from chromosome 17. The breakpoints are estimated by G-banding and fluorescence in situ hybridization (FISH) to consist of the region from 17p11.1 to proximal 17q21. The propositus displayed severe growth retardation, kyphoscoliosis, bilateral cataracts, severe calcaneovalgus deformity of the feet, dysmorphic facies, profound mental retardation, and multiple medical problems requiring ongoing medical management. These problems included a mitral valve prolapse with regurgitation, recurrent upper and lower respiratory tract infections, and severe respiratory insufficiency. The relatively long survival of this patient enabled us to describe the natural history of this rare chromosomal mutation.
- Published
- 1998
23. Strengthening cancer biology research, prevention, and control while reducing cancer disparities: student perceptions of a collaborative master's degree program in cancer biology, preventions, and control
- Author
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Jan Blancato, Carolyn Cousin, and Irene Anne Jillson
- Subjects
Adult ,Male ,Biomedical Research ,Adolescent ,education ,Medical Oncology ,Article ,Young Adult ,Nursing ,Neoplasms ,ComputingMilieux_COMPUTERSANDEDUCATION ,Medicine ,Humans ,Education, Graduate ,Cooperative Behavior ,Healthcare Disparities ,Students ,Curriculum ,Response rate (survey) ,Academic Medical Centers ,Cancer prevention ,business.industry ,Public Health, Environmental and Occupational Health ,Middle Aged ,Outreach ,Cross-Sectional Studies ,Oncology ,Coursework ,Female ,Program Design Language ,Biostatistics ,business ,Graduation - Abstract
This article provides the findings of a survey of previous and current students in the UDC/GU-LCCC master’s degree program. This master’s degree program, Cancer Biology, Prevention, and Control is administered and taught jointly by faculty of a Minority Serving Institution, the University of the District of Columbia, and the Lombardi Comprehensive Cancer Center to incorporate the strengths of a community-based school with a research intensive medical center. The program was initiated in 2008 through agreements with both University administrations and funding from the National Cancer Institute. The master’s degree program is 36 credits with a focus on coursework in biostatistics, epidemiology, tumor biology, cancer prevention, medical ethics, and cancer outreach program design. For two semesters during the second year, students work full-time with a faculty person on a laboratory or outreach project that is a requirement for graduation. Students are supported and encouraged to transition to a doctoral degree after they obtain the master’s and many of them are currently in doctorate programs. Since the inception of the program, 45 students have initiated the course of study, 28 have completed the program, and 13 are currently enrolled in the program. The survey was designed to track the students in their current activities, as well as determine which courses, program enhancements, and research experiences were the least and most useful, and to discern students’ perceptions of knowledge acquired on various aspects of Cancer Biology Prevention, and Control Master’s Program. Thirty of the 35 individuals to whom email requests were sent responded to the survey, for a response rate of 85.7 %. The results of this study will inform the strengthening of the Cancer Biology program by the Education Advisory Committee. They can also be used in the development of comparable collaborative master’s degree programs designed to address the significant disparities in prevalence of cancer, low screening awareness, and access to and outcomes of cancer prevention and treatment services. This, in turn, will contribute to the elimination of the dearth of underrepresented minority scientists who address these disparities. By far, the students were satisfied with the program and believe that it has had significant impact on their ability to contribute to cancer prevention and control. They provided both general and specific recommendations to strengthen the program.
- Published
- 2013
24. J7 Intermediate allele expansion leads to huntington’s disease expression in large kindred
- Author
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Hope Heller, Fahd Amjad, Thomas J. Cummings Jr., Jan Blancato, Ira Shoulson, Karen E. Anderson, and Fernando Pagan
- Subjects
Proband ,Genetics ,business.industry ,Genetic counseling ,Buccal swab ,Genetic disorder ,medicine.disease ,Psychiatry and Mental health ,Huntington's disease ,medicine ,Surgery ,Neurology (clinical) ,Allele ,Family history ,Trinucleotide repeat expansion ,business - Abstract
Background Huntington’s disease is a progressive autosomal dominant genetic disorder characterised by variable motor, cognitive, and psychiatric symptoms. The mean age of onset is 35–44 years.1 Clinical expression is associated with a trinucleotide repeat expansion, 36 or more CAG trinucleotide repeats in HTT gene on chromosome 4. Case history A 39 year old Caucasian female with apparent HD was referred to our clinic for diagnosis and treatment. It was reported that she experienced onset of psychiatric symptoms in her early 30s and chorea at age 35. There was no family history of HD in a sibship of 15 individuals, 5 older than her. DNA testing revealed CAG repeat alleles of 47 and 24, consistent with the clinical diagnosis of HD. The proband’s parents were DNA tested and her 72 y/o asymptomatic father had alleles of 32/17 and her 65 year old mother had CAG repeat alleles of 24/15. Two of the proband’s siblings underwent DNA testing and found to have the normal paternal allele. Conclusion These data support the hypothesis that the paternal intermediate allele (IA) of 32 CAG repeats, a moderate risk allele for disease causing expansion in offspring,2 was expanded to an unprecedented number in the fertilisation event involving our patient. Genetic counselling and DNA testing for “at risk” offspring is ongoing with two normal DNA results of 17/15 having been reported in two of the siblings. DNA testing on paternal buccal cells is ongoing to rule out somatic mosaicism. Clinical and psychological evaluation of the IA carrier is in progress. References Warby, et al. Genetests 2014 Smelka, et al.
- Published
- 2016
25. The inception and evolution of a unique masters program in cancer biology, prevention and control
- Author
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Carolyn Cousin and Jan Blancato
- Subjects
Medical education ,Academic Medical Centers ,Cancer prevention ,Biomedical Research ,business.industry ,Doctoral studies ,Control (management) ,education ,Public Health, Environmental and Occupational Health ,Medical Oncology ,Article ,Outreach ,Oncology ,Coursework ,Neoplasms ,Degree program ,District of Columbia ,Medicine ,Humans ,University medical ,Cancer biology ,Education, Graduate ,business - Abstract
The University of the District of Columbia (UDC) and the Lombardi Comprehensive Cancer Center (LCCC), Georgetown University Medical Center established a Masters Degree Program in Cancer Biology, Prevention and Control at UDC that is jointly administered and taught by UDC and LCCC faculty. The goal of the Masters Degree Program is to educate students as master-level cancer professionals capable of conducting research and service in cancer biology, prevention, and control or to further advance the education of students to pursue doctoral studies. The Program’s unique nature is reflected in its philosophy “the best cancer prevention and control researchers are those with a sound understanding of cancer biology”. This program is a full-time, 2-year, 36-credit degree in which students take half of their coursework at UDC and half of their coursework at LCCC. During the second year, students are required to conduct research either at LCCC or UDC. Unlike most cancer biology programs, this unique Program emphasizes both cancer biology and cancer outreach training.
- Published
- 2010
26. Molecular characterization of a large cohort of patients with Chronic Granulomatous Disease and identification of novel CYBB mutations: an Italian multicenter study
- Author
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Domenico De Mattia, Annamaria Ventura, Cecilia Sinibaldi, Lucia Giordani, Gigliola Di Matteo, Paolo Rossi, Claudio Pignata, Antonino Trizzino, Alessandro Plebani, Jan Blancato, Baldassarre Martire, Roberto Rondelli, Rosa Maria Dellepiane, Annarosa Soresina, Andrea Finocchi, Fausto Cossu, Maria Chiriaco, Di Matteo, G., Giordani, L., Finocchi, A., Ventura, A., Chiriaco, M., Blancato, J., Sinibaldi, C., Plebani, A., Soresina, A., Pignata, C., Dellepiane, Rm., Trizzino, A., Cossu, F., Rondelli, R., Rossi, P., De Mattia, D., and Martire, B.
- Subjects
Adult ,Male ,Genotype-phenotype correlation ,medicine.medical_specialty ,Adolescent ,Genotype ,Immunology ,Biology ,medicine.disease_cause ,Granulomatous Disease, Chronic ,Chronic Granulomatous Disease ,CYBB ,White People ,Cell Line ,Cohort Studies ,Chronic granulomatous disease ,Molecular genetics ,Gene duplication ,medicine ,Missense mutation ,Humans ,Child ,Molecular Biology ,Settore MED/38 - Pediatria Generale e Specialistica ,Genetics ,Mutation ,NADPH oxidase ,Membrane Glycoproteins ,Incidence ,Mutation analysi ,Infant, Newborn ,Infant ,NADPH Oxidases ,Middle Aged ,medicine.disease ,Molecular biology ,Mutation analysis ,Italy ,Child, Preschool ,NADPH Oxidase 2 ,biology.protein ,Female - Abstract
Chronic Granulomatous Disease (CGD) is a rare inherited disorder in which phagocytes fail to produce antimicrobial superoxide because NADPH oxidase activity is absent. In about 65% of the cases, the disease is due to mutations affecting the X-linked CYBB gene, encoding the gp91(phox) subunit of NADPH oxidase. We investigated 34 CGD male patients by DHPLC and direct sequencing. A mutation was found in the CYBB gene of 33 patients and 9 of these were novel: one non-sense mutation (c.1123 G>T), three missense mutations (c.58G>A; c.1076 G>C; c.1357 T>A), two splice site mutations (c.141+5G>T; c.142-1G>A), one duplication (c.42_45dupCATT), one deletion (c.184delT), and one rare deletion of two non-contiguous nucleotides (c.1287delT+c.1290delC). One patient had the most frequent GT homozygous deletion in exon2 of the NCF-1 gene encoding the p47(phox) subunit of NADPH oxidase. The carrier analysis was performed in 23 patients' mothers and 16 female relatives through molecular and FISH studies. No clear correlation between the severity of clinical symptoms and the type of mutation could be demonstrated. This study further supports the great heterogeneity of the disease and the notion that genetic analysis is a critical step in obtaining a definitive diagnosis for CGD.
- Published
- 2009
27. Prognostic relevance of c-MYC gene amplification and polysomy for chromosome 8 in suboptimally-resected, advanced stage epithelial ovarian cancers: a Gynecologic Oncology Group study
- Author
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William McGuire, William J. Hoskins, Michael J. Birrer, Kathleen M. Darcy, William E. Brady, Jan Blancato, and Robert B. Dickson
- Subjects
Oncology ,medicine.medical_specialty ,Paclitaxel ,Genes, myc ,Gynecologic oncology ,Article ,Internal medicine ,Antineoplastic Combined Chemotherapy Protocols ,medicine ,Humans ,Stage (cooking) ,Cyclophosphamide ,In Situ Hybridization, Fluorescence ,Aged ,Neoplasm Staging ,Ovarian Neoplasms ,Polysomy ,medicine.diagnostic_test ,Performance status ,business.industry ,Hazard ratio ,Gene Amplification ,Obstetrics and Gynecology ,Combination chemotherapy ,Middle Aged ,medicine.disease ,Treatment Outcome ,Female ,Cisplatin ,Ovarian cancer ,business ,Fluorescence in situ hybridization ,Chromosomes, Human, Pair 8 - Abstract
Objective The Gynecologic Oncology Group (GOG) examined the prognostic relevance of c-MYC amplification and polysomy 8 in epithelial ovarian cancer (EOC). Methods Women with suboptimally-resected, advanced stage EOC who participated in GOG-111, a multicenter randomized phase III trial of cyclophosphamide+cisplatin vs. paclitaxel+cisplatin, and who provided a tumor block through GOG-9404 were eligible. Fluorescence in situ hybridization (FISH) with probes for c-MYC and the centromere of chromosome 8 (CEP8) was used to examine c-MYC amplification (≥2 copies c-MYC /CEP8) and polysomy 8 (≥4 CEP8 copies). Results c-MYC amplification, defined as ≥2 copies c-MYC /CEP8, was observed in 29% (28/97) of EOCs and levels were ranged from 2.0–3.3 copies of c-MYC /CEP8. c-MYC amplification was not associated with patient age, race, GOG performance status, stage, cell type, grade, measurable disease status following surgery, tumor response or disease status following platinum-based combination chemotherapy. Women with vs. without c-MYC amplification did not have an increased risk of disease progression (hazard ratio [HR]=1.03; 95% confidence interval [CI]=0.65–1.64; p =0.884) or death (HR=1.08; 95% CI=0.68–1.72; p =0.745). c-MYC amplification was not an independent prognostic factor for progression-free survival (HR=1.03, 95% CI=0.57–1.85; p =0.922) or overall survival (HR=1.01, 95% CI=0.56–1.80; p =0.982). Similar insignificant results were obtained for c-MYC amplification categorized as ≥1.5 copies c-MYC /CEP8. Polysomy 8 was observed in 22 patients without c-MYC amplification and 3 with c-MYC amplification, and was associated with age and measurable disease status, but not other clinical covariates or outcomes. Conclusions c-MYC amplification and polysomy 8 have limited predictive or prognostic value in suboptimally-resected, advanced stage EOC treated with platinum-based combination chemotherapy.
- Published
- 2009
28. Fluorescence In Situ Hybridization Assessment of c -myc Gene Amplification in Breast Tumor Tissues
- Author
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Jan Blancato, Mary Steele Williams, and Robert B. Dickson
- Subjects
chemistry.chemical_compound ,medicine.diagnostic_test ,Chemistry ,Hybridization probe ,Centromere ,medicine ,Fluorescence microscope ,Chromosome ,In situ hybridization ,Cover slip ,Molecular biology ,DNA ,Fluorescence in situ hybridization - Abstract
This chapter details methods used for analysis of DNA copy-number changes in breast tumor tissues through the use of fluorescence in situ hybridization. The specific DNA probe described herein is the oncogene c-myc, although the tissue fluorescence in situ hybridization methodology presented is suitable for dual-color studies of most unique sequence and chromosome specific control probes. The breast tumor tissue sections are first deparaffinized in a solvent and clearing agent, and then pretreated with a protease to allow the target DNA within the breast tissue cells to be uncovered. This allows the DNA to be available for hybridization with the labeled c-myc probe. The tissue sections are then analyzed to assure that appropriate digestion of cellular material has been attained. The tissues are then denatured. The c-myc probe and control probe for the centromere of chromosome 8 are commercially available as differentially labeled and are cohybridized to the tissue and sealed beneath a cover slip in a humid chamber. They are incubated for 12-16 h. The cover slip is then removed, and the section is postwashed in 2X saline sodium citrate at 72 degrees C for 5 min and allowed to cool to room temperature in a detergent solution. The slides are then counterstained with 4',6-diamidino-2-phenylindole, and a cover slip is applied. The slides are then viewed with fluorescence microscopy using filters that allow the c-myc and chromosome 8 signals to be visualized. If possible, 50 cells are counted, and the data are expressed as number of c-myc signals/number of chromosome 8 signals.
- Published
- 2006
29. Diagnostic FISH probes for del(17)(p11.2p11.2) associated with Smith-Magenis syndrome should contain the RAI1 gene
- Author
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Christopher N, Vlangos, Meredith, Wilson, Jan, Blancato, Ann C M, Smith, and Sarah H, Elsea
- Subjects
Male ,Infant ,Proteins ,Child Behavior Disorders ,Syndrome ,Musculoskeletal Abnormalities ,Craniofacial Abnormalities ,Child, Preschool ,Intellectual Disability ,Trans-Activators ,Humans ,Abnormalities, Multiple ,Chromosome Deletion ,Child ,In Situ Hybridization, Fluorescence ,Chromosomes, Human, Pair 17 ,Transcription Factors - Abstract
Smith-Magenis syndrome (SMS) is a mental retardation syndrome with distinctive behavioral characteristics, dysmorphic features, and congenital anomalies usually associated with an interstitial deletion of chromosome 17p11.2. While high quality G-banding will identify most SMS patients, fluorescent in situ hybridization (FISH) is the recommended test for confirmation of an SMS diagnosis. Recently, haploinsufficiency of the RAI1 gene due to deletion or mutation was determined to be the likely cause of SMS. All diagnostic FISH probes available commercially contain the FLII gene and are approximately 580 kb centromeric to RAI1. We present two patients with SMS who have interstitial deletions at 17p11.2 but are not deleted for currently available commercial FISH probes that include FLII; both patients have deletions that are demonstrated with probes containing the RAI1 gene. We recommend that for diagnostic accuracy, all future FISH tests for SMS be performed with probes containing the RAI1 gene, as some atypical deletions in the region critical to the SMS phenotype will otherwise be missed.
- Published
- 2005
30. The human ovarian teratocarcinoma cell line PA-1 demonstrates a single translocation: analysis with fluorescence in situ hybridization, spectral karyotyping, and bacterial artificial chromosome microarray
- Author
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Tom Dennis, Jan Blancato, Shireen A. Sarraf, Raphael Tejada, Michael Oberst, Massih Abawi, and Kelly Claire Simon
- Subjects
Teratocarcinoma ,Cancer Research ,Chromosomes, Artificial, Bacterial ,Blotting, Western ,Chromosomes, Human, Pair 20 ,Chromosomal translocation ,Biology ,Translocation, Genetic ,Ovarian tumor ,WFDC2 ,Gene duplication ,Genetics ,medicine ,Tumor Cells, Cultured ,Humans ,Molecular Biology ,In Situ Hybridization, Fluorescence ,Chromosome Aberrations ,Ovarian Neoplasms ,Bacterial artificial chromosome ,medicine.diagnostic_test ,Microarray analysis techniques ,Spectral Karyotyping ,Karyotype ,Oncogenes ,Microarray Analysis ,Molecular biology ,Chromosome Banding ,Female ,Fluorescence in situ hybridization ,Chromosomes, Human, Pair 8 - Abstract
Cell lines derived from tumors contain numerous chromosomal aberrations and are the focus of study in tumor evolution. The ovarian teratocarcinoma cell line PA-1 demonstrates a single chromosomal aberration: a reciprocal t(15;20)(p11.2;q11.2). A complete molecular genetic analysis was undertaken to characterize this cell line. The PA-1 cell line was studied with fluorescence in situ hybridization (FISH), spectral karyotyping (SKY), bacterial artificial chromosome (BAC) microarray, and Western blotting. Amplification of 20q is frequently implicated in both breast and ovarian cancer; this region contains a number of oncogenes including MDM2, ZNF217, and the ovarian tumor marker WFDC2 (alias HE4). FISH revealed gene amplification of AIB1 (now known as NCOA3) but not STK15 (now known as AURKA). Immunoblot analysis demonstrated 3.6-fold overexpression of the AIB1 protein product, but no elevation of the STK15. BAC cancer gene microarray analysis showed gene amplification of > or =1.20 for five oncogenes. The presence of a consistent single change in PA-1, the t(15;20)(p11.2;q11.2), suggests that the aberration is significant with respect to the transformation status of the cell line. This translocation appears to cause overexpression of AIB1 (and perhaps other proteins), which may provide an immortalizing effect on this cell line.
- Published
- 2004
31. SYK Allelic Loss and the Role of Syk-Regulated Genes in Breast Cancer Survival
- Author
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Maria Moroni, Banafsheh Rashidi, Ashley Graves, Leopold Tchobe, Catalin Marian, Bhaskar Kallakury, Jan Blancato, Metin Ozdemirli, Susette C. Mueller, and Kepher H. Makambi
- Subjects
Cell ,lcsh:Medicine ,Loss of Heterozygosity ,Syk ,Kaplan-Meier Estimate ,Metastasis ,Loss of heterozygosity ,0302 clinical medicine ,Molecular Cell Biology ,Tyrosine Kinase Signaling Cascade ,Basic Cancer Research ,lcsh:Science ,Promoter Regions, Genetic ,skin and connective tissue diseases ,In Situ Hybridization, Fluorescence ,0303 health sciences ,Mammary tumor ,Multidisciplinary ,medicine.diagnostic_test ,Cancer Risk Factors ,Carcinoma, Ductal, Breast ,Intracellular Signaling Peptides and Proteins ,Chromosome Mapping ,Obstetrics and Gynecology ,hemic and immune systems ,Protein-Tyrosine Kinases ,Prognosis ,Signaling Cascades ,3. Good health ,Gene Expression Regulation, Neoplastic ,Treatment Outcome ,medicine.anatomical_structure ,Oncology ,030220 oncology & carcinogenesis ,DNA methylation ,Medicine ,Female ,Tyrosine kinase ,Research Article ,Signal Transduction ,Genetic Causes of Cancer ,Breast Neoplasms ,chemical and pharmacologic phenomena ,Biology ,Genomic Instability ,Cytogenetics ,03 medical and health sciences ,Breast cancer ,Breast Cancer ,Genetics ,Cancer Genetics ,medicine ,Humans ,Syk Kinase ,Neoplasm Invasiveness ,030304 developmental biology ,lcsh:R ,DNA Methylation ,medicine.disease ,Immunology ,Cancer research ,lcsh:Q ,Cytogenetic Techniques ,Fluorescence in situ hybridization - Abstract
Heterozygotic loss of SYK, a non-receptor tyrosine kinase, gives rise to mouse mammary tumor formation where Syk protein levels are reduced by about half; loss of SYK mRNA is correlated with invasive cell behavior in in vitro models; and SYK loss has been correlated with distant metastases in patients. Here, allelic loss of the SYK gene was explored in breast ductal carcinoma in situ (DCIS) using fluorescence in situ hybridization and pyrosequencing, respectively, and in infiltrating ductal carcinoma (IDC) using genomic data from The Cancer Genome Atlas (TCGA). Allelic loss was present in a subset of DCIS cases where adjacent IDC was present. SYK copy number loss was found in about 26% of 1002 total breast cancer cases and 30% of IDC cases. Quantitative immunofluorescence revealed Syk protein to be six-fold higher in infiltrating immune cells compared with epithelial cells. This difference distorted tumor cell mRNA and protein levels in extracts. 20% of 1002 IDC cases contained elevated immune cell infiltration as estimated by elevated immune-specific mRNAs. In cases without immune cell infiltration, loss of SYK copy number was associated with a significant reduction of SYK mRNA. Here we define a 55 Gene Set consisting of Syk interacting, motility- and invasion-related genes. We found that overall survival was significantly reduced in IDC and Luminal A+B cases where copy number and mutations of these 55 genes were affected (Kaplan-Meier, Logrank test p-value 0.007141 and Logrank test p-value 0.001198, respectively). We conclude that reduction in Syk expression and contributions of genomic instability to copy number and mutations in the 55 Syk interacting genes significantly contribute to poorer overall patient survival. A closer examination of the role of Syk interacting motility and invasion genes and their prognostic and/or causative association with metastatic disease and patient outcome is warranted.
- Published
- 2014
32. Anisomastia associated with interstitial duplication of chromosome 16, mental retardation, obesity, dysmorphic facies, and digital anomalies: molecular mapping of a new syndrome by fluorescent in situ hybridization and microsatellites to 16q13 (D16S419-D16S503)
- Author
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Jan Blancato, Rachel I Gafni, Jeanne M. Meck, Susan E. Taymans, Constantine A. Stratakis, and Antony R Lafferty
- Subjects
Adult ,Pathology ,medicine.medical_specialty ,Endocrinology, Diabetes and Metabolism ,Clinical Biochemistry ,Biology ,Biochemistry ,Molecular cytogenetics ,Fingers ,Centimorgan ,Endocrinology ,Chromosome 16 ,Gene Duplication ,Intellectual Disability ,Gene duplication ,medicine ,Humans ,Breast ,Obesity ,In Situ Hybridization, Fluorescence ,Genetics ,medicine.diagnostic_test ,Biochemistry (medical) ,Cytogenetics ,Trisomy 16 ,DNA ,medicine.disease ,Face ,Karyotyping ,Female ,Trisomy ,DNA Probes ,Chromosomes, Human, Pair 16 ,Fluorescence in situ hybridization ,Microsatellite Repeats - Abstract
Anisomastia is a common problem among developing adolescent girls. We recently evaluated a 22-yr-old female patient who had severe anisomastia (which had been repaired by surgery), associated with moderate to severe mental retardation, a stocky body habitus with mild obesity, dysmorphic facies (prominent, upslanting palpebral fissures, beaked nose, and a prominent philtrum), webbed neck, low hairline, and severe bilateral clinodactyly of the third, fourth, and fifth fingers with acral (but not large joint) flexion contractures. A peripheral blood high resolution karyotype revealed additional chromosomal material within the long arm of chromosome 16. Densitometric analysis of amplified polymorphic sequence-tagged sites (STS) mapping to 16q suggested that the duplication is defined by the noninvolved markers D16S419 [16q12-cen, 66 centimorgan (cM) from 16p terminus] and D16S421 (16q13-q21, 84.4 cM), encompassing a maximum of 18.4 cM of genetic distance. The STS analysis showed that the duplication was on the maternally derived chromosome 16, resulting in two maternal (and one paternal) copies of that region of chromosome 16. The location was further confirmed by bacterial artificial chromosomes (BACs) that were obtained from a commercially available library, labeled, and used for fluorescence in situ hybridization. The BACs containing STSs D16S408, D16S3137, and D16S3032 (markers that correspond to 16q13) showed two regions of hybridization, indicating that these sites were duplicated, whereas a BAC containing the STS D16S512 (which corresponds to 16q21-q22) revealed one hybridization signal per 16q, indicating that the corresponding region was not involved in the duplication. The distance between the probe signals suggested a tandem duplication. We conclude that even though trisomy 16 is the most common autosomal trisomy in spontaneous abortions, few patients with unbalanced chromosome 16 abnormalities survive to adulthood; in this report we describe one such patient with an interstitial chromosome 16 duplication (at 16q13), who had a specific phenotype associated with abnormal breast size. There are clinical similarities between this patient and patients with other 16q abnormalities, although the breast findings were unique. Molecular cytogenetics, including fluorescence in situ hybridization and densitometric analysis of amplified STSs, provided useful tools for the precise mapping of the syndrome to 16q13, where the gene(s) responsible for this phenotype might be localized.
- Published
- 2000
33. The Syk tyrosine kinase suppresses malignant growth of human breast cancer cells
- Author
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Peter Coopman, Mara Barth, Michael Tri H. Do, Jan Blancato, Andrew Hayes, Phyllis R. Vezza, Eugenia Basyuk, Emma T. Bowden, Susette C. Mueller, Paul Mangeat, and Sandra W. McLeskey
- Subjects
Tumor suppressor gene ,Syk ,Mice, Nude ,chemical and pharmacologic phenomena ,Apoptosis ,Breast Neoplasms ,Biology ,medicine.disease_cause ,Transfection ,environment and public health ,Catalysis ,Mice ,hemic and lymphatic diseases ,medicine ,In Situ Nick-End Labeling ,Tumor Cells, Cultured ,Animals ,Humans ,Syk Kinase ,Genes, Tumor Suppressor ,Breast ,RNA, Messenger ,Enzyme Precursors ,Multidisciplinary ,Cell growth ,Intracellular Signaling Peptides and Proteins ,Cancer ,hemic and immune systems ,Cell cycle ,Protein-Tyrosine Kinases ,medicine.disease ,Cell Transformation, Neoplastic ,Cancer cell ,Immunology ,Cancer research ,Female ,biological phenomena, cell phenomena, and immunity ,Carcinogenesis ,Tyrosine kinase ,Cell Division ,Neoplasm Transplantation - Abstract
Syk is a protein tyrosine kinase that is widely expressed in haematopoietic cells. It is involved in coupling activated immunoreceptors to downstream signalling events that mediate diverse cellular responses including proliferation, differentiation and phagocytosis1,2,3,4. Syk expression has been reported in cell lines of epithelial origin5, but its function in these cells remains unknown. Here we show that Syk is commonly expressed in normal human breast tissue, benign breast lesions and low-tumorigenic breast cancer cell lines. Syk messenger RNA and protein, however, are low or undetectable in invasive breast carcinoma tissue and cell lines. Transfection of wild-type Syk into a Syk-negative breast cancer cell line markedly inhibited its tumour growth and metastasis formation in athymic mice. Conversely, overexpression of a kinase-deficient Syk in a Syk-positive breast cancer cell line significantly increased its tumour incidence and growth. Suppression of tumour growth by the reintroduction of Syk appeared to be the result of aberrant mitosis and cytokinesis. We propose that Syk is a potent modulator of epithelial cell growth and a potential tumour suppressor in human breast carcinomas.
- Published
- 2000
34. Detection of chromosomal aberrations by a whole-genome microsatellite screen
- Author
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Louanne Hudgins, Kenneth N. Rosenbaum, Richard M. Pauli, David S. Wargowski, Jeanne M. Meck, Yi Ning, Cynthia J. Tifft, David Vaske, James L. Weber, Christina Killoran, Marjorie J. Rosenberg, Leslie G. Biesecker, and Jan Blancato
- Subjects
Male ,Genotype ,Pilot Projects ,Biology ,Nuclear Family ,symbols.namesake ,Gene Duplication ,medicine ,Genetics ,Humans ,Genetics(clinical) ,Abnormalities, Multiple ,Deletions ,Genetic Testing ,Allele ,Child ,Genetics (clinical) ,Alleles ,In Situ Hybridization, Fluorescence ,Genetic testing ,Sequence Deletion ,Chromosome Aberrations ,medicine.diagnostic_test ,Genome, Human ,Duplications ,Reproducibility of Results ,Mental retardation ,Karyotype ,Articles ,Meiosis ,Genetic marker ,Segmental aneusomy ,Karyotyping ,Multiple congenital anomalies ,Mendelian inheritance ,symbols ,Genetic markers ,Microsatellite ,Human genome ,Female ,Microsatellite Repeats - Abstract
Chromosomal aberrations are a common cause of multiple anomaly syndromes that include developmental and growth retardation. Current microscopic techniques are useful for the detection of such aberrations but have a limit of resolution that is above the threshold for phenotypic effect. We hypothesized that a genomewide microsatellite screen could detect chromosomal aberrations that were not detected by standard cytogenetic techniques in a portion of these individuals. To test this hypothesis, we performed a genomewide microsatellite screen of patients, by use of a currently available genetic-marker panel that was originally designed for meiotic mapping of Mendelian traits. We genotyped approximately 400 markers on 17 pairs of parents and their children who had normal karyotypes. By using this approach, we detected and confirmed two cases of segmental aneusomy among 11 children with multiple congenital anomalies. These data demonstrate that a genomewide microsatellite scan can be used to detect chromosomal aberrations that are not detected by microscopic techniques.
- Published
- 2000
35. Highly skewed X-chromosome inactivation is associated with idiopathic recurrent spontaneous abortion
- Author
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Mark C. Lanasa, Eric P. Hoffman, C Kubik, W A Hogge, and Jan Blancato
- Subjects
Abortion, Habitual ,Abortion ,Biology ,Polymerase Chain Reaction ,law.invention ,03 medical and health sciences ,0302 clinical medicine ,Text mining ,law ,Polymorphism (computer science) ,Pregnancy ,X-linked recessive lethal inheritance ,Dosage Compensation, Genetic ,medicine ,Genetics ,Humans ,Genetics(clinical) ,Skewed X-inactivation ,Genetics (clinical) ,Polymerase chain reaction ,030304 developmental biology ,0303 health sciences ,030219 obstetrics & reproductive medicine ,Dosage compensation ,Polymorphism, Genetic ,Models, Genetic ,business.industry ,Chromosome inactivation ,medicine.disease ,Female ,business ,Recurrent spontaneous abortion ,Research Article - Published
- 1999
36. Monosomy 7 Detected by FISH at Disease Presentation Is a Marker for Non-Response to Immunosuppression
- Author
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Edward Fenlon, Jan Blancato, Phillip Scheinberg, Elaine M. Sloand, Loretta Pfannes, and Neal S. Young
- Subjects
Chromosome 7 (human) ,medicine.medical_specialty ,Pathology ,medicine.medical_treatment ,Immunology ,Cytogenetics ,Bone marrow failure ,Immunosuppression ,Cell Biology ,Hematology ,Biology ,medicine.disease ,Trisomy 8 ,Biochemistry ,Granulocyte colony-stimulating factor ,medicine.anatomical_structure ,medicine ,Bone marrow ,Aplastic anemia - Abstract
About fifteen percent of patients with severe aplastic anemia (sAA) who undergo immunosuppression therapy (IST) develop clonal disease in the decade following treatment. Of 122 patients treated with horse ATG/CsA at NIH, 13 developed cytogenetic abnormalities (monosomy 7 in 9 patients, 7p deletion in one, and trisomy 8 in 2) (Rosenfeld et al: JAMA289:1130; 2003). Monosomy 7 usually occurs in patients with sAA who are unresponsive to IST. Factors responsible for clonal progression in bone marrow failure are still unclear. We and others have reported that monosomy 7 may be detected by fluorescent in situ hybridization (FISH) months before conventional cytogenetics and that high levels of GCSF foster preferential expansion of the monosomy 7 clone in vitro (Sloand E et al; Proc. Natl. Acad. Sci2006;103:14483). We hypothesized that a clone of monsomy 7 cells might indicate an underlying stem cell disorder unlikely to respond to immunosuppression or reflect more severe disease marked by higher endogenous GCSF levels. FISH was undertaken on 40 bone marrow samples obtained from aplastic anemia patients at presentation and before immunosuppressive treatment. Bone marrow mononuclear cells were hybridized with centromeric probes specific for chromosome 7 and chromosome 8 (to control for hybridization efficiency); samples were assessed on duplicate slides by three investigators who were unaware of the diagnoses and outcomes. Twenty-five healthy controls were tested concurrently. The upper limit of normal was set at 6% based on control data. Of twenty-one patients with > 6% monosomy 7 cells, response to IST was observed in 12 (57%), while 6 of 19 (84%) with normal FISH responded to IST (p=0.08). Thirteen patients had monosomy 7 frequencies of >10%. Of these 5 (38%) responded to IST, compared with 23/27 (85%) with ≤10% monosomy 7. The proportion of monosomy 7 cells in the bone marrow correlated with age (R2 =0.6, p10% monosomy 7 by FISH at presentation, but normal cytogenetics. The two responding patients tested with >6% monosomy 7 prior to IST demonstrated
- Published
- 2007
37. Telomere Shortening and Genomic Instability: Primary Cells from Patients with Telomere Repair Complex Mutations Are Susceptible to End-to-End Chromosome Fusion and Aneuploidy
- Author
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Rodrigo T. Calado, Elaine M. Sloand, Neal S. Young, Simant Shah, Jan Blancato, and Loretta Pfannes
- Subjects
Genome instability ,Telomerase ,Immunology ,Aneuploidy ,Cell Biology ,Hematology ,Biology ,medicine.disease ,Biochemistry ,Molecular biology ,Telomere ,Telomerase RNA component ,medicine ,Sister chromatids ,Telomerase reverse transcriptase ,Dyskeratosis congenita - Abstract
Telomeres that cap the ends of chromosomes, protect them from recombination, end-to-end fusion, and recognition as damaged DNA. Telomere shorteningmay result in cell cycle arrest and senescence or in genomic instability that precedes malignancy. Maintenance of the integrity of telomeres requires the telomerase ribonucleoprotein complex, mainly the telomerase reverse transcriptase (TERT) and its integral RNA template (TERC). Mutations in TERC and TERT cause progressive telomere shortening; short telomeres are found in constitutional marrow failure syndromes and also in apparently acquired aplastic anemia (Fogarty PF et al, Lancet362:1628, 2003;. Yamaguchi H et al, NEJM352:1413, 2005). Myelodysplasia and acute leukemia are often present in these kindreds and associated with pathogenic mutations. The mechanism resulting in genomic instability in TERT and TERC-deficient individuals is unclear. End-to-end chromosome fusion in TERT-”knock-out” embryonic mouse stem cells are seen after multiple passages in culture and are associated with profound telomere shortening and the development of aneuploidy (Lee HW et al. Nature 9:569–74, 1998). End-to-end fusion of sister chromatids presumably results in disruption of chromosome segregation during metaphase. In the present study, we examined blood and marrow cells from 7 patients with TERT, and TERC gene mutations; 10 healthy young controls; and 2 individuals older than age 60 years of age, as well as 6 umbilical cord bloods. Leukocytes with TERT or TERC mutations had short telomeres as previously measured by flow-fluorescent in situ hybridization (FISH) and confirmed by Southern hybridization (mean telomere shortening in comparison to age-matched controls, 3.4 ± 1.7 kb) and low telomerase enzymatic activity (144±13 vs. 270±56 respectively; P
- Published
- 2006
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