388 results on '"Adriana Heguy"'
Search Results
152. Dominance of Alpha and Iota variants in SARS-CoV-2 vaccine breakthrough infections in New York City
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Andrea B. Troxel, Adriana Heguy, Christian Marier, Ralf Duerr, Guiqing Wang, Dacia Dimartino, Jennifer Lighter, Paul Zappile, and Brian Elbel
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Adult ,Male ,medicine.medical_specialty ,COVID-19 Vaccines ,Alpha (ethology) ,medicine.disease_cause ,Iota ,Evolution, Molecular ,Immune system ,McNemar's test ,Protein Domains ,Epidemiology ,medicine ,Humans ,BNT162 Vaccine ,Dominance (genetics) ,Aged ,Immune Evasion ,Aged, 80 and over ,Mutation ,Ad26COVS1 ,business.industry ,SARS-CoV-2 ,COVID-19 ,General Medicine ,Middle Aged ,Vaccine efficacy ,Virology ,Vaccination ,Spike Glycoprotein, Coronavirus ,Female ,New York City ,business ,2019-nCoV Vaccine mRNA-1273 ,Research Article - Abstract
The efficacy of COVID-19 mRNA vaccines is high, but breakthrough infections still occur. We compared the SARS-CoV-2 genomes of 76 breakthrough cases after full vaccination with BNT162b2 (Pfizer/BioNTech), mRNA-1273 (Moderna), or JNJ-78436735 (Janssen) to unvaccinated controls (February-April 2021) in metropolitan New York, including their phylogenetic relationship, distribution of variants, and full spike mutation profiles. Their median age was 48 years; seven required hospitalization and one died. Most breakthrough infections (57/76) occurred with B.1.1.7 (Alpha) or B.1.526 (Iota). Among the 7 hospitalized cases, 4 were infected with B.1.1.7, including 1 death. Both unmatched and matched statistical analyses considering age, sex, vaccine type, and study month as covariates supported the null hypothesis of equal variant distributions between vaccinated and unvaccinated in chi-squared and McNemar tests (p>0.1) highlighting a high vaccine efficacy against B.1.1.7 and B.1.526. There was no clear association among breakthroughs between type of vaccine received and variant. In the vaccinated group, spike mutations in the N-terminal domain and receptor-binding domain that have been associated with immune evasion were overrepresented. The evolving dynamic of SARS-CoV-2 variants requires broad genomic analyses of breakthrough infections to provide real-life information on immune escape mediated by circulating variants and their spike mutations.
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- 2021
153. Disruption of Ca 2+ i Homeostasis and Connexin 43 Hemichannel Function in the Right Ventricle Precedes Overt Arrhythmogenic Cardiomyopathy in Plakophilin-2–Deficient Mice
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Marta Pérez-Hernández, Jérôme Montnach, Sun-Hee Woo, Svetlana Rajkumar Maurya, Mingliang Zhang, Alicia Lundby, Xianming Lin, Carolina Vasquez, Yandong Yin, Francisco J. Alvarado, Eli Rothenberg, Feng-Xia Liang, Adriana Heguy, Marina Cerrone, Joon-Chul Kim, Gregory E. Morley, Héctor H. Valdivia, and Mario Delmar
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Pathology ,medicine.medical_specialty ,Cardiomyopathy ,sudden death ,Connexin ,right ventricle ,030204 cardiovascular system & hematology ,Sudden death ,Right ventricular cardiomyopathy ,03 medical and health sciences ,0302 clinical medicine ,Physiology (medical) ,PLAKOPHILIN 2 ,plakophilin 2 ,medicine ,030304 developmental biology ,arrhythmogenic right ventricular cardiomyopathy ,0303 health sciences ,business.industry ,medicine.disease ,connexin43 ,medicine.anatomical_structure ,Ventricle ,Cardiology and Cardiovascular Medicine ,business ,Homeostasis ,Function (biology) - Abstract
Background: Plakophilin-2 (PKP2) is classically defined as a desmosomal protein. Mutations in PKP2 associate with most cases of gene-positive arrhythmogenic right ventricular cardiomyopathy. A better understanding of PKP2 cardiac biology can help elucidate the mechanisms underlying arrhythmic and cardiomyopathic events consequent to PKP2 deficiency. Here, we sought to capture early molecular/cellular events that can act as nascent arrhythmic/cardiomyopathic substrates. Methods: We used multiple imaging, biochemical and high-resolution mass spectrometry methods to study functional/structural properties of cells/tissues derived from cardiomyocyte-specific, tamoxifen-activated, PKP2 knockout mice (PKP2cKO) 14 days post-tamoxifen injection, a time point preceding overt electrical or structural phenotypes. Myocytes from right or left ventricular free wall were studied separately. Results: Most properties of PKP2cKO left ventricular myocytes were not different from control; in contrast, PKP2cKO right ventricular (RV) myocytes showed increased amplitude and duration of Ca 2+ transients, increased Ca 2+ in the cytoplasm and sarcoplasmic reticulum, increased frequency of spontaneous Ca 2+ release events (sparks) even at comparable sarcoplasmic reticulum load, and dynamic Ca 2+ accumulation in mitochondria. We also observed early- and delayed-after transients in RV myocytes and heightened susceptibility to arrhythmias in Langendorff-perfused hearts. In addition, ryanodine receptor 2 in PKP2cKO-RV cells presented enhanced Ca 2+ sensitivity and preferential phosphorylation in a domain known to modulate Ca 2+ gating. RNAseq at 14 days post-tamoxifen showed no relevant difference in transcript abundance between RV and left ventricle, neither in control nor in PKP2cKO cells. Instead, we found an RV-predominant increase in membrane permeability that can permit Ca 2+ entry into the cell. Connexin 43 ablation mitigated the membrane permeability increase, accumulation of cytoplasmic Ca 2+ , increased frequency of sparks and early stages of RV dysfunction. Connexin 43 hemichannel block with GAP19 normalized [Ca 2+ ] i homeostasis. Similarly, protein kinase C inhibition normalized spark frequency at comparable sarcoplasmic reticulum load levels. Conclusions: Loss of PKP2 creates an RV-predominant arrhythmogenic substrate (Ca 2+ dysregulation) that precedes the cardiomyopathy; this is, at least in part, mediated by a Connexin 43-dependent membrane conduit and repressed by protein kinase C inhibitors. Given that asymmetric Ca 2+ dysregulation precedes the cardiomyopathic stage, we speculate that abnormal Ca 2+ handling in RV myocytes can be a trigger for gross structural changes observed at a later stage.
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- 2019
154. The bone marrow microenvironment at single-cell resolution
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Edlira Hoxha, Catherine Diefenbach, Yutong Zhang, Adriana Heguy, Anastasia N. Tikhonova, Stavroula Kousteni, Kishor K. Sivaraj, Aristotelis Tsirigos, Hai Hu, Ralf H. Adams, Igor Dolgalev, Aris N. Economides, Matthew T. Witkowski, Ilseyar Akhmetzyanova, Michael C. Gutkin, Jason M. Butler, Maria Guillamot, Álvaro Cuesta-Domínguez, Jie Gao, Sandra Pinho, Hua Zhong, Paul S. Frenette, Rahul Satija, David R. Fooksman, Christian Marier, and Iannis Aifantis
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Male ,0301 basic medicine ,Myeloid ,Cellular differentiation ,Biology ,Article ,Transcriptome ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Single-cell analysis ,Bone Marrow ,Stress, Physiological ,Adipocytes ,medicine ,Animals ,Cell Lineage ,Myeloid Cells ,RNA-Seq ,Stem Cell Niche ,Adaptor Proteins, Signal Transducing ,Regulation of gene expression ,Osteoblasts ,Multidisciplinary ,Receptors, Notch ,Calcium-Binding Proteins ,Cell Differentiation ,Hematopoietic Stem Cells ,Hematopoiesis ,Cell biology ,Haematopoiesis ,030104 developmental biology ,medicine.anatomical_structure ,Cellular Microenvironment ,Gene Expression Regulation ,030220 oncology & carcinogenesis ,Female ,Endothelium, Vascular ,Bone marrow ,Single-Cell Analysis ,Stem cell - Abstract
The molecular complexity of the bone marrow (BM) microenvironment and its response to stress are incompletely understood, despite its key role in the regulation of hematopoiesis. Here we map the transcriptional landscape of BM vascular, perivascular, and osteoblast niche populations at single-cell resolution at both homeostasis and under stress hematopoiesis. This analysis revealed a previously unappreciated level of cellular heterogeneity within the BM niche, identified novel cellular subsets, and resolved cellular sources of pro-hematopoietic growth factors, chemokines, and membrane-bound ligands. Under conditions of stress, our studies revealed a significant transcriptional remodeling of these niche elements, including an adipocytic skewing of the perivascular cells. Among the stress-induced changes, we observed that vascular Notch ligand delta-like ligands (Dll1,4) were downregulated. In the absence of vascular Dll4, hematopoietic stem cells (HSC) prematurely induced a myeloid transcriptional program. These findings refine our understanding of the cellular architecture of the BM niche, reveal a dynamic and heterogeneous molecular landscape that is highly sensitive to stress, and illustrate the utility of single cell transcriptomic data in systematically evaluating the regulation of hematopoiesis by discrete niche populations.
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- 2019
155. Microglandular adenosis is an advanced precursor breast lesion with evidence of molecular progression to matrix-producing metaplastic carcinoma
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Eleazar C. Vega-Saenz de Miera, Farbod Darvishian, George Jour, Esther Yoon, Adriana Heguy, Christopher J Schwartz, Iman Osman, Diana Nimeh, and Igor Dolgalev
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0301 basic medicine ,Metaplastic carcinoma ,Breast Neoplasms ,Locus (genetics) ,Biology ,Pathology and Forensic Medicine ,03 medical and health sciences ,ADAMTS Proteins ,0302 clinical medicine ,Cyclin D1 ,Breast cancer ,medicine ,Humans ,Fibrocystic Breast Disease ,Exome sequencing ,Carcinoma in situ ,Carcinoma, Ductal, Breast ,medicine.disease ,Immunohistochemistry ,030104 developmental biology ,030220 oncology & carcinogenesis ,Disease Progression ,Cancer research ,Female ,Tumor Suppressor Protein p53 ,Trisomy ,Precancerous Conditions - Abstract
Summary Microglandular adenosis (MGA) is a rare breast lesion reported to be associated with invasive carcinoma in up to 20% to 30% of cases and has been proposed as a nonobligate precursor to basal-like breast cancers. We identified a case of matrix-producing metaplastic carcinoma with morphologic and immunohistochemical evidence of progression from MGA to atypical MGA, carcinoma in situ, and invasive carcinoma. We performed whole-exome sequencing of each component (MGA, atypical MGA, carcinoma in situ, and cancer) to characterize the mutational landscape of these foci. There was a significant copy number overlap between all foci, including a segmental amplification of the CCND1 locus (partial chromosome 11 trisomy) and MYC (8q24.12-13). Using a bioinformatics approach, we were able to identify 3 putative mutational clusters and recurrent, stop-gain nonsynonymous mutations in both ZNF862 and TP53 that were shared across all foci. Finally, we identified a novel deleterious splice-acceptor site mutation of chr5:5186164 G>T (chromosome 5p15) encoding the gene, ADAMTS16, in the invasive component.
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- 2019
156. Correction to: Profiling Basal Forebrain Cholinergic Neurons Reveals a Molecular Basis for Vulnerability Within the Ts65Dn Model of Down Syndrome and Alzheimer’s Disease
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Melissa J. Alldred, Sai C. Penikalapati, Sang Han Lee, Adriana Heguy, Panos Roussos, and Stephen D. Ginsberg
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Cellular and Molecular Neuroscience ,Neurology ,Neuroscience (miscellaneous) - Published
- 2021
157. Airway Microbiota Is Associated with Upregulation of the PI3K Pathway in Lung Cancer
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Daniel H. Sterman, Richard Bonneau, E. Olsen, Jose C. Clemente, Martin J. Blaser, Peter Meyn, Adriana Heguy, Benjamin G. Wu, Jun Chieh J. Tsay, Nan Shen, Leopoldo N. Segal, Michael D. Weiden, Aristotelis Tsirigos, Michelle H. Badri, Yonghua Li, Vivek Murthy, Ting An Yie, Imran Sulaiman, William N. Rom, Harvey I. Pass, Tenzin Lhakhang, and Gaetane Michaud
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Adult ,Male ,0301 basic medicine ,Pulmonary and Respiratory Medicine ,Lung Neoplasms ,Respiratory System ,Critical Care and Intensive Care Medicine ,Phosphatidylinositol 3-Kinases ,03 medical and health sciences ,0302 clinical medicine ,Downregulation and upregulation ,Bronchoscopy ,medicine ,Humans ,Prospective Studies ,Microbiome ,Lung cancer ,PI3K/AKT/mTOR pathway ,Aged ,Original Research ,business.industry ,Cell growth ,Microbiota ,Middle Aged ,respiratory system ,medicine.disease ,respiratory tract diseases ,Up-Regulation ,Cross-Sectional Studies ,030104 developmental biology ,030220 oncology & carcinogenesis ,Host-Pathogen Interactions ,Cancer research ,Tissue invasion ,Female ,business ,Airway - Abstract
Rationale: In lung cancer, upregulation of the PI3K (phosphoinositide 3-kinase) pathway is an early event that contributes to cell proliferation, survival, and tissue invasion. Upregulation of this pathway was recently described as associated with enrichment of the lower airways with bacteria identified as oral commensals. Objectives: We hypothesize that host–microbe interactions in the lower airways of subjects with lung cancer affect known cancer pathways. Methods: Airway brushings were collected prospectively from subjects with lung nodules at time of diagnostic bronchoscopy, including 39 subjects with final lung cancer diagnoses and 36 subjects with noncancer diagnoses. In addition, samples from 10 healthy control subjects were included. 16S ribosomal RNA gene amplicon sequencing and paired transcriptome sequencing were performed on all airway samples. In addition, an in vitro model with airway epithelial cells exposed to bacteria/bacterial products was performed. Measurements and Main Results: The composition of the lower airway transcriptome in the patients with cancer was significantly different from the control subjects, which included up-regulation of ERK (extracellular signal–regulated kinase) and PI3K signaling pathways. The lower airways of patients with lung cancer were enriched for oral taxa (Streptococcus and Veillonella), which was associated with up-regulation of the ERK and PI3K signaling pathways. In vitro exposure of airway epithelial cells to Veillonella, Prevotella, and Streptococcus led to upregulation of these same signaling pathways. Conclusions: The data presented here show that several transcriptomic signatures previously identified as relevant to lung cancer pathogenesis are associated with enrichment of the lower airway microbiota with oral commensals.
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- 2018
158. Role of plakophilin-2 expression on exercise-related progression of arrhythmogenic right ventricular cardiomyopathy: a translational study
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Marina Cerrone, Grecia M Marrón-Liñares, Chantal J M van Opbergen, Sarah Costa, Mimount Bourfiss, Marta Pérez-Hernández, Florencia Schlamp, Fabian Sanchis-Gomar, Kabir Malkani, Kamelia Drenkova, Mingliang Zhang, Xianming Lin, Adriana Heguy, Birgitta K Velthuis, Niek H J Prakken, Andre LaGerche, Hugh Calkins, Cynthia A James, Anneline S J M Te Riele, Mario Delmar, and Basic and Translational Research and Imaging Methodology Development in Groningen (BRIDGE)
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Mice, Knockout ,Plakophilin-2 ,PLAKOGLOBIN ,Myocardium ,Desmosomes ,Mice ,Physical Conditioning, Animal ,Translational Research ,Mutation ,ARVC ,Animals ,Humans ,Myocytes, Cardiac ,Cardiology and Cardiovascular Medicine ,Arrhythmogenic right ventricular cardiomyopathy ,ADAPTATION ,Exercise ,Plakophilins ,Arrhythmogenic Right Ventricular Dysplasia - Abstract
Aims Exercise increases arrhythmia risk and cardiomyopathy progression in arrhythmogenic right ventricular cardiomyopathy (ARVC) patients, but the mechanisms remain unknown. We investigated transcriptomic changes caused by endurance training in mice deficient in plakophilin-2 (PKP2cKO), a desmosomal protein important for intercalated disc formation, commonly mutated in ARVC and controls. Methods and results Exercise alone caused transcriptional downregulation of genes coding intercalated disk proteins. The changes converged with those in sedentary and in exercised PKP2cKO mice. PKP2 loss caused cardiac contractile deficit, decreased muscle mass and increased functional/transcriptomic signatures of apoptosis, despite increased fractional shortening and calcium transient amplitude in single myocytes. Exercise accelerated cardiac dysfunction, an effect dampened by pre-training animals prior to PKP2-KO. Consistent with PKP2-dependent muscle mass deficit, cardiac dimensions in human athletes carrying PKP2 mutations were reduced, compared to matched controls. Conclusions We speculate that exercise challenges a cardiomyocyte “desmosomal reserve” which, if impaired genetically (e.g., PKP2 loss), accelerates progression of cardiomyopathy.
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- 2021
159. Molecular analysis of encapsulated papillary carcinoma of the breast with and without invasion
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George Jour, Paolo Cotzia, Farbod Darvishian, Alireza Khodadadi-Jamayran, Adriana Heguy, Matija Snuderl, Christopher J Schwartz, and Amir Momeni Boroujeni
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0301 basic medicine ,Male ,Somatic cell ,Class I Phosphatidylinositol 3-Kinases ,Breast Neoplasms ,Biology ,Pathology and Forensic Medicine ,Collagen fibril organization ,Transcriptome ,Extracellular matrix ,03 medical and health sciences ,0302 clinical medicine ,Gene expression ,Biomarkers, Tumor ,Humans ,Cell adhesion ,neoplasms ,Aged ,Retrospective Studies ,Aged, 80 and over ,Carcinoma, Ductal, Breast ,Myoepithelial cell ,Middle Aged ,Carcinoma, Papillary ,030104 developmental biology ,KMT2A ,030220 oncology & carcinogenesis ,Mutation ,Cancer research ,biology.protein ,Female - Abstract
Encapsulated papillary carcinomas (EPCs) of the breast are a unique variant of papillary carcinoma confined to a cystic space with absent or attenuated myoepithelial cell layer. Although staged as an in situ lesion, it can be associated with invasive ductal carcinoma (IDC). We sought to compare the genomic characteristics of pure EPC and EPC with associated invasive carcinoma (EPCi) at the genomic level. All cases of EPCi harbored recurrent hotspot mutations in PIK3CA. PIK3CA, KMT2A, and CREBBP deleterious somatic events were found across both tumor groups, irrespective of invasion status. At the whole transcriptomic level, EPCi cases displayed remarkably similar mRNA profiles when compared to EPC. When EPCi cases were compared with their corresponding IDC, despite significant overlap, we identified differential gene expression in 39 genes with enrichment of multiple pathways including extracellular matrix regulation, cell adhesion, and collagen fibril organization. Despite morphologic, genotypic, and transcriptomic overlap between pure EPC and EPCi, the latter tumors are likely advanced lesions with PIK3CA activating mutations and enrichment of stromal-related genes implicated in the switch to IDC.
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- 2021
160. Transcriptomic Coupling of PKP2 With Inflammatory and Immune Pathways Endogenous to Adult Cardiac Myocytes
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Mingliang Zhang, Chantal J. M. van Opbergen, Florencia Schlamp, Marta Pérez-Hernández, Grecia M. Marrón-Liñares, Mario Delmar, Marina Cerrone, Valeria Mezzano, Adriana Heguy, and Feng-Xia Liang
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plakophilin-2 ,Gene knockdown ,lcsh:QP1-981 ,Physiology ,Inflammation ,Biology ,arrhythmogenic cardiomyopathy ,inflammation/immune response ,lcsh:Physiology ,Cell biology ,Transcriptome ,medicine.anatomical_structure ,Immune system ,Downregulation and upregulation ,Desmosome ,Physiology (medical) ,medicine ,biology.protein ,Myocyte ,GTEx ,medicine.symptom ,Antibody ,transcriptome ,Original Research - Abstract
Plakophilin-2 (PKP2) is classically defined as a component of the desmosome. Besides its role in cell–cell adhesion, PKP2 can modulate transcription through intracellular signals initiated at the site of cell–cell contact. Mutations in PKP2 associate with arrhythmogenic right ventricular cardiomyopathy (ARVC). Recent data demonstrate that inflammation plays a key role in disease progression; other results show an abundance of anti-heart antibodies in patients with confirmed diagnosis of ARVC. Here, we test the hypothesis that, in adult cardiac myocytes, PKP2 transcript abundance is endogenously linked to the abundance of transcripts participating in the inflammatory/immune response. Cardiac-specific, tamoxifen (TAM)-activated PKP2-knockout mice (PKP2cKO) were crossed with a RiboTag line to allow characterization of the ribosome-resident transcriptome of cardiomyocytes after PKP2 knockdown. Data were combined with informatics analysis of human cardiac transcriptome using GTEx. Separately, the presence of non-myocyte cells at the time of analysis was assessed by imaging methods. We identified a large number of transcripts upregulated consequent to PKP2 deficiency in myocytes, inversely correlated with PKP2 abundance in human transcriptomes, and part of functional pathways associated with inflammatory/immune responses. Our data support the concept that PKP2 is transcriptionally linked, in cardiac myocytes, to genes coding for host-response molecules even in the absence of exogenous triggers. Targeted anti-inflammatory therapy may be effective in ARVC.
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- 2021
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161. Multimodal single-cell analysis of cutaneous T-cell lymphoma reveals distinct subclonal tissue-dependent signatures
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Anthony Cheng, Terkild B. Buus, Marshall E. Kadin, Jose U. Scher, Kenneth B. Hymes, Zhengqing Ouyang, Sergei B. Koralov, Alberto Herrera, Larisa J. Geskin, Dean David George, Peter Smibert, Jo-Ann Latkowski, Kelly V. Ruggles, Niels Ødum, Angelina Seffens, Michal Bar-Natan, Adriana Heguy, and Eleni P. Mimitou
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Skin Neoplasms ,Immunology ,Biology ,Biochemistry ,Somatic evolution in cancer ,Single-cell analysis ,Gene expression ,medicine ,Tumor Cells, Cultured ,Sezary Syndrome ,Humans ,Receptor ,Cells, Cultured ,Mycosis fungoides ,Lymphoid Neoplasia ,Cluster of differentiation ,Cutaneous T-cell lymphoma ,Cell Biology ,Hematology ,medicine.disease ,Flow Cytometry ,Lymphoma ,Lymphoma, T-Cell, Cutaneous ,Cancer research ,Single-Cell Analysis ,Transcriptome - Abstract
Cutaneous T-cell lymphoma (CTCL) is a heterogeneous group of mature T-cell neoplasms characterized by the accumulation of clonal malignant CD4+ T cells in the skin. The most common variant of CTCL, mycosis fungoides (MF ), is confined to the skin in early stages but can be accompanied by extracutaneous dissemination of malignant T cells to the blood and lymph nodes in advanced stages of disease. Sézary syndrome (SS), a leukemic form of disease, is characterized by significant blood involvement. Little is known about the transcriptional and genomic relationship between skin- and blood-residing malignant T cells in CTCL. To identify and interrogate malignant clones in matched skin and blood from patients with leukemic MF and SS, we combine T-cell receptor clonotyping with quantification of gene expression and cell surface markers at the single cell level. Our data reveal clonal evolution at a transcriptional and genetic level within the malignant populations of individual patients. We highlight highly consistent transcriptional signatures delineating skin- and blood-derived malignant T cells. Analysis of these 2 populations suggests that environmental cues, along with genetic aberrations, contribute to transcriptional profiles of malignant T cells. Our findings indicate that the skin microenvironment in CTCL promotes a transcriptional response supporting rapid malignant expansion, as opposed to the quiescent state observed in the blood, potentially influencing efficacy of therapies. These results provide insight into tissue-specific characteristics of cancerous cells and underscore the need to address the patients’ individual malignant profiles at the time of therapy to eliminate all subclones.
- Published
- 2021
162. Profiling Basal Forebrain Cholinergic Neurons Reveals a Molecular Basis for Vulnerability Within the Ts65Dn Model of Down Syndrome and Alzheimer's Disease
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Melissa J. Alldred, Sai C. Penikalapati, Panos Roussos, Adriana Heguy, Sang Han Lee, and Stephen D. Ginsberg
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Male ,Basal Forebrain ,Population ,Neuroscience (miscellaneous) ,Mice, Transgenic ,Biology ,Article ,Cellular and Molecular Neuroscience ,Mice ,Alzheimer Disease ,medicine ,Animals ,Cognitive Dysfunction ,Gene Regulatory Networks ,Cholinergic neuron ,Cognitive decline ,KEGG ,education ,Laser capture microdissection ,Medial septal nucleus ,education.field_of_study ,Basal forebrain ,Mice, Inbred C3H ,Sequence Analysis, RNA ,Computational Biology ,Cholinergic Neurons ,Mice, Inbred C57BL ,medicine.anatomical_structure ,Neurology ,Cholinergic ,Female ,Down Syndrome ,Neuroscience - Abstract
BACKGROUND: Basal forebrain cholinergic neuron (BFCN) degeneration is a hallmark of Down syndrome (DS) and Alzheimer’s disease (AD). Current therapeutics have been unsuccessful in slowing disease progression, likely due to complex pathological interactions and dysregulated pathways that are poorly understood. The Ts65Dn trisomic mouse model recapitulates both cognitive and morphological deficits of DS and AD, including BFCN degeneration. METHODS: We utilized Ts65Dn mice to understand mechanisms underlying BFCN degeneration to identify novel targets for therapeutic intervention. We performed high-throughput, single population RNA sequencing (RNA-seq) to interrogate transcriptomic changes within medial septal nucleus (MSN) BFCNs, using laser capture microdissection to individually isolate ~500 choline acetyltransferase-immunopositive neurons in Ts65Dn and normal disomic (2N) mice at 6 months of age (MO). RESULTS: Ts65Dn mice had unique MSN BFCN transcriptomic profiles at ~6 MO clearly differentiating them from 2N mice. Leveraging Ingenuity Pathway Analysis and KEGG analysis, we linked differentially expressed gene (DEG) changes within MSN BFCNs to several canonical pathways and aberrant physiological functions. The dysregulated transcriptomic profile of trisomic BFCNs provides key information underscoring selective vulnerability within the septohippocampal circuit. CONCLUSIONS: We propose both expected and novel therapeutic targets for DS and AD, including specific DEGs within cholinergic, glutamatergic, GABAergic, and neurotrophin pathways, as well as select targets for repairing oxidative phosphorylation status in neurons. We demonstrate and validate an interrogative quantitative bioinformatic analysis of a key dysregulated neuronal population linking single population transcript changes to an established pathological hallmark associated with cognitive decline for therapeutic development in human DS and AD.
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- 2020
163. Lower airway dysbiosis affects lung cancer progression
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Daniel H. Sterman, Jose C. Clemente, Peter Meyn, Sergei B. Koralov, Kevin Felner, R. Schluger, Luisannay Perez, Linchen He, Adriana Heguy, J. Carpenito, Richard Bonneau, Mariam El-Ashmawy, Yonghua Li, B. Franca, William N. Rom, Ting An Yie, Imran Sulaiman, James T. Morton, Anastasia Maria Zavitsanou, Valeria Mezzano, Benjamin G. Wu, Katherine Gershner, Cynthia Loomis, Robert L. Smith, Samaan Rafeq, Andre L. Moreira, Gaetane Michaud, Huilin Li, Leopoldo N. Segal, Harald Sauthoff, William Moore, Tadasu Iizumi, Ray Pillai, Aristotelis Tsirigos, Harvey I. Pass, Jamie L. Bessich, Thales Papagiannakopoulos, E. Olsen, Kwok-Kin Wong, Jun Chieh J. Tsay, Michelle H. Badri, and Nan Shen
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0301 basic medicine ,Lung Neoplasms ,medicine.medical_treatment ,New York ,Mice, Transgenic ,Adenocarcinoma ,Gut flora ,medicine.disease_cause ,Article ,Cohort Studies ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,medicine ,Animals ,Humans ,Neoplasm Metastasis ,Lung cancer ,Lung ,Neoplasm Staging ,Proportional Hazards Models ,biology ,business.industry ,Microbiota ,Cancer ,Immunotherapy ,respiratory system ,medicine.disease ,biology.organism_classification ,Survival Analysis ,respiratory tract diseases ,Gastrointestinal Microbiome ,Disease Models, Animal ,030104 developmental biology ,Oncology ,Tumor progression ,030220 oncology & carcinogenesis ,Immunology ,Disease Progression ,Dysbiosis ,Female ,Carcinogenesis ,business - Abstract
In lung cancer, enrichment of the lower airway microbiota with oral commensals commonly occurs, and ex vivo models support that some of these bacteria can trigger host transcriptomic signatures associated with carcinogenesis. Here, we show that this lower airway dysbiotic signature was more prevalent in the stage IIIB–IV tumor–node–metastasis lung cancer group and is associated with poor prognosis, as shown by decreased survival among subjects with early-stage disease (I–IIIA) and worse tumor progression as measured by RECIST scores among subjects with stage IIIB–IV disease. In addition, this lower airway microbiota signature was associated with upregulation of the IL17, PI3K, MAPK, and ERK pathways in airway transcriptome, and we identified Veillonella parvula as the most abundant taxon driving this association. In a KP lung cancer model, lower airway dysbiosis with V. parvula led to decreased survival, increased tumor burden, IL17 inflammatory phenotype, and activation of checkpoint inhibitor markers. Significance: Multiple lines of investigation have shown that the gut microbiota affects host immune response to immunotherapy in cancer. Here, we support that the local airway microbiota modulates the host immune tone in lung cancer, affecting tumor progression and prognosis. See related commentary by Zitvogel and Kroemer, p. 224. This article is highlighted in the In This Issue feature, p. 211
- Published
- 2020
164. SARS-CoV-2 genomic characterization and clinical manifestation of the COVID-19 outbreak in Uruguay
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Veronica Estevez, Matilde Nin, Paul Zappile, Maria Noel Zubillaga, Susana Boschi, Carolina Beloso, Natalia Mazza, Andrea Santini, Fabiana Ross, Veronica Perez, Silvana Ifran, Batsirai Mabvakure, Adriana Heguy, Gonzalo Manrique, Leticia Perez, Gordon William Harkins, Ralf Duerr, Matthew T. Maurano, Sabine Smidt, M. Fernández, Christian Marier, Victoria Elizondo, Victoria Perez, Raquel Balleste, and Simon Dellicour
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0301 basic medicine ,Male ,Latin Americans ,Epidemiology ,viruses ,Clinical manifestation ,Pharmacologie ,Disease Outbreaks ,Drug Discovery ,Immunologie ,Pandemic ,Pathologie maladies infectieuses ,Clade ,Phylogeny ,clinical correlations ,Dominance (genetics) ,Aged, 80 and over ,Parasitologie ,virus diseases ,General Medicine ,Middle Aged ,Epidémiologie ,Microbiologie et protistologie [entomologie,phytoparasitolog.] ,Infectious Diseases ,full genome amplicon sequencing ,Cohort ,Female ,phylogeographic BEAST analysis ,Research Article ,Adult ,Microbiologie et protistologie [parasitologie hum. et anim.] ,medicine.medical_specialty ,COVID-19 outbreak ,Coronavirus disease 2019 (COVID-19) ,Adolescent ,Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) ,030106 microbiology ,Immunology ,Biology ,Microbiology ,Polymorphism, Single Nucleotide ,World health ,Article ,Virus ,03 medical and health sciences ,Young Adult ,Virology ,medicine ,Humans ,spike D614G genetic mutation ,Intensive care medicine ,Aged ,Respiratory illness ,business.industry ,SARS-CoV-2 ,Public health ,COVID-19 ,Outbreak ,South America ,030104 developmental biology ,Mutation ,Uruguay ,Parasitology ,business ,Microbiologie et protistologie [bacteriol.virolog.mycolog.] ,Demography - Abstract
COVID-19 is a respiratory illness caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and declared by the World Health Organization a global public health emergency. Among the severe outbreaks across South America, Uruguay has become known for curtailing SARS-CoV-2 exceptionally well. To understand the SARS-CoV-2 introductions, local transmissions, and associations with genomic and clinical parameters in Uruguay, we sequenced the viral genomes of 44 outpatients and inpatients in a private healthcare system in its capital, Montevideo, from March to May 2020. We performed a phylogeographic analysis using sequences from our cohort and other studies that indicate a minimum of 23 independent introductions into Uruguay, resulting in five major transmission clusters. Our data suggest that most introductions resulting in chains of transmission originate from other South American countries, with the earliest seeding of the virus in late February 2020, weeks before the borders were closed to all non-citizens and a partial lockdown implemented. Genetic analyses suggest a dominance of S and G clades (G, GH, GR) that make up >90% of the viral strains in our study. In our cohort, lethal outcome of SARS-CoV-2 infection significantly correlated with arterial hypertension, kidney failure, and ICU admission (FDR < 0.01), but not with any mutation in a structural or non-structural protein, such as the spike D614G mutation. Our study contributes genetic, phylodynamic, and clinical correlation data about the exceptionally well-curbed SARS-CoV-2 outbreak in Uruguay, which furthers the understanding of disease patterns and regional aspects of the pandemic in Latin America., SCOPUS: ar.j, info:eu-repo/semantics/published
- Published
- 2020
165. Immune Response and Microbiota Profiles during Coinfection with Plasmodium vivax and Soil-Transmitted Helminths
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Adriana Heguy, Ana Rodriguez, Erica Ruan, María Fernanda Yasnot, P'ng Loke, Mayra Raciny-Aleman, Christian Marier, Victor Liu, and Alice V. Easton
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Male ,Plasmodium vivax ,Helminthiasis ,Parasitemia ,Gut flora ,Clinical Science and Epidemiology ,Feces ,Soil ,0302 clinical medicine ,fluids and secretions ,RNA-Seq ,Child ,0303 health sciences ,education.field_of_study ,biology ,Coinfection ,QR1-502 ,Child, Preschool ,Female ,Research Article ,Adolescent ,030231 tropical medicine ,Population ,malaria ,Colombia ,digestive system ,Microbiology ,03 medical and health sciences ,Virology ,Helminths ,parasitic diseases ,medicine ,microbiota ,Malaria, Vivax ,Animals ,Humans ,education ,Trichuris trichiura ,soil-transmitted helminths ,030304 developmental biology ,Bacteria ,Gene Expression Profiling ,medicine.disease ,biology.organism_classification ,Gastrointestinal Microbiome ,Cross-Sectional Studies ,Immunology ,STH ,Malaria - Abstract
Plasmodium (malaria) and helminth parasite coinfections are frequent, and both infections can be affected by the host gut microbiota. However, the relationship between coinfection and the gut microbiota is unclear. By performing comprehensive analyses on blood/stool samples from 130 individuals in Colombia, we found that the gut microbiota may have a stronger relationship with the number of P. vivax (malaria) parasites than with the number of helminth parasites infecting a host. Microbiota analysis identified more predictors of the P. vivax parasite burden, whereas analysis of blood samples identified predictors of the helminth parasite burden. These results were unexpected, because we expected each parasite to be associated with greater differences in its biological niche (blood for P. vivax and the intestine for helminths). Instead, we find that bacterial taxa were the strongest predictors of P. vivax parasitemia levels, while circulating TGF-β levels were the strongest predictor of helminth parasite burdens., The role of the gut microbiota during coinfection with soil-transmitted helminths (STH) and Plasmodium spp. is poorly understood. We examined peripheral blood and fecal samples from 130 individuals who were either infected with Plasmodium vivax only, coinfected with P. vivax and STH, infected with STH alone, or not infected with either P. vivax or STH. In addition to a complete blood count (CBC) with differential, transcriptional profiling of peripheral blood samples was performed by transcriptome sequencing (RNA-Seq), fecal microbial communities were determined by 16S rRNA gene sequencing, and circulating cytokine levels were measured by bead-based immunoassays. Differences in blood cell counts, including an increased percentage of neutrophils, associated with a transcriptional signature of neutrophil activation, were driven primarily by P. vivax infection. P. vivax infection was also associated with increased levels of interleukin 6 (IL-6), IL-8, and IL-10; these cytokine levels were not affected by STH coinfection. Surprisingly, P. vivax infection was more strongly associated with differences in the microbiota than STH infection. Children infected with only P. vivax exhibited elevated Bacteroides and reduced Prevotella and Clostridiaceae levels, but these differences were not observed in individuals coinfected with STH. We also observed that P. vivax parasitemia was higher in the STH-infected population. When we used machine learning to identify the most important predictors of the P. vivax parasite burden (among P. vivax-infected individuals), bacterial taxa were the strongest predictors of parasitemia. In contrast, circulating transforming growth factor β (TGF-β) was the strongest predictor of the Trichuris trichiura egg burden. This study provides unexpected evidence that the gut microbiota may have a stronger link with P. vivax than with STH infection.
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- 2020
166. Ontogeny and Vulnerabilities of Drug-Tolerant Persisters in HER2+ Breast Cancer
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Chewei Anderson Chang, Jayu Jen, Shaowen Jiang, Azin Sayad, Arvind Singh Mer, Kevin R. Brown, Allison M.L. Nixon, Avantika Dhabaria, Kwan Ho Tang, David Venet, Christos Sotiriou, Jiehui Deng, Kwok-kin Wong, Sylvia Adams, Peter Meyn, Adriana Heguy, Jane A. Skok, Aristotelis Tsirigos, Beatrix Ueberheide, Jason Moffat, Abhyudai Singh, Benjamin Haibe-Kains, Alireza Khodadadi-Jamayran, and Benjamin G. Neel
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Receptor, ErbB-2 ,Breast Neoplasms ,mTORC1 ,Biology ,Lapatinib ,Article ,Metastasis ,Phosphatidylinositol 3-Kinases ,03 medical and health sciences ,0302 clinical medicine ,Breast cancer ,Cell Line, Tumor ,medicine ,Humans ,Protein kinase B ,PI3K/AKT/mTOR pathway ,030304 developmental biology ,0303 health sciences ,Cancer ,medicine.disease ,respiratory tract diseases ,3. Good health ,Oncology ,Drug Resistance, Neoplasm ,030220 oncology & carcinogenesis ,Cancer cell ,Cancer research ,Female ,Signal Transduction ,medicine.drug - Abstract
Resistance to targeted therapies is an important clinical problem in HER2-positive (HER2+) breast cancer. “Drug-tolerant persisters” (DTP), a subpopulation of cancer cells that survive via reversible, nongenetic mechanisms, are implicated in resistance to tyrosine kinase inhibitors (TKI) in other malignancies, but DTPs following HER2 TKI exposure have not been well characterized. We found that HER2 TKIs evoke DTPs with a luminal-like or a mesenchymal-like transcriptome. Lentiviral barcoding/single-cell RNA sequencing reveals that HER2+ breast cancer cells cycle stochastically through a “pre-DTP” state, characterized by a G0-like expression signature and enriched for diapause and/or senescence genes. Trajectory analysis/cell sorting shows that pre-DTPs preferentially yield DTPs upon HER2 TKI exposure. Cells with similar transcriptomes are present in HER2+ breast tumors and are associated with poor TKI response. Finally, biochemical experiments indicate that luminal-like DTPs survive via estrogen receptor–dependent induction of SGK3, leading to rewiring of the PI3K/AKT/mTORC1 pathway to enable AKT-independent mTORC1 activation. Significance: DTPs are implicated in resistance to anticancer therapies, but their ontogeny and vulnerabilities remain unclear. We find that HER2 TKI-DTPs emerge from stochastically arising primed cells (“pre-DTPs”) that engage either of two distinct transcriptional programs upon TKI exposure. Our results provide new insights into DTP ontogeny and potential therapeutic vulnerabilities. This article is highlighted in the In This Issue feature, p. 873
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- 2020
167. Hippocampal metabolite concentrations in schizophrenia vary in association with rare gene variants in the TRIO gene
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Haley Rhodes, Adriana Heguy, Moses V. Chao, Julie Walsh-Messinger, Kevin W. Hoffman, Oded Gonen, Thorsten M. Kranz, and Dolores Malaspina
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Genetics ,Metabolite ,Hippocampal formation ,Biology ,medicine.disease ,Hippocampus ,Polymorphism, Single Nucleotide ,Psychiatry and Mental health ,chemistry.chemical_compound ,chemistry ,Schizophrenia ,medicine ,Autism ,Humans ,Association (psychology) ,Gene ,Biological Psychiatry - Published
- 2020
168. WITHDRAWN: ASSOCIATION OF INITIAL VIRAL LOAD IN SARS-CoV-2 PATIENTS WITH OUTCOME AND SYMPTOMS
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Matija Snuderl, Lawrence Hsu Lin, George Jour, Adriana Heguy, Andrea B. Troxel, Antonio Serrano, Guomiao Shen, Brendan Belovarac, Melissa Call, Paolo Cotzia, Una Moran, Xiaojun Feng, Jiyuan Hu, Min Jae Kim, Margaret Black, Kimon V. Argyropoulos, Theodore Vougiouklakis, Iman Osman, and Andrew Lytle
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2019-20 coronavirus outbreak ,Coronavirus disease 2019 (COVID-19) ,business.industry ,Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) ,Medicine ,business ,Virology ,Viral load ,Pathology and Forensic Medicine - Abstract
The Publisher regrets that this article is an accidental duplication of an article that has already been published, https://doi.org/10.1016/j.ajpath.2020.07.001. The duplicate article has therefore been withdrawn. The full Elsevier Policy on Article Withdrawal can be found at https://www.elsevier.com/about/our-business/policies/article-withdrawal.
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- 2020
169. Genome-Wide DNA Methylation Profiles in Community Members Exposed to the World Trade Center Disaster
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Y. Zhang, Anne Zeleniuch-Jacquotte, Stephanie Tuminello, Nedim Durmus, Joan Reibman, Yongzhao Shao, Adriana Heguy, Matija Snuderl, Lei Yang, and Alan A. Arslan
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exposure assessment ,Health, Toxicology and Mutagenesis ,environmental exposure ,lcsh:Medicine ,Pilot Projects ,Biology ,Genome ,Article ,Bioconductor ,03 medical and health sciences ,0302 clinical medicine ,Peptide Elongation Factor 1 ,9/11 ,Humans ,Epigenetics ,Prospective Studies ,KEGG ,030304 developmental biology ,Genetics ,0303 health sciences ,lcsh:R ,Public Health, Environmental and Occupational Health ,Dust ,Environmental exposure ,Methylation ,DNA Methylation ,epigenome-wide association study ,humanities ,World Trade Center ,pathway analysis ,CpG site ,030220 oncology & carcinogenesis ,DNA methylation ,Environmental Pollutants ,Female ,methylation ,September 11 Terrorist Attacks ,Rho Guanine Nucleotide Exchange Factors - Abstract
The primary goal of this pilot study was to assess feasibility of studies among local community members to address the hypothesis that complex exposures to the World Trade Center (WTC) dust and fumes resulted in long-term epigenetic changes. We enrolled 18 WTC-exposed cancer-free women from the WTC Environmental Health Center (WTC EHC) who agreed to donate blood samples during their standard clinical visits. As a reference WTC unexposed group, we randomly selected 24 age-matched cancer-free women from an existing prospective cohort who donated blood samples before 11 September 2001. The global DNA methylation analyses were performed using Illumina Infinium MethylationEpic arrays. Statistical analyses were performed using R Bioconductor package. Functional genomic analyses were done by mapping the top 5000 differentially expressed CpG sites to the Kyoto Encyclopedia of Genes and Genomes (KEGG) Pathway database. Among cancer-free subjects, we observed substantial methylation differences between WTC-exposed and unexposed women. The top 15 differentially methylated gene probes included BCAS2, OSGIN1, BMI1, EEF1A2, SPTBN5, CHD8, CDCA7L, AIDA, DDN, SNORD45C, ZFAND6, ARHGEF7, UBXN8, USF1, and USP12. Several cancer-related pathways were enriched in the WTC-exposed subjects, including endocytosis, mitogen-activated protein kinase (MAPK), viral carcinogenesis, as well as Ras-associated protein-1 (Rap1) and mammalian target of rapamycin (mTOR) signaling. The study provides preliminary data on substantial differences in DNA methylation between WTC-exposed and unexposed populations that require validation in further studies.
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- 2020
170. Distinct Transcriptomic Profiles in the Dorsal Hippocampus and Prelimbic Cortex Are Transiently Regulated following Episodic Learning
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Adriana Heguy, Xiaojing Ye, Cristina M. Alberini, Aaron Katzman, Alireza Khodadadi-Jamayran, Dana Kapeller-Libermann, and Aristotelis Tsirigos
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0301 basic medicine ,Male ,Memory, Episodic ,Infralimbic cortex ,Hippocampus ,Prefrontal Cortex ,Biology ,Axonogenesis ,03 medical and health sciences ,0302 clinical medicine ,Cortex (anatomy) ,medicine ,Avoidance Learning ,Animals ,Gene Regulatory Networks ,Rats, Long-Evans ,Synapse organization ,Research Articles ,General Neuroscience ,Fear ,Synaptic vesicle cycle ,Cell biology ,Rats ,030104 developmental biology ,medicine.anatomical_structure ,Lipid modification ,Transcriptome ,030217 neurology & neurosurgery ,Vesicle localization - Abstract
A fundamental, evolutionarily conserved biological mechanism required for long-term memory formation is rapid induction of gene transcription upon learning in relevant brain areas. For episodic types of memories, two regions undergoing this transcription are the dorsal hippocampus (dHC) and prelimbic (PL) cortex. Whether and to what extent these regions regulate similar or distinct transcriptomic profiles upon learning remain to be understood. Here, we used RNA sequencing in the dHC and PL cortex of male rats to profile their transcriptomes in untrained conditions (baseline) and at 1 h and 6 d after inhibitory avoidance learning. We found that, of 33,713 transcripts, >14,000 were significantly expressed at baseline in both regions and ∼3000 were selectively enriched in each region. Gene Ontology biological pathway analyses indicated that commonly expressed pathways included synapse organization, regulation of membrane potential, and vesicle localization. The enriched pathways in the dHC were gliogenesis, axon development, and lipid modification, while in the PL cortex included vesicle localization and synaptic vesicle cycle. At 1 h after learning, 135 transcripts changed significantly in the dHC and 478 in the PL cortex; of these, only 34 were shared. Biological pathways most significantly regulated by learning in the dHC were protein dephosphorylation, glycogen and glucan metabolism, while in the PL cortex were axon development and axonogenesis. The transcriptome profiles returned to baseline by 6 d after training. Thus, a significant portion of dHC and PL cortex transcriptomic profiles is divergent, and their regulation upon learning is largely distinct and transient.SIGNIFICANCE STATEMENTLong-term episodic memory formation requires gene transcription in several brain regions, including the hippocampus and PFC. The comprehensive profiles of the dynamic mRNA changes that occur in these regions following learning are not well understood. Here, we performed RNA sequencing in the dorsal hippocampus and prelimbic cortex, a PFC subregion, at baseline, 1 h, and 6 d after episodic learning in rats. We found that, at baseline, dorsal hippocampus and prelimbic cortex differentially express a significant portion of mRNAs. Moreover, learning produces a transient regulation of region-specific profiles of mRNA, indicating that unique biological programs in different brain regions underlie memory formation.
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- 2020
171. Hippocampal Metabolite Concentrations in A Schizophrenia Case Series Vary in Association With Rare Gene Variants For Schizophrenia Versus Autism
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Thorsten M. Kranz, Kevin W. Hoffmann, Dolores Malaspina, Oded Gonen, Haley Rhodes, Moses V. Chao, and Adriana Heguy
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Genetics ,business.industry ,Metabolite ,Hippocampal formation ,medicine.disease ,chemistry.chemical_compound ,chemistry ,Schizophrenia ,mental disorders ,medicine ,Autism ,Association (psychology) ,business ,Gene - Abstract
BackgroundRare variants in the TRIO gene are associated with schizophrenia and autism spectrum disorders (ASD), which are commonly comorbid. ASD may define a specific schizophrenia subtype. This study examined person-specific hippocampal metabolite concentrations for 4 schizophrenia cases harboring rare variants in TRIO or its interaction partner ARMS/KIDINS220 and 5 cases with other rare variants.MethodsNine of 19 cases from a prior imaging study underwent targeted exome sequencing. Multi-voxel 1H-MRS imaging of the entire 3-dimensional hippocampus found only increased Creatine [Cr] (cellular energy use) concentration, distinguished cases at the group level. However, concentrations of N-acetyl-aspartate [NAA] (neuronal integrity) and choline [Cho] (membrane turnover/myelination) concentrations had a greater variance in cases than controls.ResultsFour cases with rare, missense-coding mutations or non-frameshift deletions in TRIO or ARMS/KIDINS220 had significantly lower [Cho] than the other 5 (1.78 ± 0.18 mM versus 2.67 ± 0.8 mM: t = 3.55, p = 0.005) but similar [NAA]. Two cases harboring rare variants in the SLC39A1 zinc transporter had the lowest [NAA]. (8.41 ± 0 80 mM versus 10.35 ± 2.03 mM, t = 4.52, p = 0.001). The highest [Cho] accompanied rare variants in SORCS2 and SORT, associated with schizophrenia and Alzheimer’s Disease.LimitationsThis preliminary study of a small sample of exceptionally well characterized cases requires replication in larger samples for clinical utility.ConclusionsThe hippocampus is vulnerable to more than one pathology causing schizophrenia. TRIO rare variants predicted significantly lower Cho, indicating reduced myelin. [Cho] and [NAA] may have importance for choosing and monitoring schizophrenia treatment.
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- 2020
172. Sequencing identifies multiple early introductions of SARS-CoV-2 to the New York City Region
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Manon Ragonnet-Cronin, John Cadley, Lawrence Hsu Lin, Megan S. Hogan, Ludovic Boytard, Antonio Serrano, Dacia Dimartino, Marie I. Samanovic, Melissa Call, Iman Osman, Matija Snuderl, Vanessa Raabe, Mark J. Mulligan, Brendan Belovarac, Xiaojun Feng, Christian Marier, George Jour, Maria E Aguero-Rosenfeld, Amy Rapkiewicz, Peter Meyn, Raquel Ordoñez, Nicholas A. Vulpescu, Carolina Arguelles-Grande, Jared Pinnell, Paul Zappile, Lily Geidelberg, Kimon V. Argyropoulos, Tatyana Gindin, Theodore Vougiouklakis, Guomiao Shen, André M. Ribeiro-dos-Santos, Andrew Lytle, Emily P. Huang, Min Jae Kim, Margaret Black, Adriana Heguy, Gael Westby, Paolo Cotzia, Erik M. Volz, Raven Luther, John J. Chen, Yutong Zhang, Emily Guzman, Matthew T. Maurano, and Sitharam Ramaswami
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Male ,medicine.medical_specialty ,Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) ,Genome, Viral ,Biology ,Article ,DNA sequencing ,03 medical and health sciences ,0302 clinical medicine ,Epidemiology ,Pandemic ,Genetics ,medicine ,Humans ,Genetics(clinical) ,Pandemics ,Genetics (clinical) ,Phylogeny ,030304 developmental biology ,Whole genome sequencing ,0303 health sciences ,Whole Genome Sequencing ,SARS-CoV-2 ,Transmission (medicine) ,Research ,COVID-19 ,Outbreak ,Metropolitan area ,3. Good health ,City region ,Geography ,Evolutionary biology ,Female ,New York City ,030217 neurology & neurosurgery - Abstract
Effective public response to a pandemic relies upon accurate measurement of the extent and dynamics of an outbreak. Viral genome sequencing has emerged as a powerful approach to link seemingly unrelated cases, and large-scale sequencing surveillance can inform on critical epidemiological parameters. Here, we report the analysis of 236 SARS-CoV2 sequences from cases in the New York City metropolitan area during the initial stages of the 2020 COVID-19 outbreak. The majority of cases throughout the region had no recent travel history or known exposure, and genetically linked cases were spread throughout the region. Comparison to global viral sequences showed that the majority were most related to cases from Europe. Our data are consistent with numerous seed transmissions from multiple sources and a prolonged period of unrecognized community spreading. This work highlights the complementary role of real-time genomic surveillance in addition to traditional epidemiological indicators.
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- 2020
173. iCellR: Combined Coverage Correction and Principal Component Alignment for Batch Alignment in Single-Cell Sequencing Analysis
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Joseph Pucella, Nicole A. Doudican, Adriana Heguy, Alireza Khodadadi-Jamayran, Boris Reizis, John A. Carucci, Aristotelis Tsirigos, and Hua Zhou
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Single cell sequencing ,business.industry ,Computer science ,Dimensionality reduction ,Principal component analysis ,RNA ,Pattern recognition ,Artificial intelligence ,business ,Cluster analysis ,Imputation (genetics) ,k-nearest neighbors algorithm - Abstract
SUMMARYUnder-sampling RNA molecules and low-coverage sequencing in some single cell sequencing technologies introduce zero counts (also known as drop-outs) into the expression matrices. This issue may complicate the processes of dimensionality reduction and clustering, often forcing distinct cell types to falsely resemble one another, while eliminating subtle, but important differences. Considering the wide range in drop-out rates from different sequencing technologies, it can also affect the analysis at the time of batch/sample alignment and other downstream analyses. Therefore, generating an additional harmonized gene expression matrix is important. To address this, we introduce two separate batch alignment methods: Combined Coverage Correction Alignment (CCCA) and Combined Principal Component Alignment (CPCA). The first method uses a coverage correction approach (analogous to imputation) in a combined or joint fashion between multiple samples for batch alignment, while also correcting for drop-outs in a harmonious way. The second method (CPCA) skips the coverage correction step and uses k nearest neighbors (KNN) for aligning the PCs from the nearest neighboring cells in multiple samples. Our results of nine scRNA-seq PBMC samples from different batches and technologies shows the effectiveness of both these methods. All of our algorithms are implemented in R, deposited into CRAN, and available in the iCellR package.
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- 2020
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174. Interleukin-17 Inhibition in Spondyloarthritis Is Associated With Subclinical Gut Microbiome Perturbations and a Distinctive Interleukin-25-Driven Intestinal Inflammation
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Soumya M. Reddy, Matthew Stapylton, Drew R. Jones, Richard Bonneau, Alexander A. Aksenov, Carles Ubeda, Pieter C. Dorrestein, Monica Guma, Michelle H. Badri, Andrea L. Neimann, Jose C. Clemente, Gary Solomon, Giuliana Guggino, Jose U. Scher, Francesco Ciccia, Adriana Heguy, Leopoldo N. Segal, Roxana Coras, Julia Manasson, Parvathy V. Girija, and David S. Wallach
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Male ,0301 basic medicine ,Arthritis ,Psoriatic ,0302 clinical medicine ,Interleukin 25 ,Monoclonal ,2.1 Biological and endogenous factors ,Immunology and Allergy ,Medicine ,Aetiology ,Intestinal Mucosa ,Candida albicans ,Humanized ,Subclinical infection ,biology ,Interleukin-17 ,Innate lymphoid cell ,Middle Aged ,Intestines ,Public Health and Health Services ,Female ,Tumor necrosis factor alpha ,Interleukin 17 ,Clinical Sciences ,Immunology ,Antibodies, Monoclonal, Humanized ,Autoimmune Disease ,Antibodies ,Article ,03 medical and health sciences ,Rheumatology ,Clinical Research ,Spondylarthritis ,Humans ,Microbiome ,Inflammation ,030203 arthritis & rheumatology ,business.industry ,Inflammatory and immune system ,Arthritis, Psoriatic ,biology.organism_classification ,medicine.disease ,Arthritis & Rheumatology ,Gastrointestinal Microbiome ,030104 developmental biology ,Tumor Necrosis Factor Inhibitors ,Digestive Diseases ,business - Abstract
Objective To characterize the ecological effects of biologic therapies on the gut bacterial and fungal microbiome in psoriatic arthritis (PsA)/spondyloarthritis (SpA) patients. Methods Fecal samples from PsA/SpA patients pre- and posttreatment with tumor necrosis factor inhibitors (TNFi; n = 15) or an anti-interleukin-17A monoclonal antibody inhibitor (IL-17i; n = 14) underwent sequencing (16S ribosomal RNA, internal transcribed spacer and shotgun metagenomics) and computational microbiome analysis. Fecal levels of fatty acid metabolites and cytokines/proteins implicated in PsA/SpA pathogenesis or intestinal inflammation were correlated with sequence data. Additionally, ileal biopsies obtained from SpA patients who developed clinically overt Crohn's disease (CD) after treatment with IL-17i (n = 5) were analyzed for expression of IL-23/Th17-related cytokines, IL-25/IL-17E-producing cells, and type 2 innate lymphoid cells (ILC2s). Results There were significant shifts in abundance of specific taxa after treatment with IL-17i compared to TNFi, particularly Clostridiales (P = 0.016) and Candida albicans (P = 0.041). These subclinical alterations correlated with changes in bacterial community co-occurrence, metabolic pathways, IL-23/Th17-related cytokines, and various fatty acids. Ileal biopsies showed that clinically overt CD was associated with expansion of IL-25/IL-17E-producing tuft cells and ILC2s (P < 0.05), compared to pre-IL-17i treatment levels. Conclusion In a subgroup of SpA patients, the initiation of IL-17A blockade correlated with features of subclinical gut inflammation and intestinal dysbiosis of certain bacterial and fungal taxa, most notably C albicans. Further, IL-17i-related CD was associated with overexpression of IL-25/IL-17E-producing tuft cells and ILC2s. These results may help to explain the potential link between inhibition of a specific IL-17 pathway and the (sub)clinical gut inflammation observed in SpA.
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- 2020
175. Human blastocysts of normal and abnormal karyotypes display distinct transcriptome profiles
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Aristotelis Tsirigos, Frederick Licciardi, Yael Kramer, Adriana Heguy, Tenzin Lhakhang, and Yutong Zhang
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0301 basic medicine ,Monosomy ,animal structures ,Transcription, Genetic ,Abnormal Karyotype ,lcsh:Medicine ,Trisomy ,Biology ,Article ,Transcriptome ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Humans ,RNA, Messenger ,lcsh:Science ,Gene ,Genetics ,030219 obstetrics & reproductive medicine ,Multidisciplinary ,Ploidies ,lcsh:R ,Chromosome ,Embryo ,medicine.disease ,Embryonic stem cell ,Embryo transfer ,030104 developmental biology ,Blastocyst ,Gene Expression Regulation ,embryonic structures ,lcsh:Q - Abstract
Unveiling the transcriptome of human blastocysts can provide a wealth of important information regarding early embryonic ontology. Comparing the mRNA production of embryos with normal and abnormal karyotypes allows for a deeper understanding of the protein pathways leading to viability and aberrant fetal development. In addition, identifying transcripts specific for normal or abnormal chromosome copy number could aid in the search for secreted substances that could be used to non-invasively identify embryos best suited for IVF embryo transfer. Using RNA-seq, we characterized the transcriptome of 71 normally developing human blastocysts that were karyotypically normal vs. trisomic or monosomic. Every monosomy and trisomy of the autosomal and sex chromosomes were evaluated, mostly in duplicate. We first mapped the transcriptome of three normal embryos and found that a common core of more than 3,000 genes is expressed in all embryos. These genes represent pathways related to actively dividing cells, such as ribosome biogenesis and function, spliceosome, oxidative phosphorylation, cell cycle and metabolic pathways. We then compared transcriptome profiles of aneuploid embryos to those of normal embryos. We observed that non-viable embryos had a large number of dysregulated genes, some showing a hundred-fold difference in expression. On the contrary, sex chromosome abnormalities, XO and XXX displayed transcriptomes more closely mimicking those embryos with 23 normal chromosome pairs. Intriguingly, we identified a set of commonly deregulated genes in the majority of both trisomies and monosomies. This is the first paper demonstrating a comprehensive transcriptome delineation of karyotypic abnormalities found in the human pre-implantation embryo. We believe that this information will contribute to the development of new pre-implantation genetic screening methods as well as a better understanding of the underlying developmental abnormalities of abnormal embryos, fetuses and children.
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- 2018
176. Axon TRAP reveals learning-associated alterations in cortical axonal mRNAs in the lateral amygdala
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Robert M. Sears, Adriana Heguy, Eric Klann, Zachary Deane, Rahul N. Kanadia, Aristotelis Tsirigos, Joseph E. LeDoux, Tenzin Lhakhang, Linnaea E. Ostroff, and Emanuela Santini
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QH301-705.5 ,Science ,translatome ,Biology ,Amygdala ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,local translation ,0302 clinical medicine ,medicine ,Protein biosynthesis ,Animals ,Learning ,RNA, Messenger ,Biology (General) ,Axon ,030304 developmental biology ,Cerebral Cortex ,0303 health sciences ,General Immunology and Microbiology ,Basolateral Nuclear Complex ,Sequence Analysis, RNA ,General Neuroscience ,Translation (biology) ,amygdala ,General Medicine ,axonal translation ,Axons ,Rats ,Cortex (botany) ,memory consolidation ,medicine.anatomical_structure ,nervous system ,Protein Biosynthesis ,Synaptic plasticity ,Forebrain ,Rat ,Medicine ,Memory consolidation ,Neuroscience ,030217 neurology & neurosurgery ,Research Article - Abstract
Local translation can support memory consolidation by supplying new proteins to synapses undergoing plasticity. Translation in adult forebrain dendrites is an established mechanism of synaptic plasticity and is regulated by learning, yet there is no evidence for learning-regulated protein synthesis in adult forebrain axons, which have traditionally been believed to be incapable of translation. Here, we show that axons in the adult rat amygdala contain translation machinery, and use translating ribosome affinity purification (TRAP) with RNASeq to identify mRNAs in cortical axons projecting to the amygdala, over 1200 of which were regulated during consolidation of associative memory. Mitochondrial and translation-related genes were upregulated, whereas synaptic, cytoskeletal, and myelin-related genes were downregulated; the opposite effects were observed in the cortex. Our results demonstrate that axonal translation occurs in the adult forebrain and is altered after learning, supporting the likelihood that local translation is more a rule than an exception in neuronal processes.
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- 2019
177. GENE-62. IDENTIFICATION OF FGFR4 p.G388R VARIANT IN CEREBELLAR HEMANGIOBLASTOMAS
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Kasthuri Kannan, Jean-Pierre Gagner, Elad Mashiach, Adriana Heguy, Matija Snuderl, David Zagzag, and Matthias A. Karajannis
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Genetics and Epigenetics ,Cancer Research ,Pathology ,medicine.medical_specialty ,Cerebellum ,Mutation ,Fibroblast growth factor receptor 4 ,Biology ,medicine.disease ,medicine.disease_cause ,Germline mutation ,medicine.anatomical_structure ,Oncology ,Hemangioblastoma ,medicine ,Neurology (clinical) ,Exome ,Gene ,Exome sequencing - Abstract
BACKGROUND While most hemangioblastomas (~70%) are sporadic and occur predominantly in the cerebellum, they may present as well as familial form associated with von Hippel-Lindau (VHL) syndrome, an autosomal dominant disorder caused by germline mutations of the VHL gene that trigger nuclear translocation of hypoxia-inducible factor (HIF)-1α and angiogenesis. Although inactivation of VHL, a tumor suppressor gene, has been observed in hemangioblastomas, the underlying pathogenic mechanisms responsible for familial and sporadic hemangioblastomas remain incompletely understood. METHODS Whole exome sequencing of cerebellar hemangioblastoma tumors and matched blood leukocytes from 24 patients, age 24–63, was performed. After preparation and amplification of barcoded libraries, exomes were captured using Kapa Biosystems methodology and paired-end sequenced on Illumina HiSeq 2500 to an average 100-fold coverage. Following read alignment to hg19 genome, tumor and germline (leukocyte) sequences were compared, and pathogenic single nucleotide variants (SNVs) identified and validated by re-sequencing followed by pathway analysis. Additionally, tumor RNA isolated using Maxwell Promega was sequenced on Illumina instrument and the expression counts determined and normalized. RESULTS We found 314 pathogenic and/or highly deleterious mutations (both germline and somatic) with a median of 13 mutations per patient. Five patients had VHL syndrome (germline VHL mutation) and 4 carried somatic VHL mutations. Among the VHL tumors, 82 mutations were identified, including HNF1B, NOTCH1 and TCF7L1, suggesting a potential contribution of altered RNA metabolism based upon pathway analysis. Among all hemangioblastomas, germline growth factor receptor variants (FGFR4 p.G388R (14/23 (61%) patients), IGF1R, PDGFRA and TYK2) known to activate STAT3 signaling and induce HIF-1α and angiogenesis, were identified. Non-hierarchical clustering of RNA sequencing data revealed two transcriptionally-distinct subtypes of hemangioblastomas. CONCLUSIONS Our findings indicate that hemangioblastomas can also occur by germline mutations known to activate STAT3 signaling, which may have significant implication in genetic testing and counseling of patients with hemangioblastomas.
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- 2019
178. Platelet Transcriptome Profiling in HIV and ATP-Binding Cassette Subfamily C Member 4 (ABCC4) as a Mediator of Platelet Activity
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Adriana Heguy, Benjamin Kim, Nicole Allen, Rebecca Dann, Michael Cammer, Tilla S. Worgall, Tenzin Lhakhang, Aristotelis Tsirigos, Matthew Cambria, Emanuela Marcantoni, Jeffrey S. Berger, and Meagan O’Brien
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0301 basic medicine ,lcsh:Diseases of the circulatory (Cardiovascular) system ,CLINICAL RESEARCH ,PBS, phosphate-buffered saline ,PAH, pulmonary artery hypertension ,ATP-binding cassette transporter ,platelet activity ,ABCC4 ,CVD, cardiovascular disease ,HUVEC, human umbilical vein endothelial cell(s) ,RNA-Seq, RNA sequencing ,030204 cardiovascular system & hematology ,Pharmacology ,Proinflammatory cytokine ,Transcriptome ,ART, antiretroviral therapy ,03 medical and health sciences ,Cyclic nucleotide ,chemistry.chemical_compound ,0302 clinical medicine ,Mediator ,cardiovascular disease ,Medicine ,Platelet ,Platelet activation ,S1P, sphingosine-1-phosphate ,biology ,business.industry ,HIV ,IL, interleukin ,3. Good health ,cAMP, cyclic adenosine monophosphate ,HIV, human immunodeficiency virus ,030104 developmental biology ,chemistry ,lcsh:RC666-701 ,biology.protein ,qPCR, quantitative polymerase chain reaction ,RT, room temperature ,BSA, bovine serum albumin ,NSAID, nonsteroidal anti-inflammatory drug ,Cardiology and Cardiovascular Medicine ,business ,VASP, vasodilator-stimulated phosphoprotein ,ABCC4, ATP binding cassette subfamily C member 4 - Abstract
Visual Abstract, Highlights • Platelet activity and its effector cell properties are increased in persons with virologically suppressed HIV on antiretroviral therapy. • The platelet transcriptome is differentially expressed in participants with HIV compared with healthy individuals. • ABCC4 expression and translation was enhanced in HIV-infected subjects compared with healthy individuals. • ABCC4 is a membrane transporter that plays an important role in regulating several cardiovascular processes, including platelet activation and aggregation. • Platelet ABCC4 inhibition in HIV attenuated platelet activation and platelet effector cell function by regulating cyclic nucleotide homeostasis and the extrusion of platelet proinflammatory mediators., Summary An unbiased platelet transcriptome profile identified ATP binding cassette subfamily C member 4 (ABCC4) as a novel mediator of platelet activity in virologically suppressed human immunodeficiency virus (HIV)-infected subjects on antiretroviral therapy. Using ex vivo and in vitro cellular and molecular assays we demonstrated that ABCC4 regulated platelet activation by altering granule release and cyclic nucleotide homeostasis through a cAMP-protein kinase A (PKA)–mediated mechanism. Platelet ABCC4 inhibition attenuated platelet activation and effector cell function by reducing the release of inflammatory mediators, such as sphingosine-1-phosphate. ABCC4 inhibition may represent a novel antithrombotic strategy in HIV-infected subjects on antiretroviral therapy.
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- 2018
179. Gut Microbiota Perturbations in Reactive Arthritis and Postinfectious Spondyloarthritis
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Adriana Heguy, Luis R. Espinoza, Leopoldo N. Segal, Helga Raquel Garcia Ferrer, Joan M. Von Feldt, Jose C. Clemente, Isa Iraheta, Alexis Ogdie, Abraham Garcia Kutzbach, Julia Manasson, Nan Shen, Carles Ubeda, and Jose U. Scher
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Adult ,Male ,0301 basic medicine ,Adolescent ,Immunology ,Arthritis ,Disease ,Biology ,Gut flora ,medicine.disease_cause ,Arthritis, Reactive ,Article ,Feces ,Young Adult ,03 medical and health sciences ,0302 clinical medicine ,Rheumatology ,RNA, Ribosomal, 16S ,Prohibitins ,Spondylarthritis ,medicine ,Humans ,Immunology and Allergy ,Reactive arthritis ,Microbiome ,030203 arthritis & rheumatology ,HLA-A Antigens ,Campylobacter ,Case-control study ,Sacroiliitis ,Middle Aged ,medicine.disease ,biology.organism_classification ,Gastrointestinal Microbiome ,030104 developmental biology ,Case-Control Studies ,Female - Abstract
Objective Reactive arthritis (ReA) is an inflammatory disorder occurring several weeks after gastrointestinal or genitourinary infections. HLA-B27 positivity is considered a risk factor, though not necessarily predictive of disease incidence. Among non-genetic factors, the intestinal microbiome may play a role in disease susceptibility. The objective of this study was to characterize the gut microbiota and host gene interactions in ReA and post-infectious spondyloarthritis. Methods Adult peripheral spondyloarthritis and control subjects with preceding infections that did not develop arthritis were prospectively recruited from a highly prevalent geographic region. Clinical variables, HLA status, and 16S rRNA gene sequencing of intestinal microbiota were analyzed. Results ReA subjects showed no significant differences from controls in gut bacterial richness or diversity. However, there was a significantly higher abundance of Erwinia and Pseudomonas, and increased prevalence of typical enteropathogens associated with ReA. Subjects with ultrasound evidence of enthesitis were enriched in Campylobacter, while subjects with uveitis and radiographic sacroiliitis were respectively enriched in Erwinia and unclassified Ruminococcaceae, and both enriched in Dialister. Host genetics, particularly HLA-A24, were associated with differences in gut microbiota diversity irrespective of disease status. We determined several co-occurring taxa that were also predictive of HLA-A24 status. Conclusion This is the first culture-independent study characterizing the gut microbial community of post-infectious arthritis. Although bacterial factors correlated with disease presence and clinical features of ReA, host genetics also appeared to be a major independent driver of intestinal community composition. Understanding of these gut microbiota host-genetic relationships may further clarify the pathogenesis of post-infectious spondyloarthropathies. This article is protected by copyright. All rights reserved.
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- 2018
180. Keap1 loss promotes Kras-driven lung cancer and results in dependence on glutaminolysis
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Angela Davies, Sarah E. LeBoeuf, Francisco J. Sánchez-Rivera, Triantafyllia R. Karakousi, Igor Dolgalev, Tyler Jacks, Matthew G. Vander Heiden, Volkan I. Sayin, Adriana Heguy, Justin R. Prigge, Lakshmipriya Subbaraj, Donald C Ellis, John T. Poirier, Edward E. Schmidt, Roderick T. Bronson, Arjun Bhutkar, Craig J. Thomas, Rodrigo Romero, Viola Allaj, Shawn M. Davidson, Andre L. Moreira, Britney Martinez, Thales Papagiannakopoulos, Matthew R. Bauer, Chandra Goparaju, Harvey I. Pass, Simranjit X. Singh, and Charles M. Rudin
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0301 basic medicine ,Lung Neoplasms ,Glutamine ,Adenocarcinoma of Lung ,Adenocarcinoma ,Biology ,medicine.disease_cause ,Article ,General Biochemistry, Genetics and Molecular Biology ,Mice ,03 medical and health sciences ,Glutaminase ,medicine ,Animals ,Humans ,Clustered Regularly Interspaced Short Palindromic Repeats ,Lung cancer ,Kelch-Like ECH-Associated Protein 1 ,Glutaminolysis ,Hydrolysis ,General Medicine ,respiratory system ,medicine.disease ,KEAP1 ,digestive system diseases ,NFE2L2 ,respiratory tract diseases ,3. Good health ,Genes, ras ,030104 developmental biology ,Immunology ,Cancer research ,KRAS ,Carcinogenesis - Abstract
Treating KRAS-mutant lung adenocarcinoma (LUAD) remains a major challenge in cancer treatment given the difficulties associated with directly inhibiting the KRAS oncoprotein. One approach to addressing this challenge is to define mutations that frequently co-occur with those in KRAS, which themselves may lead to therapeutic vulnerabilities in tumors. Approximately 20% of KRAS-mutant LUAD tumors carry loss-of-function mutations in the KEAP1 gene encoding Kelch-like ECH-associated protein 1 (refs. 2, 3, 4), a negative regulator of nuclear factor erythroid 2-like 2 (NFE2L2; hereafter NRF2), which is the master transcriptional regulator of the endogenous antioxidant response. The high frequency of mutations in KEAP1 suggests an important role for the oxidative stress response in lung tumorigenesis. Using a CRISPR-Cas9-based approach in a mouse model of KRAS-driven LUAD, we examined the effects of Keap1 loss in lung cancer progression. We show that loss of Keap1 hyperactivates NRF2 and promotes KRAS-driven LUAD in mice. Through a combination of CRISPR-Cas9-based genetic screening and metabolomic analyses, we show that Keap1- or Nrf2-mutant cancers are dependent on increased glutaminolysis, and this property can be therapeutically exploited through the pharmacological inhibition of glutaminase. Finally, we provide a rationale for stratification of human patients with lung cancer harboring KRAS/KEAP1- or KRAS/NRF2-mutant lung tumors as likely to respond to glutaminase inhibition.
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- 2017
181. A Dramatic Difference in Global Gene Expression between TCDD-Treated Atlantic Tomcod Larvae from the Resistant Hudson River and a Nearby Sensitive Population
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Isaac Wirgin, Adriana Heguy, A. Goradia, Y. L. Wang, K. Vitale, Stuart M. Brown, Yuhan Hao, Paul Zappile, Nirmal K. Roy, R. C. Chambers, and Hao Chen
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0301 basic medicine ,environmental toxicants ,Population ,Zoology ,RNA-Seq ,010501 environmental sciences ,Biology ,01 natural sciences ,Transcriptome ,03 medical and health sciences ,Genetics ,education ,Gene ,Ecology, Evolution, Behavior and Systematics ,0105 earth and related environmental sciences ,geography ,Larva ,education.field_of_study ,geography.geographical_feature_category ,draft genome ,Ecology ,aryl hydrocarbon receptor ,Estuary ,030104 developmental biology ,bioaccumulation ,Bioaccumulation ,Bay ,transcriptome ,Research Article - Abstract
Atlantic tomcod in the Hudson River Estuary bioaccumulate high hepatic burdens of environmental toxicants. Previously, we demonstrated that Hudson River tomcod developed resistance to TCDD and PCB toxicity probably through strong natural selection during their early life-stages for a variant of the Aryl Hydrocarbon Receptor2 (AHR2). Here, we evaluated the genomic consequences of the resistant genotype by comparing global gene expression in larval tomcod from the Hudson River with expression in larvae from a nearby sensitive population (Shinnecock Bay). We developed an annotated draft tomcod genome to explore the effects of multigenerational exposure to toxicants and a functionally impaired AHR2 on the transcriptome. We used the tomcod genome as a reference in RNA-Seq to compare global gene expression in tomcod larvae from the Hudson River and Shinnecock Bay after experimental exposure of larvae to graded doses of TCDD. We found dramatic differences between offspring from the two populations in the number of genes that were differentially expressed at all doses (0.01, 0.1, and 1 ppb) and even in the vehicle controls. At the two lowest TCDD doses, 250 and 1,141 genes were differentially expressed in Shinnecock Bay larvae compared with 14 and 12, respectively, in Hudson River larvae. At the highest dose (1.0 ppb), 934 genes were differentially expressed in Shinnecock Bay larvae and 173 in Hudson River larvae, but only 28 (16%) of affected genes were shared among both populations. Given the large difference between the two populations in the number and identity of differentially expressed genes, it is likely that the AHR2 pathway interacts directly or indirectly with many genes beyond those known in the AHR2 battery and that other regulatory systems may also respond to TCDD exposure. The effects of chronic multi-generational exposure to environmental toxicants on the genome of Hudson River tomcod are much greater than previously expected.
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- 2017
182. Plakophilin-2 is required for transcription of genes that control calcium cycling and cardiac rhythm
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Adriana Heguy, Marina Cerrone, Eli Rothenberg, Joanne J.A. Van Bavel, Esperanza Agullo-Pascual, Mingliang Zhang, Alejandra Leo-Macias, Kabir Malkani, Hua Qian Yang, Jérôme Montnach, Thomas V. Karathanos, William A. Coetzee, Yan-Ting Zhao, Gregory E. Morley, Héctor H. Valdivia, Feng-Xia Liang, Natalia A. Trayanova, Michael J. Ackerman, Xianming Lin, Francisco J. Alvarado, David J. Tester, Toon A.B. van Veen, Carolina Vasquez, Mario Delmar, Chantal J. M. van Opbergen, Steven J. Fowler, and Igor Dolgalev
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0301 basic medicine ,medicine.medical_specialty ,Chemistry(all) ,Transcription, Genetic ,Science ,Blotting, Western ,General Physics and Astronomy ,Gene Expression ,030204 cardiovascular system & hematology ,Biology ,Physics and Astronomy(all) ,Ryanodine receptor 2 ,Biochemistry ,General Biochemistry, Genetics and Molecular Biology ,Calcium in biology ,Article ,03 medical and health sciences ,0302 clinical medicine ,Desmosome ,ANK2 ,Internal medicine ,Gene expression ,medicine ,Animals ,Humans ,Myocytes, Cardiac ,Calcium metabolism ,Mice, Knockout ,Multidisciplinary ,Microscopy, Confocal ,Biochemistry, Genetics and Molecular Biology(all) ,Reverse Transcriptase Polymerase Chain Reaction ,Myocardium ,Arrhythmias, Cardiac ,Heart ,General Chemistry ,3. Good health ,Cell biology ,Mice, Inbred C57BL ,030104 developmental biology ,Endocrinology ,medicine.anatomical_structure ,Triadin ,Knockout mouse ,Calcium ,Plakophilins ,Genetics and Molecular Biology(all) - Abstract
Plakophilin-2 (PKP2) is a component of the desmosome and known for its role in cell–cell adhesion. Mutations in human PKP2 associate with a life-threatening arrhythmogenic cardiomyopathy, often of right ventricular predominance. Here, we use a range of state-of-the-art methods and a cardiomyocyte-specific, tamoxifen-activated, PKP2 knockout mouse to demonstrate that in addition to its role in cell adhesion, PKP2 is necessary to maintain transcription of genes that control intracellular calcium cycling. Lack of PKP2 reduces expression of Ryr2 (coding for Ryanodine Receptor 2), Ank2 (coding for Ankyrin-B), Cacna1c (coding for CaV1.2) and Trdn (coding for triadin), and protein levels of calsequestrin-2 (Casq2). These factors combined lead to disruption of intracellular calcium homeostasis and isoproterenol-induced arrhythmias that are prevented by flecainide treatment. We propose a previously unrecognized arrhythmogenic mechanism related to PKP2 expression and suggest that mutations in PKP2 in humans may cause life-threatening arrhythmias even in the absence of structural disease., It is believed that mutations in desmosomal adhesion complex protein plakophilin 2 (PKP2) cause arrhythmia due to loss of cell-cell communication. Here the authors show that PKP2 controls the expression of proteins involved in calcium cycling in adult mouse hearts, and that lack of PKP2 can cause arrhythmia in a structurally normal heart.
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- 2017
183. Author response: Axon TRAP reveals learning-associated alterations in cortical axonal mRNAs in the lateral amygdala
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Robert M. Sears, Tenzin Lhakhang, Linnaea E. Ostroff, Adriana Heguy, Rahul N. Kanadia, Aristotelis Tsirigos, Joseph E. LeDoux, Eric Klann, Zachary Deane, and Emanuela Santini
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Trap (computing) ,medicine.anatomical_structure ,medicine ,Axon ,Biology ,Amygdala ,Neuroscience - Published
- 2019
184. Disruption of Ca
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Joon-Chul, Kim, Marta, Pérez-Hernández, Francisco J, Alvarado, Svetlana R, Maurya, Jerome, Montnach, Yandong, Yin, Mingliang, Zhang, Xianming, Lin, Carolina, Vasquez, Adriana, Heguy, Feng-Xia, Liang, Sun-Hee, Woo, Gregory E, Morley, Eli, Rothenberg, Alicia, Lundby, Hector H, Valdivia, Marina, Cerrone, and Mario, Delmar
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Mice, Knockout ,Desmosomes ,Article ,Disease Models, Animal ,Mice ,Connexin 43 ,Mutation ,Animals ,Homeostasis ,Humans ,Calcium ,Myocytes, Cardiac ,Calcium Signaling ,Plakophilins ,Arrhythmogenic Right Ventricular Dysplasia ,Cells, Cultured - Abstract
BACKGROUND: Plakophilin-2 (PKP2) is classically defined as a desmosomal protein. Mutations in PKP2 associate with most cases of gene-positive arrhythmogenic right ventricular cardiomyopathy (ARVC). A better understanding of PKP2 cardiac biology can help elucidate the mechanisms underlying arrhythmic and cardiomyopathic events consequent to PKP2 deficiency. Here, we sought to capture early molecular/cellular events that can act as nascent arrhythmic/cardiomyopathic substrates. METHODS: We used multiple imaging, biochemical and high-resolution mass spectrometry methods to study functional/structural properties of cells/tissues derived from cardiomyocyte-specific, tamoxifen-activated, PKP2 knockout mice (“PKP2cKO”) 14 days post-tamoxifen (post-TAM) injection, a time point preceding overt electrical or structural phenotypes. Myocytes from right or left ventricular free wall were studied separately. RESULTS: Most properties of PKP2cKO left ventricular (PKP2cKO-LV) myocytes were not different from control; in contrast, PKP2cKO right ventricular (PKP2cKO-RV) myocytes showed increased amplitude and duration of Ca(2+) transients, increased [Ca(2+)] in the cytoplasm and sarcoplasmic reticulum (SR), increased frequency of spontaneous Ca(2+) release events (sparks) even at comparable SR load, and dynamic Ca(2+) accumulation in mitochondria. We also observed early- and delayed-after transients in RV myocytes and heightened susceptibility to arrhythmias in Langendorff-perfused hearts. In addition, RyR2 in PKP2cKO-RV cells presented enhanced Ca(2+) sensitivity and preferential phosphorylation in a domain known to modulate Ca(2+) gating. RNAseq at 14 days post-TAM showed no relevant difference in transcript abundance between RV and LV, neither in control nor in PKP2cKO cells. Instead, we found an RV-predominant increase in membrane permeability that can permit Ca(2+) entry into the cell. Cx43 ablation mitigated the membrane permeability increase, accumulation of cytoplasmic Ca(2+), increased frequency of sparks and early stages of RV dysfunction. Cx43 hemichannel block with GAP19 normalized [Ca(2+)](i) homeostasis. Similarly, PKC inhibition normalized spark frequency at comparable SR load levels. CONCLUSIONS: Loss of PKP2 creates an RV-predominant arrhythmogenic substrate (Ca(2+) dysregulation) that precedes the cardiomyopathy; this is, at least in part, mediated by a Cx43-dependent membrane conduit and repressed by PKC inhibitors. Given that asymmetric Ca(2+) dysregulation precedes the cardiomyopathic stage, we speculate that abnormal Ca(2+) handling in RV myocytes can be a trigger for gross structural changes observed at a later stage.
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- 2019
185. Revisiting multifocal breast cancer: a clonality study of ductal carcinoma using whole exome sequencing
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Matija Snuderl, Farbod Darvishian, Varshini Vasudevaraja, Paolo Cotzia, Adriana Heguy, Christopher J Schwartz, George Jour, Stephen Kelly, and Igor Dolgalev
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0301 basic medicine ,Pathology ,medicine.medical_specialty ,Multifocal breast cancer ,Focus (geometry) ,Breast Neoplasms ,Biology ,Pathology and Forensic Medicine ,Pathogenesis ,Neoplasms, Multiple Primary ,03 medical and health sciences ,0302 clinical medicine ,Predictive Value of Tests ,Exome Sequencing ,polycyclic compounds ,medicine ,Biomarkers, Tumor ,Humans ,Genetic Predisposition to Disease ,Neoplasm Invasiveness ,skin and connective tissue diseases ,Gene ,Exome sequencing ,Laser capture microdissection ,Aged ,Nodal metastasis ,Carcinoma, Ductal, Breast ,Ductal carcinoma ,Middle Aged ,Prognosis ,Clone Cells ,030104 developmental biology ,Phenotype ,030220 oncology & carcinogenesis ,Lymphatic Metastasis ,Mutation ,Female - Abstract
Multifocal breast cancer (MFBC), ductal type, has been hypothesized to arise by one of two mechanisms: either through intramammary/intralymphatic spread from a single index tumor (MBC-1), or as multiple independent tumors with each focus carrying its corresponding ductal carcinoma in-situ (MBC-2). In order to improve our understanding of MFBC pathogenesis, we employed laser capture microdissection coupled with whole-exome sequencing to study clonal origin in MFBC. We selected three cases of MBC-1 (C1 to C3) and MBC-2 (C4 to C6) and analyzed three foci from each case. MBC-1 cases were histologically similar and showed a strong predilection for satellite foci, vascular invasion and nodal metastasis when compared to MBC-2. Our bioinformatics approach provided strong evidence for clonal relationships in MBC-1, as demonstrated by distinct clusters of genes conserved across all tumor foci. Conversely, no gene clusters were shared across all the foci in MBC-2, suggesting multiple independent tumors. These findings provide further support for the two distinct pathogenetic mechanisms in MFBC.
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- 2019
186. Near full genome characterization of HIV‐1 unique recombinant forms in Cameroon reveals dominant CRF02_AG and F2 recombination patterns
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Ralf Duerr, Andrew N. Banin, I. O. Okonko, Jude S. Bimela, Dora Mbanya, Aubin Nanfack, Paul Zappile, Xiao-Hong Wang, Adriana Heguy, Jeanne Ngogang, Alireza Khodadadi-Jamayran, Marcel Tongo, Josephine Meli, Michael Tuen, Charles Fokunang, and Miroslaw K. Gorny
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Anti-HIV Agents ,Context (language use) ,HIV Infections ,Genome, Viral ,Genome ,Polymerase Chain Reaction ,DNA sequencing ,03 medical and health sciences ,0302 clinical medicine ,Genotype ,HIV Seropositivity ,Medicine ,Humans ,030212 general & internal medicine ,Cameroon ,Longitudinal Studies ,Clade ,Env epitopes and drug resistance mutations ,Research Articles ,Phylogeny ,Whole genome sequencing ,Genetics ,030505 public health ,Phylogenetic tree ,business.industry ,Public Health, Environmental and Occupational Health ,virus diseases ,unique recombinant forms ,third‐generation sequencing ,3. Good health ,intra‐patient viral diversity ,Infectious Diseases ,Mutation ,HIV-1 ,Female ,Primer (molecular biology) ,0305 other medical science ,business ,near full genome sequencing ,Reassortant Viruses ,Research Article - Abstract
Introduction In Cameroon, a manifold diversity of HIV strains exists with CRF02_AG and unique recombinant forms (URFs) being the predominant strains. In recent years, a steady increase in URFs and clade F2 viruses has been monitored through partial genome sequencing. There is an information gap in the characterization of emerging URFs along the full genome, which is needed to address the challenges URFs pose towards diagnosis, treatment and HIV‐1 vaccine design. Method Eighteen Cameroonian URFs from samples collected between the years 2000 and 2015 were studied using a newly developed near full genome sequencing (NFGS) protocol based on variable nested RT‐PCRs with a versatile primer set. Near full genomes were characterized for recombination patterns and sequence signatures with possible impact on antiretroviral treatment or Env‐directed immune responses. Third‐generation sequencing (3GS) of near full or half genomes (HGs) gave insight into intra‐patient URF diversity. Results The characterized URFs were composed of a broad variety of subtypes and recombinants including A, F, G, CRF01_AE, CRF02_AG and CRF22_01A1. Phylogenetic analysis unveiled dominant CRF02_AG and F2 recombination patterns. 3GS indicated a high intra‐patient URF diversity with up to four distinct viral sub‐populations present in plasma at the same time. URF pol genomic analysis revealed a number of accessory drug resistance mutations (DRMs) in the ART‐naïve participants. Genotypic env analysis suggests CCR5 usage in 14/18 samples and identified deviations at residues, critical for gp120/gp41 interphase and CD4 binding site broadly neutralizing antibodies in more than half of the studied URFs. V1V2 sites of immune pressure in the human RV144 vaccine study varied in more than a third of URFs. Conclusions This study identified novel mosaic patterns in URFs in Cameroon. In line with the regional predominance of CRF_02AG and the increased prevalence of clade F2, prominent CRF_02AG and F2 background patterns were observed underlying the URFs. In the context of the novel mosaic genomes, the impact of the identified accessory DRMs and Env epitope variations on treatment and immune control remains elusive. The evolving diversity of HIV‐1 URFs in Cameroon requires continuous monitoring to respond to the increasing challenges for diagnosis, antiretroviral treatment and prevention.
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- 2019
187. Transcriptomic profiles conducive to immune-mediated tumor rejection in human breast cancer skin metastases treated with Imiquimod
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Davide Bedognetti, Cynthia A. Loomis, Adriana Heguy, Sylvia Adams, Luis Chiriboga, Karina Ray, Sandra Demaria, Farbod Darvishian, Francesco M. Marincola, Mikala Egeblad, Wouter Hendrickx, and Mariya Rozenblit
- Subjects
Adult ,0301 basic medicine ,Skin Neoplasms ,Antigen presentation ,lcsh:Medicine ,Breast Neoplasms ,Imiquimod ,CD8-Positive T-Lymphocytes ,Article ,03 medical and health sciences ,Breast cancer ,0302 clinical medicine ,Immune system ,medicine ,Humans ,Cytotoxic T cell ,Basal cell carcinoma ,Neoplasm Metastasis ,lcsh:Science ,Aged ,Immunity, Cellular ,Multidisciplinary ,business.industry ,Gene Expression Profiling ,Melanoma ,lcsh:R ,Cancer ,Diagnostic markers ,Middle Aged ,Th1 Cells ,medicine.disease ,030104 developmental biology ,Cancer research ,lcsh:Q ,Female ,business ,030217 neurology & neurosurgery ,medicine.drug - Abstract
Imiquimod is a topical toll-like-receptor-7 agonist currently used for treating basal cell carcinoma. Recently, imiquimod has demonstrated tumor regression in melanoma and breast cancer skin metastases. However, the molecular perturbations induced by imiquimod in breast cancer metastases have not been previously characterized. Here, we describe transcriptomic profiles associated with responsiveness to imiquimod in breast cancer skin metastases. Baseline and post-treatment tumor samples from patients treated with imiquimod in a clinical trial were profiled using Nanostring technology. Through an integrative analytic pipeline, we showed that tumors from patients who achieved a durable clinical response displayed a permissive microenvironment, substantiated by the upregulation of transcripts encoding for molecules involved in leukocyte adhesion and migration, cytotoxic functions, and antigen presentation. In responding patients, Imiquimod triggered a strong T-helper-1 (Th-1)/cytotoxic immune response, characterized by the coordinated upregulation of Th-1 chemokines, migration of Th-1 and cytotoxic T cells into the tumor, and activation of immune-effector functions, ultimately mediating tumor destruction. In conclusion, we have shown that topical imiquimod can induce a robust immune response in breast cancer metastases, and this response is more likely to occur in tumors with a pre-activated microenvironment. In this setting, imiquimod could be utilized in combination with other targeted immunotherapies to increase therapeutic efficacy.
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- 2019
188. Lower Airway Dysbiosis Leads to A Pro-Tumor Inflammatory State and Worsens Non-Small Cell Lung Cancer Prognosis in a Preclinical Model
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K. Yie, Kwok K. Wong, S. Ma, L. Perez, Peter Meyn, Benjamin G. Wu, Harvey I. Pass, E. Olsen, Adriana Heguy, Yi Li, Mariam El-Ashmawy, Thales Papagiannakopoulos, Ting-An Yie, Imran Sulaiman, Daniel H. Sterman, Jun-Chieh J. Tsay, Leopoldo N. Segal, B. Franca, and A. Gonzalez
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business.industry ,medicine ,Cancer research ,Non small cell ,medicine.disease ,Lung cancer ,Airway ,business ,Dysbiosis - Published
- 2019
189. Experimental and pan-cancer genome analyses reveal widespread contribution of acrylamide exposure to carcinogenesis in humans
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James McKay, Vincent Cahais, Michael Korenjak, Claire Renard, Adriana Heguy, Monica Hollstein, Liacine Bouaoun, Alexis Robitaille, Stephanie Villar, Martha R. Stampfer, Maude Ardin, Steven G. Rozen, Maria Zhivagui, Mona I. Churchwell, Kathryn Z. Guyton, Alvin Wei Tian Ng, Magali Olivier, Jiri Zavadil, Manuraj Pandey, and Frederick A. Beland
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Carcinogenesis ,medicine.disease_cause ,Genome ,Medical and Health Sciences ,Tobacco smoke ,chemistry.chemical_compound ,Mice ,0302 clinical medicine ,Neoplasms ,2.2 Factors relating to the physical environment ,2.1 Biological and endogenous factors ,Aetiology ,Lung ,Cells, Cultured ,Genetics (clinical) ,Cancer ,0303 health sciences ,Mutation ,Cultured ,Liver Disease ,Lung Cancer ,Environmental exposure ,Biological Sciences ,Acrylamide ,Human ,Bioinformatics ,Cells ,Biology ,03 medical and health sciences ,Rare Diseases ,Tobacco ,medicine ,Genetics ,Animals ,Humans ,Carcinogen ,030304 developmental biology ,Acrylamides ,Tobacco Smoke and Health ,Genome, Human ,Research ,Prevention ,Human Genome ,Environmental Exposure ,medicine.disease ,chemistry ,Cancer research ,Epoxy Compounds ,Tumor Suppressor Protein p53 ,Digestive Diseases ,030217 neurology & neurosurgery ,Mutagens - Abstract
Humans are frequently exposed to acrylamide, a probable human carcinogen found in commonplace sources such as most heated starchy foods or tobacco smoke. Prior evidence has shown that acrylamide causes cancer in rodents, yet epidemiological studies conducted to date are limited and, thus far, have yielded inconclusive data on association of human cancers with acrylamide exposure. In this study, we experimentally identify a novel and unique mutational signature imprinted by acrylamide through the effects of its reactive metabolite glycidamide. We next show that the glycidamide mutational signature is found in a full one-third of approximately 1600 tumor genomes corresponding to 19 human tumor types from 14 organs. The highest enrichment of the glycidamide signature was observed in the cancers of the lung (88% of the interrogated tumors), liver (73%), kidney (>70%), bile duct (57%), cervix (50%), and, to a lesser extent, additional cancer types. Overall, our study reveals an unexpectedly extensive contribution of acrylamide-associated mutagenesis to human cancers.
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- 2019
190. Radiotherapy enhances responses of lung cancer to CTLA-4 blockade
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Abraham Chachoua, Adriana Heguy, Silvia C. Formenti, Erik Wennerberg, Kai W. Wucherpfennig, Tuo Zhang, Naoko Imai, Sacha Gnjatic, Encouse B. Golden, Xi Kathy Zhou, Kent Friedman, Benjamin T. Cooper, Sandra Demaria, Ryan O. Emerson, Nils Rudqvist, Claire Lhuillier, Lucas Ferrari de Andrade, and Claire Vanpouille-Box
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0301 basic medicine ,Oncology ,Male ,Cancer Research ,Lung Neoplasms ,medicine.medical_treatment ,CD8-Positive T-Lymphocytes ,0302 clinical medicine ,Non-small cell lung cancer ,Interferon ,Cytotoxic T cell ,Immunology and Allergy ,Medicine ,CTLA-4 Antigen ,Aged, 80 and over ,Abscopal effect ,Antibodies, Monoclonal ,General Medicine ,Middle Aged ,lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,Combined Modality Therapy ,3. Good health ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Molecular Medicine ,Female ,medicine.drug ,medicine.medical_specialty ,T cell ,Immunology ,Ipilimumab ,CTLA-4 blockade ,lcsh:RC254-282 ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,Immune system ,Internal medicine ,Cell Line, Tumor ,Humans ,Lung cancer ,Aged ,Pharmacology ,Radiotherapy ,business.industry ,Immunotherapy ,medicine.disease ,Blockade ,Radiation therapy ,030104 developmental biology ,CTLA-4 ,Drug Resistance, Neoplasm ,Cancer research ,Commentary ,business - Abstract
Focal radiation therapy enhances systemic responses to anti-CTLA-4 antibodies in preclinical studies and in some patients with melanoma1-3, but its efficacy in inducing systemic responses (abscopal responses) against tumors unresponsive to CTLA-4 blockade remained uncertain. Radiation therapy promotes the activation of anti-tumor T cells, an effect dependent on type I interferon induction in the irradiated tumor4-6. The latter is essential for achieving abscopal responses in murine cancers6. The mechanisms underlying abscopal responses in patients treated with radiation therapy and CTLA-4 blockade remain unclear. Here we report that radiation therapy and CTLA-4 blockade induced systemic anti-tumor T cells in chemo-refractory metastatic non-small-cell lung cancer (NSCLC), where anti-CTLA-4 antibodies had failed to demonstrate significant efficacy alone or in combination with chemotherapy7,8. Objective responses were observed in 18% of enrolled patients, and 31% had disease control. Increased serum interferon-β after radiation and early dynamic changes of blood T cell clones were the strongest response predictors, confirming preclinical mechanistic data. Functional analysis in one responding patient showed the rapid in vivo expansion of CD8 T cells recognizing a neoantigen encoded in a gene upregulated by radiation, supporting the hypothesis that one explanation for the abscopal response is radiation-induced exposure of immunogenic mutations to the immune system.
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- 2019
191. Draft Genome Sequence of Streptococcus halitosis sp. nov., Isolated from the Dorsal Surface of the Tongue of a Patient with Halitosis
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Adriana Heguy, Igor Dolgalev, Aristotelis Tsirigos, Paul Zappile, Daria Vikina, George Tetz, Stuart M. Brown, and Victor Tetz
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Whole genome sequencing ,0303 health sciences ,030306 microbiology ,Streptococcus ,Genome Sequences ,Virulence ,Biology ,biology.organism_classification ,medicine.disease_cause ,Genome ,Microbiology ,03 medical and health sciences ,medicine.anatomical_structure ,Immunology and Microbiology (miscellaneous) ,Tongue ,Genetics ,medicine ,Molecular Biology ,Gene ,GC-content ,Bacteria ,030304 developmental biology - Abstract
Here, we report the draft genome of Streptococcus halitosis sp. nov. strain VT-4, a novel bacterium isolated from the dorsal part of the tongue of a patient with halitosis. The genome comprised 1,880,608 bp with a GC content of 41.0%., Here, we report the draft genome of Streptococcus halitosis sp. nov. strain VT-4, a novel bacterium isolated from the dorsal part of the tongue of a patient with halitosis. The genome comprised 1,880,608 bp with a GC content of 41.0%. There were 1,782 predicted protein-coding genes, including those associated with virulence and antibiotic resistance.
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- 2019
192. Correction: The fecal, oral, and skin microbiota of children with Chagas disease treated with benznidazole
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Adriana Heguy, Marina Hoashi, Carlos Robello, Valentina Montacutti, Doris Patricia Maldonado, Anna Hevia, Igor Dolgalev, Maria Gloria Dominguez-Bello, Paola Frattaroli, Gregorio Iraola, and Maricruz Mojica
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Chagas disease ,Multidisciplinary ,business.industry ,lcsh:R ,lcsh:Medicine ,medicine.disease ,Benznidazole ,Immunology ,Medicine ,lcsh:Q ,business ,lcsh:Science ,Feces ,medicine.drug - Abstract
[This corrects the article DOI: 10.1371/journal.pone.0212593.].
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- 2019
193. The fecal, oral, and skin microbiota of children with Chagas disease treated with benznidazole
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Adriana Heguy, Gregorio Iraola, Doris Patricia Maldonado, Valentina Montacutti, Carlos Robello, Anna Hevia, Maricruz Mojica, Maria Gloria Dominguez-Bello, Paola Frattaroli, Marina Hoashi, and Igor Dolgalev
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0301 basic medicine ,Bacterial Diseases ,Male ,Rural Population ,Physiology ,Skin infection ,medicine.disease_cause ,Pediatrics ,Feces ,0302 clinical medicine ,fluids and secretions ,Medicine and Health Sciences ,Child ,Skin ,Protozoans ,Trypanosoma Cruzi ,Multidisciplinary ,biology ,Streptococcus ,Microbiota ,Eukaryota ,Genomics ,Infectious Diseases ,Benznidazole ,Medical Microbiology ,Nitroimidazoles ,Child, Preschool ,Medicine ,Female ,Roseburia ,Pediatric Infections ,medicine.drug ,Research Article ,Neglected Tropical Diseases ,Chagas disease ,Skin Infections ,Trypanosoma ,Bolivia ,Adolescent ,Science ,030231 tropical medicine ,Microbial Genomics ,Dermatology ,Microbiology ,03 medical and health sciences ,Streptococcal Infections ,Prevotella Infection ,medicine ,Genetics ,Parasitic Diseases ,Humans ,Chagas Disease ,Microbiome ,Protozoan Infections ,Bacteria ,business.industry ,Organisms ,Mouth Mucosa ,Biology and Life Sciences ,Correction ,medicine.disease ,biology.organism_classification ,Tropical Diseases ,Parasitic Protozoans ,stomatognathic diseases ,030104 developmental biology ,business ,Dysbiosis - Abstract
BackgroundChagas disease is still prevalent in rural areas of South America. In endemic areas of Bolivia, school children are screened for the program of Chagas disease eradication of the Ministry of Health, and positive children are treated. Here, we compared the fecal, oral and skin microbiomes of children with or without Chagas disease, and before and after benznidazol treatment of infected children.MethodsA total of 543 Bolivian children (5-14 years old) were tested for Chagas disease, and 20 positive children were treated with Benznidazole. Fecal samples and oral and skin swabs were obtained before and after treatment, together with samples from a group of 35 uninfected controls. The 16S rRNA genes were sequenced and analyzed using QIIME to determine Alpha diversity differences and community distances, and linear discriminant analyses to determine marker taxa by infection status or treatment.ResultsTwenty out of 543 children screened were seropositive for Chagas disease (3.7%) and were included in the study, together with 35 control children that were seronegative for the disease. Fecal samples, oral and skin swabs were taken at the beginning of the study and after the anti-protozoa therapy with Benznidazole to the chagasic children. Infected children had higher fecal Firmicutes (Streptococcus, Roseburia, Butyrivibrio, and Blautia), and lower Bacteroides and also showed some skin -but not oral- microbiota differences. Treatment eliminated the fecal microbiota differences from control children, increasing Dialister (class Clostridia) and members of the Enterobacteriaceae, and decreasing Prevotella and Coprococcus, with minor effects on the oral and skin bacterial diversity.ConclusionsThe results of this study show differences in the fecal microbiota associated with Chagas disease in children, and also evidence that treatment normalizes fecal microbiota (makes it more similar to that in controls), but is associated with oral and skin microbiota differences from control children. Since microbiota impacts in children, it is important to determine the effect of drugs on the children microbiota, since dysbiosis could lead to physiological effects which might be avoidable with microbiota restoration interventions.
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- 2019
194. The translatome of adult cortical axons is regulated by learning in vivo
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Adriana Heguy, Linnaea E. Ostroff, Joseph E. LeDoux, Emanuela Santini, Tenzin Lhakhang, Eric Klann, Zachary Deane, Aristotelis Tsirigos, and Robert M. Sears
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0303 health sciences ,Translation (biology) ,Biology ,Amygdala ,Cortex (botany) ,03 medical and health sciences ,0302 clinical medicine ,medicine.anatomical_structure ,nervous system ,Synaptic plasticity ,Forebrain ,medicine ,Protein biosynthesis ,Memory consolidation ,Cytoskeleton ,Neuroscience ,030217 neurology & neurosurgery ,030304 developmental biology - Abstract
SummaryLocal translation can support memory consolidation by supplying new proteins to synapses undergoing plasticity. Translation in adult forebrain dendrites is an established mechanism of synaptic plasticity and is regulated by learning, yet there is no evidence for learning-regulated protein synthesis in adult forebrain axons, which have traditionally been believed to be incapable of translation. Here we show that axons in the adult rat amygdala contain translation machinery, and use translating ribosome affinity purification (TRAP) with RNASeq to identify mRNAs in cortical axons projecting to the amygdala, over 1200 of which were regulated during consolidation of associative memory. Mitochondrial and translation-related genes were upregulated, whereas synaptic, cytoskeletal, and myelin-related genes were downregulated; the opposite effects were observed in the cortex. Our results demonstrate that learning-regulated axonal translation occurs in the adult forebrain, and support the likelihood that local translation is more a rule than an exception in neuronal processes.
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- 2018
195. Huntington’s Disease Protein Huntingtin Associates with its own mRNA
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Brady P. Culver, Man-Shan Yu, Igor Dolgalev, Josh DeClercq, Adriana Heguy, Naoko Tanese, and Bin Ma
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Research Report ,0301 basic medicine ,Untranslated region ,congenital, hereditary, and neonatal diseases and abnormalities ,RNA-protein interaction ,mRNA translation ,Huntingtin ,animal diseases ,Mice, Transgenic ,RNA-binding protein ,Biology ,Cell Line ,Mice ,03 medical and health sciences ,Cellular and Molecular Neuroscience ,Exon ,huntingtin exon 1 ,mental disorders ,Huntingtin Protein ,Animals ,RNA, Messenger ,Fmrp ,Visual Cortex ,Messenger RNA ,Sequence Analysis, RNA ,Intron ,RNA ,Molecular biology ,nervous system diseases ,3. Good health ,Huntington Disease ,030104 developmental biology ,nervous system ,Neurology (clinical) ,Huntington’s disease - Abstract
Background: The Huntington’s disease (HD) protein huntingtin (Htt) plays a role in multiple cellular pathways. Deregulation of one or more of these pathways by the mutant Htt protein has been suggested to contribute to the disease pathogenesis. Our recent discovery-based proteomics studies have uncovered RNA binding proteins and translation factors associated with the endogenous Htt protein purified from mouse brains, suggesting a potential new role for Htt in RNA transport and translation. Objective: To investigate how Htt might affect RNA metabolism we set out to purify and analyze RNA associated with Htt. Methods: RNA was extracted from immunopurified Htt-containing protein complexes and analyzed by microarrays and RNA-Seq. Results: Surprisingly, the most enriched mRNA that co-purified with Htt was Htt mRNA itself. The association of Htt protein and Htt mRNA was detected independent of intact ribosomes suggesting that it is not an RNA undergoing translation. Furthermore, we identified the recently reported mis-spliced Htt mRNA encoding a truncated protein comprised of exon 1 and a portion of the downstream intron in the immunoprecipitates containing mutant Htt protein. We show that Htt protein co-localizes with Htt mRNA and that wild-type Htt reduces expression of a reporter construct harboring the Htt 3’ UTR. Conclusions: HD protein is found in a complex with its own mRNA and RNA binding proteins and translation factors. Htt may be involved in modulating its expression through post-transcriptional pathways. It is possible that Htt shares mechanistic properties similar to RNA binding proteins such as TDP-43 and FUS implicated in other neurodegenerative diseases.
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- 2016
196. Abstract P1-06-06: Mammary stem cell modulation of wildtype and Trp53 null stem cells by CAPE (caffeic acid phenethyl ester), a potential therapeutic agent
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Adriana Heguy, Kurunthachalam Kannan, M Patel, Coral Omene, and Mary Helen Barcellos-Hoff
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Cancer Research ,biology ,Aryl hydrocarbon receptor ,Chromatin ,chemistry.chemical_compound ,Oncology ,SOX2 ,chemistry ,Biochemistry ,Downregulation and upregulation ,biology.protein ,Cancer research ,Epigenetics ,Progenitor cell ,Stem cell ,Caffeic acid phenethyl ester - Abstract
CAPE is the major active component of propolis, a widely available, non-toxic, honeybee natural product with anti-inflammatory, antioxidant, and antitumor properties. We have previously shown that CAPE inhibits growth of breast cancer cells and the tumorigenic potential of breast cancer stem cells. We have identified inhibition of histone deacetylase (HDAC) as one mechanism of action, which suggests that it mediates its effects through epigenetic modifications. We postulated that CAPE may be useful in chemoprevention for women at high risk for triple-negative breast cancers since the cell-of-origin hypothesis states that these cancers likely arise from transformation of mammary stem or progenitor cells, whose self-renewal is maintained via epigenetic states. We tested the effect of CAPE on wildtype (WT) and Trp53 null mammary stem cell (MaSC) self-renewal from BALB/c mice cultured as mammospheres (MMS). Primary mammary epithelial cells were cultured as MMS for 7 days, dissociated into single cells, re-cultured in the presence of CAPE for 7 days and passaged in secondary and tertiary passages without CAPE. MMS frequency and differentiation potential was analyzed using immunofluorescence detection of luminal marker, cytokeratin 18, basal marker, cytokeratin 14, and progesterone receptor (PR). Chromatin states were identified using ATAC-seq and open chromatin areas unique to CAPE treated murine MMS were used for pathway analysis performed by Ingenuity Pathway Analysis, Gene Set Enrichment Analysis and confirmed by Integrative Genome Viewer. CAPE treatment resulted in a dose dependent decrease in both WT and p53 null mammosphere forming efficiency that persisted in secondary and tertiary passages, suggesting reduced self-renewal. CAPE treatment also shifted differentiation from predominantly basal K14 to luminal K18-positive in both WT and p53 null MMS and increased PR expression in WT MMS. ATAC-seq of CAPE treated WT MMS showed significant pathway enrichment for p53 signaling, SOX2 signaling, and enrichment of open chromatin for several genes including the SMARCA4 gene, which regulates transcription of genes involved in stem cell renewal. ATAC-seq of CAPE treated Trp53 null MMS showed that genes defining early and late response to estrogen were particularly important. Significant canonical pathways included Aryl hydrocarbon receptor signaling, whose upregulation results in inhibition of self renewal and has been targeted as a potential drug target for estrogen receptor negative breast cancer. The integrin signaling pathway was also highly enriched. These data suggest that CAPE both inhibits MaSC self-renewal and shifts the lineage commitment to a luminal, ER + lineage. ATAC-seq demonstrated genomic effects that are important in differentiation, SC renewal and adhesion. These data suggest that CAPE may have an effect on lineage commitment in support of our chemoprevention strategy to reduce triple-negative breast cancer. Citation Format: Omene C, Patel M, Kannan K, Heguy A, Barcellos-Hoff MH. Mammary stem cell modulation of wildtype and Trp53 null stem cells by CAPE (caffeic acid phenethyl ester), a potential therapeutic agent. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P1-06-06.
- Published
- 2016
197. NF2 Loss Promotes Oncogenic RAS-Induced Thyroid Cancers via YAP-Dependent Transactivation of RAS Proteins and Sensitizes Them to MEK Inhibition
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Yogindra Persaud, Jeffrey A. Knauf, Maria E.R. Garcia-Rendueles, Ronald Ghossein, Vicki Smith, Agnes Viale, Adriana Heguy, Kety Huberman, Julio C. Ricarte-Filho, Yuqiang Fang, Barry S. Taylor, Suresh C. Jhanwar, Iňigo Landa, Brian R. Untch, Gisele Oler, Ian Ganly, Francesca Voza, Filippo G. Giancotti, and James A. Fagin
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Transcriptional Activation ,MAPK/ERK pathway ,DNA Copy Number Variations ,Chromosomes, Human, Pair 22 ,Cell Cycle Proteins ,Mice, Transgenic ,Biology ,Bioinformatics ,medicine.disease_cause ,Models, Biological ,Mice ,Transactivation ,Cell Line, Tumor ,Anti-apoptotic Ras signalling cascade ,Gene Order ,medicine ,Animals ,Humans ,Position-Specific Scoring Matrices ,Thyroid Neoplasms ,HRAS ,Nucleotide Motifs ,Promoter Regions, Genetic ,Protein Kinase Inhibitors ,Transcription factor ,Neoplasm Staging ,Neurofibromin 2 ,Binding Sites ,Nuclear Proteins ,Gene Expression Regulation, Neoplastic ,Merlin (protein) ,Disease Models, Animal ,Cell Transformation, Neoplastic ,Genes, ras ,Oncology ,Drug Resistance, Neoplasm ,Gene Targeting ,Cancer research ,Chromosome Deletion ,Mitogen-Activated Protein Kinases ,Signal transduction ,Carcinogenesis ,Gene Deletion ,Protein Binding ,Signal Transduction ,Transcription Factors - Abstract
Ch22q LOH is preferentially associated with RAS mutations in papillary and in poorly differentiated thyroid cancer (PDTC). The 22q tumor suppressor NF2, encoding merlin, is implicated in this interaction because of its frequent loss of function in human thyroid cancer cell lines. Nf2 deletion or Hras mutation is insufficient for transformation, whereas their combined disruption leads to murine PDTC with increased MAPK signaling. Merlin loss induces RAS signaling in part through inactivation of Hippo, which activates a YAP–TEAD transcriptional program. We find that the three RAS genes are themselves YAP–TEAD1 transcriptional targets, providing a novel mechanism of promotion of RAS-induced tumorigenesis. Moreover, pharmacologic disruption of YAP–TEAD with verteporfin blocks RAS transcription and signaling and inhibits cell growth. The increased MAPK output generated by NF2 loss in RAS-mutant cancers may inform therapeutic strategies, as it generates greater dependency on the MAPK pathway for viability. Significance: Intensification of mutant RAS signaling through copy-number imbalances is commonly associated with transformation. We show that NF2/merlin inactivation augments mutant RAS signaling by promoting YAP/TEAD-driven transcription of oncogenic and wild-type RAS, resulting in greater MAPK output and increased sensitivity to MEK inhibitors. Cancer Discov; 5(11); 1178–93. ©2015 AACR. This article is highlighted in the In This Issue feature, p. 1111
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- 2015
198. Immunosuppressive milieu of high-risk localized prostate cancer
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Jiansheng Wu, William C. Huang, Victor Ricardo Adorno Febles, Arjun Vasant Balar, Qinghu Ren, Fang-Ming Deng, Adriana Heguy, Aristotelis Tsirigos, Dennis Adeegbe, Samir S. Taneja, Kwok-Kin Wong, Jonathan Melamed, Alireza Khodadad-Jamayran, Donald Kufe, David R. Wise, Aarif Ahsan, James S. Wysock, and Herbert Lepor
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Cancer Research ,Prostate cancer ,Oncology ,business.industry ,medicine ,Cancer research ,medicine.disease ,business ,Peripheral blood ,Immune Factors - Abstract
344 Background: The immune factors that modulate the aggressiveness of localized treatment-naïve prostate cancer remain poorly understood. Methods: Fresh tumor and peripheral blood were collected at the time of radical prostatectomy in patients with localized prostate cancer. We evaluated the immune cell composition of 22 patient samples employing multi-parametric flow cytometry. Samples were grouped by histological grade into intermediate (INTPCA) and high-grade (HIGHPCA) prostate cancers based on standard NCCN criteria and immune cell abundances were quantified by mean +/- SEM. Statistical significance was assessed using the Mann-Whitney test. Results: INTPCA and HIGHPCA tumors harbored a similar increase in CD8+ T cells ( p < 0.0005 and p < 0.05, respectively) and CD11b+CD68-CD16+ myeloid-derived cells (p < 0.05) relative to the peripheral blood. Other cell types were similarly decreased in both INTPCA and HIGHPCA, including CD11b+CD68+CD14+ ( p < 0.005 and p < 0.05, respectively). By contrast, regulatory T cells were the only cell type in our analysis to be uniquely enriched in HIGHPCA rather than INTPCA ( p < 0.05). The most unique feature found in phenotypic profiling of the immune repertoire of HIGHPCA relative to INTPCA was an increase in the immune inhibitory receptor ligand, PD-L1, in the tumor associated macrophages (CD11b+CD68+CD14+) compared to the periphery (p < 0.05). Conclusions: Collectively, our findings reveal that HIGHPCA harbors a distinct immunological landscape. Although effector CD8+ T cells are preferentially expressed in the tumor, these are met with an increased proportion of regulatory T cells as well as PD-L1 expressing macrophages that contribute to the inert tumor microenvironment. These are key features of aggressive prostate cancer that may serve as potential biomarkers and therapeutic targets.
- Published
- 2020
199. Abstract OT1-01-03: Impact of RANKL inhibition on tumor microenvironment of early-stage breast cancer, a pre-surgical trial
- Author
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Nancy Tray, Karen Hiotis, Deborah Axelrod, Victor Ty, Judith D. Goldberg, Farbod Darvishian, Maryann Kwa, Yelena Novik, Sylvia Adams, Amber A. Guth, James L. Speyer, Adriana Heguy, and Douglas K. Marks
- Subjects
Oncology ,Cancer Research ,medicine.medical_specialty ,Tumor microenvironment ,biology ,medicine.diagnostic_test ,business.industry ,Tumor-infiltrating lymphocytes ,medicine.medical_treatment ,medicine.disease ,Denosumab ,Breast cancer ,Tumor progression ,RANKL ,Internal medicine ,Biopsy ,biology.protein ,Medicine ,business ,Neoadjuvant therapy ,medicine.drug - Abstract
BACKGROUND: Receptor activator of NF-kB (RANK) and its ligand (RANKL) have a well-established role in osteoclast-driven bone remodeling, however RANK/RANKL expression can also be seen on mammary epithelial cells and immune cells (IC). In breast cancer (BC) mouse models, RANKL is a paracrine effector of the mitogenic activity of progesterone and inhibition of RANKL can attenuate development of BC as well as the incidence of metastases. Pre-clinical evidence demonstrates that RANKL exerts effects on IC such as regulatory T cells (Tregs) and tumor associated macrophages (TAMs) within the tumor microenvironment (TME), thereby promoting tumor progression and metastases. Clinical studies also support a deleterious role for these IC subsets specifically in breast cancer, with high density of either Tregs or TAMs associated with increased risk for distant recurrence. Despite this preclinical support for targeting the RANK/RANKL pathway, clinical trials in early stage BC have had discordant results regarding the clinical benefit of denosumab (RANKL antagonist). As such, it is of significant clinical relevance to identify a drug response signature following treatment as this may suggest biomarkers predictive of benefit from adjuvant denosumab. This window-of-opportunity study was designed to evaluate the pharmacodynamic biomarkers of RANKL inhibition, as well as denosumab’s impact on the TME in paired tumor specimens from patients with BC following treatment with denosumab. Defining the immunomodulatory effects of denosumab may additionally provide biologic rationale for synergistic combination with specific immunotherapy agents. TRIAL DESIGN: This study is a single-arm open label Phase 0 (pre-surgical/window of opportunity) trial. Following diagnostic core needle biopsy, patients receive one dose of denosumab 120 mg given subcutaneously prior to surgical resection. ELIGIBILITY CRITERIA: ≥18 years of age with histologically-confirmed invasive BC >1 cm by radiographic or clinical criteria with available tissue from the core biopsy and planned surgery. Patients are ineligible for this study if neoadjuvant therapy is being considered. STUDY OBJECTIVES: The primary objective is to identify pharmacodynamics markers of RANKL inhibition in early BC. Planned analyses include protein and transcriptomic analyses of known targets of RANKL and compare treatment effect in both RANK/RANKL+ and RANK/RANKL- BC. As a secondary objective, the effect of RANKL inhibition on host immune response will be assessed by comparing pre-treatment biopsy specimens and surgical excision specimens from patients treated with denosumab and untreated controls matched for BC subtype and menopausal status. Planned analyses include RANK/RANKL expression on IC subsets, enumeration of tumor infiltrating lymphocytes by H&E, immunophenotype by immunohistochemistry (e.g. cytotoxic T lymphocytes, Tregs, TAMs) and expression of immune related genes (nanoString Pancancer Immune Panel). STATISTICAL METHODS: As RANK/RANKL protein expression is expected in 25-30% of BC, we plan to enroll 35 subjects to ensure at least 10 subjects with RANK/RANKL+ tumors will be included in our study cohort. With 10 subjects with tumors with high RANK/RANKL expression at baseline, a change from baseline of a single gene of >|1.7| standard deviations between subjects with and without RANK/RANKL expression is detectable based on a 2-sided two sample t-test with power of 80% and false discovery rate of 10%. Differential gene expression analysis will be performed comparing denosumab treated patients with RANK/RANKL+ vs. RANK/RANKL- BC, and between treated patients and untreated controls. PRESENT ACCRUAL: 29 of the planned 35 patients have been enrolled to date. Citation Format: Douglas Kanter Marks, Deborah Axelrod, Maryann Kwa, Nancy Tray, Karen Hiotis, Amber Guth, Yelena Novik, James Speyer, Victor Ty, Adriana Heguy, Judith Goldberg, Farbod Darvishian, Sylvia Adams. Impact of RANKL inhibition on tumor microenvironment of early-stage breast cancer, a pre-surgical trial [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr OT1-01-03.
- Published
- 2020
200. Transposon insertion profiling by sequencing (TIPseq) for mapping LINE-1 insertions in the human genome
- Author
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Cheng Ran Lisa Huang, Lindsay M. Payer, Adriana Heguy, Sitharam Ramaswami, David Fenyö, Pedro A. F. Galante, Jef D. Boeke, Fernanda Rego, Mark Grivainis, Jared P. Steranka, Zuojian Tang, Thiago Luiz Araujo Miller, and Kathleen H. Burns
- Subjects
Transposable element ,0303 health sciences ,lcsh:QH426-470 ,Methodology ,Retrotransposon ,Computational biology ,Biology ,Genome ,Targeted PCR ,DNA sequencing ,3. Good health ,Long interspersed nuclear element ,Structural variation ,lcsh:Genetics ,03 medical and health sciences ,LINE-1 ,0302 clinical medicine ,Next generation sequencing ,Human genome ,Molecular Biology ,030217 neurology & neurosurgery ,Illumina dye sequencing ,030304 developmental biology - Abstract
Background Transposable elements make up a significant portion of the human genome. Accurately locating these mobile DNAs is vital to understand their role as a source of structural variation and somatic mutation. To this end, laboratories have developed strategies to selectively amplify or otherwise enrich transposable element insertion sites in genomic DNA. Results Here we describe a technique, Transposon Insertion Profiling by sequencing (TIPseq), to map Long INterspersed Element 1 (LINE-1, L1) retrotransposon insertions in the human genome. This method uses vectorette PCR to amplify species-specific L1 (L1PA1) insertion sites followed by paired-end Illumina sequencing. In addition to providing a step-by-step molecular biology protocol, we offer users a guide to our pipeline for data analysis, TIPseqHunter. Our recent studies in pancreatic and ovarian cancer demonstrate the ability of TIPseq to identify invariant (fixed), polymorphic (inherited variants), as well as somatically-acquired L1 insertions that distinguish cancer genomes from a patient’s constitutional make-up. Conclusions TIPseq provides an approach for amplifying evolutionarily young, active transposable element insertion sites from genomic DNA. Our rationale and variations on this protocol may be useful to those mapping L1 and other mobile elements in complex genomes. Electronic supplementary material The online version of this article (10.1186/s13100-019-0148-5) contains supplementary material, which is available to authorized users.
- Published
- 2018
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