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ABSTRACT: Stridor is a common presenting symptom for children in emergency departments (EDs) and usually represents an infectious process, such as croup, or aspiration of a foreign body. We present the case of an otherwise healthy 5-year-old girl with episodic increased work of breathing for several months that was initially diagnosed as asthma by her primary care physician. She subsequently presented to the ED with acutely worsening noisy breathing and dyspnea. Patient and parent denied any recent foreign body ingestions or choking episodes. We gave multiple doses of racemic epinephrine in the ED without symptom improvement. A lateral neck x-ray showed an occlusive subglottic airway mass. Otolaryngology (ENT) evaluation demonstrated an 85% airway occlusion. The mass was partially resected, resolving all of her respiratory symptoms. Although primary airway tumors in children are rare, they must be considered on the differential diagnosis of new noisy breathing or respiratory distress. Failure to diagnose these tumors in a timely manner can be life-threatening.
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Obstrucción de las Vías Aéreas , Asma , Crup , Cuerpos Extraños , Obstrucción de las Vías Aéreas/diagnóstico , Obstrucción de las Vías Aéreas/etiología , Niño , Preescolar , Femenino , Cuerpos Extraños/diagnóstico , Cuerpos Extraños/diagnóstico por imagen , Humanos , Ruidos Respiratorios/etiologíaRESUMEN
OBJECTIVES: Head and neck lymphatic malformations (HNLM) are caused by gain-of-function somatic mutations in PIK3CA. Acetylsalicylic acid (ASA/aspirin) is thought to limit growth in PIK3CA-mutated neoplasms through PI3K pathway suppression. We sought to determine if ASA could be beneficial for HNLM. METHODS: Retrospective case series of patients (0-18 years) offered ASA (3-5 mg/kg/day) for HNLM treatment (2010-2018). Clinical and treatment characteristics, patient-reported symptom improvement, medication tolerance, compliance, and complications were recorded. Treatment response was determined by change in patient/caregiver-reported symptoms, or HNLM size [complete (resolved), partial (decreased), or stable]. RESULTS: Fifty-three patients were offered ASA, 23 (43%) accepted (median age 10 years, IQR 6-14). Compared to patients who declined, patients receiving ASA were more likely to have extensive malformations: ex-utero intrapartum treatment procedure, bilateral malformations, oral cavity location, ≥2 invasive treatments, or tracheotomy (p < 0.05). All patients with tissue available had PIK3CA mutations (13/23). Treatment indications included oral pain/blebs (12, 52%), recurrent pain/swelling (6, 26%), or sudden/persistent swelling (5, 22%). Treatment plan was commonly one 81 mg tablet daily (19, 83%) for 3-12 months (8, 42%). Therapeutic adherence was reported by 18 patients (78%). Symptoms improved in 18 patients [78%; decreased pain (9, 39%) and swelling (8, 35%)]. Treatment resulted in partial (14, 61%) or complete response (4, 17%). Three patients developed oral bleb bleeding, which resolved with medication discontinuation. CONCLUSION: ASA seems to be a well-tolerated, low-risk medication for HNLM treatment. This pilot study suggests that it often improves symptoms and reduces HNLM size. Further prospective, randomized studies are warranted to comprehensively assess indications, safety, and efficacy. LEVEL OF EVIDENCE: Level 4.
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Anomalías Linfáticas , Fosfatidilinositol 3-Quinasas , Aspirina/uso terapéutico , Niño , Humanos , Anomalías Linfáticas/tratamiento farmacológico , Anomalías Linfáticas/genética , Proyectos Piloto , Estudios RetrospectivosRESUMEN
The risk of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) transmission to health care workers during myringotomy and tympanostomy tube (MT) insertion is unknown. To determine the need for enhanced precautions to prevent potential spread via aerosolized particles, we used an optical particle sizer to measure aerosol generation intraoperatively during a case series of MT insertion. We also discuss our institutional experience with safe pandemic-era perioperative practices. There was no measured increase in aerosol particle number during the procedure at a distance of 30 cm from the external auditory canal. These initial data are reassuring regarding the risk of SARS-CoV-2 transmission to the operating room team due to aerosol generation, but further study is necessary before making definitive recommendations.
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Aerosoles , COVID-19/prevención & control , COVID-19/transmisión , Control de Infecciones/organización & administración , Transmisión de Enfermedad Infecciosa de Paciente a Profesional/prevención & control , Ventilación del Oído Medio/efectos adversos , COVID-19/epidemiología , Niño , Humanos , Tempo OperativoRESUMEN
INTRODUCTION: Obstructive sleep apnea (OSA) in young children is caused by upper airway obstruction and is associated with changes in cognitive development, temperament and behavior. Adenotonsillectomy (AT) is often utilized as first line therapy for pediatric OSA. Children with laryngomalacia (LM) have a high chance of residual OSA after AT. There is paucity of literature regarding surgically naïve young children with OSA and laryngomalacia. Our study aimed to compare demographics, comorbidities and outcomes associated with OSA in surgically naïve young children with and without laryngomalacia. METHODS: Retrospective chart review of surgically naïve young children (<2-year-old) with polysomnogram (PSG) diagnosed OSA. All young children underwent pre-operative PSG followed by drug-induced sleep endoscopy (DISE) directed intervention. Variables documented included demographics, comorbidities, history of adenotonsillectomy, DISE directed surgical interventions and pre and post PSG findings. Laryngomalacia was defined as presence of obstruction (Chan Parikh score ≥ 2) at the supraglottic level on DISE evaluation. Demographics and prevalence of comorbidities of those with and without LM were compared using t-test (continuous) and Chi Square (categorical). P value is significant for <0.05. RESULTS: 79 surgically naïve young children with PSG diagnosed OSA performed between 2015 and 2019 were included in the study. Children with LM were younger in age (11 months) and had a higher OSA-18 score (35) compared to children without LM (17 months) (OSA-18 score: 5). No significant difference was noted in the pre-DISE PSG parameters in the 2 groups. 56.1% of children with OSA and laryngomalacia did not need AT. Both children with and without laryngomalacia showed improvement in AHI on post-DISE PSG. Higher baseline AHI was associated with greater improvement in AHI. CONCLUSION: Surgically naïve young children with OSA and LM present earlier than those without LM and report a significantly worse quality of life Young children with a higher baseline AHI were found to have greater improvement in severity of OSA post-DISE and surgical intervention. Majority of children with OSA and laryngomalacia did not need adenotonsillectomy after a preprocedural DISE assessment yet showed similar improvement in AHI. More studies are needed to determine which patients with OSA and laryngomalacia will need multilevel interventions.
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Endoscopía , Laringomalacia/complicaciones , Laringomalacia/cirugía , Apnea Obstructiva del Sueño/etiología , Adenoidectomía , Endoscopía/efectos adversos , Femenino , Humanos , Lactante , Recién Nacido , Masculino , Polisomnografía , Calidad de Vida , Estudios Retrospectivos , Índice de Severidad de la Enfermedad , TonsilectomíaRESUMEN
We present a case of two siblings born to nonconsanguineous parents that presented with hypotonia, respiratory insufficiency, and auditory neuropathy spectrum disorder (ANSD) correlated with NFASC (MIM: 609145) and the homozygous loss of function variant p.P924RfsX35. This appears to be the first two reported cases of NFASC correlated with ANSD. NFASC encodes for neurofascin which plays an important role in the formation, function and maintenance of axon initial segments and nodes of Ranvier. Due to the rarity of this gene variation, reports are sparse in the literature leading to delays in diagnosis which can impact patient's language acquisition and spoken language skills.
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Moléculas de Adhesión Celular/genética , Pérdida Auditiva Central/genética , Mutación con Pérdida de Función , Factores de Crecimiento Nervioso/genética , Femenino , Homocigoto , Humanos , Lactante , Trastornos del Desarrollo del Lenguaje/genética , Masculino , Hipotonía Muscular/genética , HermanosRESUMEN
OBJECTIVE: The U.S. Food and Drug Administration guidelines for cochlear implantation (CI) include age greater than 12 months. Studies have suggested that implantation in children younger than 12 months with congenital deafness may be associated with better spoken language outcomes. Compare auditory comprehension (AC) outcomes for children with congenital deafness who received CI less than 12 months of age to those implanted at 12 to 24 months of age. METHODS: Retrospective review of prospectively collected data in consecutively implanted patients under 2 years of age who received CI and had post-CI Preschool Language Scale (PLS)-AC scores. Receptive language was assessed with the AC subtest of the PLS. Patients without pre-CI PLS-AC scores were excluded. The association between age at implantation and post-CI PLS-AC scores up to 2 years after CI surgery was modeled using a linear mixed-effects model. Time from CI surgery, number of implants, risk factors for language delay, pre-CI PLS-AC score, and sex were included in the model. Patients implanted less than 12 months of age were compared to those implanted between 12 and 24 months. RESULTS: Twenty-nine patients who had CI surgery by 12 months and 82 who had CI surgery between 12 and 24 months were included in the analysis. Younger age at implantation and better pre-CI PLS-AC scores were significantly associated with better post-CI PLS-AC scores. CONCLUSION: Cochlear implantation in children with congenital deafness less than 12 months of age was associated with better PLS-AC than in children implanted over 12 months of age up to 2 years after implantation. LEVEL OF EVIDENCE: 4 Laryngoscope, 130:776-781, 2020.
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Implantación Coclear , Implantes Cocleares , Sordera/congénito , Sordera/cirugía , Audición , Factores de Edad , Preescolar , Implantación Coclear/normas , Femenino , Humanos , Lactante , Masculino , Estudios Retrospectivos , Resultado del TratamientoRESUMEN
Purpose: Head and neck squamous cell carcinoma (HNSCC) is the sixth most common cancer worldwide, with high mortality and a lack of targeted therapies. To identify and prioritize druggable targets, we performed genome analysis together with genome-scale siRNA and oncology drug profiling using low-passage tumor cells derived from a patient with treatment-resistant HPV-negative HNSCC.Experimental Design: A tumor cell culture was established and subjected to whole-exome sequencing, RNA sequencing, comparative genome hybridization, and high-throughput phenotyping with a siRNA library covering the druggable genome and an oncology drug library. Secondary screens of candidate target genes were performed on the primary tumor cells and two nontumorigenic keratinocyte cell cultures for validation and to assess cancer specificity. siRNA screens of the kinome on two isogenic pairs of p53-mutated HNSCC cell lines were used to determine generalizability. Clinical utility was addressed by performing drug screens on two additional HNSCC cell cultures derived from patients enrolled in a clinical trial.Results: Many of the identified copy number aberrations and somatic mutations in the primary tumor were typical of HPV(-) HNSCC, but none pointed to obvious therapeutic choices. In contrast, siRNA profiling identified 391 candidate target genes, 35 of which were preferentially lethal to cancer cells, most of which were not genomically altered. Chemotherapies and targeted agents with strong tumor-specific activities corroborated the siRNA profiling results and included drugs that targeted the mitotic spindle, the proteasome, and G2-M kinases WEE1 and CHK1 We also show the feasibility of ex vivo drug profiling for patients enrolled in a clinical trial.Conclusions: High-throughput phenotyping with siRNA and drug libraries using patient-derived tumor cells prioritizes mutated driver genes and identifies novel drug targets not revealed by genomic profiling. Functional profiling is a promising adjunct to DNA sequencing for precision oncology. Clin Cancer Res; 24(12); 2828-43. ©2018 AACR.
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Biomarcadores de Tumor , Neoplasias de Cabeza y Cuello/tratamiento farmacológico , Terapia Molecular Dirigida , Medicina de Precisión , Antineoplásicos/farmacología , Antineoplásicos/uso terapéutico , Biomarcadores de Tumor/antagonistas & inhibidores , Biomarcadores de Tumor/genética , Hibridación Genómica Comparativa , Biología Computacional/métodos , Perfilación de la Expresión Génica , Genómica/métodos , Neoplasias de Cabeza y Cuello/diagnóstico , Neoplasias de Cabeza y Cuello/genética , Humanos , Masculino , Persona de Mediana Edad , Terapia Molecular Dirigida/métodos , Mutación , Tomografía de Emisión de Positrones , Medicina de Precisión/métodos , ARN Interferente Pequeño/genética , Tomografía Computarizada por Rayos X , Transcriptoma , Secuenciación del ExomaRESUMEN
OBJECTIVE: To evaluate the association between craniofacial phenotype and hearing loss in children with craniofacial microsomia. DESIGN: Retrospective cohort study. SETTING: Tertiary care children's hospital. PATIENTS: Individuals with craniofacial microsomia. MAIN OUTCOME MEASURES: Ear-specific audiograms and standardized phenotypic classification of facial characteristics. RESULTS: A total of 79 participants were included in the study. The mean age was 9 years (range, 1 to 23 years) and approximately 60% were boys. Facial anomalies were bilateral in 39 participants and unilateral in 40 participants (24 right, 16 left). Microtia (hypoplasia of the ear) was the most common feature (94%), followed by mandibular hypoplasia (76%), soft tissue deficiency (60%), orbital hypoplasia or displacement (53%), and facial nerve palsy (32%). Sixty-five individuals had hearing loss (12 bilateral and 53 unilateral). Hearing loss was conductive in 73% of affected ears, mixed in 10%, sensorineural in 1%, and indeterminate in 16%. Hypoplasia of the ear or mandible was frequently associated with ipsilateral hearing loss, although contralateral hearing loss occurred in 8% of hemifaces. CONCLUSIONS: Hearing loss is strongly associated with malformations of the ipsilateral ear in craniofacial microsomia and is most commonly conductive. Hearing loss can occur contralaterally to the side with malformations in children with apparent hemifacial involvement. Children with craniofacial microsomia should receive early diagnostic hearing assessments.
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Síndrome de Goldenhar/complicaciones , Pérdida Auditiva/etiología , Adolescente , Niño , Preescolar , Femenino , Humanos , Lactante , Masculino , Fenotipo , Estudios Retrospectivos , Índice de Severidad de la Enfermedad , Adulto JovenRESUMEN
Tonsillectomy is a commonly performed procedure with an accepted risk of posttonsillectomy hemorrhage (PTH) approaching 5%, but catastrophic effects of hemorrhage are exceedingly rare. A variety of surgical techniques and hemostatic agents have been used to reduce the rate of hemorrhage, although none eliminate the risk. Numerous patient, surgical, and postoperative care factors have been studied for an association with PTH. The most consistent risk factors for PTH seem to be patient age and coagulopathies. Surgeon skill and surgical technique are most consistently associated with primary PTH.
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Hemostasis Quirúrgica/métodos , Técnicas Hemostáticas , Hemostáticos/farmacología , Cuidados Posoperatorios/métodos , Hemorragia Posoperatoria , Tonsilectomía/efectos adversos , Trastornos de la Coagulación Sanguínea/epidemiología , Competencia Clínica , Humanos , Hemorragia Posoperatoria/etiología , Hemorragia Posoperatoria/terapia , Factores de Riesgo , Tonsilectomía/métodosRESUMEN
IMPORTANCE: Surgical cure of head and neck squamous cell carcinoma (HNSCC) remains hampered by inadequately resected tumors and poor recognition of lesions with malignant potential. BLZ-100 is a chlorotoxin-based, tumor-targeting agent that has not yet been studied in HNSCC. OBJECTIVE: To evaluate BLZ-100 uptake in models of HNSCC and oral dysplasia. DESIGN, SETTING, AND PARTICIPANTS: This was an observational study (including sensitivity and specificity analysis) of BLZ-100 uptake in an orthotopic xenograft mouse model of HNSCC and a carcinogen-induced dysplasia model of hamster cheek pouches. INTERVENTIONS: Various HNSCC xenografts were established in the tongues of NOD-scid IL2Rgammanull (NSG) mice. BLZ-100 was intravenously injected and fluorescence uptake was measured. To induce dysplasia, the carcinogen 7,12-dimethylbenz(a)anthracene (DMBA) was applied to the cheek pouch of Golden Syrian hamsters for 9 to16 weeks. BLZ-100 was subcutaneously injected, and fluorescence uptake was measured. MAIN OUTCOMES AND MEASURES: The signal-to-background ratio (SBR) of BLZ-100 was measured in tumor xenografts. To calculate the sensitivity and specificity of BLZ-100 uptake, a digital grid was placed over tissue sections and correlative histologic sections to discretely measure fluorescence intensity and presence of tumor; a receiver operating characteristic (ROC) curve was then plotted. In the hamster dysplasia model, cheeks were graded according to dysplasia severity. The SBR of BLZ-100 was compared among dysplasia grades. RESULTS: In HNSCC xenografts, BLZ-100 demonstrated a mean (SD) SBR of 2.51 (0.47). The ROC curve demonstrated an area under the curve (AUC) of 0.89; an SBR of 2.50 corresponded to 92% sensitivity and 74% specificity. When this analysis was focused on the tumor and nontumor interface, the AUC increased to 0.97; an SBR of 2.50 corresponded to 95% sensitivity and 91% specificity. DMBA treatment of hamster cheek pouches generated lesions representing all grades of dysplasia. The SBR of high-grade dysplasia was significantly greater than that of mild-to-moderate dysplasia (2.31 [0.71] vs 1.51 [0.34], P = .006). CONCLUSIONS AND RELEVANCE: BLZ-100 is a sensitive and specific marker of HNSCC and can distinguish high-risk from low-risk dysplasia. BLZ-100 has the potential to serve as an intraoperative guide for tumor margin excision and identification of premalignant lesions.
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Carcinoma de Células Escamosas/diagnóstico , Neoplasias de Cabeza y Cuello/diagnóstico , Verde de Indocianina/análogos & derivados , Verde de Indocianina/farmacocinética , Neoplasias de la Boca/diagnóstico , Neoplasias Experimentales , Venenos de Escorpión/farmacología , Venenos de Escorpión/farmacocinética , Lengua/patología , Animales , Carcinoma de Células Escamosas/metabolismo , Línea Celular Tumoral , Colorantes/farmacología , Cricetinae , Neoplasias de Cabeza y Cuello/metabolismo , Xenoinjertos , Humanos , Procesamiento de Imagen Asistido por Computador , Radioisótopos de Yodo , Mesocricetus , Ratones , Ratones Endogámicos NOD , Neoplasias de la Boca/metabolismo , Curva ROC , Carcinoma de Células Escamosas de Cabeza y CuelloRESUMEN
BACKGROUND: Laryngeal angiosarcoma is rare and the prognosis is poor. The purpose of this study was to describe the first case of cutaneous angiosarcoma metastatic to the larynx and systematically review all cases of laryngeal angiosarcoma. METHODS: A 61-year-old man presented with an alar lesion diagnosed as angiosarcoma and was treated with wide local resection and radiation. Six years later, he presented with a laryngeal mass histologically similar to the initial tumor. A systematic review of reported cases of angiosarcoma of the larynx was performed. RESULTS: Eighteen cases were identified. Mean age of presentation was 64.3 years. Men represented 66.7%. Mean follow-up was 34.1 months. Forty-seven percent died with disseminated disease at a mean of 18.4 months. Our patient did well with serial resection. CONCLUSION: To the best of our knowledge, this case represents the first documented case of cutaneous angiosarcoma metastatic to the larynx and suggests that serial resection with long-term surveillance may be of benefit in some cases.
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Hemangiosarcoma/secundario , Neoplasias Laríngeas/secundario , Neoplasias Nasales/patología , Humanos , Masculino , Persona de Mediana EdadRESUMEN
IMPORTANCE: Evidence supports short courses of perioperative antibiotics for patients receiving minor head and neck procedures. Few studies have addressed antibiotic prophylaxis for patients undergoing free flap reconstruction of head and neck defects. OBJECTIVE: To determine ideal antibiotic prophylaxis in patients undergoing head and neck free flap reconstruction. DESIGN, SETTING, AND PARTICIPANTS: Retrospective cohort study of 427 adults receiving free flap reconstruction of head and neck defects at 2 affiliated tertiary care academic hospitals between January 1, 2006, and January 28, 2013. EXPOSURES: Prophylactic antibiotic type and duration were recorded from patient records. MAIN OUTCOMES AND MEASURES: Outcome data were abstracted from patients' medical records including infection at the surgical sites and distant nonsurgical sites and flap site complications including flap compromise, dehiscence, or fistula. Multivariate logistic regression was used to determine the association of risk factors with the primary outcome of any infection within 30 days of surgery. RESULTS: Ninety-six patients (22.5%) received prophylactic antibiotics for 24 hours or less, and 331 patients received prolonged courses of prophylactic antibiotics. The majority of patients received ampicillin-sulbactam alone for prophylaxis (53.2%), while 36.5% received clindamycin alone and 10.3% received an alternative regimen. Postoperative infections occurred in 46% of patients, and 22% of patients had an infection at the flap inset site or neck incision. The use of clindamycin (odds ratio [OR], 2.54; 95% CI, 1.25-5.14 [P = .01]) was associated with an increased risk of postoperative infection; extended duration of antibiotics (OR, 0.63; 95% CI, 0.34-1.19 [P = .18]) was not associated with increased risk of postoperative infection. By multivariate analysis, use of clindamycin (OR, 6.71; 95% CI, 1.83-24.60 [P = .004]) and oral tobacco use (OR, 1.20; 95% CI, 1.04-1.39 [P = .02]), but not extended course of prophylactic antibiotics (OR, 0.75; 95% CI, 0.30-1.86 [P = .53]), were associated with a higher risk of postoperative flap or neck infections. CONCLUSIONS AND RELEVANCE: The choice of antibiotic appears to affect the rate of all postoperative infections and flap site infections more than the duration of antibiotics following head and neck free flap reconstruction. At our institutions, ampicillin-sulbactam is the preferred prophylactic antibiotic for major clean-contaminated head and neck procedures when possible.
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Profilaxis Antibiótica , Colgajos Tisulares Libres , Neoplasias de Cabeza y Cuello/cirugía , Procedimientos de Cirugía Plástica/métodos , Complicaciones Posoperatorias/prevención & control , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Factores de RiesgoRESUMEN
Cochlear implantation has been performed safely several hundred thousand times, although a variety of complications have been reported including surgical site infection, device failure, migration of the receiver/stimulator or electrode array. Here we report the asymptomatic migration of the ground electrode through the squamous temporal bone, which was discovered at the time of device explantation. The incidence of this phenomenon is not known since it may remain asymptomatic and delayed postoperative imaging is not routinely performed. This finding should motivate implant surgeons to consider placement of the ground electrode into a subperiosteal pocket in a region with greater skull thickness.
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Implantes Cocleares/efectos adversos , Electrodos Implantados , Migración de Cuerpo Extraño/diagnóstico , Duramadre , Femenino , Humanos , Lactante , CráneoRESUMEN
INTRODUCTION: H63D HFE polymorphisms increase the risk of neurodegenerative disorders and, specifically, may increase amyotrophic lateral sclerosis (ALS) risk. Investigating the physiological alterations induced by H63D polymorphisms in ALS patients may elucidate mechanisms by which this genotype alters disease. METHODS: Clinical measures and muscle biopsies were available from patients previously diagnosed with ALS who underwent HFE genotyping. Clinical outcomes and SOD1 protein expression were analyzed using standard statistical analyses. RESULTS: ALS patients harboring H63D HFE (n = 16) had 28.1 months longer average disease duration and 39.3% lower muscle SOD1 protein than ALS patients with wild-type HFE (n = 22). CONCLUSIONS: Combined with previous reports suggesting the H63D polymorphism is associated with ALS, these results support a model wherein the H63D polymorphism is involved in ALS by means of pathways involving SOD1 but may limit cellular damage in individuals who develop disease. The association between HFE genotype and disease duration has important implications for clinical care and treatment trials.
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Esclerosis Amiotrófica Lateral , Regulación de la Expresión Génica/genética , Antígenos de Histocompatibilidad Clase I/genética , Proteínas de la Membrana/genética , Músculo Esquelético/enzimología , Polimorfismo de Nucleótido Simple/genética , Superóxido Dismutasa/metabolismo , Adulto , Anciano , Anciano de 80 o más Años , Esclerosis Amiotrófica Lateral/enzimología , Esclerosis Amiotrófica Lateral/genética , Esclerosis Amiotrófica Lateral/patología , Femenino , Estudios de Asociación Genética , Genotipo , Proteína de la Hemocromatosis , Histidina/genética , Humanos , Masculino , Persona de Mediana Edad , Fenilalanina/genética , Superóxido Dismutasa-1RESUMEN
Head and neck squamous cell carcinoma (HNSCC) is characterized by significant genomic instability that could lead to clonal diversity. Intratumor clonal heterogeneity has been proposed as a major attribute underlying tumor evolution, progression, and resistance to chemotherapy and radiation. Understanding genetic heterogeneity could lead to treatments specific to resistant and metastatic tumor cells. To characterize the degree of intratumor genetic heterogeneity within a single tumor, we performed whole-genome sequencing on three separate regions of an human papillomavirus (HPV)-positive oropharyngeal squamous cell carcinoma and two separate regions from one corresponding cervical lymph node metastasis. This approach achieved coverage of approximately 97.9% of the genome across all samples. In total, 5701 somatic point mutations (SPMs) and 4347 small somatic insertions and deletions (indels)were detected in at least one sample. Ninety-two percent of SPMs and 77% of indels were validated in a second set of samples adjacent to the discovery set. All five tumor samples shared 41% of SPMs, 57% of the 1805 genes with SPMs, and 34 of 55 cancer genes. The distribution of SPMs allowed phylogenetic reconstruction of this tumor's evolutionary pathway and showed that the metastatic samples arose as a late event. The degree of intratumor heterogeneity showed that a single biopsy may not represent the entire mutational landscape of HNSCC tumors. This approach may be used to further characterize intratumor heterogeneity in more patients, and their sample-to-sample variations could reveal the evolutionary process of cancer cells, facilitate our understanding of tumorigenesis, and enable the development of novel targeted therapies.
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Carcinoma de Células Escamosas/genética , Neoplasias Orofaríngeas/genética , Anciano , Carcinoma de Células Escamosas/secundario , Análisis Mutacional de ADN , Genes Relacionados con las Neoplasias , Genoma Humano , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Mutación INDEL , Metástasis Linfática , Masculino , Anotación de Secuencia Molecular , Sistemas de Lectura Abierta , Neoplasias Orofaríngeas/patologíaRESUMEN
HFE gene variants are relatively common genetic variants in Caucasians. The H63D HFE genetic variant has been repeatedly associated with a number of neurodegenerative diseases. We developed neuroblastoma cell lines expressing different HFE polymorphisms to explore the mechanisms behind these associations. Here we tested the hypothesis that cells with the H63D variant have a phenotype that promotes glutamate toxicity. In support of this hypothesis, expression of H63D HFE is associated with increased calcium-induced glutamate secretion and decreased cellular glutamate uptake. The polymorphism-associated changes in glutamate secretion were mimicked by altering cellular iron. Additionally, intracellular calcium is altered in a genotype-specific manner which could further impact glutamate secretion. HFE-dependent effects on glutamate uptake were confirmed in astrocytoma cell lines with endogenous expression of HFE. The ability of minocycline and the antioxidant Trolox to increase glutamate uptake differed by HFE genotype and implicate oxidative stress in glutamate regulation. This study demonstrates HFE cellular effects that extend beyond iron regulation, and suggests that H63D HFE may promote glutamate toxicity.
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Ácido Glutámico/metabolismo , Antígenos de Histocompatibilidad Clase I/genética , Proteínas de la Membrana/genética , Polimorfismo Genético/genética , Análisis de Varianza , Calcio/metabolismo , Línea Celular Tumoral , Deferoxamina/farmacología , Inhibidores Enzimáticos/farmacología , Compuestos Férricos/farmacología , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Proteínas de Transporte de Glutamato en la Membrana Plasmática/metabolismo , Glutaminasa/metabolismo , Proteína de la Hemocromatosis , Humanos , Líquido Intracelular/efectos de los fármacos , Líquido Intracelular/metabolismo , Hierro/metabolismo , Minociclina/farmacología , Neuroblastoma/patología , Compuestos de Amonio Cuaternario/farmacología , Sideróforos/farmacología , Sodio/metabolismo , Tacrina/farmacología , Transfección/métodos , Tritio/metabolismo , Proteína 1 de Transporte Vesicular de Glutamato/metabolismoRESUMEN
HFE is a protein that impacts cellular iron uptake. HFE gene variants are identified as risk factors or modifiers for multiple diseases. Using HFE stably transfected human neuroblastoma cells, we found that cells carrying the C282Y HFE variant do not differentiate when exposed to retinoic acid. Therefore, we hypothesized HFE variants would impact response to therapeutic agents. Both the human neuroblastoma and glioma cells that express the C282Y HFE variant are resistant to Temodar, geldanamycin and γ-radiation. A gene array analysis revealed that p16INK4A (p16) expression was increased in association with C282Y expression. Decreasing p16 protein by siRNA resulted in increased vulnerability to all of the therapeutic agents suggesting that p16 is responsible for the resistance. Decreasing HFE expression by siRNA resulted in a 85% decrease in p16 expression in the neuroblastoma cells but not the astrocytoma cells. These data suggest a potential direct relationship between HFE and p16 that may be cell specific or mediated by different pathways in the different cell types. In conclusion, the C282Y HFE variant impacts the vulnerability of cancer cells to current treatment strategies apparently by increasing expression of p16. Although best known as a tumor suppressor, there are multiple reports that p16 is elevated in some forms of cancer. Given the frequency of the HFE gene variants, as high as 10% of the Caucasian population, these data provide compelling evidence that the C282Y HFE variant should be part of a pharmacogenetic strategy for evaluating treatment efficacy in cancer cells.
Asunto(s)
Antineoplásicos/farmacología , Inhibidor p16 de la Quinasa Dependiente de Ciclina/genética , Antígenos de Histocompatibilidad Clase I/genética , Proteínas de la Membrana/genética , Neoplasias/genética , Polimorfismo Genético , Miembro 1 de la Subfamilia B de Casetes de Unión a ATP/genética , Miembro 1 de la Subfamilia B de Casetes de Unión a ATP/metabolismo , Antineoplásicos/uso terapéutico , Ciclo Celular , Línea Celular Tumoral , Inhibidor p16 de la Quinasa Dependiente de Ciclina/metabolismo , Metilación de ADN , Metilasas de Modificación del ADN/genética , Metilasas de Modificación del ADN/metabolismo , Enzimas Reparadoras del ADN/genética , Enzimas Reparadoras del ADN/metabolismo , Dacarbazina/análogos & derivados , Dacarbazina/farmacología , Doxorrubicina/farmacología , Resistencia a Antineoplásicos/genética , Expresión Génica/efectos de los fármacos , Expresión Génica/efectos de la radiación , Perfilación de la Expresión Génica , Glioma/tratamiento farmacológico , Glioma/genética , Proteínas del Choque Térmico HSP72/genética , Proteína de la Hemocromatosis , Humanos , NAD(P)H Deshidrogenasa (Quinona)/genética , NAD(P)H Deshidrogenasa (Quinona)/metabolismo , Neoplasias/tratamiento farmacológico , Regiones Promotoras Genéticas , Tolerancia a Radiación/genética , Temozolomida , Proteínas Supresoras de Tumor/genética , Proteínas Supresoras de Tumor/metabolismoRESUMEN
Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease with complicated pathogenesis with variable presentation and disease progression. There is a critical need for a panel of biomarkers to provide clinicians and researchers with additional information. In this study, multiplex immunoassays were used to screen a number of cytokines, growth factors, and iron-related proteins. ALS patients had significantly higher plasma levels of L-ferritin and lower concentrations of transferrin when compared to healthy controls and together classified a test group of subjects with 82% accuracy. Duration of ALS symptoms correlated positively with levels of monocyte chemoattractant protein 1 (MCP-1) and negatively with levels of granulocyte-macrophage colony stimulating factor (GM-CSF). The biomarker profile suggests iron homeostasis is disrupted in ALS patients, and changes in ferritin and transferrin (Tf) appear to be indicators of ongoing inflammatory processes. The data demonstrate a plasma biomarker profile in ALS patients that may differ from published reports of cerebrospinal fluid biomarkers.