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ABSTRACT: BACKGROUND: Heavy alcohol consumption increases risk of developing squamous cell carcinoma of the head and neck (SCCHN). Alcohol metabolism to cytotoxic and mutagenic intermediates acetaldehyde and reactive oxygen species is critical for alcohol-drinking-associated carcinogenesis. We hypothesized that polymorphisms in alcohol metabolism-related and antioxidant genes influence SCCHN survival. METHODS: Interview and genotyping data (64 polymorphisms in 12 genes) were obtained from 1227 white and African-American cases from the Carolina Head and Neck Cancer Epidemiology study, a population-based case-control study of SCCHN conducted in North Carolina from 2002 to 2006. Vital status, date and cause of death through 2009 were obtained from the National Death Index. Kaplan-Meier log-rank tests and adjusted hazard ratios were calculated to identify alleles associated with survival. RESULTS: Most tested SNPs were not associated with survival, with the exception of the minor alleles of rs3813865 and rs8192772 in CYP2E1. These were associated with poorer cancer-specific survival (HRrs3813865, 95%CI=2.00, 1.33-3.01; HRrs8192772, 95%CI=1.62, 1.17-2.23). Hazard ratios for 8 additional SNPs in CYP2E1, GPx2, SOD1, and SOD2, though not statistically significant, were suggestive of differences in allele hazards for all-cause and/or cancer death. No consistent associations with survival were found for SNPs in ADH1B, ADH1C, ADH4, ADH7, ALDH2, GPx2, GPx4, and CAT. CONCLUSIONS: We identified some polymorphisms in alcohol and oxidative stress metabolism genes that influence survival in subjects with SCCHN. Previously unreported associations of SNPs in CYP2E1 warrant further investigation.
Cancer epidemiology. 04/2013;
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ABSTRACT: PURPOSE: Tobacco and alcohol use are well-known risk factors for squamous cell carcinoma of the head and neck (SCCHN), but there has been little examination of disparities in SCCHN and racial patterns of tobacco and alcohol use, especially for African-Americans. The Carolina Head and Neck Cancer Study, a population-based case-control study, was utilized to determine whether relationships between tobacco and alcohol use and SCCHN differed by race. METHODS: Using a rapid case ascertainment system, cases were recruited from 46 contiguous counties in North Carolina from 2002 to 2006. Controls, selected from motor vehicle records, were frequency-matched to cases on age, sex, and race. This analysis was based on 989 white and 351 African-American cases and 1,114 white and 264 African-American controls. Analyses were performed using unconditional logistic regression, adjusting for age, sex, race, education, and fruit and vegetable consumption. RESULTS: The association between SCCHN and ever tobacco use among African-Americans (odds ratio (OR), 9.68; 95 % confidence interval (CI), 4.70, 19.9) was much greater than that observed in whites (OR, 1.94; 95 % CI, 1.51, 2.50). Smaller differences were observed when examining ever alcohol use (African-Americans: OR, 3.71; CI, 1.65, 8.30, and Whites: OR, 1.31: CI 0.96, 1.78). African-Americans consistently had greater effect measure estimates when examining common levels of duration and intensity metrics of tobacco and alcohol use, both independently and jointly. No racial differences in the effects of environmental (passive) tobacco smoke were observed. CONCLUSIONS: These findings suggest racial differences in SCCHN are not solely explained by differences in consumption patterns, and tobacco and alcohol may have greater impact in African-Americans.
Cancer Causes and Control 06/2012; · 2.88 Impact Factor
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Juneko E Grilley-Olson,
D Neil Hayes,
Dominic T Moore,
Kevin O Leslie,
Matthew D Wilkerson,
Bahjat F Qaqish,
Michele C Hayward,
Christopher R Cabanski,
Xiaoying Yin,
Mark A Socinski, [......],
Georgean G Deblois,
Göran Elmberger,
Paolo Graziano,
Craig F Hart,
Kirk D Jones,
Diane M Maia,
C Ryan Miller,
Keith V Nance,
William D Travis, William K Funkhouser
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ABSTRACT: Context.-Precise subtype diagnosis of non-small cell lung carcinoma is increasingly relevant, based on the availability of subtype-specific therapies, such as bevacizumab and pemetrexed, and based on the subtype-specific prealence of activating epidermal growth factor receptor mutations. Objectives.-To establish a baseline measure of interobserver reproducibility for non-small cell lung carcinoma diagnoses with hematoxylin-eosin for the current 2004 World Health Organization classification, to estimate interobserver reproducibility for the therapeutically relevant squamous/nonsquamous subsets, and to examine characteristics that improve interobserver reproducibility. Design.-Primary, resected lung cancer specimens were converted to digital (virtual) slides. Based on a single hematoxylin-eosin virtual slide, pathologists were asked to assign a diagnosis using the 2004 World Health Organization classification. Kappa statistics were calculated for each pathologist-pair for each slide and were summarized by classification scheme, pulmonary pathology expertise, diagnostic confidence, and neoplastic grade. Results.-The 12 pulmonary pathology experts and the 12 community pathologists each independently diagnosed 48 to 96 single hematoxylin-eosin digital slides derived from 96 cases of non-small cell lung carcinoma resection. Overall agreement improved with simplification from the comprehensive 44 World Health Organization diagnoses (κ = 0.25) to their 10 major header subtypes (κ = 0.48) and improved again with simplification into the therapeutically relevant squamous/nonsquamous dichotomy (κ = 0.55). Multivariate analysis showed that higher diagnostic agreement was associated with better differentiation, better slide quality, higher diagnostic confidence, similar years of pathology experience, and pulmonary pathology expertise. Conclusions.-These data define the baseline diagnostic agreement for hematoxylin-eosin diagnosis of non-small cell lung carcinoma, allowing future studies to test for improved diagnostic agreement with reflex ancillary tests.
Archives of pathology & laboratory medicine 05/2012; · 2.58 Impact Factor
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ABSTRACT: Few studies have examined the associations between dietary patterns and head and neck squamous cell carcinoma (SCC) or whether they differ by race. This was evaluated using data from a population-based case-control study (2002-2006) including 1,176 cases of head and neck SCC and 1,317 age-, race-, and gender-matched controls from central and eastern North Carolina whose diets had been assessed by food frequency questionnaire. Factor analysis identified 2 patterns of intake: 1) high consumption of fruits, vegetables, and lean protein and 2) high consumption of fried foods, high-fat and processed meats, and sweets. Associations were estimated using logistic regression, adjusting for matching factors and confounders. Heterogeneity by tumor site (oral/pharyngeal vs. laryngeal) and effect-measure modification were also evaluated. Reduced odds of head and neck SCC were found for the fruit, vegetable, and lean protein pattern (for highest quartile vs. lowest, odds ratio = 0.53, 95% confidence interval: 0.39, 0.71). The fried foods, high-fat and processed meats, and sweets pattern was positively associated only with laryngeal cancer (odds ratio = 2.12, 95% confidence interval: 1.21, 3.72). These findings underline the importance of a dietary pattern rich in fruits and vegetables and low in high-fat and processed meats and sweets for prevention of head and neck cancer.
American journal of epidemiology 05/2012; 175(12):1225-33. · 5.59 Impact Factor
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Matthew D Wilkerson,
Xiaoying Yin,
Vonn Walter,
Ni Zhao,
Christopher R Cabanski,
Michele C Hayward,
C Ryan Miller,
Mark A Socinski,
Alden M Parsons,
Leigh B Thorne,
Benjamin E Haithcock,
Nirmal K Veeramachaneni, William K Funkhouser,
Scott H Randell,
Philip S Bernard,
Charles M Perou,
D Neil Hayes
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ABSTRACT: BACKGROUND: Lung adenocarcinoma (LAD) has extreme genetic variation among patients, which is currently not well understood, limiting progress in therapy development and research. LAD intrinsic molecular subtypes are a validated stratification of naturally-occurring gene expression patterns and encompass different functional pathways and patient outcomes. Patients may have incurred different mutations and alterations that led to the different subtypes. We hypothesized that the LAD molecular subtypes co-occur with distinct mutations and alterations in patient tumors. METHODOLOGY/PRINCIPAL FINDINGS: The LAD molecular subtypes (Bronchioid, Magnoid, and Squamoid) were tested for association with gene mutations and DNA copy number alterations using statistical methods and published cohorts (n = 504). A novel validation (n = 116) cohort was assayed and interrogated to confirm subtype-alteration associations. Gene mutation rates (EGFR, KRAS, STK11, TP53), chromosomal instability, regional copy number, and genomewide DNA methylation were significantly different among tumors of the molecular subtypes. Secondary analyses compared subtypes by integrated alterations and patient outcomes. Tumors having integrated alterations in the same gene associated with the subtypes, e.g. mutation, deletion and underexpression of STK11 with Magnoid, and mutation, amplification, and overexpression of EGFR with Bronchioid. The subtypes also associated with tumors having concurrent mutant genes, such as KRAS-STK11 with Magnoid. Patient overall survival, cisplatin plus vinorelbine therapy response and predicted gefitinib sensitivity were significantly different among the subtypes. CONCLUSIONS/ SIGNIFICANCE: The lung adenocarcinoma intrinsic molecular subtypes co-occur with grossly distinct genomic alterations and with patient therapy response. These results advance the understanding of lung adenocarcinoma etiology and nominate patient subgroups for future evaluation of treatment response.
PLoS ONE 01/2012; 7(5):e36530. · 4.09 Impact Factor
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ABSTRACT: KRAS mutations may predict poor response to radiotherapy. Downstream events from KRAS, such as activation of BRAF, AKT and ERK, may also confer prognostic information but have not been tested in rectal cancer (RC). Our objective was to explore the relationships of KRAS and BRAF mutation status with p-AKT and p-ERK and outcomes in RC.
Pre-radiotherapy RC tumor biopsies were evaluated. KRAS and BRAF mutations were assessed by pyrosequencing; p-AKT and p-ERK expression by immunohistochemistry.
Of 70 patients, mean age was 58; 36% stage II, 56% stage III, and 9% stage IV. Responses to neoadjuvant chemoradiotherapy: 64% limited, 19% major, and 17% pathologic complete response. 64% were KRAS WT, 95% were BRAF WT. High p-ERK levels were associated with improved OS but not for p-AKT. High levels of p-AKT and p-ERK expression were associated with better responses. KRAS WT correlated with lower p-AKT expression but not p-ERK expression. No differences in OS, residual disease, or tumor downstaging were detected by KRAS status.
KRAS mutation was not associated with lesser response to chemoradiotherapy or worse OS. High p-ERK expression was associated with better OS and response. Higher p-AKT expression was correlated with better response but not OS.
Radiation Oncology 09/2011; 6:114. · 2.32 Impact Factor
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Bert H O'Neil, William K Funkhouser,
Benjamin F Calvo,
Michael O Meyers,
Hong Jin Kim,
Richard M Goldberg,
Stephen A Bernard,
Laura Caskey,
Allison M Deal,
Fred Wright,
Albert S Baldwin,
Joel E Tepper
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ABSTRACT: Rectal cancer is often clinically resistant to radiotherapy (RT) and identifying molecular markers to define the biologic basis for this phenomenon would be valuable. The nuclear factor κ-light chain-enhancer of activated B cells (NF-κB) is a potential anti-apoptotic transcription factor that has been associated with resistance to RT in model systems. The present study was designed to evaluate NF-κB activation in patients with rectal cancer undergoing chemoradiotherapy to determine whether NF-κB activity correlates with the outcome in rectal cancer patients.
A total of 22 patients underwent biopsy at multiple points in a prospective study and the data from another 50 were analyzed retrospectively. The pretreatment tumor tissue was analyzed for multiple NF-κB subunits by immunohistochemistry. Serial tumor biopsy cores were analyzed for NF-κB-regulated gene expression using reverse transcriptase polymerase chain reaction and for NF-κB subunit nuclear localization using immunohistochemistry.
Several NF-κB target genes (Bcl-2, cellular inhibitor of apoptosis protein [cIAP]2, interleukin-8, and tumor necrosis factor receptor-associated-1) were significantly upregulated by a single fraction of RT at 24 h, demonstrating for the first time that NF-κB is activated by RT in human rectal tumors. The baseline NF-κB p50 nuclear expression did not correlate with the pathologic response to RT. However, an increasing baseline p50 level was prognostic for overall survival (hazard ratio, 2.15; p = .040).
NF-κB nuclear expression at baseline in rectal cancer was prognostic for overall survival but not predictive of the response to RT. Larger patient numbers are needed to assess the effect of NF-κB target gene upregulation on the response to RT. Our results suggest that NF-κB might play an important role in tumor metastasis but not to the resistance to chemoradiotherapy.
International journal of radiation oncology, biology, physics 07/2011; 80(3):705-11. · 4.59 Impact Factor
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James D McKay,
Therese Truong,
Valerie Gaborieau,
Amelie Chabrier,
Shu-Chun Chuang,
Graham Byrnes,
David Zaridze,
Oxana Shangina,
Neonila Szeszenia-Dabrowska,
Jolanta Lissowska, [......],
Doris Lechner,
Hélène Blanché,
Ivo G Gut,
Pilar Galan,
Simon Heath,
Mia Hashibe,
Richard B Hayes,
Paolo Boffetta,
Mark Lathrop,
Paul Brennan
PLoS Genetics 04/2011; 7(4). · 8.69 Impact Factor
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James D McKay,
Therese Truong,
Valerie Gaborieau,
Amelie Chabrier,
Shu-Chun Chuang,
Graham Byrnes,
David Zaridze,
Oxana Shangina,
Neonila Szeszenia-Dabrowska,
Jolanta Lissowska, [......],
Doris Lechner,
Hélène Blanché,
Ivo G Gut,
Pilar Galan,
Simon Heath,
Mia Hashibe,
Richard B Hayes,
Paolo Boffetta,
Mark Lathrop,
Paul Brennan
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ABSTRACT: Genome-wide association studies (GWAS) have been successful in identifying common genetic variation involved in susceptibility to etiologically complex disease. We conducted a GWAS to identify common genetic variation involved in susceptibility to upper aero-digestive tract (UADT) cancers. Genome-wide genotyping was carried out using the Illumina HumanHap300 beadchips in 2,091 UADT cancer cases and 3,513 controls from two large European multi-centre UADT cancer studies, as well as 4,821 generic controls. The 19 top-ranked variants were investigated further in an additional 6,514 UADT cancer cases and 7,892 controls of European descent from an additional 13 UADT cancer studies participating in the INHANCE consortium. Five common variants presented evidence for significant association in the combined analysis (p ≤ 5 × 10⁻⁷). Two novel variants were identified, a 4q21 variant (rs1494961, p = 1×10⁻⁸) located near DNA repair related genes HEL308 and FAM175A (or Abraxas) and a 12q24 variant (rs4767364, p =2 × 10⁻⁸) located in an extended linkage disequilibrium region that contains multiple genes including the aldehyde dehydrogenase 2 (ALDH2) gene. Three remaining variants are located in the ADH gene cluster and were identified previously in a candidate gene study involving some of these samples. The association between these three variants and UADT cancers was independently replicated in 5,092 UADT cancer cases and 6,794 controls non-overlapping samples presented here (rs1573496-ADH7, p = 5 × 10⁻⁸); rs1229984-ADH1B, p = 7 × 10⁻⁹; and rs698-ADH1C, p = 0.02). These results implicate two variants at 4q21 and 12q24 and further highlight three ADH variants in UADT cancer susceptibility.
PLoS Genetics 03/2011; 7(3):e1001333. · 8.69 Impact Factor
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Matthew D Wilkerson,
Xiaoying Yin,
Katherine A Hoadley,
Yufeng Liu,
Michele C Hayward,
Christopher R Cabanski,
Kenneth Muldrew,
C Ryan Miller,
Scott H Randell,
Mark A Socinski,
Alden M Parsons, William K Funkhouser,
Carrie B Lee,
Patrick J Roberts,
Leigh Thorne,
Philip S Bernard,
Charles M Perou,
D Neil Hayes
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ABSTRACT: Lung squamous cell carcinoma (SCC) is clinically and genetically heterogeneous, and current diagnostic practices do not adequately substratify this heterogeneity. A robust, biologically based SCC subclassification may describe this variability and lead to more precise patient prognosis and management. We sought to determine if SCC mRNA expression subtypes exist, are reproducible across multiple patient cohorts, and are clinically relevant.
Subtypes were detected by unsupervised consensus clustering in five published discovery cohorts of mRNA microarrays, totaling 382 SCC patients. An independent validation cohort of 56 SCC patients was collected and assayed by microarrays. A nearest-centroid subtype predictor was built using discovery cohorts. Validation cohort subtypes were predicted and evaluated for confirmation. Subtype survival outcome, clinical covariates, and biological processes were compared by statistical and bioinformatic methods.
Four lung SCC mRNA expression subtypes, named primitive, classical, secretory, and basal, were detected and independently validated (P < 0.001). The primitive subtype had the worst survival outcome (P < 0.05) and is an independent predictor of survival (P < 0.05). Tumor differentiation and patient sex were associated with subtype. The expression profiles of the subtypes contained distinct biological processes (primitive: proliferation; classical: xenobiotic metabolism; secretory: immune response; basal: cell adhesion) and suggested distinct pharmacologic interventions. Comparison with lung model systems revealed distinct subtype to cell type correspondence.
Lung SCC consists of four mRNA expression subtypes that have different survival outcomes, patient populations, and biological processes. The subtypes stratify patients for more precise prognosis and targeted research.
Clinical Cancer Research 10/2010; 16(19):4864-75. · 7.74 Impact Factor
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Otolaryngology Head and Neck Surgery 07/2010; 143(1):167-8, 168.e1. · 1.72 Impact Factor
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ABSTRACT: Recent reports have linked oral health and periodontal disease indicators with increased risk of squamous cell carcinoma of head and neck (SCCHN). Thus far, evidence has been inconclusive; our objective was to study the association between oral health and SCCHN risk in the context of a large population-based study.
A population-based case-control study of incident SCCHN, the Carolina Head and Neck Cancer Study was carried out in 2002-2006 in 46 counties in North Carolina. Controls (n = 1,361) were frequency matched with cases (n = 1,289) on age, race, and gender. Oral health was assessed using interview data on tooth loss and mobility, mouthwash use, and frequency of dental visits.
Subjects were 26-80 years old (median age = 61). The distribution of tooth loss among controls was 0-5 teeth = 60%; 5-14 = 15%; and 16-28 = 25%. After controlling for covariates, tooth loss did not yield any notable association with SCCHN (16-28 vs. 0-5 lost teeth: OR: 1.21, 95% CI: 0.94, 1.56). Self-reported history of tooth mobility was moderately associated with increased SCCHN risk (OR: 1.33, 95% CI: 1.07, 1.65); however, the association did not persist among never smokers. Routine dental visits were associated with 30% risk reduction (OR: 0.68, 95% CI: 0.53, 0.87).
These data provide support for a possible modest association of periodontal disease, as measured by self-reported tooth loss indicators, but not tooth loss per se, with SCCHN risk.
Cancer Causes and Control 04/2010; 21(4):567-75. · 2.88 Impact Factor
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Clinical Chemistry 03/2010; 56(5):698-701. · 7.91 Impact Factor
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ABSTRACT: NF-kappaB is an antiapoptotic transcription factor that has been shown to be a mediator of treatment resistance. Bcl-3 is a regulator of NF-kappaB that may play a role in oncogenesis. The goal of this study was to correlate the activation status of NF-kappaB and Bcl-3 with clinical outcome in a group of patients with metastatic colorectal cancer (CRC).
A retrospective study of 23 patients who underwent surgical resection of CRC at the University of North Carolina (UNC). Activation of NF-kappaB was defined by nuclear expression of select components of NF-kappaB (p50, p52, p65) and Bcl-3. Tissue microarrays were created from cores of normal mucosa, primary tumor, lymph node metastases and liver metastases in triplicate from disparate areas of the blocks, and an intensity score was generated by multiplying intensity (0-3+) by percent of positive tumor cells. Generalized estimating equations were used to note differences in intensity scores among normal mucosa and nonnormal tissues. Cox regression models were fit to see if scores were significantly associated with overall survival.
p65 NE was significantly higher in primary tumor and liver metastases than normal mucosa (both p < 0.01). p50 nuclear expression was significantly higher for all tumor sites than for normal mucosa (primary tumor and lymph node metastases p < 0.0001, liver metastases p < 0.01). Bcl-3 nuclear expression did not differ significantly between normal mucosa and tumor; however, nuclear expression in primary tumor for each of these components was strongly associated with survival: the increase in hazard for each 50-point increase in nuclear expression was 91% for Bcl-3, 66% for p65, and 52% for p50 (all p < 0.05).
Activation of canonical NF-kappaB subunits p50 and p65 as measured by nuclear expression is strongly associated with survival suggesting NF-kappaB as a prognostic factor in this disease. Primary tumor nuclear expression appears to be as good as, or better than, metastatic sites at predicting prognosis. Bcl-3 nuclear expression is also negatively associated with survival and deserves further study in CRC.
Oncology 01/2010; 78(3-4):181-8. · 2.27 Impact Factor
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ABSTRACT: Hypoxia-inducible factor-1alpha (HIF-1alpha) and the neo-angiogenic factors induced as a result of hypoxia-inducible factor transcriptional activation may contribute to tumorigenesis by inducing vessel formation that in turn provides oxygen and nutrients promoting tumor expansion. In vitro studies of nasopharyngeal carcinoma (NPC), an aggressive malignancy that is nearly always infected by Epstein-Barr virus, show HIF-1alpha is upregulated by viral latent membrane protein 1 (LMP1). The current study used immunohistochemistry to examine the extent to which HIF-1alpha and LMP1 are co-expressed in naturally infected NPC tissues. Analytic procedures were optimized for sensitive localization of HIF-1alpha and LMP1 in fixed tissue sections using immunohistochemistry with sensitive fluorescent and signal amplification technologies. Vessel density was quantified by CD31 immunohistochemistry. LMP1 was expressed focally in all 18 NPCs examined, including 7/8 in situ lesions. There was no consistent co-localization with HIF-1alpha which was usually only weakly expressed in a subset of neoplastic cells. Neither LMP1 nor HIF-1alpha expression correlated with vessel density, and degree of vascularization varied widely among cases. Advanced immunohistochemical technologies reveal that LMP1 is expressed more commonly than previously reported in NPC. There is no consistent relationship between LMP1 and either HIF-1alpha expression or degree of microvasculature. The biologic basis for the wide variation in vessel density deserves further investigation.
Head and Neck Pathology 12/2009; 3(4):276-82.
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Pediatric Dermatology 11/2009; 26(6):747-8. · 1.07 Impact Factor
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ABSTRACT: Malignant rhabdoid tumors (MRTs) are poorly differentiated pediatric cancers that arise in various anatomical locations and have a very poor outcome. The large majority of these malignancies are caused by loss of function of the SNF5/INI1 component of the SWI/SNF chromatin remodeling complex. However, the mechanism of tumor development associated with SNF5 loss remains unclear. Multiple studies have demonstrated a role for SNF5 in the regulation of cyclin D1, p16(INK4A), and pRb(f) activities suggesting it functions through the SWI/SNF complex to affect transcription of genes involved in cell cycle control. Previous studies in genetically engineered mouse models (GEMM) have shown that loss of SNF5 on a p53-null background significantly accelerates tumor development. Here, we use established GEMM to further define the relationship between the SNF5 and p53 tumor suppressor pathways. Combined haploinsufficiency of p53 and Snf5 leads to decreased latency for MRTs arising in alternate anatomical locations but not for the standard facial MRTs. We also observed acceleration in the appearance of T-cell lymphomas in the p53(+/-);Snf5(+/-) mice. Our studies suggest that loss of SNF5 activity does not bestow a selective advantage on the p53 spectrum of tumors in the p53(+/-);Snf5(+/-) mice. However, reduced p53 expression specifically accelerated the growth of a subset of MRTs in these mice.
Molecular Carcinogenesis 09/2009; 48(12):1139-48. · 3.16 Impact Factor
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Giorgio Zanotti,
Monica Casiraghi,
John B Abano,
Jason R Tatreau,
Mayura Sevala,
Hilary Berlin,
Susan Smyth, William K Funkhouser,
Keith Burridge,
Scott H Randell,
Thomas M Egan
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ABSTRACT: Toll-like receptors (TLRs) of the innate immune system contribute to noninfectious inflammatory processes. We employed a murine model of hilar clamping (1 h) with reperfusion times between 15 min and 3 h in TLR4-sufficient (C3H/OuJ) and TLR4-deficient (C3H/HeJ) anesthetized mice with additional studies in chimeric and myeloid differentiation factor 88 (MyD88)- and TLR4-deficient mice to determine the role of TLR4 in lung ischemia-reperfusion injury. Human pulmonary microvascular endothelial monolayers were subjected to simulated warm ischemia and reperfusion with and without CRX-526, a competitive TLR4 inhibitor. Functional TLR4 solely on pulmonary parenchymal cells, not bone marrow-derived cells, mediates early lung edema following ischemia-reperfusion independent of MyD88. Activation of MAPKs and NF-kappaB was significantly blunted and/or delayed in lungs of TLR4-deficient mice as a consequence of ischemia-reperfusion injury, but edema development appeared to be independent of activation of these signaling pathways. Pretreatment with a competitive TLR4 inhibitor prevented edema in vivo and reduced actin cytoskeletal rearrangement and gap formation in pulmonary microvascular endothelial monolayers subjected to simulated warm ischemia and reperfusion. In addition to its well-accepted role to alter gene transcription, functioning TLR4 on pulmonary parenchymal cells plays a key role in very early and profound pulmonary edema in murine lung ischemia-reperfusion injury. This may be due to a novel mechanism: regulation of endothelial cell cytoskeleton affecting microvascular endothelial cell permeability.
AJP Lung Cellular and Molecular Physiology 05/2009; 297(1):L52-63. · 3.66 Impact Factor
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Gordon P Flake,
M Patricia Rivera, William K Funkhouser,
Susan J Maygarden,
Kellen L Meadows,
Elizabeth H Long,
Pat S Stockton,
Tina C Jones,
Hyeon Woo Yim,
Robbert J C Slebos,
Jack A Taylor
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ABSTRACT: Lung cancer is the leading cause of cancer deaths in both men and women in the United States. The LIFE (Light Induced Fluorescence Endoscopy) Project was initiated at the University of North Carolina Medical Center in November, 1999, for the dual purposes of (1) detecting pre-invasive lung cancer in high-risk patients and (2) studying the molecular biology of pre-invasive lesions of the bronchus for possible development of molecular biomarkers. Of the 47 patients enrolled, all were current or former tobacco smokers, except for 1. Fluorescence endoscopy was utilized, in addition to white light bronchoscopy, to increase the detection of intraepithelial lesions. Adjacent biopsies were submitted for permanent and frozen sections, respectively, from four predetermined sites as well as from any abnormal areas. The snap-frozen specimens were cryostat sectioned, and the mucosal epithelial cells laser capture microdissected for DNA analysis. The great majority of specimens yielded sufficiently abundant and intact DNA to accomplish the molecular objectives. Histologic concordance of adjacent permanent and frozen sections was equivalent to the concordance of adjacent permanent sections, suggesting that frozen section diagnosis was adequate for the research purpose of correlating histology with molecular analysis.
Toxicologic Pathology 02/2007; 35(1):65-74. · 1.91 Impact Factor
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ABSTRACT: Recently, in vitro models of coagulation have called into question the traditional conception of Factor IX as an intrinsic pathway protein, essential to propagation of coagulation but not central to the initiation of hemostatic plug, which has been thought instead to involve TF/FVIIa interactions with factor X and platelets. We hypothesized that the activation of factor IX, and its role in a factor IXa/FVIIa "tenase" complex leading to thrombin generation, plays a more important role than that of TF/FVIIa complex activation of factor X in the early hemostatic response to vascular injury. In vivo modeling is possible because of the generation of factor IX(-/-) mice.
We used two models of arterial vascular injury, histological examination following mechanical carotid artery disruption and intravital microscopy of a mesenteric arteriole subsequent to ferric chloride arteriolar injury to examine mice having complete deficiency of factor IX (FIX(-/-)).
Both injury models demonstrate that platelet rich thrombi /hemostatic plug in FIX(-/-) mice is dramatically reduced as compared to wild type mice under conditions of high shear; in fact, no platelet thrombi (>20 mum) were observed in the intravital experiments. Interestingly, the platelet defect is more striking than that described in mice lacking fibrinogen and/or von Willebrand factor.
The results suggest TF/FVIIa-->FX pathway is insufficient for effective platelet aggregation in the presence of high flow, requiring factor IX at the convergence of both intrinsic and extrinsic pathways. Following platelet adhesion, factor IX is required for normal platelet aggregation in vivo, as well as thrombin generation and propagation of occlusive thrombus at the site of vascular injury.
Thrombosis Research 01/2007; 121(2):225-34. · 2.44 Impact Factor
