TNM Visual Manual

This thesis presents a series of illustrations intended to make the TNM (Tumor, Node, Metastasis) staging system more useful for physicians in predicting cancer spread patterns. The illustrations are color coded and clearly organized, and are designed to be compiled into a visual manual to be used as a quick referenc for physicians to use when diagnosing patients. The thesis focuses on the illustrations, diagrams, and TNM staging system for the head and neck

Circulating tumor cells: approaches to isolation and characterization

Circulating tumor cells (CTCs) shed from primary and metastatic cancers are admixed with blood components and are thus rare, making their isolation and characterization a major technological challenge. CTCs hold the key to understanding the biology of metastasis and provide a biomarker to noninvasively measure the evolution of tumor genotypes during treatment and disease progression. Improvements in technologies to yield purer CTC populations amenable to better cellular and molecular characterization will enable a broad range of clinical applications, including early detection of disease and the discovery of biomarkers to predict treatment responses and disease progression

Understanding Radiation Resistance in Head and Neck Tumor Xenografts Using Diffuse Reflectance and Raman Spectroscopy

Each year, 800,000 new patients are diagnosed with head and neck squamous cell carcinoma (HNSCC), a majority of whom are treated with a combination of daily fractions of radiation and weekly chemotherapy sessions for up to seven weeks. Current methods to evaluate treatment response of individual patients are limited to anatomical measurements of tumor burden using CT scan or MRI 4-8 weeks after completion of treatment. However, earlier knowledge of radiation-response prior to or at early days after commencement of therapy can aid oncologist with escalating and de-escalating treatment plans for exceptionally non-responding and responding patients. Such a knowledge can be only be gained if proper understanding of radiation-induced physiological and biomolecular changes is established and associated with treatment response. This dissertation presents two quantitative optical spectroscopic methods that can provide snapshots of tumor physiology and biomolecular content which can be used as biomarkers of treatment response. Because tumor hypoxia has been linked to poor treatment outcome, we employed diffuse reflectance spectroscopy to measure vascular oxygen saturation. In chapter 2, we first investigated the sensitivity of diffuse reflectance spectroscopy to tumor hypoxia and determined that optical measurement of tumor vascular oxygen saturation is negatively correlated with tumor hypoxia. In chapter 3, we utilized this technique to study radiation-induced kinetics of tumor oxygenation among radiation-resistant and -sensitive tumors. We established tumor xenografts from two human head and neck cancer cell lines in mice which were treated with 4 doses of 2 Gy twice weekly for two weeks. We observed greater rate of reoxygenation in radiation-resistant tumors which was accompanied with greater content of hypoxia inducible factor-1α (HIF-1α). Our results indicate that reduced oxygen consumption rate can potentially play a significant role in promoting radiation resistance. In addition, the radiation-induced changes in tumor optical properties were used to train a logistic regression model which successfully differentiated local-control and treatment-failure tumors. In addition to changes in reoxygenation, radiation treatment has also been known to induce microenvironmental changes within tumor. Thus in chapter 4, we used Raman spectroscopy to investigate early radiation-induced biomolecular changes in tumor microenvironment of radiation-resistant and -sensitive tumors. Raman spectra of head and neck tumor xenografts 1, 24, and 48 hours after radiation was collected and the spectra were analyzed using multivariate curve resolution-alternating least squares (MCR-ALS) and pure spectral profiles of biological specimen were extracted. We observed higher contributions to Raman spectra from lipid- and collagen-like species respectively in radiation-sensitive and -resistant tumors. Our results indicate the sensitivity of Raman spectroscopy to radiation-induced microenvironmental changes at early time points after radiation. The association between the observed functional and biomolecular changes with the radiosensitivity of the utilized tumors motivate further clinical studies to investigate whether such changes can be used as potential biomarkers of radiation response

Revolutionary impact of PET and PET-CT on the day-to-day practice of medicine and its great potential for improving future health care

In this communication, we present an overview of the impact and advantages of PET and PET-CT fusion imaging in the practice of medicine. We also discuss the evolution of this promising molecular imaging technique since its inception and the future prospects of the combined structure-function approach. Superior contrast resolution, accurate quantification and above all optimal image quality aid in improved diagnosis of many serious disorders including cancer. We speculate that this powerful imaging approach will almost completely replace most other conventional methods in the future. Currently, 18[F]-fluorode- -oxyglucose (FDG) is the main radiopharmaceutical employed for PET studies around the globe. With the availability of high quality PET images on a routine basis in most centres around the world and the likelihood that several other useful PET tracers will be approved in the near future for routine clinical applications, this technique will likely become essential in almost any medical disorder

Proteomic analysis in nasopharyngeal carcinoma

Nasopharyngeal carcinoma (NPC) is a particular type of head and neck cancer with a strong ethnic and enigmatic epidemiology that long puzzles the scientific society. Despite present all over the world, around 80% of all cases are in Asia where it is endemic, followed by low incidence regions in all other continents. Portugal has the second incidence in Europe for women, and third for men (Globocan 2012) among all European countries. Many efforts have been made to clarify and understand this unique geographic and epidemiologic distribution. The onset and evolution of nasopharyngeal carcinoma is a complex multi-stage process and may take a long time to occur. Although the molecular basis remains uncertain, we know that environmental factors, Epstein-Barr virus (EBV) infection and genetic susceptibility are considered to be the three major contributors. Further examination of these genes’ expression in each tumor revealed that around 93 oncogenes are up regulated in each tumor, while the mean number of TSGs down regulated was 109. In Portugal, the works of Souza & Breda have identified important polymorphism markers that may play a role in the onset and NPC development on the Portuguese northern region population. Despite the efforts made, the molecular mechanisms of NPC carcinogenesis and progression remains to be understood. In this regard, current omics methodologies offer a different approach to identify unique miRNAs and proteins that expression signatures associated with the cancer phenotype, reflecting the biological and pathological grade of the disease. Thus, the discovery of useful NPC biomarkers will lead to new diagnostic and prognostic tools. Meanwhile, the treatment of NPC has evolved in the past two decades. Since 2000, intensity modulated radiation therapy (IMRT) has been widely used to treat nasopharyngeal carcinoma. IMRT provides better dose delivery to the target while sparing the surrounding normal tissues while local control reaches 98% at 4 years24-28. At least one prospective randomized controlled trial showed its benefit in salivary protection in HNC. However, despite the excellent local control, 43% of patients will develop distant metastasis before 5 years and die25-26. In Portugal, IMRT is used at the Instituto Português de Oncologia de Lisboa (IPOLFG) since 2009 and represents the current standard of care for HNC RT, including NPC. Until now, several prognostic markers were identified and investigated for screening, and prognostic tools for NPC, have been purposed, particularly in Asia. However, to our knowledge, there are no validated identified markers to predict distant metastasis or outcome. Moreover, the studies exploring this subject have identified a population-based variety of biomarkers stressing an omic translation of NPC ethnic distribution. At the present work, this research explored formalin-fixed paraffin-embedded samples of biopsied nasopharyngeal carcinoma tumors via proteomic analysis. The aim is to describe a tumor profiling from the studied cohort and discover biomarkers to predict distant metastasis. Secondary endpoints are the discovery of biomarkers related to tumor radioresistance and treatment toxicity. Label-free quantitative mass spectrometry was able to identify 12 up-regulated proteins on early-stage primary tumors that were not present on advanced-stage primaries. Moreover, EBV and HSV co-infection was detected. No signs of HPV-related proteins were seen. Although the clinical outcomes of early and advanced primary tumors were identical, this can be attributed to the known effectiveness of intensive chemoradiation. Moreover, the difference in tumor profiling may reflect that those earlystage tumors could benefit from de-escalation protocols. We were able to detect the presence of a pool of 10 proteins related to distant metastases. Among them, the significant presence of interferon and tyrosine kinase proteins generates the hypothesis that they may represent therapeutic targets. Validation is needed.O carcinoma da nasofaringe (NPC) é um tipo particular de cancro da cabeça e pescoço com um epidemiologia enigmática e forte cariz étnico que há muito intriga a comunidade científica. Apesar de presente em todo o mundo, 805 dos casos encontram-se na Ásia, onde é endémico, seguido de regiões de media e baixa incidência em todos os outros continentes. Portugal tem a segunda incidência em mulheres e a terceira em homens dentre todos os países da Europa (Globocan 2012). Muitos esforços foram realizados para esclarecer e compreender esta distribuição epidemiológica e geográfica. A instalação e evolução do NPC é um processo com múltiplas fases e pode demorar um longo período de tempo para ocorrer. Apesar da base molecular da sua origem permanecer incerta, sabe-se que factores ambientais, a infecção pelo vírus Epstein-Barr e a susceptibilidade genética do hospedeiro são os três maiores contribuidores. Análise aprofundada da expressão genética revelou que cerca de 93 oncogenes estão sobre regulados enquanto que o número médio de genes supressores de tumor down regulated é de cerca de 109. Em Portugal, o trabalho de Souza & Breda identificaram um importante marcador de polimorfismo que pode estar relacionado com a instalação do NPC na população portuguesa na região norte. À despeito dos esforços realizados, o mecanismo molecular da carcinogênese do NPC e sua progressão permanecem por ser esclarecidos. A este respeito, as actuais tecnologias de ômica oferecem uma abordagem diferente capaz de identificar miRNAs e proteínas únicos que expressam assinatura com um fenótipo do cancro, refletindo o grau biológico e patológico da doença. Assim, a descoberta de biomarcadores úteis levará a novas ferramentas prognósticas. Ao mesmo tempo, o tratamento do NPC evoluiu consideravelmente nas últimas duas décadas. Desde 2000, a radioterapia com intensidade modulada do feixe (IMRT) tem sido amplamente utilizada para o tratamento do carcinoma da nasofaringe. IMRT fornece melhor entrega da dose ao alvo enquanto poupa os tecidos adjacentes sadios ao mesmo tempo que o controlo local alcança 98% aos 4 anos (Lee et al, 2002). Pelo menos uma meta-análise demonstrou o benefício na capacidade de proteção salivar em carcinomas da cabeça e pescoço. Entretanto, apesar do excelente controlo local, até 43% dos doentes vão morrer devido às metástases à distância antes dos 5 anos. Em Portugal, IMRT é utilizada no Instituto Português de Oncologia de Lisboa (IPOLFG) desde 2009 e representa o actual estado da arte no tratamento da maioria dos carcinomas da cabeça e pescoço incluindo NPC. Até o momento, diversos marcadores prognósticos de NPC foram propostos, particularmente na Ásia. Contudo, até o momento não são utilizados marcadores validades para predizer metástases ou evolução do doente. Estudos avaliando esta questão identificaram uma população variada de biomarcadores distintos a outras regiões geográficas, ressaltando a diversidade étnica da população dos doentes com NPC. Neste trabalho, esta pesquisa explorou amostras de carcinoma da nasofaringe provenientes de biopsias fixadas em formalina e embebidas em parafina através de análise proteómica. O objectivo é descrever o perfil tumoral da coorte estudada e descobrir biomarcadores capazes de predizer metástases à distância. Objectivos secundários é a descoberta de biomarcadores relacionados ao tumor que determinem radioresistência ou toxicidade ao tratamento. Espectroscopia de massa por ressonância magnética foi capaz de identificar 12 proteínas sobre expressadas nas em tumores primários iniciais que não estão presentes em tumores primários avançados. Além disso, foi detectada uma coinfecção EBV e HSV sem sinais de proteínas relacionadas ao HPV. Apesar dos resultados clínicos terem sido idênticos em ambos os grupos, isto pode ser explicado pela efectividade e intensidade do tratamento combinado. Além disso, a diferença em perfil tumorais podem refletir um perfil de doentes que poderia se beneficiar de protocolos de desintensificação. Esta pesquisa foi capaz de detectar a presença de 10 proteínas relacionadas aos doentes com metástases à distância. Entre elas, a presença de interferon e inibidores da tirosina cinase geram a hipótese de que podem representar potenciais alvos terapêuticos

Head and Neck Cancer Invasion: Contributions of Actin Regulatory Proteins and the Microenvironment

Metastasis of primary tumor lesions is the leading cause of cancer-related death. In head and neck cancer, a local-regional disease, metastasis is achieved mainly through invasion into surrounding tissue and spreads to cervical lymph nodes. Movement from the initial tumor site requires dynamic reorganization of the actin cytoskeleton, which utilizes the coordinated action of many actin regulatory proteins. However, there is increasing evidence that the tumor microenvironment is also a driver of invasion. This work aims to determine the contributions of proteins which regulate the actin cytoskeleton during head and neck cancer invasion both in vitro and in vivo, and provide details on how the HNSCC tumor microenvironment influences progression. This was accomplished, by the following Studies. In Study one, the actin binding protein coronin 1B is found to be amplified and overexpressed in invasive HNSCC patient samples, and a novel function in the regulation of protrusive membrane structures called invadopodia is described. Study two defines an in vivo role for the actin regulatory protein cortactin, which has been previously associated with more aggressive cancers in vitro and in patients. This work finds that cortactin expression is dispensable for tongue tumor invasion in a transgenic model of oral cancer, implicating the tumor microenvironment as being the major contributor to driving oral cancer invasion. Study three describes a technique for monitoring and biopsying cervical lymph nodes of mice using high frequency ultrasound. By using this technique, alterations in cervical lymph node size and blood flow were discovered in mice given the carcinogen 4-NQO to induce oral carcinogenesis. Collectively, these studies shed light on the importance of choosing comprehensive model systems for studying roles of actin binding proteins in cancer invasion

Theranostic Potential of Oncolytic Vaccinia Virus

Biological cancer therapies, such as oncolytic, or replication-selective viruses have advantages over traditional therapeutics as they can employ multiple different mechanisms to target and destroy cancers (including direct cell lysis, immune activation and vascular collapse). This has led to their rapid recent clinical development. However this also makes their pre-clinical and clinical study complex, as many parameters may affect their therapeutic potential and so defining reason for treatment failure or approaches that might enhance their therapeutic activity can be complicated. The ability to non-invasively image viral gene expression in vivo both in pre-clinical models and during clinical testing will considerably enhance the speed of oncolytic virus development as well as increasing the level and type of useful data produced from these studies. Further, subsequent to future clinical approval, imaging of reporter gene expression might be used to evaluate the likelihood of response to oncolytic viral therapy prior to changes in tumor burden. Here different reporter genes used in conjunction with oncolytic viral therapy are described, along with the imaging modalities used to measure their expression, while their applications both in pre-clinical and clinical testing are discussed. Possible future applications for reporter gene expression from oncolytic viruses in the phenotyping of tumors and the personalizing of treatment regimens are also discussed

Gene expression profiling of head and neck cancer

MDThe purpose of this study was to classify oral squamous cell carcinomas (OSCCs) based on their gene expression profiles, to identify differentially expressed genes in these cancers, and to correlate genetic deregulation with clinical-histopathological data and patient outcome. After conducting proof of principle experiments utilizing six head and neck squamous cell carcinomas (HNSCCs) cell lines, the gene expression profiles of 20 OSCCs and subsequently an additional 8 OSCCs were determined using cDNA microarrays containing 19,200 sequences and the Binary Tree-Structured Vector Quantization (BTSVQ) method of data analysis. Two sample clusters were identified in the group of 20 tumors that correlated with T3-T4 category of disease (P=0.035) and nodal metastasis( p=0.035). Samplec lustering of 28 OSCCsa nd the 6 cell lines revealed a correlation with disease free survival. BTSVQ analysis identified a subset of 23 differentially expressed genes with the lowest quantization error scores in the cluster containing more advanceds taget umors from the 20 OSCC dataset.T he expressiono f six of these differentially expressedg enesw as validated by quantitative real-time RT-PCR. Statistical analysis of quantitative real-time RT-PCR data was performed and, after Bonferroni correction, CLDNI (p = 0.007) over-expressionw as significantly correlated with the cluster containing more advanced stage tumors. Despite the clinical heterogeneity of OSCC, molecular subtyping by cDNA microarray analysis was able to identify distinct patternso f genee xpressiona ssociatedw ith relevant clinical parameters. The application of this methodology represents an advance in the classification of oral cavity tumors, and may ultimately aid in the development of more tailored therapies for oral carcinoma

Focal Spot, Fall/Winter 2003/2004

https://digitalcommons.wustl.edu/focal_spot_archives/1095/thumbnail.jp

Nasopharyngeal carcinoma in Finland : Epstein-Barr virus, human papillomaviruses, and toll-like receptors as prognostic factors

Nasopharyngeal carcinoma (NPC) is a malignant tumour arising from the surface epithelium of the nasopharynx. NPC is a rare cancer type in Northern Europe, and approximately ten new cases are diagnosed in Finland yearly. NPC carcinogenesis has been linked to Epstein-Barr virus (EBV) infection, but human papillomaviruses (HPVs) have also been found in NPC tumours. Presumably, the host’s innate immunological properties also have an impact on pathological processes. Toll-like receptors (TLRs) are transmembrane proteins, which recognize both microbial and host’s own structures released in tissue damage in order to regulate innate immunity. The expression of TLRs has been observed in many cancer types, but their actual function is still unknown. TLR stimulation may have both pro- or anti-tumoural effects. The aim of this nationwide study was to evaluate NPC incidence, histological subgroups, provided treatments and their adverse effects, and outcome in NPC patients diagnosed and treated in Finland from 1990 to 2009. A total of 207 patients were identified from the Finnish Cancer Registry database during the 20-year study period. In addition, we examined the samples of 150/207 patients for the presence of EBV, high-risk HPVs, and TLR expression (TLR1, TLR2, TLR4, TLR5, TLR7, and TLR9), and compared the results with survival data. EBV was found in 62% and HPVs in 14% of the tumour cases, while 24% of the tumours were EBV/HPV-negative. The patients with EBV-positive tumours had the best 5-year survival rates, and the patients with HPV-positive tumours had a significantly better overall survival than those with EBV/HPV-negative tumours. Moreover, positive TLR7 expression was found to be an independent prognostic factor for favourable outcome