Emerging tumor spheroids technologies for 3D in vitro cancer modeling

"Article in Press, Available online 31 October 2017" ; "S0163-7258(17)30268-1"Cancer is a leading cause of mortality and morbidity worldwide. Around 90% of deaths are caused by metastasis and just 10% by primary tumor. The advancement of treatment approaches is not at the same rhythm of the disease; making cancer a focal target of biomedical research. To enhance the understanding and promts the therapeutic delivery; concepts of tissue engineering are applied in the development of in vitro models that can bridge between 2D cell culture and animal models, mimicking tissue microenvironment. Tumor spheroid represents highly suitable 3D organoid-like framework elucidiating the intra and inter cellular signaling of cancer, like that formed in physiological niche. However, spheroids are of limited value in studying critical biological phenomenon such as tumor-stroma interactons involving extra cellular matrix or immune system. Therefore, a compelling need of tailoring spheroid technologies with physiologically relevant biomaterials or in silico models, is ever emerging. The diagnostic and prognostic role of spheroids rearrangements within biomaterials or microfluidic channel is indicative of patient management; particularly for the decision of targated therapy. Fragmented information on available in vitro spheroid models and lack of critical analysis on transformation aspects of these strategies; pushes the urge to comprehensively overview the recent technological advancements (e.g. bioprinting, micro-fluidic technologies or use of biomaterials to attain the third dimension) in the shed of tranlationable cancer research. In present article, relationships between current models and their possible exploitation in clinical success is explored with the highlight of existing challenges in defining therapeutic targets and screening of drug efficacy.The authors are thankful to European Union (Horizon 2020) funded project FoReCaST (No. 668983), the FCT fellowship to J. Silva-Correia (Grant No. SFRH/BPD/100590/2014), distinctions to J.M.O. under the Investigator FCT program (IF/00423/2012) and V.M.C. under the Investigator FCT program (IF/01214/2014) for supporting this work financially.info:eu-repo/semantics/publishedVersio

Establishment of a Multi-Analyte Serum Biomarker Panel to Identify Lymph Node Metastases in Non-small Cell Lung Cancer

IntroductionIn non-small cell lung cancer (NSCLC), the presence of locoregional lymph node metastases remains the most important prognostic factor and significantly guides treatment regimens. Unfortunately, currently-available noninvasive staging modalities have limited accuracy. The objective of this study was to create a multianalyte blood test capable of discriminating a patient's true (pathologic) nodal status preoperatively.MethodsPretreatment serum specimens collected from 107 NSCLC patients with localized disease were screened with 47 biomarkers implicated in disease presence or progression. Multivariate statistical algorithms were then used to identify the optimal combination of biomarkers for accurately discerning each patient's nodal status.ResultsWe identified 15 candidate biomarkers that met our criteria for statistical relevance in discerning a patient's preoperative nodal status. A ‘random forest’ classification algorithm was used with these parameters to define a 6-analyte panel, consisting of macrophage inflammatory protein-1α, carcinoembryonic antigen, stem cell factor, tumor necrosis factor-receptor I, interferon-γ, and tumor necrosis factor-α, that was the optimum combination of biomarkers for identifying a patient's pathologic nodal status. A Classification and Regression Tree analysis was then created with this panel that was capable of correctly classifying 88% of the patients tested, relative to the pathologic assessments. This value is in contrast to our observed 85% classification rate using conventional clinical methods.ConclusionsThis study establishes a serum biomarker panel with efficacy in discerning preoperative nodal status. With further validation, this blood test may be useful for assessing nodal status (including occult disease) in NSCLC patients facing tumor resection therapy

Molecular Targets of CNS Tumors

Molecular Targets of CNS Tumors is a selected review of Central Nervous System (CNS) tumors with particular emphasis on signaling pathway of the most common CNS tumor types. To develop drugs which specifically attack the cancer cells requires an understanding of the distinct characteristics of those cells. Additional detailed information is provided on selected signal pathways in CNS tumors

Markers of Bone Turnover In Preclinical Development of Drugs for Skeletal Diseases

Skeletal tissue is constantly remodeled in a process where osteoclasts resorb old bone and osteoblasts form new bone. Balance in bone remodeling is related to age, gender and genetic factors, but also many skeletal diseases, such as osteoporosis and cancer-induced bone metastasis, cause imbalance in bone turnover and lead to decreased bone mass and increased fracture risk. Biochemical markers of bone turnover are surrogates for bone metabolism and may be used as indicators of the balance between bone resorption and formation. They are released during the remodeling process and can be conveniently and reliably measured from blood or urine by immunoassays. Most commonly used bone formation markers include N-terminal propeptides of type I collagen (PINP) and osteocalcin, whereas tartrate-resistant acid phosphatase isoform 5b (TRACP 5b) and C-terminal cross-linked telopeptide of type I collagen (CTX) are common resorption markers. Of these, PINP has been, until recently, the only marker not commercially available for preclinical use. To date, widespread use of bone markers is still limited due to their unclear biological significance, variability, and insufficient evidence of their prognostic value to reflect long term changes. In this study, the feasibility of bone markers as predictors of drug efficacy in preclinical osteoporosis models was elucidated. A non-radioactive PINP immunoassay for preclinical use was characterized and validated. The levels of PINP, N-terminal mid-fragment of osteocalcin, TRACP 5b and CTX were studied in preclinical osteoporosis models and the results were compared with the results obtained by traditional analysis methods such as histology, densitometry and microscopy. Changes in all bone markers at early timepoints correlated strongly with the changes observed in bone mass and bone quality parameters at the end of the study. TRACP 5b correlated strongly with the osteoclast number and CTX correlated with the osteoclast activity in both in vitro and in vivo studies. The concept “resorption index” was applied to the relation of CTX/TRACP 5b to describe the mean osteoclast activity. The index showed more substantial changes than either of the markers alone in the preclinical osteoporosis models used in this study. PINP was strongly associated with bone formation whereas osteocalcin was associated with both bone formation and resorption. These results provide novel insight into the feasibility of PINP, osteocalcin, TRACP 5b and CTX as predictors of drug efficacy in preclinical osteoporosis models. The results support clinical findings which indicate that short-term changes of these markers reflect long-term responses in bone mass and quality. Furthermore, this information may be useful when considering cost-efficient and clinically predictive drug screening and development assays for mining new drug candidates for skeletal diseases.Luun biokemialliset merkkiaineet luustosairauksien prekliinisessä lääkekehityksessä Luun uudismuodostusta tapahtuu koko elämän ajan. Tässä prosessissa osteoklastit hajottavat vanhaa luuta ja osteoblastit muodostavat uutta luuta. Tasapainoon vaikuttaa ikä, sukupuoli ja perinnöllisyys, mutta myös monissa luustosairauksissa, kuten osteoporoosissa ja syövän luustometastaaseissa, tämä tasapaino on järkkynyt johtaen vähentyneeseen luun määrään ja lisääntyneeseen murtumaherkkyyteen. Luun biokemialliset merkkiaineet kertovat luun aineenvaihdunnasta eli hajotuksen ja muodostuksen välisestä tasapainosta. Merkkiaineita vapautuu luun uudismuodostuksessa ja niitä voidaan helposti ja luotettavasti mitata seerumista tai virtsasta immunomääritysmenetelmillä. Yleisesti käytettyjä luun muodostuksen merkkiaineita ovat tyypin I kollageenin aminoterminaalinen propeptidi (PINP) ja osteokalsiini, sekä luun hajotuksen merkkiaineita tartraatti-resistentti hapan fosfataasi alatyyppi 5b (TRACP 5b) ja tyypin I kollageenin karboksiterminaalinen telopeptidi (CTX). Näistä PINP on ainoa merkkiaine, jolle ei ole aiemmin ollut saatavilla prekliiniseen käyttöön soveltuvaa kaupallista immunomääritysmenetelmää. Tällä hetkellä biokemiallisten merkkiaineiden laajamittainen käyttö on vielä rajoittunutta, koska niihin liittyy paljon biologista ja analyyttistä variaatiota eikä niiden merkitsevyydestä ja käyttökelpoisuudesta pitkän aikavälin muutosta ennustavana tekijänä ole riittävästi näyttöä. Tämän tutkimuksen tavoitteena oli selvittää luuston biokemiallisten merkkiaineiden soveltuvuutta lääkemolekyylien tehokkuuden ennustajina prekliinisissä osteoporoositutkimusmalleissa. Tutkimuksessa karakterisoitiin ja validoitiin PINP:lle prekliiniseen käyttöön kehitetty immunomääritysmenetelmä. PINP:n, osteokalsiinin N-terminaalisen keskifragmentin, TRACP 5b:n ja CTX:n tasoja tutkittiin prekliinisissä osteoporoosimalleissa, ja saatuja tuloksia verrattiin perinteisillä menetelmillä kuten histologialla, tiheysmittauksilla ja mikroskopialla saatuihin tuloksiin. Kaikkien tutkittujen luuston merkkiaineiden alkuvaiheen muutosten havaittiin korreloivan kokeen lopussa nähtyihin luuston rakenteellisiin muutoksiin. TRACP 5b korreloi osteoklastien lukumäärään ja CTX osteoklastien aktiivisuuteen sekä in vitro että in vivo kokeissa. CTX/TRACP 5b suhteelle luotiin termi resorptio-indeksi, joka kuvaa osteoklastien keskimääräistä aktiivisuutta. Indeksi antoi tarkempaa tietoa kuin kumpikaan merkkiaine erikseen käytetyissä prekliinisissä osteoporoosimalleissa. PINP:n havaittiin korreloivan vahvasti luun muodostukseen, kun taas osteokalsiini kuvasi sekä luun muodostusta että hajotusta. Tämän tutkimuksen tulokset antavat uutta tietoa luun biokemiallisten merkkiaineiden soveltuvuudesta lääkemolekyylien tehokkuuden ennustajina prekliinisissä osteoporoosimalleissa. Havainnot tukevat kliinisiä tutkimuksia, joissa merkkiaineiden on havaittu korreloivan myöhemmin nähtävien luuston rakenteellisten muutosten kanssa. Tutkimuksessa saatu tieto auttaa suunniteltaessa kliinisen ennustettavuuden kannalta parempia määritysmenetelmiä, joiden avulla voidaan nopeammin ja tehokkaammin löytää uusia toimivia lääkemolekyylejä luustosairauksien hoitoon.Siirretty Doriast

Research on Melanoma

The book Research on Melanoma: A Glimpse into Current Directions and Future Trends, is divided into sections to represent the most cutting-edge topics in melanoma from around the world. The emerging epigenetics of disease, novel therapeutics under development and the molecular signaling aberrations are explained in detail. Since there are a number of areas in which unknowns exist surrounding the complex development of melanoma and its response to therapy, this book illuminates and comprehensively discusses such aspects. It is relevant for teaching the novice researcher who wants to initiate projects in melanoma and the more senior researcher seeking to polish their existing knowledge in this area. Many chapters include visuals and illustrations designed to easily guide the reader through the ideas presented

Evolution of the Molecular Biology of Brain Tumors and the Therapeutic Implications

A dramatic increase in knowledge regarding the molecular biology of brain tumors has been established over the past few years. In particular recent new avenues regarding the role of stem cells and microRNAs along with further understanding of the importance of angiogenesis, immunotherapy and explanations for the resistance of the tumors to chemotherapeutic agents and radiation therapy has been developed. It is hopeful that this new information will lead to efficacious treatment strategies for these tumors which remain a challenge. In this book a review of the latest information on these topics along with a variety of new therapeutic treatment strategies with an emphasis on molecular targeted therapies is provided

Factors involved in the pathogenesis of human uveal melanoma

Trabalho final de mestrado integrado em Medicina (Otorrinolaringologia), apresentado à Faculdade de Medicina da Universidade de Coimbra.Introdução: A presbiacusia é a causa mais comum de défice auditivo em adultos e está na origem de muitas outras alterações que comprometem a qualidade de vida dos doentes, entre as quais se destaca a maior incidência de patologia do foro depressivo e a correlação com perda de funções cognitivas que pode vir a culminar em quadros demenciais. No entanto, a maior parte dos indivíduos desvaloriza a situação e são poucos os que procuram ajuda médica. A vida social do indivíduo desempenha um papel essencial neste processo e na decisão de utilizar ou não uma prótese auditiva. Objetivos: Precisar o número de doentes seguidos no Centro Hospitalar e Universitário de Coimbra (CHUC) por presbiacusia e qual a percentagem destes que utiliza prótese auditiva. Simultaneamente, pretendemos apreciar o impacto que esta patologia tem na vertente social destes doentes. Materiais e métodos: Foram analisados todos os processos clínicos do serviço de Otorrinolaringologia (ORL) do CHUC que datassem do ano civil de 2010 em diante. Paralelamente, foi aplicado um questionário a 50 destes doentes. As perguntas em causa foram selecionadas a partir do Hearing Handicap Inventory for the Elderly (HHIE) Portuguese short version. Resultados: Este estudo revelou um total de 2643 indivíduos com diagnóstico de presbiacusia corroborado por audiograma tonal típico. Apenas 3,14% utilizam prótese auditiva, dos quais 67,45% são do sexo feminino. A análise dos questionários mostrou que 74% dos indivíduos apresenta scores que indicam que a patologia em causa prejudica a sua qualidade de vida. Conclusões: Perante um elevado número de pacientes com queixas de hipoacusia, só uma pequena fração recorre a medidas de reabilitação auditiva, sendo a utilização de próteses substancialmente superior no sexo feminino. A perda auditiva da maior parte dos indivíduos em estudo tem repercussões na sua vida social. A gravidade dessas repercussões é maior no sexo feminino, principalmente para idades inferiores a 75 anos.Introduction: Presbycusis is the most common cause of auditory deficit in adults and it’s also the root of many other pathological processes that compromise the patient’s quality of life, such as the augmented incidence of depression and the connection to the loss of cognitive functions that may culminate in dementia. However, the majority of these individuals devalues the situation and only a few seek medical help. The social life of the individual plays an essential role in this process and in the decision to use or not a hearing aid. Objectives: Specify the number of patients followed at the Centro Hospitalar e Universitário de Coimbra (CHUC) due to presbycusis and the percentage of those who use hearing aids. Simultaneously, we aim to assess the impact that this disease has on the social life of these patients. Methods: All clinical files present in the department of Otorhinolaryngology (ORL) dating from 2010 onwards were analyzed. Simultaneously, a questionnaire was administered to 50 of these patients. The questions composing this questionnaire were selected from the Hearing Handicap Inventory for the Elderly (HHIE) Portuguese short version. Results: This study revealed a total of 2643 individuals diagnosed with presbycusis corroborated by a typical tonal audiogram. Only 3.14% use hearing aids, of which 67.45% are female. Analysis of the questionnaires showed that 74% of the individuals present scores that indicate this pathology adversely affects their quality of life. Conclusions: Given the large number of patients with complaints of hearing loss, only a small fraction uses measures of auditory rehabilitation, with the use of hearing aids being substantially higher in females. The hearing loss of most of the individuals in this study has repercussions in their social life. The severity of these effects is higher in females, especially for the age group ‘younger than 75 years’