Cell microarray (CMA) : a novel screening method for prognostic factors in head and neck squamous cell carcinoma

Pään ja kaulan alueen levyepiteelisyöpä on yksi yleisimmistä syöpätyypeistä, muta sillä on silti yhä verrattain huono ennuste muihin yleisiin syöpätyyppeihin verrattuna. Hoitosuunnitelma ja ennuste määräytyvät pääasiassa syövän tarkemman anatomisen sijainnin sekä TNM-luokituksen perusteella. Sädehoito on yksi yleinen hoitomuoto, mutta ongelmana on syöpien vaihteleva herkkyys sädehoidolle. Prekliinisissä tutkimuksissa ennusteen ja sädeherkkyyden on todettu korreloivan syöpäsolujen tiettyjen biokemiallisten markkereiden ilmentymiseen. Näitä ovat muun muassa tuumorisuppressori PP2A:n toimintaan vaikuttavat endogeeniset proteiinit CIP2A, PME-1 sekä SET. Tutkimuksessa selvitettiin näiden kolmen proteiinin ilmentymistä pään ja kaulan alueen syövissä solumikrosirun avulla ja tavoitteena oli löytää uusia hoitovastetta ennustavia tekijöitä. Solumikrosiru kehitettiin yhdistämällä 30 pään ja kaulan alueen levyepiteelikarsinooman solulinjan solumateriaalia yhteen yksittäiseen parafiiniblokkiin. Solulinjat ovat peräisin eri pään ja kaulan alueilta ja osassa solulinjoista CIP2A:n ilmentyminen oli hiljennetty shRNA:n avulla. Suuri määrä kasvatettuja soluja kerättiin ja fiksoitiin formaliinilla, jonka jälkeen nämä sulautettiin parafiiniin. Kaikkien solulinjojen fiksoidut solulieriöt sisälsivät vain solumateriaalia ja ne liitettiin samaan parafiiniblokkiin niin että muodostui yhtenäinen solumikrosiru. Mikrosiruun liitettiin kontrolliksi lisäksi kuutta eri hiirikudosmateriaalia. Mikrosirusta leikatut leikkeet värjättiin immunohistokemiallisin menetelmin. Värjäyksiin käytettiin CIP2A-, PME-1- sekä SET-vasta-aineita. Värjäykset pisteytettiin kvantitatiivisen pisteytysmenetelmän mukaisesti arvioimalla mikroskoopilla manuaalisesti värjäytyneiden solujen määrää sekä värjäyksen intensiteettiä. Projektissa kehitimme uuden työkalun, jolla solujen proteiiniekspressio voidaan selvittää immunohistokemiallisesti tehokkaasti suuresta määrästä eri solulinjoja yksittäisellä värjäyksellä. Tällä menetelmällä immunohistokemiallisia värjäyksiä voidaan suorittaa pelkälle fiksoidulle solumateriaalille ilman solunulkoista komponenttia ja tuottaa luotettavan sekä laadukkaasti tulkittavan värjäystuloksen. Värjäysten perusteella CIP2A:n, PME-1:n sekä SET:n ilmentyminen vaihteli eri solulinjoissa, ja CIP2A-shRNA solulinjoissa CIP2A:n ilmentyminen oli merkittävästi alentunut, mikä viittaa onnistuneeseen CIP2A:n hiljentymiseen sekä metodin kvantitatiivisuuteen. Tutkimuksemme perusteella solumikrosiru on luotettava työkalu proteiinien ilmentymisen määrittämiseksi immunohistokemiallisesti suuresta solunäytemäärästä, ja se voisi tulevaisuudessa olla varteenotettava vaihtoehto kudosmikrosirun käytölle

Early-Onset Hypertension – Clinical characteristics and relation to adverse outcomes

ABSTRACT Established hypertension is a well-known risk factor for cardiovascular disease and dementia. However, limited evidence exists on the relation between the age of hypertension onset and adverse events. The aim of this thesis was to study the impact of early-onset hypertension on end-organ damage and cognitive function in midlife. Additional aims were to examine risk factors for early-onset hypertension and the feasibility of different methods for detecting hypertension onset age. This thesis is based on the Coronary Artery Risk Development in Young Adults (CARDIA), which is a biracial prospective follow-up study. The CARDIA study was initiated in 1985–1986 and included a sample of 5115 American young adults. The study participants underwent up to 30 years of follow-up with regularly conducted follow-up exams. We also included a separate study sample from the Health 2000 survey, a population-based study carried out in 2000–2001 that examined 8028 Finns. Both study samples collected either objective or self-reported data on the participants’ age of hypertension onset as well as an assessment of adverse outcomes. In the CARDIA study sample, we observed that an early age of hypertension onset was most robustly associated with echocardiographic left ventricular hypertrophy (LVH), diastolic dysfunction, and coronary calcification in midlife. Additionally, early-onset hypertension in these individuals was related to midlife cognitive impairment. In contrast, late-onset hypertension was not associated with these adverse outcomes. Self-reported hypertension onset age was not related to electrocardiographic LVH in the Health 2000 study sample. However, self-reported early-onset hypertension was associated with end-organ damage in the CARDIA participants. We also identified African American ethnicity, diabetes, and obesity as the most potent correlates of early-onset hypertension. In conclusion, early-onset hypertension, but not late-onset hypertension, is associated with having end-organ damage by midlife. Our findings demonstrate the importance of assessing the age of hypertension onset, even by patients’ self-report, in order to feasibly assess the lifetime burden of hypertension and to improve risk stratification of individuals with hypertension in clinical practice. TIIVISTELMÄ Verenpainetauti on merkittävä dementian sekä sydän- ja verisuonisairauksien riskitekijä. Aiempia tutkimuksia verenpainetaudin puhkeamisiän merkityksestä haittatapahtumien kehittymiselle on kuitenkin vähän. Tämän väitöskirjatutkimuksen tavoitteena oli tutkia varhain puhkeavan verenpainetaudin vaikutusta pääteelinvaurioihin sekä kognitiiviseen suorituskykyyn keski-iässä. Lisäksi selvitimme varhain puhkeavalle verenpainetaudille altistavia tekijöitä sekä eri menetelmien soveltuvuutta verenpainetaudin puhkeamisiän arvioimiseksi. Tutkimusaineistona käytettiin ensisijaisesti yhdysvaltalaista prospektiivista CARDIA-seurantatutkimusta, johon osallistui 5115 nuorta amerikkalaista miestä ja naista vuosina 1985–1986. Tutkimushenkilöt ovat osallistuneet 30 vuoden ajan säännöllisesti suoritettuihin seurantatutkimuksiin. Toinen tutkimusväestö koostui 8028 väestörekisteristä satunnaisotannalla valitusta suomalaisesta, jotka osallistuivat Terveys 2000-tutkimukseen vuosina 2000–2001. Molempien tutkimusotoksien osallistujilta määritettiin verenpainetaudin puhkeamisikä joko objektiivisesti tai itseilmoitettuna sekä mitattiin verenpainetaudin pääte-elintapahtumia. Ensimmäisessä tutkimusaineistossa varhaisella iällä puhjennut verenpainetauti oli voimakkaimmin yhteydessä vasemman kammion hypertrofiaan, sydämen diastoliseen toimintahäiriöön sekä sepelvaltimoiden kalkkeutumiseen keski-iässä. Varhainen verenpainetauti altisti myös kognitiivisten toimintojen heikentymiselle. Myöhäinen verenpainetauti ei ollut yhteydessä vastaaviin haittatapahtumiin. Myös itseilmoitettu varhain puhjennut verenpainetauti oli yhteydessä pääte-elinvaurioihin ensimmäisessä tutkimusotoksessa, mutta ei elektrokardiografisesti määritettyyn vasemman kammion hypertrofiaan toisessa tutkimusotoksessa. Afroamerikkalainen syntyperä, diabetes ja ylipaino olivat riskitekijöitä varhaiselle verenpainetaudille. Löydökset osoittavat, että erityisesti varhain puhkeava verenpainetauti altistaa verenpainetaudin pääte-elinvaurioille jo keski-iässä. Verenpainetaudin alkamisiän määrittäminen osana verenpainetaudin hoitoa voisi edesauttaa lääkäreitä huomioimaan korkean verenpaineen elinikäistä vaikutusta elimistöön ja mahdollisesti edistää näiden potilaiden tulevien haittatapahtumien ennaltaehkäisyä

Age of Hypertension Onset: Overview of Research and How to Apply in Practice

Purpose of review: To review the current evidence on research related to age of hypertension onset-its definition, correlates, heritability, and association with adverse outcomes. We also propose a framework for implementing assessment of hypertension onset age into clinical practice.Recent findings: Prior studies have used both objective measurements and self-report to determine age of hypertension onset or early-onset hypertension. Yet, no criterion for standard definition currently exists for either. Data from epidemiological and clinical studies demonstrate that early-onset hypertension is a highly heritable trait that confers an increased risk for cardiovascular death and end-organ damage compared with late-onset hypertension. Literature to date suggests that (parental) age of hypertension onset can be feasibly assessed for estimating (1) risk of future hypertension in non-hypertensive persons; and (2) the propensity for cardiovascular disease in individuals with established hypertension.Keywords: Age of hypertension onset; Blood pressure; Clinical implications; Hypertension; Hypertension and cardiovascular disease; Hypertension heritability.</p

Longitudinal blood pressure patterns and cardiovascular disease risk

Observational and interventional studies have unequivocally demonstrated that "present", i.e. single-occasion, blood pressure is one of the key determinants of cardiovascular disease risk. Over the past two decades, however, numerous publications have suggested that longitudinal blood pressure data and assessment of long-term blood pressure exposure provide incremental prognostic value over present blood pressure. These studies have used several different indices to quantify the overall exposure to blood pressure, such as time-averaged blood pressure, cumulative blood pressure, blood pressure trajectory patterns, and age of hypertension onset. This review summarises existing research on the association between these indices and hard cardiovascular outcomes, outlines the strengths and weaknesses of these indices, and provides an overview of how longitudinal blood pressure changes can be measured and used to improve cardiovascular disease risk prediction.KEY MESSAGES Numerous recent publications have examined the relation between cardiovascular disease and long-term blood pressure (BP) exposure, quantified using indices such as time-averaged BP, cumulative BP, BP trajectory patterns, and age of hypertension onset. This review summarises existing research on the association between these indices and hard cardiovascular outcomes, outlines the strengths and weaknesses of these indices, and provides an overview of how longitudinal BP changes can be measured and used to improve cardiovascular disease risk prediction. Although longitudinal BP indices seem to predict cardiovascular outcomes better than present BP, there are considerable differences in the clinical feasibility of these indices along with a limited number of prospective data.</p

Clinical Correlates of Early-Onset Hypertension

BACKGROUNDEarly-onset hypertension has been established as a heritable trait and a risk factor for cardiovascular disease outcomes. However, the clinical correlates of early-onset hypertension remain unidentified.METHODSIn this study, we assessed the demographic characteristics and lifestyle factors related to hypertension onset age in a sample of 3,286 Coronary Artery Risk Development in Young Adults (CARDIA) study participants (mean baseline age 25 4 years, 57% women). We examined the association between the participants' baseline characteristics and age of hypertension onset subgroups (= 45 years) using a multinomial logistic regression model with those who did not develop hypertension as the reference group. Hypertension onset was defined as blood pressure >= 140/90 mm Hg or antihypertensive medication use on 2 consecutively attended follow-up visits.RESULTSIn the multinomial logistic regression model, individuals who were black (odds ratio [OR], 5.08; 95% confidence interval [CI], 3.17-8.14), were more obese (OR, 1.57; 95% CI, 1.32-1.88), or had higher total cholesterol (OR, 1.34; 95% CI, 1.13-1.60 per SD) had increased odds of early-onset hypertension (onset at CONCLUSIONSOur findings suggest that individuals who are black, obese, have higher total cholesterol, or have lower HDL-cholesterol level, are potentially at an increased risk of having early-onset hypertension.</p

Cancer cell line microarray as a novel screening method for identification of radioresistance biomarkers in head and neck squamous cell carcinoma

Background: Currently, no clinically useful biomarkers for radioresistance are available in head and neck squamous cell carcinoma (HNSCC). This study assesses the usefulness of Cell Line Microarray (CMA) method to enhance immunohistochemical screening of potential immunohistochemical biomarkers for radioresistance in HNSCC cell lines.Methods: Twenty-nine HNSCC cell lines were cultured, cell pellets formalin-fixed, paraffin-embedded, and arrayed. Radioresistance features of the cell lines were combined to immunohistochemical stains for p53, NDFIP1, EGFR, stem cell marker Oct4, and PP2A inhibitor CIP2A.Results: Expression of p53, EGFR or CIP2A did not indicate intrinsic radioresistance in vitro. Stem cell marker Oct4 nuclear positivity and NDFIP1 nuclear positivity was correlated with increased intrinsic radioresistance.Conclusion: The usefulness of CMA in analysis of HNSCC cell lines and discovery of biomarkers is demonstrated. CMA is very well adapted to both testing of antibodies in a large panel of cell lines as well as correlating staining results with other cell line characteristics. In addition, CMA-based antibody screening proved an efficient and relatively simple method to identify potential radioresistance biomarkers in HNSCC.</p

Polygenic Risk Scores Predict Hypertension Onset and Cardiovascular Risk

Although genetic risk scores have been used to predict hypertension, their utility in the clinical setting remains uncertain. Our study comprised N=218 792 FinnGen participants (mean age 58 years, 56% women) and N=22 624 well-phenotyped FINRISK participants (mean age 50 years, 53% women). We used public genome-wide association data to compute polygenic risk scores (PRSs) for systolic and diastolic blood pressure (BP). Using time-to-event analysis, we then assessed (1) the association of BP PRSs with hypertension and cardiovascular disease (CVD) in FinnGen and (2) the improvement in model discrimination when combining BP PRSs with the validated 4- and 10-year clinical risk scores for hypertension and CVD in FINRISK. In FinnGen, compared with having a 20 to 80 percentile range PRS, a PRS in the highest 2.5% conferred 2.3-fold (95% CI, 2.2-2.4) risk of hypertension and 10.6 years (95% CI, 9.9-11.4) earlier hypertension onset. In subgroup analyses, this risk was only 1.6-fold (95% CI, 1.5-1.7) for late-onset hypertension (age >= 55 years) but 2.8-fold (95% CI, 2.6-2.9) for early-onset hypertension (agePeer reviewe

Self-reported Age of Hypertension Onset and Hypertension-Mediated Organ Damage in Middle-Aged Individuals

BackgroundObjectively defined early onset hypertension, based on repeated blood pressure measurements, is a strong risk factor for cardiovascular disease (CVD). We aimed to assess if also self-reported hypertension onset age is associated with hypertension-mediated organ damage (HMOD). Additionally, we evaluated the agreement between self-reported and objectively defined hypertension onset age.MethodsWe studied 2,649 participants (50 4 years at the time of outcome assessment, 57% women) of the Coronary Artery Risk Development in Young Adults (CARDIA) study who underwent measurements for echocardiographic left ventricular hypertrophy (LVH), left ventricular diastolic dysfunction (LVDD), coronary calcification, and albuminuria. We divided the participants into groups according to self-reported hypertension onset age (= 45 years, and no hypertension). We used multivariable-adjusted logistic regression models to assess the relation between self-reported hypertension onset age with the presence of HMOD, with those who did not report hypertension as the referent group.ResultsCompared with individuals without self-reported hypertension, self-reported hypertension onset at = 45 years was only associated with LVDD (OR, 1.81; 95% CI, 1.06-3.08). The agreement between self-reported and objectively defined hypertension onset age groups was 78-79%.ConclusionsOur findings suggest that self-reported hypertension onset age, a pragmatically feasible assessment in clinical practice, is a reasonable method for assessing risk of HMOD and CVD