Tapaus-verrokkitutkimus Serpina3 c.918-1G>C muunnoksen rintasyöpärikin arvioimiseksi Etelä-Suomen väestössä

Rintasyöpä on yleisin naisilla esiintyvä syöpä ja vuonna 2020 se oli viidenneksi yleisin kuolemaan johtava syöpä. Suomessa diagnosoitiin vuonna 2019 yli 5000 uutta rintasyöpä tapausta, joista 94 % esiintyi naisilla ja 6 % miehillä. Tähän mennessä korkean riskin rintasyöpägeenit kuten BRCA1, BRCA2 ja TP53 on tunnistettu sekä lisäksi monia keskikorkean riskin geenejä. Silti yhdessä kaikki löydetyt geenit selittävät vain noin puolet familiaarisista rintasyöpätapauksista. Lisäksi kaikki löydetyt rintasyövän alttiusgeenit ovat yhteydessä DNA:n korjausmekanismiin. Serpina3 erottuu joukosta ei-DNA-korjausmekanismigeeninä vain geeninä, joka koodaa Serpin-superperheeseen kuuluvaa proteaasi-inhibiittoria. Serpina3 on yhdistetty moniin sairauksiin ja erityisesti muutokset sen ilmentymistasossa on liitetty kasvaimen etenemiseen monissa syövissä, myös rintasyövässä. Aikaisempi tutkimus kuitenkin esittää, että Serpina3 c.918- 1G>C variantti on rintasyövälle altistava Pohjois-Suomen väestössä. Tässä pro gradu- työssä tutkitaan tapaus-verrokkitutkimuksena Serpina3 c.918-1G>C variantin liittymistä rintasyöpään Etelä- Suomen väestössä. Lisäksi Serpina3 c.918-1G>C muutoksen kantajien kasvaimen histologia ja solumarkkerit analysoitiin. Tutkimuksessa käytettiin rintasyöpäpotilailta kerättyä DNA:ta sekä DNA:ta veren luovuttajilta ja biopankista. Rintasyöpäpotilaat kattoivat sekä familiaariset että valikoimattomat tapaukset. Serpina3 c.918-1G>C variantin esiintyvyyttä kartoitettiin genotyypittämällä potilaat ja kontrollit. Genotyypitys tehtiin käyttämällä TaqMan reaaliaikaista PCR:ää ja kantajat varmistettiin Sanger sekvensoinnilla. Tilastollisia testejä käytettiin aineiston analysoinnissa. Tutkittua Serpina3 c.918-1G>C varianttia ei löydetty tilastollisesti merkittävästi enemmän (p>0.05) rintasyöpätapauksista kuin kontrolleista. Varianttia löydettiin 0.23 % familiaarista tapauksista ja 0.36 % valikoimattomista tapauksista, yhteensä 0.28 % kaikista rintasyöpätapauksista. Yleisyys väestökontrolleissa oli 0.27 %. Kasvaimen histologia oli duktaalinen 73 % kantajilla ja 9 % kasvain oli lobulaarinen. Toisin sanoen kantajien kasvainten histologia mukaili tavallista jakaumaa. Kaikki kantajat olivat HER2 negatiivisia ja kaikki paitsi yksi olivat sekä ER- että PR-positiivisia. Solun jakautumisen markkeriainetta, Ki-67, löydettiin noin puolelta kantajista. Yhteenvetona tämän tutkimuksen perusteella voidaan esittää, että Serpina3 c.918-1G>C ei ole rintasyövälle altistava geenimuutos ainakaan Etelä-Suomen väestössä.Breast cancer is the most prevalent cancer in women worldwide and in 2020 it was the fifth deadliest. In Finland 2019 more than 5000 breast cancer cases were diagnosed, 94% in women and 6% in men. Until now, the high-risk breast cancer susceptibility genes have been identified including BRCA1, BRCA2 and TP53 as well as many of the moderate risk genes. Still, together all the identified genes explain only approximately half of the familial breast cancer cases. Furthermore, all the known breast cancer susceptibility genes are linked to the DNA repair mechanism. Serpina3 stands out as a non-DNA repair gene but as a gene that encodes a protease inhibitor which belongs to the serpin superfamily. Serpina3 has been associated with various diseases before and especially changes in its expression levels are linked to the tumor prognosis in many cancers including breast cancer. However, a previous study proposed that Serpina3 c.918-1G>C is a susceptibility variant for breast cancer in the Northern Finland population. This thesis a case-control study to investigate whether Serpina3 c.918-1G>C variant is associated with breast cancer in the Southern Finland population. In addition, the tumor histology and cellular markers of Serpina3 c.918-1G>C carriers were examined. This study utilized DNA collected from breast cancer patients as well as DNA from blood donors and healthy biobank controls. Breast cancer patients included both familial and unselected cases. The prevalence of Serpina3 c.918- 1G<C variant was studied by genotyping the cases and controls. Genotyping was done by TaqMan real-time PCR and carriers were further confirmed by Sanger sequencing. Moreover, statistical tests were used in the data analyses. The studied Serpina3 c.918-1G>C variant was not found to be significantly (p>0.05) enriched in the breast cancer cases. The variant was found in 0.23 % of familial and 0.36 % of unselected cases, altogether in 0.28 % of all studied breast cancer cases, the frequency in controls was 0.27 %. The tumor histology was found to be ductal in 73 % of the Serpina3 c.918- 1G>C variant carriers and only 9 % had lobular tumor. In other words, the tumor histology followed the usual distribution. All the carriers had a HER2 negative tumor and all except one case were both ER and PR positive. About half of the carriers expressed the cellular proliferation marker Ki67. As a conclusion, the results from this study do not suggest Serpina3 c.918-1G>C as a breast cancer risk variant at least in the Southern Finland population

TAS-116, a Well-Tolerated Hsp90 Inhibitor, Prevents the Activation of the NLRP3 Inflammasome in Human Retinal Pigment Epithelial Cells

Chronic inflammation has been associated with several chronic diseases, such as age-related macular degeneration (AMD). The NLRP3 inflammasome is a central proinflammatory signaling complex that triggers caspase-1 activation leading to the maturation of IL-1 beta. We have previously shown that the inhibition of the chaperone protein, Hsp90, prevents NLRP3 activation in human retinal pigment epithelial (RPE) cells; these are cells which play a central role in the pathogenesis of AMD. In that study, we used a well-known Hsp90 inhibitor geldanamycin, but it cannot be used as a therapy due to its adverse effects, including ocular toxicity. Here, we have tested the effects of a novel Hsp90 inhibitor, TAS-116, on NLRP3 activation using geldanamycin as a reference compound. Using our existing protocol, inflammasome activation was induced in IL-1 alpha-primed ARPE-19 cells with the proteasome and autophagy inhibitors MG-132 and bafilomycin A1, respectively. Intracellular caspase-1 activity was determined using a commercial caspase-1 activity kit and the FLICA assay. The levels of IL-1 beta were measured from cell culture medium samples by ELISA. Cell viability was monitored by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test and lactate dehydrogenase (LDH) measurements. Our findings show that TAS-116 could prevent the activation of caspase-1, subsequently reducing the release of mature IL-1 beta. TAS-116 has a better in vitro therapeutic index than geldanamycin. In summary, TAS-116 appears to be a well-tolerated Hsp90 inhibitor, with the capability to prevent the activation of the NLRP3 inflammasome in human RPE cells.Peer reviewe

Kapeneeko kuilu? : Tilannekatsaus terveyserojen kaventamiseen Suomessa 2007-2010

Kansallinen terveyserojen kaventamisen toimintaohjelma 2008-201

An internet-based photography therapy intervention into the grief of persons who have lost a loved one

Persons who have lost a loved one by death are increasingly searching for support from online peer groups to process their grief. One approach involves sharing photographs with peers online. The purpose of this study was to assess the impact of a four-week internet-based photography therapy intervention concerning participants’ grief reactions using the Hogan Grief Reaction Checklist (HGRC). Another aim was to assess the role of the moderator and describe development targets in the internet-based photography therapy group based on the participants’ experiences. The participants were individuals who had lost a loved one divided into an intervention group (n = 101) and a comparison group (n = 55). The changes in the dimensions of grief of the intervention group members were slightly greater than in the comparison group. The dimension of panic behaviour declined statistically significantly more in the intervention than in the comparison group during the follow-up period. While the participants from the intervention group valued the expertise of the moderator, the moderator’s role could have been more visible. The participants also wished that the therapeutic photography group would have included more active involvement. Additionally, the participants made suggestions on modifying intervention content and developing a platform better suited for sharing photographs.Peer reviewe

Permutation-based significance analysis reduces the type 1 error rate in bisulfite sequencing data analysis of human umbilical cord blood samples

DNA methylation patterns are largely established in-utero and might mediate the impacts of in-utero conditions on later health outcomes. Associations between perinatal DNA methylation marks and pregnancy-related variables, such as maternal age and gestational weight gain, have been earlier studied with methylation microarrays, which typically cover less than 2% of human CpG sites. To detect such associations outside these regions, we chose the bisulphite sequencing approach. We collected and curated clinical data on 200 newborn infants; whose umbilical cord blood samples were analysed with the reduced representation bisulphite sequencing (RRBS) method. A generalized linear mixed-effects model was fit for each high coverage CpG site, followed by spatial and multiple testing adjustment of P values to identify differentially methylated cytosines (DMCs) and regions (DMRs) associated with clinical variables, such as maternal age, mode of delivery, and birth weight. Type 1 error rate was then evaluated with a permutation analysis. We discovered a strong inflation of spatially adjusted P values through the permutation analysis, which we then applied for empirical type 1 error control. The inflation of P values was caused by a common method for spatial adjustment and DMR detection, implemented in tools comb-p and RADMeth. Based on empirically estimated significance thresholds, very little differential methylation was associated with any of the studied clinical variables, other than sex. With this analysis workflow, the sex-associated differentially methylated regions were highly reproducible across studies, technologies, and statistical models.Peer reviewe

Umbilical cord blood DNA methylation in children who later develop type 1 diabetes

Aims/hypothesis Distinct DNA methylation patterns have recently been observed to precede type 1 diabetes in whole blood collected from young children. Our aim was to determine whether perinatal DNA methylation is associated with later progression to type 1 diabetes. Methods Reduced representation bisulphite sequencing (RRBS) analysis was performed on umbilical cord blood samples collected within the Finnish Type 1 Diabetes Prediction and Prevention (DIPP) Study. Children later diagnosed with type 1 diabetes and/or who tested positive for multiple islet autoantibodies (n = 43) were compared with control individuals (n = 79) who remained autoantibody-negative throughout the DIPP follow-up until 15 years of age. Potential confounding factors related to the pregnancy and the mother were included in the analysis. Results No differences in the umbilical cord blood methylation patterns were observed between the cases and controls at a false discovery rate Conclusions/interpretation Based on our results, differences between children who progress to type 1 diabetes and those who remain healthy throughout childhood are not yet present in the perinatal DNA methylome. However, we cannot exclude the possibility that such differences would be found in a larger dataset.Peer reviewe

Permutation-based significance analysis reduces the type 1 error rate in bisulfite sequencing data analysis of human umbilical cord blood samples

DNA methylation patterns are largely established in-utero and might mediate the impacts of in-utero conditions on later health outcomes. Associations between perinatal DNA methylation marks and pregnancy-related variables, such as maternal age and gestational weight gain, have been earlier studied with methylation microarrays, which typically cover less than 2% of human CpG sites. To detect such associations outside these regions, we chose the bisulphite sequencing approach. We collected and curated clinical data on 200 newborn infants; whose umbilical cord blood samples were analysed with the reduced representation bisulphite sequencing (RRBS) method. A generalized linear mixed-effects model was fit for each high coverage CpG site, followed by spatial and multiple testing adjustment of P values to identify differentially methylated cytosines (DMCs) and regions (DMRs) associated with clinical variables, such as maternal age, mode of delivery, and birth weight. Type 1 error rate was then evaluated with a permutation analysis. We discovered a strong inflation of spatially adjusted P values through the permutation analysis, which we then applied for empirical type 1 error control. The inflation of P values was caused by a common method for spatial adjustment and DMR detection, implemented in tools comb-p and RADMeth. Based on empirically estimated significance thresholds, very little differential methylation was associated with any of the studied clinical variables, other than sex. With this analysis workflow, the sex-associated differentially methylated regions were highly reproducible across studies, technologies, and statistical models.</p

Umbilical cord blood DNA methylation in children who later develop type 1 diabetes

Aims/hypothesis: Distinct DNA methylation patterns have recently been observed to precede type 1 diabetes in whole blood collected from young children. Our aim was to determine whether perinatal DNA methylation is associated with later progression to type 1 diabetes.Methods: Reduced representation bisulphite sequencing (RRBS) analysis was performed on umbilical cord blood samples collected within the Finnish Type 1 Diabetes Prediction and Prevention (DIPP) Study. Children later diagnosed with type 1 diabetes and/or who tested positive for multiple islet autoantibodies (n = 43) were compared with control individuals (n = 79) who remained autoantibody-negative throughout the DIPP follow-up until 15 years of age. Potential confounding factors related to the pregnancy and the mother were included in the analysis.Results: No differences in the umbilical cord blood methylation patterns were observed between the cases and controls at a false discovery rate Conclusions/interpretation: Based on our results, differences between children who progress to type 1 diabetes and those who remain healthy throughout childhood are not yet present in the perinatal DNA methylome. However, we cannot exclude the possibility that such differences would be found in a larger dataset.</p

KIELLETYN VÄRINEN TYTTÖ. Naiselliset piirteet Aila Meriluodon fantasiaromaanissa Vihreä tukka

Hakutermit: satu, fantasiakirjallisuus, lastenkirjallisuus, feministinen kirjallisuudentutkimus, Julia Kristeva, myyttinen tarina, stereotypiat, interpellaatio, dikotomiat, erillistäminen Tutkielmassa käsitellään Aila Meriluodon romaanin Vihreä tukka (1982) naisellisia piirteitä sekä niitä seikkoja, jotka tekevät romaanista feministisen. Teoreettisena viitekehyksenä tutkielmassa on käytetty useiden eri naistutkijoiden ajatuksia niin ranskalaisen kuin anglosaksisenkin tutkimuksen alueelta. Teoksen naisellisia piirteitä tarkastellaan teoksen tarinan, teemojen ja henkilöhahmojen kautta. Keskeiset tutkimusongelmat ovat naisellisuuden teemojen ohella tutkittavan teoksen lajityypin määrittely, teoksen luokittelun ongelmien tarkasteleminen sekä aikuisten- ja lastenkirjallisuuden välimaaston hahmotteleminen. Tutkielmassa keskeiseksi näin ollen nousee myös kysymys lastenkirjallisuuden ja feminismin sekä feministisen kirjallisuudentutkimuksen suhteesta. Teoksen lajityypin määrittelyn kohdalla tukeudutaan pääasiassa perinteisiin sadun ja fantasiakirjallisuuden teorioihin. Aikuisten- ja lastenkirjallisuuden rajamaastoa ja teoksen luokittelun ongelmia käsiteltäessä analyysi pohjaa pääasiassa tutkijan omaan tulkintaan. Tutkielman vahvasti tulkinnallisin osuus pohjaa Julia Kristevan ajatusten soveltamiseen fantasiakirjallisuuden analysoinnissa. Kristevan käsitteistöä ei tiettävästi aikaisemmin ole käytetty fantasiallisen aineksen tulkinnan välineenä. Esiin nousevat keskeiset kristevalaiset käsitteet geno- ja fenoteksti, semioottinen ja symbolinen sekä khora. Tutkielmassa päädytään siihen tulokseen, että Vihreä tukka kuuluu sadun lajityypin sijasta fantasiakirjallisuuden lajityyppiin. Teosta esitetään huomioitavaksi niihin konteksteihin, joissa puhutaan suomalaisen modernin lastenfantasian alkujuurista. Tulokset viittaavat myös siihen suuntaan, että teos on aikanaan luokiteltu virheellisesti pelkästään lastenkirjaksi. Tutkielman mukaan se soveltuu feministisen ja yhteiskuntakriittisen tematiikkansa ansiosta hyvin myös nuorten ja aikuisten luettavaksi. Lopuksi pohditaan kysymystä siitä, miksi Meriluodon teos on unohdettu suomalaisessa kirjallisuusinstituutiossa. Vastausta kysymykseen haetaan niin teoksen lajityypin, luokittelun, naisellisten piirteiden, kuin sen suoranaisen feministisen sanomankin tiimoilta