TGF-β family signaling in the regulation of cell plasticity in lung cells and mesothelioma

The aim of this thesis was to extend the understanding of molecular mechanisms of asbestos exposure related diseases asbestosis (lung fibrosis), lung cancer and mesothelioma. Epithelial-to-mesenchymal transition (EMT) is implicated in fibrosis and in cancer, both asbestos induced diseases. The first of the three studies investigated whether asbestos fibers induced EMT. In the next two studies, the focus was on mesothelioma. In these two studies, the aim was to recognize novel potential therapeutic target molecules and pathways involved in chemoresistance and invasion. We found that exposure to asbestos led to loss of epithelial characteristics when lung epithelial cells retained the type II airway epithelial cell phenotype. This loss of epithelial characteristics was found not to depend on TGF-β or Smad signaling cascades. The MAPK/ERK pathway was found to mediate epithelial plasticity in response to asbestos exposure. A screen for new gremlin-1 interacting proteins discovered fibrillin-2. This finding was validated in mesothelioma tumor samples in which gremlin-1 and fibrillin-2 co-localized in the tumors. Primary mesothelioma cells were harvested from pleural effusion samples of mesothelioma patients. These cells recapitulated primary tumor characteristics and overexpressed gremlin-1, fibrillin-2 and transcription factor slug. Silencing of gremlin-1 downregulated slug, reverted the EMT-phenotype and sensitized the cells to the cytotoxic effect of chemotherapeutic drugs. The concomitant overexpression of gremlin-1 and fibrillin-2 enables gremlin targeting to fibrillin-2 containing fibers in mesothelioma extracellular matrix, where it supports the chemoresistant EMT phenotype through transcription factor slug. Mesothelioma tumors as well as primary mesothelioma cells and mesothelioma cell lines were found to overexpress activin-A and activin-B. Canonical Smad3 response to activin stimulation was attenuated in mesothelioma cells. Attenuation of the Smad3 response associated with migratory and invasive phenotype. Mesothelioma cells switch from canonical Smad3 signaling to non-canonical MAPK/ERK pathway, and this promotes migration and invasion. Invasion, migration and ERK activation was impaired by sequestering extracellular activins by soluble activin-receptor. Likewise, inhibiting MAPK/ERK upstream kinase impaired migration and invasion The results presented in this thesis support the concept that pathological EMT is a central mechanism in the development and progression of asbestos induced fibrotic and malignant diseases. They also suggest gremlin-1 and activin-A as new potential therapeutic targets in mesothelioma.Väitöskirjatyö tutki asbestin aiheuttamien sairauksien molekyylimekanismeja. Asbestialtistus aiheuttaa keuhkojen sidekudostumista (asbestoosi), keuhkosyöpää sekä keuhkopussin syöpää (mesoteliooma). Epiteeli-mesenkyymi-transitio (EMT) on tärkeä mekanismi syövän ja sidekudossairauksien etenemisessä. Ensimmäisessä osatyössä tutkittiin aiheuttaako asbesti EMT:tä. Kaksi seuraavaa osatyötä keskittyivät mesotelioomaan ja niissä etsittiin molekyylejä ja solunsisäisiä viestireittejä, jotka vaikuttavat mesoteliooman kykyyn tunkeutua ympäröiviin kudoksiin (invasiivinen kasvu) ja kykyyn vastustaa solunsalpaajahoitoja. Havaitsimme, että tyypin II keuhkoepiteelisolut, menettävät epiteliaalisia ominaisuuksiaan asbestialtistuksen seurauksena. Tällainen plastisuus viittaa EMT tapahtumiin. Nämä tapahtumat eivät olleet riippuvaisia TGF-β tai Smad- viestireiteistä. MAPK/ERK viestireitti osoittautui olevan se, joka aktivoituu asbestialtistuksen seurauksena ja joka sai aikaan epiteeli-mesenkyymi-transitioon viittaavaa plastisuutta keuhkojen epiteelisoluissa. Systemaattisessa etsinnässä fibrilliini-2 paljastui uudeksi gremliini-1:teen sitoutuvaksi proteiiniksi. Gremliiniä ja fibrilliiniä ilmennettiin mesotelioomapotilaiden syöpäkudoksessa ja proteiinit lokalisoituivat hyvin lähelle toisiaan. Mesotelioomapotilaiden pleuranesteestä eristetyt ja kasvatetut primaarit syöpäsolut yli-ilmensivät gremliiniä, fibrilliiniä sekä transkriptiofaktori slugia syöpäkasvaimen tapaan. Gremliinin ilmenemistasojen lasku johti slugin ilmenemistasojen laskuun, EMT-fenotyypin peruuntumiseen ja lisäsi solujen herkkyyttä solunsalpaajille. Gremliinin ja fibrilliinin samanaikainen yli-ilmentäminen mesotelioomassa mahdollistaa gremliini-1:n lokalisaation soluväliaineeseen mesotelioomassa. Gremliini-1 ylläpitää lääkeresistenttiä EMT- fenotyyppiä slugin välityksellä. Aktiviini-A:ta ja aktiviini-B:tä yli-ilmennetään mesotelioomasyöpäkudoksessa sekä mesotelioomasoluissa. Havaittiin, että mesotelioomasoluissa kanoninen Smad3 vaste aktiviineille on heikentynyt ja tämä piirre kytkeytyi solun kykyyn invasiiviseen kasvuun ja migraatioon. Muutos Smad3- viestireitiltä MAPK/ERK-viestireitille edisti mesotelioomasolujen invasiivisuutta ja migraatiota. Kun aktiviinien sitoutuminen reseptoreihin estettiin, heikkeni mesotelioomasolujen migraatio ja invaasio. Samoin teki MAPK/ERK-viestireitin salpaus. Tämän väitöskirjatyön tulokset tukevat näkemystä, että patologinen EMT on keskeinen mekanismi asbestisairauksien kehittymisessä ja etenemisessä. Tutkimuksen valossa gremliini-1 ja aktiviini-A ovat uusia potentiaalisia kohdemolekyylejä mesoteliooman hoidossa

Quantitative online survey of self-perceived knowledge and knowledge gaps of medicines research and development among Finnish general public

Objectives This study explored self-reported knowledge and interest to learn more about medicines research, development and health technology assessment among Finnish general public. It also aimed to define possible knowledge gaps and needs for public education regarding these topics. Design Online survey with 503 participants. The questionnaire was originally developed as a part of the Needs Assessment Work Package of the European Patients' Academy on Therapeutic Innovation Project. The survey was carried out in Finland in 2019. Methods The survey was conducted as an online survey by Kantar TNS Gallup Forum online panel. The data were analysed by using the freely available programming language R. Relationships between the demographic characteristics (such as age, gender and education level) of respondents and their knowledge or interest in medicines research and development were determined using Pearson's chi(2) tests. Statistically significant responses of demographic characteristics in the respondents' knowledge or interest in medicines research were determined by logistic regression. Results Of the 503 respondents (age 16-64) only 12% reported having good or very good knowledge of medicines research and development in general. Regarding health technology assessment, pharmacoeconomics and regulation, the percentage of respondents reporting good or very good knowledge was as low as 8%. Respondents were most interested in learning more about predictive and personalised medicine (47%) and least interested in medicines regulation (30%) and pharmacoeconomics (31%). Conclusions Self-reported knowledge about medicines research and development and health technology assessment appears to be very low in Finland. Patient and public participation is recognised as an important and essential element in up-to-date medical research and assessment of new treatments. In order to participate as an active and equal partner in these processes, the public needs more information and education in these topics.Peer reviewe

Finanssipolitiikan päätösperäisyyden arvioiminen – vaihtoehtoisten mittareiden esittely

Tässä artikkelissa kuvataan vaihtoehtoisia tapoja arvioida finanssipolitiikan viritystä. Tarkastelun pääpaino on EU-yhteyksissä käytettyjen indikaattoreiden vertailussa. Keskitymme rakenteelliseen alijäämään ja sen muutokseen, menosääntöön, yksittäisten harkinnanvaraisten toimien tarkasteluun kokonaisuutena, sekä päätösperäistä finanssipolitiikkaa kuvaavaan DFE-mittariin. Lisäksi arvioimme väestön ikääntymisen menoja automaattisesti kasvattavan vaikutuksen suuruutta ja sen vaikutusta eri mittareihin. Vertailumme tärkein tulos on, että kaikissa mittareissa on puutteensa. Ne myös mittaavat osin eri asioita. Tarkastelun perusteella eri mittareiden antama kuva finanssipolitiikan mitoituksesta on pääsääntöisesti samansuuntainen, vaikka arvio vaikutuksen suuruudesta vaihtelee toisinaan paljon. Siten useamman eri mittarin käyttö on perusteltua. Ikääntymisen vaikutus esimerkiksi rakenteellisen jäämän muutokseen on keskimäärin noin 0,3 prosenttiyksikköä vuodessa (elvyttävä). Vaikka ikääntyminen vaikuttaa eri mittareihin eri tavoin, se ei täysin selitä mittareissa havaittuja eroj

Ecology determines how low antibiotic concentration impacts community composition and horizontal transfer of resistance genes

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Host range of antibiotic resistance genes in wastewater treatment plant influent and effluent

10.1093/femsec/fiy038Wastewater treatment plants (WWTPs) collect wastewater from various sources for a multi-step treatment process. By mixing a large variety of bacteria and promoting their proximity, WWTPs constitute potential hotspots for the emergence of antibiotic resistant bacteria. Concerns have been expressed regarding the potential of WWTPs to spread antibiotic resistance genes (ARGs) from environmental reservoirs to human pathogens. We utilized epicPCR (Emulsion, Paired Isolation and Concatenation PCR) to detect the bacterial hosts of ARGs in two WWTPs. We identified the host distribution of four resistance-associated genes (tetM, int1, qacE Delta 1 and bla(OXA-58)) in influent and effluent. The bacterial hosts of these resistance genes varied between the WWTP influent and effluent, with a generally decreasing host range in the effluent. Through 16S rRNA gene sequencing, it was determined that the resistance gene carrying bacteria include both abundant and rare taxa. Our results suggest that the studied WWTPs mostly succeed in decreasing the host range of the resistance genes during the treatment process. Still, there were instances where effluent contained resistance genes in bacterial groups not carrying these genes in the influent. By permitting exhaustive profiling of resistance-associated gene hosts in WWTP bacterial communities, the application of epicPCR provides a new level of precision to our resistance gene risk estimates.Peer reviewe

Construction and characterization of synthetic bacterial community for experimental ecology and evolution

Experimental microbial ecology and evolution have yielded foundational insights into ecological and evolutionary processes using simple microcosm setups and phenotypic assays with one- or two-species model systems. The fields are now increasingly incorporating more complex systems and exploration of the molecular basis of observations. For this purpose, simplified, manageable and well-defined multispecies model systems are required that can be easily investigated using culturing and high-throughput sequencing approaches, bridging the gap between simpler and more complex synthetic or natural systems. Here we address this need by constructing a completely synthetic 33 bacterial strain community that can be cultured in simple laboratory conditions. We provide whole-genome data for all the strains as well as metadata about genomic features and phenotypic traits that allow resolving individual strains by amplicon sequencing and facilitate a variety of envisioned mechanistic studies. We further show that a large proportion of the strains exhibit coexistence in co-culture over serial transfer for 48 days in the absence of any experimental manipulation to maintain diversity. The constructed bacterial community can be a valuable resource in future experimental work.Peer reviewe

Quantitative online survey of self-perceived knowledge and knowledge gaps of medicines research and development among Finnish general public

Objectives This study explored self-reported knowledge and interest to learn more about medicines research, development and health technology assessment among Finnish general public. It also aimed to define possible knowledge gaps and needs for public education regarding these topics.Design Online survey with 503 participants. The questionnaire was originally developed as a part of the Needs Assessment Work Package of the European Patients’ Academy on Therapeutic Innovation Project. The survey was carried out in Finland in 2019.Methods The survey was conducted as an online survey by Kantar TNS Gallup Forum online panel. The data were analysed by using the freely available programming language R. Relationships between the demographic characteristics (such as age, gender and education level) of respondents and their knowledge or interest in medicines research and development were determined using Pearson’s χ2 tests. Statistically significant responses of demographic characteristics in the respondents’ knowledge or interest in medicines research were determined by logistic regression.Results Of the 503 respondents (age 16–64) only 12% reported having good or very good knowledge of medicines research and development in general. Regarding health technology assessment, pharmacoeconomics and regulation, the percentage of respondents reporting good or very good knowledge was as low as 8%. Respondents were most interested in learning more about predictive and personalised medicine (47%) and least interested in medicines regulation (30%) and pharmacoeconomics (31%).Conclusions Self-reported knowledge about medicines research and development and health technology assessment appears to be very low in Finland. Patient and public participation is recognised as an important and essential element in up-to-date medical research and assessment of new treatments. In order to participate as an active and equal partner in these processes, the public needs more information and education in these topics.</p

Overexpression of activin-A and -B in malignant mesothelioma - Attenuated Smad3 signaling responses and ERK activation promote cell migration and invasive growth

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Perinnöllinen sairastumisalttius kiinnostaa kansalaisia

Vertaisarvioitu. English summaryJOHDANTO : Selvitimme kansalaisten asenteita ja odotuksia yksilön perimästä saatavan tiedon, genomitiedon, keräämisestä ja käytöstä lääketieteellisiin tutkimustarkoituksiin. MENETELMÄT : Valtakunnallinen kyselytutkimus toteutettiin tammikuussa 2019. TULOKSET : Kansalaiset suhtautuvat genomitiedon keräämiseen ja käyttöön myönteisesti ja haluaisivat tietää terveydentilaansa mahdollisesti vaikuttavista perinnöllisistä alttiuksista. Erityisesti naiset olisivat valmiita tekemään saamansa tiedon perusteella myös elintapamuutoksia. PÄÄTELMÄT : Tutkimus vahvistaa käsitystä siitä, että suomalaiset ovat tutkimusmyönteisiä ja valmiita luovuttamaan näytteensä biopankkiin. Toisaalta useat kyselytutkimuksen vastaajista eivät osanneet sanoa kantaansa heille esitettyihin kysymyksiin, mikä viittaa osaltaan siihen, että kansalaiset kaipaisivat lisää tietoa sekä genomitiedon käytöstä että siihen liittyvistä ongelmista ja mahdollisuuksista.Peer reviewe