Risk factors and biomarkers for metastatic cutaneous squamous cell carcinoma

The incidence of cutaneous squamous cell carcinoma (cSCC), the most common skin cancer with metastatic potential, continues to increase. Although proportion of cSCCs metastasize and cause mortality, sufficient means to identify the metastasis-prone tumors are not available. In this thesis the metastatic cSCCs from the area served by Turku University Hospital were identified and characterized revealing that the rate of metastasis in the study region was 2.3%. Further, it was discovered that metastasis occurs rapidly and that there was no history of cSCC in 85% of patients with metastatic cSCC. Invasion depth, tumor diameter, age and location on lower lip or forehead were associated with increased risk of metastasis. On the other hand, usage of isosorbide mono-/dinitrate and aspirin as well as comorbidity with premalignant lesions or basal cell carcinoma were associated with lower risk of metastasis. With multiplexed immunohistochemistry, it was demonstrated that the activity and phenotype of cancer-associated fibroblasts (CAFs) evolve during the progression of cSCC. Elevation of α-smooth muscle actin (αSMA), secreted protein acidic and rich in cysteine (SPARC) and fibroblast activating protein (FAP) expression was associated with invasion and expression of FAP and platelet-derived growth factor receptor-β (PDGFRβ) with metastasis. High expression of stromal PDGFRβ and periostin were associated with worse prognosis. CAF107 (PDGFRα-/PDGFRβ+/FAP+) subset was associated with invasion and metastasis, and predicted poor prognosis of cSCC. A deep learning algorithm was harnessed to distinguish primary tumors that metastasize rapidly from non-metastatic cSCCs with slide level area under the receiver operating characteristic curve (AUROC) of 0.747 on whole slide images representing primary cSCCs. Furthermore, a risk factor model, that utilized prediction by AI, was created and provided staging systems and comparative risk factor models surpassing classification and prognostivity. These results characterize features associated with the metastasis risk of cSCC and indicate that CAF-markers and AI could provide clinical tools for the metastasis risk assessment and thus improve the prognosis of patient with metastatic cSCC.Etäpesäkkeitä lähettävän okasolusyövän riskitekijät ja biomarkkerit Yleisimmän etäpesäkkeitä lähettävän ihosyövän, okasolusyövän, ilmaantuvuus jatkaa kasvuaan. Vaikka osa okasolusyövistä lähettää etäpesäkkeitä ja aiheuttaa kuolleisuutta, ei etäpesäkkeitä lähettämään tulevien okasolusyöpien tunnistamiseksi ole toistaiseksi riittäviä keinoja. Tässä väitöskirjassa karakterisoitiin Turun yliopistollisen keskussairaalan vastuualueen metastasoituneet okasolusyövät ja osoitettiin että tutkimusalueen okasolusyövistä 2.3% etenee etäpesäkkeitä lähettäväksi. Metastasoituminen tapahtui nopeasti ja valtaosassa tapauksista (85%) etäpesäkkeen lähetti ensimmäinen potilaalla todettu okasolusyöpä. Ikä, kasvaimen invaasiosyvyys, halkaisija ja sijainti alahuulessa tai otsalla yhdistyivät kohonneeseen metastaasiriskiin. Isosorbidinitraatin ja aspiriinin käyttö sekä esiasteiden ja tyvisolusyövän esiintyminen taas liittyivät alentuneeseen metastaasiriskiin. Multiplex-immunohistokemiaa hyödyntäen osoitettin, että syöpään liittyvien fibroblastien (CAF) aktiviteetti ja ilmiasu muuttuu okasolusyövän edetessä. Kohonnut sileälihasaktiini alfan (αSMA), osteonektiinin ja fibroblastia aktivoivan proteiinin (FAP) ilmentyminen liittyi invaasioon ja FAP:n sekä verihiutaleista johdetun kasvutekijäreseptori β:n (PDGFRβ) etäpesäkkeiden lähettämiseen. PDGFRβ:n ja periostiinin ilmentyminen taas yhdistyi huonoon ennusteeseen. CAF107 (PDGFRα-/PDGFRβ+/FAP+) alatyyppi liittyi invaasioon, metastasointiin ja huonoon ennusteeeseen. Etäpesäkkeitä lähettämään tulevien okasolusyöpien tunnistamiseen valjastettu syväoppimisalgoritmi erotti okasolusyöpiä edustavista digitalisoiduista mikroskopiakuvista nopeasti etäpesäkkeitä lähettävät okasolusyövät okasolusyövistä, jotka eivät lähetä etäpesäkkeitä, leiketason AUROC-arvolla 0.747. Tekoälyarviota hyödyntävä riskitekijämalli voitti luokittelujärjestelmät ja kilpailevat riskitekijämallit okasolusyöpien luokittelussa ja ennusteen arvioinnissa. Tulokset antavat lisätietoa metastasoituvan okasolusyövän luonteesta ja osoittavat CAF-markkereiden sekä tekoälyn voivan tarjota kliinisiä työkaluja okasolusyövän metastaasiriskin arviointiin ja täten voivan parantaa etäpesäkkeitä lähettävän okasolusyöpäpotilaan ennustetta tulevaisuudessa

Risk Factors and Prognosis for Metastatic Cutaneous Squamous Cell Carcinoma: A Cohort Study

Cutaneous squamous cell carcinoma (cSCC) has me-tastatic potential. The aims of this study were to identify the risk factors for metastasis of primary cSCC and for poor prognosis in metastatic cSCC. Retrospective primary tumour cohorts of metastatic cSCCs (n = 85) and non-metastatic cSCCs (n = 218) were analysed. The mean annual rate of metastasis for primary cSCCs was 2.28%. In 49.4% of patients with metastatic cSCC, metastasis was detected within 6 months of diagnosis of the primary cSCC. There was no prior history of cSCC in 84.7% of metastatic cSCCs. Risk factors for metastasis included Clark's level 5, tumour diameter 20-29.9 mm, age at diagnosis = 3 nodal metastases and extranodal extension of metastasis. These results characterise new risk factors for metastatic cSCC

Ympäristövahingot Suomessa vuosina 2006-2012

Selvityksen tavoitteena oli tuottaa tietoa ympäristövahinkojen lukumäärästä, vahinkojen seurannasta ja merkityksestä Suomessa vuosina 2006–2012. Tarkastelu kohdistui ympäristövastuudirektiivin tarkoittamiin merkittäviin ympäristövahinkoihin sekä öljy- ja kemikaalivahinkoihin. Lisäksi selvitettiin, onko tapahtunut isännättömiä vahinkoja. Selvityksen ulkopuolelle rajattiin säteilyn, hajakuormituksen ja luonnonmullistusten aiheuttamat vahingot. Selvitys on jatkoa kolmelle aiemmalle selvitykselle vuosien 1989–1994, 1995–1999 ja 2000–2005 ympäristöonnettomuuksista. Selvitys tehtiin Suomen ympäristökeskuksessa. Aineistoon valittiin vuosilta 2006–2012 yhteensä noin 900 päästöjen ja niiden vaikutusten perusteella huomattavinta ympäristövahinkoa noin 20 000 öljy- ja kemikaalivahingon kokonaismäärästä. Selvityksen ulkopuolelle rajattiin tapaukset, joissa ympäristön pilaantumista ei ollut tapahtunut ja joissa päästö on ollut vähäinen. Aineistoa koottiin sisäasiainministeriön PRONTO-järjestelmästä, Turvallisuus- ja kemikaalivirasto Tukesin VARO-rekisteristä, Suomen ympäristökeskuksen vahinkopäivystyksestä sekä tiedostusvälineistä. Lisäksi tehtiin kysely elinkeino-, liikenne- ja ympäristökeskuksille ympäristövastuudirektiivissä tarkoitettujen vahinkotapausten kartoittamiseksi. Eri lähteiden kattavuus vaihteli, mutta PRONTO-järjestelmä on kattavin ja luotettavin yksittäinen tiedonlähde. Selvityksen aineistosta ilmenee, että noin 90 % ympäristövahingoista on öljyvahinkoja ja loput lähinnä teollisuuskemikaalien käytössä syntyneitä. Onnettomuuksia tapahtuu varsinkin liikenteessä, kuljetustoiminnassa sekä jätevedenpuhdistamoilla, ja syynä niihin ovat erityisesti tekniset viat ja inhimilliset erehdykset. Öljyn aiheuttamia isännättömiä vahinkoja tapahtui vuosittain 15–25 kappaletta. Muiden kemikaalien aiheuttamia isännättömiä vahinkoja tapahtui vain kaksi. Vahinkotapauksissa syntyy jonkun verran uusia pilaantuneita maa-alueita. Öljyvahinkojen yhteydessä syntyy myös saastuneita maamassoja, mutta niiden aiheuttama kuormitus jätehuollolle on kokonaisuudessaan vähäistä. Mahdollisia ympäristövahinkodirektiivissä tarkoitettuja merkittäviä ympäristövahinkoja havaittiin kaksi, kummatkin kaivostoiminnan yhteydessä

Pathways affected by asbestos exposure in normal and tumour tissue of lung cancer patients

<p>Abstract</p> <p>Background</p> <p>Studies on asbestos-induced tumourigenesis have indicated the role of, e.g., reactive oxygen/nitrogen species, mitochondria, as well as NF-κB and MAPK signalling pathways. The exact molecular mechanisms contributing to asbestos-mediated carcinogenesis are, however, still to be characterized.</p> <p>Methods</p> <p>In this study, gene expression data analyses together with gene annotation data from the Gene Ontology (GO) database were utilized to identify pathways that are differentially regulated in lung and tumour tissues between asbestos-exposed and non-exposed lung cancer patients. Differentially regulated pathways were identified from gene expression data from 14 asbestos-exposed and 14 non-exposed lung cancer patients using custom-made software and Iterative Group Analysis (iGA). Western blotting was used to further characterize the findings, specifically to determine the protein levels of UBA1 and UBA7.</p> <p>Results</p> <p>Differences between asbestos-related and non-related lung tumours were detected in pathways associated with, e.g., ion transport, NF-κB signalling, DNA repair, as well as spliceosome and nucleosome complexes. A notable fraction of the pathways down-regulated in both normal and tumour tissue of the asbestos-exposed patients were related to protein ubiquitination, a versatile process regulating, for instance, DNA repair, cell cycle, and apoptosis, and thus being also a significant contributor of carcinogenesis. Even though UBA1 or UBA7, the early enzymes involved in protein ubiquitination and ubiquitin-like regulation of target proteins, did not underlie the exposure-related deregulation of ubiquitination, a difference was detected in the UBA1 and UBA7 levels between squamous cell carcinomas and respective normal lung tissue (p = 0.02 and p = 0.01) without regard to exposure status.</p> <p>Conclusion</p> <p>Our results indicate alterations in protein ubiquitination related both to cancer type and asbestos. We present for the first time pathway analysis results on asbestos-associated lung cancer, providing important insight into the most relevant targets for future research.</p

Complement Factor D Is a Novel Biomarker and Putative Therapeutic Target in Cutaneous Squamous Cell Carcinoma

Cutaneous squamous cell carcinoma (cSCC) is the most prevalent metastatic skin cancer. Previous studies have demonstrated the autocrine role of complement components in cSCC progression. We have investigated factor D (FD), the key enzyme of the alternative complement pathway, in the development of cSCC. RT-qPCR analysis of cSCC cell lines and normal human epidermal keratinocytes (NHEKs) demonstrated significant up-regulation of FD mRNA in cSCC cells compared to NHEKs. Western blot analysis also showed more abundant FD production by cSCC cell lines. Significantly higher FD mRNA levels were noted in cSCC tumors than in normal skin. Strong tumor cell-associated FD immunolabeling was detected in the invasive margin of human cSCC xenografts. More intense tumor cell-specific immunostaining for FD was seen in the tumor edge in primary and metastatic cSCCs, in metastases, and in recessive dystrophic epidermolysis bullosa-associated cSCCs, compared with cSCC in situ, actinic keratosis and normal skin. FD production by cSCC cells was dependent on p38 mitogen-activated protein kinase activity, and it was induced by interferon-γ and interleukin-1β. Blocking FD activity by Danicopan inhibited activation of extracellular signal-regulated kinase 1/2 and attenuated proliferation of cSCC cells. These results identify FD as a novel putative biomarker and therapeutic target for cSCC progression

Miksi kato leviää? hd-yhtymän katovariantin diffuusion syyt Helsingin puhekielessä

Peer reviewe

Super Enhancer-Regulated LINC00094 (SERLOC) Upregulates the Expression of MMP-1 and MMP-13 and Promotes Invasion of Cutaneous Squamous Cell Carcinoma

Simple Summary Cutaneous squamous cell carcinoma (cSCC) is the most common metastatic skin cancer, and its incidence is increasing worldwide. The prognosis of the metastatic disease is poor, and there are no established biomarkers for the assessment of metastasis risk or specific therapeutic targets for advanced or metastatic cSCCs. The role of long non-coding RNAs (lncRNAs) in the progression of cSCC has recently been emphasized. Super enhancers (SE) have been shown to play a role in tumorigenesis and regulate the expression of specific lncRNAs. In this study, we evaluated the role of SE-regulated BRD3OS (lncRNA LINC00094) in the progression of cSCC. Based on the results, we named this lncRNA SERLOC. The results identify SERLOC as a biomarker for invasion and metastasis of cSCC and as a putative therapeutic target in advanced cSCC. Long non-coding RNAs (lncRNAs) have emerged as important regulators of cancer progression. Super enhancers (SE) play a role in tumorigenesis and regulate the expression of specific lncRNAs. We examined the role of BRD3OS, also named LINC00094, in cutaneous squamous cell carcinoma (cSCC). Elevated BRD3OS (LINC00094) expression was detected in cSCC cells, and expression was downregulated by SE inhibitors THZ1 and JQ1 and via the MEK1/ERK1/2 pathway. Increased expression of BRD3OS (LINC00094) was noted in tumor cells in cSCCs and their metastases compared to normal skin, actinic keratoses, and cSCCs in situ. Higher BRD3OS (LINC00094) expression was noted in metastatic cSCCs than in non-metastatic cSCCs. RNA-seq analysis after BRD3OS (LINC00094) knockdown revealed significantly regulated GO terms Cell-matrix adhesion, Basement membrane, Metalloendopeptidase activity, and KEGG pathway Extracellular matrix-receptor interaction. Among the top-regulated genes were MMP1, MMP10, and MMP13. Knockdown of BRD3OS (LINC00094) resulted in decreased production of MMP-1 and MMP-13 by cSCC cells, suppressed invasion of cSCC cells through collagen I, and growth of human cSCC xenografts in vivo. Based on these observations, BRD3OS (LINC00094) was named SERLOC (super enhancer and ERK1/2-Regulated Long Intergenic non-protein coding transcript Overexpressed in Carcinomas). These results reveal the role of SERLOC in cSCC invasion and identify it as a potential therapeutic target in advanced cSCC.</p

Gene expression profiles in asbestos-exposed epithelial and mesothelial lung cell lines

BACKGROUND: Asbestos has been shown to cause chromosomal damage and DNA aberrations. Exposure to asbestos causes many lung diseases e.g. asbestosis, malignant mesothelioma, and lung cancer, but the disease-related processes are still largely unknown. We exposed the human cell lines A549, Beas-2B and Met5A to crocidolite asbestos and determined time-dependent gene expression profiles by using Affymetrix arrays. The hybridization data was analyzed by using an algorithm specifically designed for clustering of short time series expression data. A canonical correlation analysis was applied to identify correlations between the cell lines, and a Gene Ontology analysis method for the identification of enriched, differentially expressed biological processes. RESULTS: We recognized a large number of previously known as well as new potential asbestos-associated genes and biological processes, and identified chromosomal regions enriched with genes potentially contributing to common responses to asbestos in these cell lines. These include genes such as the thioredoxin domain containing gene (TXNDC) and the potential tumor suppressor, BCL2/adenovirus E1B 19kD-interacting protein gene (BNIP3L), GO-terms such as "positive regulation of I-kappaB kinase/NF-kappaB cascade" and "positive regulation of transcription, DNA-dependent", and chromosomal regions such as 2p22, 9p13, and 14q21. We present the complete data sets as Additional files. CONCLUSION: This study identifies several interesting targets for further investigation in relation to asbestos-associated diseases

Complement System in Cutaneous Squamous Cell Carcinoma

Epidermal keratinocyte-derived cutaneous squamous cell carcinoma (cSCC) is the most common metastatic skin cancer with high mortality rates in the advanced stage. Chronic inflammation is a recognized risk factor for cSCC progression and the complement system, as a part of innate immunity, belongs to the microenvironment of tumors. The complement system is a double-edged sword in cancer, since complement activation is involved in anti-tumor cytotoxicity and immune responses, but it also promotes cancer progression directly and indirectly. Recently, the role of several complement components and inhibitors in the regulation of progression of cSCC has been shown. In this review, we will discuss the role of complement system components and inhibitors as biomarkers and potential new targets for therapeutic intervention in cSCC