Sekvenciranje RNK na ravni posameznih celic: revolucionarna tehnologija, ki nadgrajuje razumevanje kompleksnih bolezni in spodbuja oseben pristop k zdravljenju – primer melanoma kože

Tehnologija sekvenciranja RNK na ravni posameznih celic (scRNAseq) nam omogoča, da z visoko ločljivostjo in natančnostjo naenkrat določimo nabor vseh molekul RNK v vsaki posamezni celici, ki se nahaja v določenem vzorcu oz. tkivu. Danes je scRNAseq pomembno orodje predvsem za proučevanje kompleksnih bioloških sistemov in tkiv, kot je tumorsko tkivo, kjer je velika celična raznolikost ključnega pomena. V članku navajamo primer melanoma kože, ki je eden najpogostejših in najbolj agresivnih rakov v razvitem svetu. Čeprav se je v zadnjem času z uvedbo imunske terapije napoved izida melanoma bistveno izboljšala, pa je še vedno približno 30–40 % bolnikov, pri katerih tovrstno zdravljenje ni uspešno. Novi podatki, pridobljeni z uporabo scRNAseq, so razkrili, da je mehanizem odpornosti na zdravljenje z zaviralci imunskih nadzornih točk zelo kompleksen, da na to poleg prisotnosti in fenotipa izčrpanih limfocitov T CD8+ vpliva tudi mutacija v genu BRAF, fenotip melanocitov, prisotnost in fenotip celic mieloičnega izvora, prisotnost fibroblastov različnega fenotipa ter interakcije med vsemi celicami, ki tvorijo tumorsko mikrookolje. V prihodnosti bo torej vse bolj pomemben oseben pristop zdravljenja, ki bo temeljil na molekularni in celični opredelitvi tumorja in njegovega mikrookolja ter na napovednih bioloških označevalcih. Z uporabo tehnologije scRNAseq se bomo lahko cilju osebne medicine zelo približali, saj nam omogoča identifikacijo posameznih celic in celičnih označevalcev, ki bi lahko napovedali odziv bolnika na zdravljenje in omogoča bolj ciljano odločitev za vrsto zdravljenja za posameznega bolnika. Na ta način bi se izognili principu zdravljenja, ki temelji na “poskusu in napaki” ter tako bistveno izboljšali učinkovitost zdravljenja. Zaenkrat pa se tehnologija scRNAseq uporablja zgolj v raziskovalne namene, zato zaradi določenih omejitev ni uvedena v dejansko klinično prakso

GENETIKA SINDROMA POLICISTIČNIH JAJNIKA

Polycystic ovary syndrome (PCOS), a heterogeneous complex disease, is presented in the light of its multifactorial pathogenesis. In particular, the genetic background of the syndrome and the most promising PCOS candidate genes are discussed in more detail.Prikazan je sindrom policističnih jajnika (PCOS) kao heterogena kompleksna bolest, uglavnom iz vidika njegove multifaktorske patogeneze. Genetska pozadina ovog sindroma i najvjerojatniji PCOS geni kandidati detaljno su obrazloženi

The effect of micro-sized titanium dioxide on WM-266-4 metastatic melanoma cell line

Titanium dioxide (TiO2) is widely used as an inorganic UV-filter in cosmetic products; however, it has been classified as possibly carcinogenic to humans. While numerous studies demonstrated cytotoxic and genotoxic effects of nano-sized TiO2 in different cell lines, including human skin cells, studies investigating the effects of micro-TiO2 on human keratinocytes and melanocytes, in healthy and cancer cells, are scarce. Adenosine triphosphate (ATP) binding cassette subfamily B member 5 (ABCB5) is a plasma membrane protein known for its role in the tumorigenicity, progression, and recurrence of melanoma. Here, we investigated the effect of micro-TiO2 (average particle size ≤5 µm) on the metabolic activity, cytotoxicity and ABCB5 mRNA expression in metastatic melanoma cells. Metastatic melanoma cell line WM-266-4 was treated with different concentrations of micro-TiO2 for different incubation times to obtain dose- and time-dependent responses. Untreated WM-266-4 cells, cultured under the same conditions, were used as control. The cell metabolic activity was determined by MTT assay. Cytotoxicity of micro-TiO2 was analyzed by lactate dehydrogenase (LDH) cytotoxicity assay. The ABCB5 mRNA expression in melanoma cells was analyzed using quantitative reverse transcription polymerase chain reaction (RT-qPCR). After 120 hours of exposure to micro-TiO2 the metabolic activity of melanoma cells decreased, especially at the two highest micro-TiO2 concentrations. Comparably, the cytotoxicity of micro-TiO2 on melanoma cells increased after 48 and 120 hours of exposure, in a time-dependent manner. The ABCB5 mRNA expression in micro-TiO2-treated melanoma cells also decreased significantly after 24 and 48 hours, in a time-dependent manner. Overall, our results suggest inhibitory effects of micro-TiO2 on the metabolic activity and ABCB5 mRNA expression in metastatic melanoma cells, indicating its potential use as an anticancer agent

Cannabinoids in cancer treatment: Therapeutic potential and legislation

The plant Cannabis sativa L. has been used as an herbal remedy for centuries and is the most important source of phytocannabinoids. The endocannabinoid system (ECS) consists of receptors, endogenous ligands (endocannabinoids) and metabolizing enzymes, and plays an important role in different physiological and pathological processes. Phytocannabinoids and synthetic cannabinoids can interact with the components of ECS or other cellular pathways and thus affect the development/progression of diseases, including cancer. In cancer patients, cannabinoids have primarily been used as a part of palliative care to alleviate pain, relieve nausea and stimulate appetite. In addition, numerous cell culture and animal studies showed antitumor effects of cannabinoids in various cancer types. Here we reviewed the literature on anticancer effects of plant-derived and synthetic cannabinoids, to better understand their mechanisms of action and role in cancer treatment. We also reviewed the current legislative updates on the use of cannabinoids for medical and therapeutic purposes, primarily in the EU countries. In vitro and in vivo cancer models show that cannabinoids can effectively modulate tumor growth, however, the antitumor effects appear to be largely dependent on cancer type and drug dose/concentration. Understanding how cannabinoids are able to regulate essential cellular processes involved in tumorigenesis, such as progression through the cell cycle, cell proliferation and cell death, as well as the interactions between cannabinoids and the immune system, are crucial for improving existing and developing new therapeutic approaches for cancer patients. The national legislation of the EU Member States defines the legal boundaries of permissible use of cannabinoids for medical and therapeutic purposes, however, these legislative guidelines may not be aligned with the current scientific knowledge

Genetic Polymorphisms of INS, INSR and IRS-1 Genes Are Not Associated with Polycystic Ovary Syndrome in Croatian Women

Obesity and insulin resistance is a common finding in patients with polycystic ovary syndrome (PCOS). Significant number of PCOS women experience insulin resistance that is irrespective of the degree of obesity suggesting possible genetic basis. Therefore, several polymorphisms of the genes encoding for the insulin (INS), insulin receptor (INSR) or insulin receptor substrates (IRS) involved in postreceptor signaling have been explored for their association with abnormal sensitivity to insulin in PCOS. The aim of the present study was to determine whether selected polymorphisms of INS, INSR and IRS-1 are associated with the development of PCOS as well as with increased insulin resistance in Croatian women with PCOS. The study enrolled 150 women with PCOS and 175 control women. The diagnosis of PCOS was based on Rotterdam consensus criteria. Each subject underwent an evaluation of body mass index (BMI), hirsutism, acne and menstrual cycle abnormalities as well as follicular stimulating hormone (FSH), luteinizing hormone (LH), total and free testosterone, androstendione, dehydroepiandrosterone sulphate (DHEAS), sex hormone binding globulin (SHBG), fasting glucose and fasting insulin. Insulin resistance (IR) was quantified using the homeostatic model assessment of IR (HOMA-IR). Molecular analyses for the genetic polymorphisms were preformed. There was a significant difference in clinical and biochemical characteristics of the studied groups except for BMI and fasting glucose levels. No significant differences were observed in the genotype and allele distribution of the VNTR INS, C/T INSR, Gly792Arg IRS-1 polymorphisms between cases and controls. Moreover, no association was found between VNTR INS, C/T INSR and Gly792Arg IRS-1 polymorphism and parameters of insulin resistance in PCOS patients. In conclusion, our data does not support an association between VNTR INS, C/T INSR and Gly792Arg IRS-1 polymorphism and susceptibility to PCOS or insulin resistance in Croatian women with PCOS

Systems Biology in ELIXIR: modelling in the spotlight

In this white paper, we describe the founding of a new ELIXIR Community - the Systems Biology Community - and its proposed future contributions to both ELIXIR and the broader community of systems biologists in Europe and worldwide. The Community believes that the infrastructure aspects of systems biology - databases, (modelling) tools and standards development, as well as training and access to cloud infrastructure - are not only appropriate components of the ELIXIR infrastructure, but will prove key components of ELIXIR\u27s future support of advanced biological applications and personalised medicine. By way of a series of meetings, the Community identified seven key areas for its future activities, reflecting both future needs and previous and current activities within ELIXIR Platforms and Communities. These are: overcoming barriers to the wider uptake of systems biology; linking new and existing data to systems biology models; interoperability of systems biology resources; further development and embedding of systems medicine; provisioning of modelling as a service; building and coordinating capacity building and training resources; and supporting industrial embedding of systems biology. A set of objectives for the Community has been identified under four main headline areas: Standardisation and Interoperability, Technology, Capacity Building and Training, and Industrial Embedding. These are grouped into short-term (3-year), mid-term (6-year) and long-term (10-year) objectives

Community-driven ELIXIR activities in single-cell omics

Single-cell omics (SCO) has revolutionized the way and the level of resolution by which life science research is conducted, not only impacting our understanding of fundamental cell biology but also providing novel solutions in cutting-edge medical research. The rapid development of single-cell technologies has been accompanied by the active development of data analysis methods, resulting in a plethora of new analysis tools and strategies every year. Such a rapid development of SCO methods and tools poses several challenges in standardization, benchmarking, computational resources and training. These challenges are in line with the activities of ELIXIR, the European coordinated infrastructure for life science data. Here, we describe the current landscape of and the main challenges in SCO data, and propose the creation of the ELIXIR SCO Community, to coordinate the efforts in order to best serve SCO researchers in Europe and beyond. The Community will build on top of national experiences and pave the way towards integrated long-term solutions for SCO research. Keywor

GENETIKA SINDROMA POLICISTIČNIH JAJNIKA

Polycystic ovary syndrome (PCOS), a heterogeneous complex disease, is presented in the light of its multifactorial pathogenesis. In particular, the genetic background of the syndrome and the most promising PCOS candidate genes are discussed in more detail.Prikazan je sindrom policističnih jajnika (PCOS) kao heterogena kompleksna bolest, uglavnom iz vidika njegove multifaktorske patogeneze. Genetska pozadina ovog sindroma i najvjerojatniji PCOS geni kandidati detaljno su obrazloženi

Variability in pharmacological response to metformin treatment

Izhodišča: Metformin je peroralni antidiabetik in je po slovenskih smernicah zdravilo prve izbire za zdravljenje sladkorne bolezni tipa 2 (SB2). Svoje mesto si utira tudi pri zdravljenju sindroma policističnih jajčnikov (PCOS), čeprav PCOS (še) ni uradno med indikacijami za zdravljenje z metforminom. Študije kažejo, da metformin izboljša klinično in biokemično sliko PCOS ter zveča stopnjo ovulacije in s tem verjetnost za zanositev pri ženskah s tem sindromom. Farmakološki učinki metformina se ob enakem režimu odmerjanja med bolniki razlikujejo, kar poskušamo med drugim razložiti tudi z vplivom polimorfnih različic v genih, povezanih s farmakodinamiko in farmakokinetiko metformina. Zaključki: Rezultati raziskav kažejo, da polimorfne različice v genih, povezanih s farmakodinamiko in farmakokinetiko metformina, lahko prispevajo k raznolikosti v odzivu na zdravljenje z metforminom tako pri bolnikih s SB2 kot pri bolnicah s PCOS.Background: According to the Slovenian guidelines, metformin, an oral antidiabetic drug, is a drug of choice for the treatment of type 2 diabetes. Additionally, metformin is paving its way in polycystic ovary syndrome (PCOS) treatment, although the drug has not (yet) been officially indicated to treat PCOS. Studies have shown that metformin improves clinical and biochemical features of PCOS as well as the rate of ovulation and consequently the likelihood of conception in PCOS women. At the same dosage regimen, pharmacological effects of metformin show interindividual variability in metformin responseefforts have been made to at least partly explain the variability with polymorphic variants in genes related to metformin pharmacodynamics and pharmacokinetics. Conclusions: The results of the current studies indicate that polymorphic variants in genes related to metformin pharmacodynamics and pharmacokinetics may contribute to the interindividual variability in pharmacological response to metformin treatment in diabetic as well as in PCOS patients

Drug interactions with angiotensin-converting enzyme inhibitors

V članku predstavljamo pregled potencialno klinično pomembnih interakcij zdravil (DDIs) z zaviralci angiotenzinske konvertaze (ACE). Na osnovi pregleda dveh knjig Stockleyʼs Drug Interactions in dveh preko spleta dostopnih podatkovnih zbirk o DDIs smo opisali več kot deset takih DDIs, razložili njihove mehanizme, opredelili njihov klinični pomen ter navedli možnosti za njihovo preprečevanje oz. ustrezno obravnavo. Večina DDIs z zaviralci ACE je farmakodinamičnih, le nekaj jih je na nivoju farmakokinetike. Največ DDIs z zaviralci ACE se odraža kot prekomerno znižanje krvnega tlaka, hiperkaliemija ali nefrotoksičnost. Dokazali so tudi, da zaviralci ACE zvečajo toksičnost litija in da lahko vplivajo na pojav hudih preobčutljivostnih reakcij pri bolnikih, ki so prejeli parenteralne pripravke železa, ali pri bolnikih, ki sočasno jemljejo alopurinol. Prav tako je zaviralce ACE zaradi njihove lastne hepatotoksičnosti potrebno previdno kombinirati z drugimi hepatoksičnimi zdravili. Zaradi velike porabe zaviralcev ACE je pogostost neželenih učinkov zdravil zaradi DDIs z zaviralci ACE visoka. Ugodno pa je, da lahko v večini primerov teh DDIs ustrezno ukrepamo že s primernim kliničnim in laboratorijskim spremljanjem bolnikov oz. s prilagoditvijo odmerkov sočasno predpisanih zdravil