Oxysterols, oxidized derivatives of cholesterol, act in breast cancer (BC) as selective estrogen receptor modulators and affect cholesterol homeostasis, drug transport, nuclear and cell receptors, and other signaling proteins. Using data from three highly overlapping sets of patients (N = 162 in total) with early-stage estrogen-receptor-positive luminal BC-high-coverage targeted DNA sequencing (113 genes), mRNA sequencing, and full micro-RNA (miRNA) transcriptome microarrays-we describe complex oxysterol-related interaction (correlation) networks, with validation in public datasets (n = 538) and 11 databases. The ESR1-CH25H-INSIG1-ABCA9 axis was the most prominent, interconnected through miR-125b-5p, miR-99a-5p, miR-100-5p, miR-143-3p, miR-199b-5p, miR-376a-3p, and miR-376c-3p. Mutations in SC5D, CYP46A1, and its functionally linked gene set were associated with multiple differentially expressed oxysterol-related genes. STARD5 was upregulated in patients with positive lymph node status. High expression of hsa-miR-19b-3p was weakly associated with poor survival. This is the first study of oxysterol-related genes in BC that combines DNA, mRNA, and miRNA multiomics with detailed clinical data. Future studies should provide links between intratumoral oxysterol signaling depicted here, circulating oxysterol levels, and therapy outcomes, enabling eventual clinical exploitation of present findings.

• MeSH
• Humans MeSH
• RNA, Messenger genetics   MeSH
• MicroRNAs * genetics   metabolism   MeSH
• Breast Neoplasms * pathology   MeSH
• Oxysterols * MeSH
• Transcriptome genetics   MeSH
• Check Tag
• Humans MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

The immune microenvironment in breast cancer (BCa) is controlled by a complex network of communication between various cell types. Here, we find that recruitment of B lymphocytes to BCa tissues is controlled via mechanisms associated with cancer cell-derived extracellular vesicles (CCD-EVs). Gene expression profiling identifies the Liver X receptor (LXR)-dependent transcriptional network as a key pathway that controls both CCD-EVs-induced migration of B cells and accumulation of B cells in BCa tissues. The increased accumulation oxysterol ligands for LXR (i.e., 25-hydroxycholesterol and 27-hydroxycholesterol) in CCD-EVs is regulated by the tetraspanin 6 (Tspan6). Tspan6 stimulates the chemoattractive potential of BCa cells for B cells in an EV- and LXR-dependent manner. These results demonstrate that tetraspanins control intercellular trafficking of oxysterols via CCD-EVs. Furthermore, tetraspanin-dependent changes in the oxysterol composition of CCD-EVs and the LXR signaling axis play a key role in specific changes in the tumor immune microenvironment.

• MeSH
• B-Lymphocytes metabolism   MeSH
• Liver X Receptors metabolism   MeSH
• Humans MeSH
• Tumor Microenvironment MeSH
• Breast Neoplasms * genetics   MeSH
• Oxysterols * pharmacology   MeSH
• Tetraspanins MeSH
• Check Tag
• Humans MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

Congenital disorders of glycosylation (CDG) and Niemann-Pick type C (NPC) disease are inborn errors of metabolism that can both present with infantile-onset severe liver disease and other multisystemic manifestations. Plasma bile acid and N-palmitoyl-O-phosphocholineserine (PPCS) are screening biomarkers with proposed improved sensitivity and specificity for NPC. We report an infant with ATP6AP1-CDG who presented with cholestatic liver failure and elevated plasma oxysterols and bile acid, mimicking NPC clinically and biochemically. On further investigation, PPCS, but not the bile acid derivative N-(3β,5α,6β-trihydroxy-cholan-24-oyl) glycine (TCG), were elevated in plasma samples from individuals with ATP6AP1-, ALG1-, ALG8-, and PMM2-CDG. These findings highlight the importance of keeping CDG within the diagnostic differential when evaluating children with early onset severe liver disease and elevated bile acid or PPCS to prevent delayed diagnosis and treatment.

• MeSH
• Child MeSH
• Glycosylation MeSH
• Hydrolases MeSH
• Infant MeSH
• Humans MeSH
• Niemann-Pick Disease, Type C * MeSH
• Oxysterols * MeSH
• Vacuolar Proton-Translocating ATPases * MeSH
• Congenital Disorders of Glycosylation * MeSH
• Bile Acids and Salts MeSH
• Check Tag
• Child MeSH
• Infant MeSH
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Research Support, N.I.H., Extramural MeSH

Oxysterols, oxidized derivatives of cholesterol, have been implicated in multiple pathologies, including cancer. In breast cancer, the link is especially strong due to interactions between oxysterols and estrogen receptor activity. Here, we provide the first dedicated study of 113 oxysterol-related genes in breast cancer patients of the luminal subtype, in terms of both their somatic and germline variability, using targeted high-throughput DNA sequencing of 100 normal-tumor pairs with very high coverage. In the full cohort, or subsets of patients stratified by therapy, we found 12 germline variants in ABCA1, ABCA8, ABCC1, GPR183, LDLR, MBTPS1, NR1I2, OSBPL2, OSBPL3, and OSBPL5 to associate with poor survival of patients and variants in ABCA8, ABCG2, and HSD3B7 (three in total) associated with better survival. However, no associations remained significant after correction for multiple tests. Analysis of somatic variants revealed significantly (after FDR correction) poorer survival in patients mutated in CYP46A1 and 9 interacting (according to STRING analysis) genes, as well as in OSBPL3 and a set of 20 genes that collectively associated with the progesterone receptor status of patients. We propose further exploration of these genes in an integrative manner together with gene expression and epigenomic data.

• MeSH
• Cohort Studies MeSH
• Humans MeSH
• Breast Neoplasms * genetics   pathology   MeSH
• Oxysterols * metabolism   MeSH
• Receptors, Steroid * genetics   MeSH
• High-Throughput Nucleotide Sequencing MeSH
• Germ Cells metabolism   MeSH
• Check Tag
• Humans MeSH
• Female MeSH
• Publication type
• Journal Article MeSH

Hlavním cílem tohoto projektu je studium genetických a fenotypických markerů oxysterolové signalizace z hlediska jejich významu pro hormonální terapii karcinomu prsu. V případě, že se podaří spojit tyto markery s prognózou pacientů, případně je navrhnout jako nástroje k individualizované terapii lze, delším časovém horizontu, očekávat významný socio-ekonomický přínos pro pacienty i systém zdravotní péče v ČR. Tento projekt má rovněž za cíl odhalit mechanismus působení markerů a navrhnout cenově dostupnou metodiku k jejich sledování. Projekt svým zaměřením přispěje k řešení několika priorit Programu na podporu zdravotnického aplikovaného výzkumu a vývoje, zejména podoblasti 1.3 Nádorová onemocnění a dílčích cílů 1.3.1 Nádorová biologie a 1.3.2 Individualizace diagnostiky a terapie. Výsledky řešení projektu budou publikovány v impaktovaných odborných časopisech, prezentovány na specializovaných vědeckých setkáních a implementovány do programů celoživotního vzdělávání lékařských pracovníků. Hlavními příjemci výsledků budou vědečtí pracovníci, pedagogové, lékaři v onkologii a pacienti.; In the frame of this project, we plan to evaluate importance of genetic and phenotypic markers of oxysterol signaling for endocrine therapy of breast carcinoma. In the long-term perspective, we expect significant socio-economic benefit for both patients and health care system with the use of these markers for prognosis and individualized therapy. This project also aims to elucidate mechanisms of action of validated markers and to determine cost-effective method for their screening. Project will contribute to the solution of several priorities of the Czech Program for Medical Research and Innovation, particularly of the domain 1.3 Cancer (subdomains 1.3.1 tumor biology and 1.3.2 individualization of diagnostics and therapy). Results of the project will be published in impacted scientific journals, presented at specialized scientific meetings, and implemented into whole-life education programs for medical personnel. Scientists, lecturers and physicians in the area of experimental and clinical oncology and patients will be major users of the project results.

• MeSH
• Hormone Replacement Therapy MeSH
• Humans MeSH
• Biomarkers, Tumor analysis   MeSH
• Breast Neoplasms diagnosis   drug therapy   MeSH
• Oxysterols analysis   MeSH
• Prognosis MeSH
• Check Tag
• Humans MeSH
• Female MeSH

• Conspectus
• Patologie. Klinická medicína
• NML Fields
• onkologie
• genetika, lékařská genetika
• NML Publication type
• závěrečné zprávy o řešení grantu AZV MZ ČR

Oxysterols are oxidized derivatives of cholesterol, both endogenous and exogenous. They have been implicated in numerous pathologies, including cancer. In addition to their roles in carcinogenesis, proliferation, migration, apoptosis, and multiple signalling pathways, they have been shown to modulate cancer therapy. They are known to affect therapy of hormonally positive breast cancer through modulating oestrogen receptor activity. Oxysterols have also been shown in various in vitro models to influence efficacy of chemotherapeutics, such as doxorubicin, vincristine, cisplatin, 5-fluorouracil, and others. Their effects on the immune system should also be considered in immunotherapy. Selective anti-cancer cytotoxic properties of some oxysterols make them candidates for new therapeutic molecules. Finally, differences in oxysterol levels in blood of cancer patients in different stages or versus healthy controls, and in tumour versus non-tumour tissues, show potential of oxysterols as biomarkers for cancer management and patient stratification for optimization of therapy. LINKED ARTICLES: This article is part of a themed issue on Oxysterols, Lifelong Health and Therapeutics. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v178.16/issuetoc.

• MeSH
• Biomarkers MeSH
• Immune System MeSH
• Immunotherapy MeSH
• Humans MeSH
• Neoplasms * drug therapy   MeSH
• Oxysterols * MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Review MeSH

Oxygenated metabolites of cholesterol (oxysterols) have been previously demonstrated to contribute to progression of various cancers and to modulate resistance to breast cancer endocrine therapy in vitro. We measured prognostic roles of circulating levels of seven major oxysterols in the progression of luminal subtype breast carcinoma. Liquid chromatography coupled with tandem mass spectrometry was used for determination of levels of non-esterified 25-hydroxycholesterol, 27-hydroxycholesterol, 7α-hydroxycholesterol, 7-ketocholesterol, cholesterol-5α,6α-epoxide, cholesterol-5β,6β-epoxide, and cholestane-3β,5α,6β-triol in plasma samples collected from patients (n = 58) before surgical removal of tumors. Oxysterol levels were then associated with clinical data of patients. All oxysterols except cholesterol-5α,6α-epoxide were detected in patient plasma samples. Circulating levels of 7α-hydroxycholesterol and 27-hydroxycholesterol were significantly lower in patients with small tumors (pT1) and cholesterol-5β,6β-epoxide and cholestane-3β,5α,6β-triol were lower in patients with stage IA disease compared to larger tumors or more advanced stages. Patients with higher than median cholestane-3β,5α,6β-triol levels had significantly worse disease-free survival than patients with lower levels (p = 0.037 for all patients and p = 0.015 for subgroup treated only with tamoxifen). In conclusion, this study shows, for the first time, that circulating levels of oxysterols, especially cholestane-3β,5α,6β-triol, may have prognostic roles in patients with luminal subtype breast cancer.

• MeSH
• Carcinoma, Ductal, Breast blood   pathology   MeSH
• Neoplasm Invasiveness MeSH
• Middle Aged MeSH
• Humans MeSH
• Survival Rate MeSH
• Biomarkers, Tumor analysis   MeSH
• Breast Neoplasms blood   pathology   MeSH
• Follow-Up Studies MeSH
• Oxysterols blood   MeSH
• Prognosis MeSH
• Receptor, ErbB-2 metabolism   MeSH
• Receptors, Estrogen metabolism   MeSH
• Receptors, Progesterone metabolism   MeSH
• Check Tag
• Middle Aged MeSH
• Humans MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Research Support, N.I.H., Extramural MeSH

Cílem sdělení je shrnutí současného přístupu k diagnostice Niemann-Pickovy choroby typu C (NP-C) s důrazem na nové laboratorní techniky. NP-C je závažné autozomálně recesivní neuroviscerální onemocnění a aktuální dostupnost terapie ovlivňující průběh nemoci zvyšuje důležitost její včasné diagnostiky. Dědičně podmíněný deficit transportního proteinu pro cholesterol (NPC1 nebo NPC2) u NP-C vede k poruše transportu lipidů uvnitř buňky. Ověření klinického podezření na NP-C se proto opírá o biochemické a/nebo molekulárně genetické metody. Nové metody využívajících analýzu biomarkerů v krevním séru nebo plazmě a pokročilé sekvenační techniky mají nyní v diagnostice NP-C důležitou roli. U části pacientů je pro ověření diagnózy nutné použít více vzájemně se doplňujících vyšetření, vč. v článku diskutovaných pokročilých buněčných a biochemických technik. Ty proto musí být k disposici ve specializované laboratoři.

This review provides a summary of current approaches to Niemann-Pick disease type C (NP-C) dianostics with an emphasis on novel laboratory techniques. NP-C is a severe autosomal recessive neurovisceral disorder and the recent availability of disease-modifying therapies increases the importance of its timely diagnosis. The hereditary deficiency of cholesterol transporter proteins (NPC1 or NPC2) in NP-C leads to abnormal intracellular lipid trafficking. Clinical suspicion for NP-C has to be confirmed by biochemical and/or molecular genetic methods. Novel biomarkers in serum or plasma and advanced sequencing techniques now have a prominent role in NP-C diagnostics. In a subset of patients, it is necessary to use several complementary techniques for confirmation of NP-C diagnosis, including advanced biochemical and cellular assays discussed in the paper. These methods therefore have to be available in a specialized laboratory.

• Keywords
• filipinový test, lyzosfingolipidy, intracelulární transport cholesterolu,
• MeSH
• Biomarkers MeSH
• Humans MeSH
• Niemann-Pick Disease, Type C * diagnosis   MeSH
• Oxysterols MeSH
• Check Tag
• Humans MeSH
• Publication type
• Research Support, Non-U.S. Gov't MeSH
• Review MeSH

Oxysteroly jsou oxidované deriváty cholesterolu, které mají pleiotropní účinky. V tomto článku se zaměříme zejména na funkci oxysterolů v organismu, která je dána vazbou na velké množství receptorů, jimiž jsou oxysteroly ligandy, a dále pak na roli oxysterolů u vybraných lidských onemocnění. Oxysteroly se podílejí na udržování homeostázy cholesterolu, a to zejména vazbou na LXR či SREBPs, čímž se spouští kaskáda dějů podílejících se na regulaci metabolismu cholesterolu. Pomocí modulace estrogenových receptorů a Hedgehog signalizace ovlivňují morfogenezi, reprodukci, imunitní reakce a zánětlivou odpověď. V článku je dále uveden vztah oxysterolů a jednotlivých typů nádorů, zejména karcinomu prsu, prostaty, kolorektálního karcinomu a plicního karcinomu. Popisována je také účast oxysterolů v patogenezi aterosklerózy a Alzheimerovy choroby.

Oxysterols are oxidized cholesterol derivatives that have pleiotropic effects. We will focus on the function of oxysterols in the body, because oxysterols are ligands of various receptors. Thus, we will try to explain the role of oxysterols in selected human diseases. Oxysterols have several functions in the body: they are involved in maintaining cholesterol homeostasis, especially by binding to LXR or SREBPs triggering a cascade of events involved in the regulation of cholesterol metabolism. By modulating estrogen receptors and Hedgehog signaling, they affect morphogenesis, reproduction, immune response, and inflammatory response. Furthermore, the relationship between oxysterols and individual tumour types, in particular breast, prostate, colorectal and lung cancer, is discussed. The involvement of oxysterols in the pathogenesis of atherosclerosis and Alzheimer's disease is also described.

• Keywords
• LXR,
• MeSH
• Alzheimer Disease pathology   MeSH
• Arteriosclerosis pathology   MeSH
• Carcinogenesis pathology   MeSH
• Humans MeSH
• Oxysterols * metabolism   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Research Support, Non-U.S. Gov't MeSH
• Review MeSH

• MeSH
• Atherosclerosis prevention & control   MeSH
• Liver X Receptors MeSH
• Cardiovascular Diseases prevention & control   MeSH
• Humans MeSH
• Metabolic Syndrome * diagnosis   etiology   drug therapy   complications   MeSH
• Oxidative Stress MeSH
• Oxysterols * chemistry   metabolism   adverse effects   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Research Support, Non-U.S. Gov't MeSH