Inns, Joseph and James, Victoria (2015) Circulating microRNAs for the prediction of metastasis in breast cancer patients diagnosed with early stage disease. The Breast, 24 (4). pp. 364-369. ISSN 1532-3080 Access from the University of Nottingham repository: http://eprints.nottingham.ac.uk/36999/1/YBRST_2141_edit_report.pdf Copyright and reuse:The Nottingham ePrints service makes this work by researchers of the University of Nottingham available open access under the following conditions. This article is made available under the University of Nottingham End User licence and may be reused according to the conditions of the licence. For more details see: http://eprints.nottingham.ac.uk/end_user_agreement.pdf A note on versions:The version presented here may differ from the published version or from the version of record. If you wish to cite this item you are advised to consult the publisher's version. Please see the repository url above for details on accessing the published version and note that access may require a subscription.For more information, please contact eprints@nottingham.ac.uk IntroductionBreast cancer is the second most common malignancy diagnosed worldwide, accounting for approximately 23% of all cancer diagnoses per year and 465,000 cancer-related deaths [1]. These cases will include early stage disease, local recurrence or metastasis in patients previously treated with a curative intent, and advanced stage disease, characterised by the significant spread of the cancer within the breast or systemically [1,2]. The metastatic cascade initiates with localised invasion of surrounding tissue, followed by systemic spread via the blood and lymphatic system and finally dissemination of tumour cells to other organs [3]. At the point of diagnosis up to 5% of patients present with metastatic disease and of those patients who show no lymph-node involvement at diagnosis, approximately 30% will develop metastases [4,5]. This subset of individuals who potentially have metastasis of early disease (MED) represent an at risk group, who may benefit from additional monitoring and augmented treatment strategies. We currently need markers to distinguish these patients from those who do not go on to develop disease recurrence. Clinical staging and molecular classificationClinical staging is essential for establishing surgical approaches and treatment regimes. The current TNM system assess primary tumour size and extent of invasion (T), the absence or presence of palpable axillary lymph nodes and indications of local invasion (N), and evidence of distant metastasis (M) [6]. The TNM system is further supplemented by the allocation of stages I-V depending on size and metastatic spread [6]. Molecular factors are also employed to assist prognosis and direct treatment strategies. Breast cancers are categorised based on the expression of oestrogen and progesterone receptors (ER and PR) and the receptor tyrosine kinase erbB-2 (ERBB2 or HER2). Further categorisation defines four molecular subtypes [Refs. 33,34]: luminal A whi...

Inflammatory responses are important in cancer, particularly in the context of monocyte-rich aggressive myeloid neoplasm. We developed a label-free cellular phenotypic drug discovery assay to identify anti-inflammatory drugs in human monocytes derived from acute myeloid leukemia (AML), by tracking several features ionizing from only 2500 cells using matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry. A proof-of-concept screen showed that the BCR-ABL inhibitor nilotinib, but not the structurally similar imatinib, blocks inflammatory responses. In order to identify the cellular (off-)targets of nilotinib, we performed thermal proteome profiling (TPP). Unlike imatinib, nilotinib and other later-generation BCR-ABL inhibitors bind to p38α and inhibit the p38α-MK2/3 signaling axis, which suppressed pro-inflammatory cytokine expression, cell adhesion, and innate immunity markers in activated monocytes derived from AML. Thus, our study provides a tool for the discovery of new anti-inflammatory drugs, which could contribute to the treatment of inflammation in myeloid neoplasms and other diseases.

Phagocytosis is a key process in innate immunity and homeostasis. After particle uptake, newly formed phagosomes mature by acquisition of endolysosomal enzymes. Macrophage activation by interferon gamma (IFN-c) increases microbicidal activity, but delays phagosomal maturation by an unknown mechanism. Using quantitative proteomics, we show that phagosomal proteins harbour high levels of typical and atypical ubiquitin chain types. Moreover, phagosomal ubiquitylation of vesicle trafficking proteins is substantially enhanced upon IFN-c activation of macrophages, suggesting a role in regulating phagosomal functions. We identified the E3 ubiquitin ligase RNF115, which is enriched on phagosomes of IFN-c activated macrophages, as an important regulator of phagosomal maturation. Loss of RNF115 protein or ligase activity enhanced phagosomal maturation and increased cytokine responses to bacterial infection, suggesting that both innate immune signalling from the phagosome and phagolysosomal trafficking are controlled through ubiquitylation. RNF115 knock-out mice show less tissue damage in response to S. aureus infection, indicating a role of RNF115 in inflammatory responses in vivo. In conclusion, RNF115 and phagosomal ubiquitylation are important regulators of innate immune functions during bacterial infections.

CYLD lysine 63 deubiquitinase (CYLD) is a ubiquitin hydrolase with important roles in immunity and cancer. Complete CYLD ablation, truncation and expression of alternate isoforms, including short CYLD, drive distinct phenotypes and offer insights into CYLD function in inflammation, cell death, cell cycle progression and cell transformation. Research in diverse model systems has shown that these are mediated via CYLD regulation of cellular pathways including the NF-κB, Wnt and TGF-β pathways. Recent biochemical advances and models have offered new insights into the regulation and function of CYLD. In addition, recent discoveries of gain-of-function germline pathogenic CYLD variants in patients with a neurodegenerative phenotype contrast with the more widely known loss-of-function mutations seen in patients with CYLD cutaneous syndrome and with sporadic cancers. Here, we provide a current review of mechanistic insights into CYLD function gained from CYLD animal models, as well as an update on the role of CYLD in human disease.

Inflammatory responses are important in cancer, particularly in the context of monocyte-rich aggressive myeloid neoplasm. We developed a label-free cellular phenotypic drug discovery assay to identify anti-inflammatory drugs in human monocytes derived from acute myeloid leukemia (AML), by tracking several biological features ionizing from only 2,500 cells using matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry. A proof-of-concept screen showed that the BCR-ABL inhibitor nilotinib, but not the structurally similar imatinib, blocks inflammatory responses. In order to identify the cellular (off-)targets of nilotinib, we performed thermal proteome profiling (TPP) using TMTpro 16plex. Unlike imatinib, nilotinib inhibits p38-alpha (MAPK14) signalling. This inhibition suppressed the expression of inflammatory cytokines, cell adhesion and innate immunity markers in activated human monocytes derived from AML. Thus, our study provides a tool for the discovery of new anti-inflammatory drugs, which could contribute to the treatment of inflammation in myeloid neoplasms and other diseases.