Lymphatic expression of CLEVER-1 in breast cancer and its relationship with lymph node metastasis

BACKGROUND Mechanisms regulating breast cancer lymph node metastasis are unclear. Staining of CLEVER-1 (common lymphatic endothelial and vascular endothelial receptor-1) in human breast tumors was used, along with in vitro techniques, to assess involvement in the metastatic process. METHODS 148 sections of primary invasive breast cancers, with 10 yr follow-up, were stained with anti-CLEVER-1. Leukocyte infiltration was assessed, along with involvement of specific subpopulations by staining with CD83 (mature dendritic cells, mDC), CD209 (immature DC, iDC) and CD68 (macrophage, Mϕ). In vitro expression of CLEVER-1 on lymphatic (LEC) and blood endothelial cells (BEC) was examined by flow cytometry. RESULTS In vitro results showed that although both endothelial cell types express CLEVER-1, surface expression was only evident on LEC. In tumour sections CLEVER-1 was expressed in blood vessels (BV, 61.4% of samples), lymphatic vessels (LV, 18.2% of samples) and in Mϕ/DCs (82.4% of samples). However, only CLEVER-1 expression in LV was associated with LN metastasis (p = 0.027) and with Mϕ indices (p = 0.021). Although LV CLEVER-1 was associated with LN positivity there was no significant correlation with recurrence or overall survival, BV CLEVER-1 expression was, however, associated with increased risk of recurrence (p = 0.049). The density of inflammatory infiltrate correlated with CLEVER-1 expression in BV (p < 0.001) and LV (p = 0.004). CONCLUSIONS The associations between CLEVER-1 expression on endothelial vessels and macrophage/leukocyte infiltration is suggestive of its regulation by inflammatory conditions in breast cancer, most likely by macrophage-associated cytokines. Its upregulation on LV, related surface expression, and association with LN metastasis suggest that it may be an important mediator of tumor cell metastasis to LN

IL-6 and IL-10 are associated with good prognosis in early stage invasive breast cancer patients

Macrophage-associated cytokines play an important role in cancer metastasis however the functions of Interleukins (IL) 6 and 10 in breast cancer (BC) progression and metastasis are not clear. In this study the roles of IL-6/IL-10 in regulating vascular invasion and their prognostic significance in BC are investigated. MDA-MB-231 and MCF-7 migration (+/- IL-6 or IL-10) was assessed by scratch wound assay. Cancer cell adhesion to IL-6/IL-10 stimulated blood and lymphatic endothelial cells (EC) was investigated. Expression of IL-6 /IL-10 was assessed using immunohistochemistry in an annotated cohort of early stage BC (n=1380) and associations with clinicopathological variables and clinical outcome evaluated. IL-6 did not alter BC cell migration however a dose-dependent inhibition in MDA-MB-231 migration with IL-10 treatment was observed (P=0.03). BC cells were more adhesive to blood vs lymphatic EC however IL-6/IL-10 had no effect on adhesion patterns. High expression of IL-6/IL-10 was associated with clinicopathological criteria (e.g. hormone receptor status, all P<0.05), improved disease free survival (DFS; P<0.05) and improved BC specific survival (BCSS; only IL-6, P=0.017). However neither IL-6 nor IL-10 expression were independent prognostic factors from multivariate analysis. In BC subgroups, IL-6 and IL-10 were good prognosticators in terms of DFS in non-basal, non-triple negative (non-TN), ER-positive, PgR-positive (only IL-10), and Her-2-negative (only IL-6) BC (all P<0.05). IL-6 was associated with improved BCSS in non-basal, ER-positive and non-TN BC (all P<0.05)

An insight into the anti-angiogenic and anti-metastatic effects of oridonin: current knowledge and future potential

Cancer is one of the leading causes of death worldwide, with a mortality rate of more than 9 million deaths reported in 2018. Conventional anti-cancer therapy can greatly improve survival however treatment resistance is still a major problem especially in metastatic disease. Targeted anti-cancer therapy is increasingly used with conventional therapy to improve patients’ outcomes in advanced and metastatic tumors. However, due to the complexity of cancer biology and metastasis, it is urgent to develop new agents and evaluate the anti-cancer efficacy of available treatments. Many phytochemicals from medicinal plants have been reported to possess anti-cancer properties. One such compound is known as oridonin, a bioactive component of Rabdosia rubescens. Several studies have demonstrated that oridonin inhibits angiogenesis in various types of cancer, including breast, pancreatic, lung, colon and skin cancer. Oridonin’s anti-cancer effects are mediated through the modulation of several signaling pathways which include upregulation of oncogenes and pro-angiogenic growth factors. Furthermore, oridonin also inhibits cell migration, invasion and metastasis via suppressing epithelial-to-mesenchymal transition and blocking downstream signaling targets in the cancer metastasis process. This review summarizes the recent applications of oridonin as an anti-angiogenic and anti-metastatic drug both in vitro and in vivo, and its potential mechanisms of action

Spatial transcriptomic analysis of tumour with high and low CAIX expression in TNBC tissue samples using GeoMx™ RNA assay

Purpose. Prognostic significance and gene signatures associated with carbonic anhydrase IX (CAIX) was investigated in triple negative breast cancer (TNBC) patients. Methods. Immunohistochemistry (IHC) for CAIX was performed in tissue microarrays (TMAs) of 136 TNBC patients.In a subset of 52 patients Digital Spatial Profiler (DSP) was performed in tumour (pan-cytokeratin+) and stroma (pan-cytokeratin-). Differentially expressed genes (DEGs) with P&lt;0.05 and fold change ≥1 or ≤-1 were identified. Four genes were validated at the protein level. Result. Cytoplasmic CAIX expression was independently associated with poor recurrence free survival in TNBC patients [hazard ratio (HR)=6.59, 95% confidence interval (CI): 1.47-29.58, P=0.014]. DEG analysis identified 4 up-regulated genes (CD68, HIF1A, pan-melanocyte, and VSIR) in the tumour region and 9 down-regulated genes in the stromal region (CD86, CD3E, MS4A1, BCL2, CCL5, NKG7, PTPRC, CD27, and FAS) when low versus high CAIX expression was explored. Employing IHC, high CD68 and HIF-1α was associated with poorer prognosis and high BCL2 and CD3 was associated with good prognosis. Conclusions. DSP technology identified DEGs in TNBC. Selected genes validated by IHC showed involvement of CD3 and BCL2 expression within stroma and HIF-1α, and CD68 expression within tumour. However, further functional analysis is warranted

A Functional and Molecular Characterisation of Tumour-Associated Lymph Endothelium

EThOS - Electronic Theses Online ServiceGBUnited Kingdo

A comparative immunohistochemical analysis of cathepsins B and S in human breast cancer

Background: Cancer progression is a complex process consisting of a series of distinct steps. Cysteine proteases, such as cathepsins (Cts), are important molecules that play a central role in cancer progression and metastasis. Previous studies on human and mouse models of pancreatic cancer showed that both Cts B and Cts S are highly expressed in malignant tissues and the infiltrating macrophages. The aim of this study was to investigate the expression pattern of Cts B and S in human breast cancer tissues. Materials and Methods: Twenty-three formalin-fixed paraffin-embedded sections of breast cancer were stained for Cts B, Cts S, and CD206 using immunohistochemistry. Results: Cytoplasmic staining of Cts B and S was observed in tumor cells, endothelial cells, and macrophages. Cts B was preferentially expressed in breast cancer tissues by the different cells types. The majority of tumor samples were Cts B-positive in tumor cells, endothelial cells and macrophages (91%, 87%, and 70%, respectively) in comparison to Cts S (39%, 48%, and 57%, respectively; P &lt; 0.001, P &lt; 0.001 and 0.002). Correlation studies indicated significant relationships between the vascular and macrophage expression of Cts B (P = 0.01) and of Cts S (P = 0.03). However, neither Cts B nor Cts S expression in tumor cells correlated with other cell types (P &gt; 0.05). Only the expression of Cts B in vascular endothelial cells correlated significantly with the tumor grade (P = 0.03). Conclusion: Results suggest that Cts B expression is more prominent than Cts S in breast cancer. Correlation studies imply different mechanisms regulating Cts B/S expression in tumor cells and other stromal components

Policy versus practice: Syrian refugee doctors in Egypt

The COVID-19 pandemic has renewed interest in streamlining processes which allow refugee doctors and other healthcare workers to make up for the shortfall in healthcare delivery, which many countries are facing increasingly. The protracted conflict in Syria is the biggest driver of forced displacement internationally with refugees, including healthcare workers seeking safety in host countries, however many face challenges to entering the workforce in a timely manner. The majority are in countries surrounding Syria (Lebanon, Jordan and Turkey) however the restrictive labour policies in these countries, particularly for healthcare workers have forced many to look further afield to Europe or the Gulf. Egypt's context is interesting in this regard, as it hosts a smaller number of registered Syrian refugees and was initially welcoming of Syrian medical students and doctors. However, recent socio-political changes have led to restrictions in training and work, leading doctors who initially considering staying in Egypt to increasingly consider it a transit country rather than a destination country. Here, we explore the processes by which Syrian doctors in Egypt can work and how documented policies may differ to practice. We do this through a document review and from the first-hand experiences of the authors

Experiences of Egypt as a destination and transit country for Syrian refugee healthcare workers: a qualitative study

Abstract Background Refugee healthcare workers (HCWs) can make important contributions in host countries, particularly in the wake of the ongoing COVID-19 pandemic, which has exacerbated existing shortages of frontline HCWs. However, refugee HCWs often face challenges entering the labour markets of such countries even where needs exist. Syria’s decade-long conflict has forced thousands of HCWs from their homes; however, data on this population are limited, impeding the formation of policies that can support them. This study explores the experiences of Syrian refugee HCWs in Egypt. Methods Key informants (KIs) were selected using purposive and snowball sampling method and semi-structured interviews were conducted in person in Cairo and remotely from the UK during July 2019. Interviews were conducted in Arabic and analysed using a combined deductive and inductive thematic analysis framework after transcription into English. Results Fifteen KI interviews were analysed. The main emerging themes from the qualitative interviews are those relating to 1. Education, training, and licensing 2. Politics and bureaucracy 3. Societal factors 4. Economic factors. Political changes in Egypt altered opportunities for Syrian HCWs over time; however, refugee HCWs broadly reported acceptance among Egyptian patients and colleagues. Bureaucratic factors which impede the ability of Syrian refugee HCWs to obtain a full license to practice and leave to remain and the absence of clearly defined policies were reported as barriers. Economic factors including the risk of economic exploitation e.g. in the informal sector and financial insecurity were noted to have a negative psychosocial impact. Conclusions This is the first qualitative research study which explores the experiences of Syrian refugee HCWs in Egypt. It adds to the sparse literature on the topic of Syrian refugee HCWs but provides evidence for further discussions on how to support refugee HCWs in Egypt and in other host countries in the region. Though interviews were conducted before the COVID-19 pandemic, the pandemic itself lends urgency to the discussion around refugee HCWs on an international level

Abstract 5210: Differential cytotoxic effects of Piperlongumine analogues and their radiotherapeutic response in breast cancer

Proceedings: AACR Annual Meeting 2017; April 1-5, 2017; Washington, DCBackground: Breast cancer is the most common cancer in women in the United Kingdom. The glutathione (GSH) system is a major redox buffering system involved in regulating radioresponse of cancer cells. Overexpression has been shown to cause multidrug and radiation resistance. Its modulation may increase radiosensitivity and improve radiotherapy efficacy. Piperlongumine (PL) can reportedly inhibit GSH system function and reduce proliferation of a variety cancer cells in vitro. The current study investigates novel PL analogues to determine efficacy in triple-negative (TNBC) and luminal breast cancer cell lines, as both single agents and in combination with ionising radiation.Methods: Two novel PL analogues (LH91 and LH92) were used along with the parental drug, PL. Cell proliferation assays were conducted in MDA-MB-231 (TNBC) and T47D (luminal) lines at various drug concentrations and times. At 48hr post treatment, cells were also assessed for clonogenic survival. Clonogenic radiosensitisation was assessed by treating with PL agents or L-buthionine-S, R-sulfoximine (BSO, a well characterised GSH radiosensitiser) for 48hr then irradiation with 0-8Gy X-rays. ROS levels were evaluated using H2DCFDA flow cytometry following IC50 drug treatment alone or with subsequent exposure to 1mM H2O2 for 1hr.Results: PL, LH91 and LH92 inhibited proliferation and decreased clonogenic survival in both lines. From proliferation assays, MDA-MB-231 cells were more sensitive to PL at 48hr (P=0.047) and 72hr (P=0.01) with an IC50 dose of approx. 4μM versus approx. 12μM in T47D�s. There was a significant difference between IC50 doses of LH91 in MDA-MB-231 (5μM) and T47D (12μM) cell lines at 72hr (P=0.006), however this was not observed at 48hr treatment (P=0.178). In contrast, there was no differential sensitivity of LH92 in cell proliferation between the two cell lines at 48 and 72hr. Interestingly, in clonogenic assays, MDA-MB-231�s were more sensitive to LH92 (P=0.01) with an IC50 of 4μM versus 11μM in T47D�s. There were no significant differences in clonogenic survival between each line after treatment with PL or LH91. No radiosensitisation was observed with PL or PL analogues in MDA-MBA-231 however BSO, as a drug comparator, enhanced radiation response with a sensitizer enhancement ratio (SER) of 1.13 at 1% iso-survival. Radiation and drug combinations in T47D cells are ongoing. There were no changes in ROS levels with the drug alone in MDA-MB-231; however, with subsequent exposure of H2O2, there was a 1.7-fold increase in ROS level with LH91 at 48hr (P=0.002).Conclusion: The TNBC cell line is more sensitive to the cytotoxic effect of PL agents (PL and LH91 in cell proliferation and LH92 in clonogenic assays) in comparison to the luminal cell line; however no altered radiosensitisation was observed. The mechanism of such differential sensitivity is yet to be determined.Note: This abstract was not presented at the meeting.Citation Format: Nurul Akmaryanti Abdullah, John Moses, Li Chen Han, Sarah Storr, Aula Ammar, Stewart G. Martin. Differential cytotoxic effects of Piperlongumine analogues and their radiotherapeutic response in breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5210. doi:10.1158/1538-7445.AM2017-521

Impact of armed conflict on health professionals’ education and training in Syria: a systematic review

Objectives To provide an overview of the holistic impact of the armed conflict on medical education and health professionals’ training (MEHPT) in Syria.Setting Syria is a country which underwent an armed conflict for 10 years and suffered from the weaponisation of health.Methods A mixed-methods systematic review including quantitative, qualitative, mixed-methods and textual literature between 2011 and 2021 including papers on the Syrian MEHPT undergraduate and postgraduate education and training personnel (including medicine, dentistry, pharmacy, nursing, midwifery and allied health professionals). The electronic search was conducted in October 2018 in Embase, Global Health, Medline, PsycINFO, Web of Science, PubMed, Scopus, CINAHL and grey literature. And an update to the search was conducted in August 2021 in PubMed, Google Scholar and Trip database.Outcomes The impact of conflict on the MEHPT system, personnel, experiences, challenges and channels of support.Results Of the 5710 citations screened, 70 met the inclusion criteria (34 quantitative, 3 qualitative, 1 mixed-method, and 32 reports and opinion papers). The two major cross-cutting themes were attacks on MEHPT and innovations (present in 41% and 44% of the papers, respectively), followed by challenges facing the MEHPT sector and attitudes and knowledge of trainees and students, and lastly health system and policy issues, and narrating experiences.Conclusion Conflict in Syria has politicised all aspects of MEHPT. Influenced by political control, the MEHPT system has been divided into two distinguished geopolitical contexts; government-controlled areas (GCAs) and non-GCAs (NGCAs), each having its characteristics and level of war impact. International and regional academic institutes collaboration and coordination efforts are needed to formulate educational platforms using innovative approaches (such as online/blended/store-and-forward/peer-training/online tutoring) to strengthen and build the capacity of the health workforce in conflict-affected areas