Pathways regulating the expression of the immunomodulatory protein glycodelin in non­small cell lung cancer.

Glycodelin [gene name, progesterone associated endometrial protein (PAEP)] was initially described as an immune system modulator in reproduction. Today, it is also known to be expressed in several types of cancer, including non­small cell lung cancer (NSCLC). In this cancer type, the feasibility of its usage as a follow­up biomarker and its potential role as an immune system modulator were described. It is assumed that NSCLC tumours secrete glycodelin to overcome immune surveillance. Therefore, targeting glycodelin might be a future approach with which to weaken the immune system defence of NSCLC tumours. In this context, it is important to understand the regulatory pathways of PAEP/glycodelin expression, as these are mostly unknown so far. In this study, we analysed the influence of several inducers and of their downstream pathways on PAEP/glycodelin expression in a human lung adenocarcinoma carcinoma (ADC; H1975) and a human lung squamous cell carcinoma (SQCC) cell line (2106T). PAEP/glycodelin expression was notably stimulated by the canonical transforming growth factor (TGF)­ß pathway in SQCC cells and the PKC signalling cascade in both cell lines. The PI3K/AKT pathway inhibited PAEP/glycodelin expression in the ADC cells and an antagonizing role towards the other investigated signalling cascades is suggested herein. Furthermore, the mitogen­activated protein kinase kinase (MEK)/extracellular­signal regulated kinases (ERK) pathway was, to a lesser extent, found to be associated with increased PAEP/glycodelin amounts. The phosphoinositide 3­kinase (PI3K)/protein kinase B (AKT), MEK/ERK pathway and TGF­ß are targets of NSCLC drugs that are already approved or are currently under investigation. On the whole, the findings of this study provide evidence that inhibiting these targets affects the expression of glycodelin and its immunosuppressive effect in NSCLC tumours. Moreover, understanding the regulation of glycodelin expression may lead to the development of novel therapeutic approaches with which to weaken the immune system defence of NSCLC tumours in the future.

Can unexplained infertility be evaluated by a new immunological four-biomarkers panel? A pilot study.

BACKGROUND: Inflammation and oxidative stress are known to be triggering factors for a decrease of the pregnancy rate like maternal immunosuppression. Under these circumstances our study was performed to verify four immunological biomarkers (IMMUNOX Panel) in terms of incidence in a sine-causa infertile population and the overall pregnancy rate when the Panel was showing some non-physiologic values. METHODS: Sera of 86 women affected by unexplained infertility were screened for the IMMUNOX panel of biomarkers composed by: tumor necrosis factor alpha (TNF-α,) glycodelin (GLY), total oxidative status (TOS), and complement activity toxic factor (CATF). When at least one of the biomarkers tested was showing values outside the physiologic range, the woman was considered IMMUNOX-Positive. RESULTS: The first data was indented to verify the incidence of the women with an IMMUNOX-positive panel. Results show that 19.8%, 18.6%, 25.6%, and 47.7% were IMMUNOX-positive for GLY, TNF-α, TOS and CATF respectively. The overall incidence of IMMUNOX-positive patients, with at least one biomarker positive was 70,9%. Subsequently we have analysed the correlation between IMMUNOX Panel positivity and the pregnancy rate. The pregnancy rate in a subgroup (N.=55) of the entire population tested (N.=86) was 2.9% and 36.6% for the IMMUNOX-positive and IMMUNOX-negative patients respectively. CONCLUSIONS: Further validation studies are needed to prove that there is a correlation between unexplained infertility and immunological disorders screened by the IMMUNOX Panel, nevertheless our data shows that this diagnostic approach may be helpful to predict and to identify women at higher risk of IVF cycles failure.

Downregulation of genes related to immune and inflammatory response in IVF implantation failure cases under controlled ovarian stimulation.

PROBLEM: Implantation failure (IF) even after the good-quality embryo transfer (ET) is main obstacle in in vitro fertilization (IVF). We aim to study the genomics of endometrial receptivity in IF patients under controlled ovarian stimulation (COS) during which ET is generally practised in IVF. METHOD OF STUDY: Endometrial gene expression profiling in IF patients (n=10) and oocyte donors (n=8) were compared during window of implantation under COS by microarray. Enrichment analysis of microarray data was performed to determine dysregulated pathways. Microarray results were validated by real-time PCR. Localization of genes related to immune response (progestagen-associated endometrial protein (PAEP), leukaemia inhibitory factor (LIF), interleukin-6 signal transducer (IL6ST) was detected by immunohistochemistry. RESULTS: The gene ontology, pathway analysis and enrichment mapping revealed significant downregulation in activation and regulation of immune and inflammation response in IF patients under COS. The lower expression of PAEP, LIF and IL6ST in cases compared to controls by real time and immunohistochemistry suggests the functional importance of these genes. CONCLUSION: Importance of immune and inflammatory response in endometrial receptivity adds on to the current knowledge of gene expression profile in IF under COS. The panel of genes involved in these pathways would be useful in determining further line of treatment for IF during IVF.