CDK4 phosphorylation status and a linked gene expression profile predict sensitivity to palbociclib.

Cyclin D-CDK4/6 are the first CDK complexes to be activated in the G1 phase in response to oncogenic pathways. The specific CDK4/6 inhibitor PD0332991 (palbociclib) was recently approved by the FDA and EMA for treatment of advanced ER-positive breast tumors. Unfortunately, no reliable predictive tools are available for identifying potentially responsive or insensitive tumors. We had shown that the activating T172 phosphorylation of CDK4 is the central rate-limiting event that initiates the cell cycle decision and signals the presence of active CDK4. Here, we report that the profile of post-translational modification including T172 phosphorylation of CDK4 differs among breast tumors and associates with their subtypes and risk. A gene expression signature faithfully predicted CDK4 modification profiles in tumors and cell lines. Moreover, in breast cancer cell lines, the CDK4 T172 phosphorylation best correlated with sensitivity to PD0332991. This gene expression signature identifies tumors that are unlikely to respond to CDK4/6 inhibitors and could help to select a subset of patients with HER2-positive and basal-like tumors for clinical studies on this class of drugs.

Diagnostic value of the UCA1 test for bladder cancer detection: a clinical study.

PURPOSE: To evaluate the efficiency of the UCA1 test as a diagnostic tool for the detection of bladder cancer. METHODS: Between October 2009 and December 2011 the UCA1 test was performed on collected urine samples from 162 patients divided into screening and follow-up groups, based on the absence or presence of prior bladder cancer. The test performance was then evaluated in each group and compared to cystoscopy and urinary cytology. RESULTS: The overall sensitivity, specificity and positive and negative predictive values for the UCA1 test were 70, 70.7, 75.6 and 64.5%, respectively. We observed no difference in performance for tumours of higher grade or stage, but sensitivity was increased in the screening population compared to patients under follow-up (83.9 vs. 59%). The UCA1 test successfully detected all 7 cases of isolated carcinoma in situ and was more sensitive in this particular setting than cystoscopy or urinary cytology. CONCLUSION: The efficiency of the UCA1 test for the detection of primary and recurring bladder cancer in our study was lower than previously reported. We confirmed the role of UCA1 as a possible adjunct to cystoscopy and cytology when a primary bladder cancer is suspected, but its role in the follow-up of recurring tumours remains limited. Further studies are needed to investigate the role of the UCA1 test in the early detection of carcinoma in situ lesions.

Expression of galectin-3 in the tumor immune response in colon cancer.

The role of tumor-associated macrophages (TAMs) is controversial. Although most studies on different cancer types associate them with a poorer prognosis, interestingly in colon cancer, most articles indicate that TAMs prevent tumor development; patients with high TAMs have better prognosis and survival rate. M1-polarized macrophages produce high level of tumor necrosis factor-alpha, interleukin-1 beta or reactive oxygen species, which can effectively kill susceptible tumor cells. In contrast, M2-polarized macrophages can secrete different factors that promote tumor cell growth and survival or favor angiogenesis and tissue invasion. Considering the beneficial role of TAMs in colon cancer, we speculated that they may not display the M2 polarization commonly observed in tumor microenvironment, but rather develop M1 properties. Therefore, we used an in vitro model to analyze the effects of supernatants from M1-polarized macrophages on DLD-1 colon cancer cells. Our data indicate that the conditioned medium from LPS-activated macrophages (CM-LAM) contains a high level of granulocyte-macrophage colony-stimulating factor, interleukins-1 beta, -6, -8 and tumor necrosis factor-alpha, and that it exerts a marked growth inhibitory activity on DLD-1 cells. Prolonged exposure to CM-LAM results in cell death by apoptosis. Such exposure to CM-LAM leads to the modulation of gal-3 expression: we observed a marked downregulation of gal-3 mRNA and protein expression following CM-LAM treatment. We also describe that the knockdown of gal-3 sensitizes DLD-1 cells to CM-LAM. These data suggest an involvement of gal-3 in the response of colon cancer cells to proinflammatory stimuli, such as the conditioned medium from activated macrophages.

Exploring the distinctive biological characteristics of pilocytic and low-grade diffuse astrocytomas using microarray gene expression profiles.

Although World Health Organization (WHO) grade I pilocytic astrocytomas and grade II diffuse astrocytomas have been classified for decades as different clinicopathologic entities, few, if any, data are available on the biologic features explaining these differences. Although more than 50 microarray-related studies have been carried out to characterize the molecular profiles of astrocytic tumors, we have identified only 11 that provide sound data on low-grade astrocytomas. We have incorporated these data into a comparative analysis for the purpose of identifying the most relevant molecular markers characterizing grade I pilocytic and grade II diffuse astrocytomas. Our analysis has identified various interesting genes that are differentially expressed in either grade I or grade II astrocytomas when compared with normal tissue and/or high-grade (WHO grade III and IV) astrocytomas. A large majority of these genes encode adhesion, extracellular matrix, and invasion-related proteins. Interestingly, a group of 6 genes (TIMP4, C1NH, CHAD, THBS4, IGFBP2, and TLE2) constitute an expression profile characteristic of grade I astrocytomas as compared with all other categories of tissue (normal brain, grade II, and high-grade astrocytomas). The end products (proteins) of these genes act as antimigratory compounds, a fact that could explain why pilocytic astrocytomas behave as compact (well-circumscribed) tumors as opposed to all the other astrocytic tumor types that diffusely invade the brain parenchyma. Having validated these molecular markers by means of real-time reverse transcriptase-polymerase chain reaction, an integrated model was proposed illustrating how and why pilocytic astrocytomas constitute a distinct biologic and pathologic entity when compared with diffuse astrocytomas.

Continuous-flow UVC irradiation: a new, effective, protein activity-preserving system for inactivating bacteria and viruses, including erythrovirus B19.

Despite the increasing number of screening tests being introduced, ensuring the inactivation of blood-borne pathogens in blood-derived therapeutic material is a major concern. Dynamic continuous-flow UVC irradiation is a new way to inactivate a large range of pathogens without adding any photosentizers. The efficacy of different methods was evaluated against the following viruses: murine parvovirus MVMp, human B19, the encephalomyocarditis virus (EMC, a picornavirus used as a model for model for hepatitis A virus), and bovine herpes virus type 1 (BHV, a model for enveloped viruses such as hepatitis B virus). We show that continuous-flow UVC irradiation is very effective, particularly against resistant pathogens (e.g. parvoviruses and bacteria) at UVC doses preserving protein activity. It may be applicable to newly emerging related viruses or variants.