Roles of the ING1 Epigenetic Regulator in Breast Cancer

ING proteins are epigenetic “readers” that can target various chromatin modifying complexes to chromatin. They are involved in various cellular processes such as DNA repair, apoptosis and cellular senescence. This study focuses on examining the potential role of ING1 as a therapeutic agent and prognostic marker for breast cancer. We began by asking whether dysregulating epigenetic pathways with different chemical inhibitors could show synergistic effects with ING1 on killing cancer cells. We tested whether ING1 could synergize better with chemotherapeutics that target the same epigenetic mechanism or a different epigenetic mechanism. Combination treatment of ING1b with LBH589 (HDAC inhibitor) showed synergy, but the combination of ING1b with 5azaC (DNMT inhibitor), thus targeting two distinct epigenetic mechanisms, was more effective. Adenoviral delivery of ING1b combined with 5azaC also inhibited cancer cell growth in a xenograft model and led to tumor regression. These data showed that targeting distinct epigenetic pathways in our model was more effective in blocking cancer cell line growth than targeting the same pathway with multiple agents. Since ING1 expression is frequently repressed in breast carcinomas, but its mechanistic role in breast cancer development and metastasis was unknown, we analyzed ING1 levels in patient samples and correlated it to patient outcome. We also studied the effects of altering ING1 levels in metastasis assays in vitro and mouse metastasis model in vivo. ING1 levels were lower in tumors compared to adjacent normal breast tissue and correlated with tumor size and distant recurrence. ING1 could also predict disease-specific and distant metastasis-free survival in these patients. Decreasing levels of ING1 increased, and increasing levels decreased migration and invasion of MDA-MB231 cells in vitro. ING1 overexpression also blocked cancer cell metastasis in vivo and eliminated tumor-induced mortality in mouse models. Lastly, we determined if ING1 expression could predict breast cancer patient outcome. We found that stromal cell expression of ING1 showed an inverse correlation with patient survival. ING1 also correlated with tumor grade in these patients and multivariate analysis showed that ING1 was an independent prognostic marker in the breast cancer cohort we tested. This study provides important pre-clinical data that could help establish ING1 as a prognostic and therapeutic agent for breast cancer.18 month

Correction of WOW and Flutter in Audio Signals using Least Mean Square Algorithm

This paper describes a system which is used to remove Wow and Flutter from audio signal using Least Mean Square Algorithm. It is impossible to completely remove Wow and Flutter from audio signal but its effect can be reduced significantly. It occurs during the process of sound reproduction. It is the group of tones created by the irregularities in turntables or tape drive speed during reproduction, duplication or recording. Wow occurs due to irregularities at low frequency whereas at high frequency irregularities, Flutter occurs. Least Mean Square Algorithm uses Adaptive Filter which adjusts their coefficient in order to minimize the required wobble effects in audio signal. Results show that LMS has comprehensively diminished the effects of wow and flutter

Comparison of Different Adaptive Speech Algorithm to Correct Wow and Flutter in Audio Signals

Abstract — This proposed research work describes a system which is used to remove Wow and Flutter from audio signal using Advanced Adaptive Speech Algorithm. It is impossible to completely remove Wow and Flutter from audio signal but its effect can be reduced significantly. It occurs during the process of sound reproduction. It is the group of tones created by the irregularities in turntables or tape drive speed during reproduction, duplication or recording. Wow occurs due to irregularities at low frequency whereas at high frequency irregularities, Flutter occurs. Wow and Flutter could be found in old gramophone recordings, wax cylinders, on the magnetic and optical sound tapes. Least Mean Square Algorithm uses Adaptive Filter which adjusts their coefficient in order to minimize the required wobble effects in audio signal. Results show tha

Additional file 1: Figure S1. of Stromal ING1 expression induces a secretory phenotype and correlates with breast cancer patient survival

Quantitation of 3-D culture colony morphological changes. Colony images from 3-D cultures of HMF3s cells expressing GFP + MCF7 cells and HMF3s cells expressing ING1a + MCF7 cells were visually scored for their levels of disorganization and aggressiveness as estimated by divergence from uniformity (** p < 0.001). (DOC 58 kb

SUMOylation of the ING1b tumour suppressor regulates gene transcription.

International audienceThe INhibitor of Growth (ING) proteins are encoded as multiple isoforms in five ING genes (ING1-5) and act as type II tumor suppressors. They are growth inhibitory when overexpressed and are frequently mislocalized or down-regulated in several forms of cancer. ING1 and ING2 are stoichiometric members of histone deacetylase (HDAC) complexes while ING3-5 are stoichiometric components of different histone acetyl-transferase (HAT) complexes. The INGs target these complexes to histone marks, thus acting as epigenetic regulators. ING proteins affect angiogenesis, apoptosis, DNA repair, metastasis and senescence, but how the proteins themselves are regulated is not yet clear. Here we find a small ubiquitin like modification (SUMOylation) of the ING1b protein and identify lysine 193 (K193) as the preferred ING1b SUMO acceptor site. We also show that PIAS4 is the E3 SUMO ligase responsible for ING1b SUMOylation on K193. Sequence alignment reveals that the SUMO consensus site on ING1b contains a phosphorylation-dependent SUMOylation motif (PDSM) and our data indicate that the SUMOylation on K193 is enhanced by the S199D phosphomimic mutant. Using an ING1b protein mutated at the major SUMOylation site (ING1b E195A), we further demonstrate that ING1b SUMOylation regulates the binding of ING1b to the ISG15 and DGCR8 promoters, consequently regulating ISG15 and DGCR8 transcription. These results suggest a role for ING1b SUMOylation in the regulation of gene transcription

Identification and in vitro validation of neoantigens for immune activation against high-risk pediatric leukemia cells

There is experimental and clinical data to indicate the contribution of immune-escape mechanisms in relapsed/refractory pediatric leukemia. Studies have shown the accumulation of mutations that translate to peptides containing tumor-specific epitopes (neoantigens). The effectiveness of neoantigen-based vaccines has been shown in several clinical trials in adults. Though the initial results are encouraging, this knowledge must be developed to account for the uniqueness of pediatric cancer biology. We have completed the initial proof-of-concept analysis on a high-risk pediatric leukemia specimen and identified usable neoantigen sequences. We describe this approach, including the bioinformatics method and experimental model to verify their function that can be further broadened for personalized neoantigen prediction and testing for the generation of anticancer vaccines against high-risk pediatric leukemias