2^nd International Conference on Molecular Biology , Nucleic Acids & Molecular Medicine August 30-31, 2017 Philadelphia, Pennsylvania, USA

Zuoren Yu is currently a Professor at Tongji University School of Medicine, Research Center for Translational, Shanghai East Hospital. After obtaining PhD degree in 2003 from Chinese Academy of Medical Sciences, he started Post-doctoral training at University of Pennsylvania School of Medicine and Thomas Jefferson University Kimmel Cancer Center. In 2009, he was assigned for a Faculty position in the track of Research Assistant Professor at Thomas Jefferson University. After joining Tongji University in Shanghai in 2012, he has been focusing on non-coding RNA regulation of breast cancer stem cells related with drug sensitivity, tumour regeneration and cancer metastasis. His main research work includes: Finding cell cycle regulator CCND1 is involved in the regulation of miRNA biogenesis and histone H3K9 tri-methylation, finding a regulatory loop between CCND1 and miR-17/20 in breast cancer, and demonstrating a microenvironment-mediated heterotypic signalling through which miR-17/20 regulate cross talking between cancer cells and inhibit breast cancer cell migration and metastasis.

Small non-coding RNA-based diagnostic and therapeutic applications for human cancer are expected soon. Knocking down the expression level or blocking the function of oncogenic miRNAs is believed to be a promising strategy for cancer treatment. miRNAs loss-of-function phenotypes are mainly induced by chemically modified antisense oligonucleotides. Here, we develop an alternative inhibitor for miRNAs, termed as small RNA zipper. It is designed to bind to the 5’-half sequences of one molecule and the 3’-half sequences of another molecule of the target miRNA through a complementary interaction. The small RNA zipper can connect the target miRNA molecules end to end forming a DNA–RNA duplex with high affinity, high specificity and high stability. To avoid self-complementarity and to enhance binding specificity, LNA nucleosides were applied to synthesize the small RNA zipper. Two miRNAs, miR-221 and miR-17, were tested in human breast cancer cell lines, demonstrating the 70%-90% knockdown of miRNA levels by 30–50 nM small RNA zippers at 24-48 h after transfection. The effect of the miRNA zipper was not limited to the target miRNA, but also to the expression of iso-miRNAs. Let-7 family members and point mutated sequence were applied to further validate the specificity of the miRNA zippers. The miR-221 zipper shows capability in rescuing the expression of target genes of miR-221 and reversing the oncogenic function of miR- 221 in breast cancer cells. In addition, we demonstrate that the miR-221 zipper attenuates doxorubicin resistance with higher efficiency than anti-miR-221 in human breast cancer cells. Taken together, small RNA zippers are a novel type of miRNA inhibitors, which can be used to induce miRNA loss-of-function phenotypes and validate miRNA target genes.

Anne Li is a Senior at Wootton High School with interest in Biological Chemistry. She takes an active role in the STEM field at her schools as the President of both Chemistry Club and Biology Club. Through her experience with the National Chemistry Olympiad and internship at the National Foundation for Cancer Research, she has been able to apply what she has learned in the classroom to help the mission to find a cure for cancer.

Methionine is one of the amino acids that are subjected to oxidation, and can subsequently lead to loss of molecular bioactivities or induce certain health conditions. Typically multiple methionine residues are present in protein molecules, and their oxidation characteristics can be dramatically different. The secondary structures, hydrophobicity, local chain flexibility, and solvent accessibility are all contributing factors to the oxidation propensity of these methionine residues. In this work, a predictive model has been developed to correlate the methionine residual oxidation propensities with their aforementioned properties. The model proteins used in this study, including monoclonal antibodies, growth hormone, interferons, etc., have all been investigated experimentally on their methionine oxidation propensity, and well resolved protein crystal structures documented in RCSB Protein Data Bank (PDB). The crystal structures of these proteins were optimized using QwikMD simulation program (UIUC) to produce well solvated structures. These properties of each methionine residue in the proteins, including hydrophobicity, local chain flexibility, and solvent accessibility, etc., were quantified and a mathematical model was developed to correlate them with the rank ordering of their oxidation propensities reported in literatures. The model, once further validated, provides a qualitative prediction tool to rank order methionine oxidation propensity in protein molecules that are complementary to experimental investigation approaches.

Reham Abo Elwafa is a Lecturer of Clinical Pathology, Faculty of Medicine in Alexandria University, Egypt. She has expertise in Research, Teaching and Administration in Hospital and Education Institution. She is expert in Molecular Techniques: PCR (conventional, and real time PCR), microarray, pyrosequencing and NGS in addition to FISH techniques including Prenatal Genetic Diagnosis (PGD) and flow cytometric immunophenotyping of different types of hematologic malignancies. She has several international publications in the field of Molecular Biology, Genetics and Epigenetics.

Background: Overexpression of the Wilms tumor 1 gene (WT1) is implicated in the prognosis of acute myeloid leukemia (AML) with high expression predicting disease progression, as well as being intensively studied as a potential molecular marker for minimal residual disease (MRD) and treatment response. Many different isoforms for WT1 are generated by alternative transcription initiation, mRNA splicing and alternative translation initiation. Recently, an alternative promoter incorporating a unique first exon, alternative WT1 transcript (AWT1), has been described. The AWT1 expression and the underlying epigenetic alterations associated with its expression in AML are still unknown. Objectives: We studied the AWT1 gene specific methylation changes and its relationship with other clinicopathological features. We also integrated the corresponding gene expression profile to explore the role of methylation in regulating gene expression. Materials & Methods: Bisulfite PCR followed by pyrosequencing were done to determine the methylation status of AWT1 gene promotor CPG islands in 50 newly diagnosed AML patients and 50 healthy subjects as a control group. The level of AWT1 expression was assessed using RQ-PCR. Results: AWT1 expression level was significantly higher in the AML patients in comparison to the control group (P<0.001) and it was surprising to find robust hypermethylation of the AWT1 promoter in AML patients compared to the controls (P<0.001). A statistically significant negative correlation between AWT1 expression and methylation level was found (r=0.67, P<0.001). At a cutoff value of 45.2% AWT1 promoter hypermethylation was found to be a highly specific marker for AML (specificity 95% and sensitivity 97.5%) Conclusion: We described an expression methylation signature of the AWT1 that are promising markers for diagnosis and MRD assessment in AML.