microRNAs as Biomarkers and Diagnostics
8:40-8:45 Chairperson's Opening Remarks
8:45-9:15 microRNA Biomarkers for Cancer Treatment
Glen J. Weiss, M.D., Co-Head, Lung Cancer Unit, The Translational Genomics Research Institute (TGen); Director, Thoracic Oncology, Virginia G. Piper Cancer Center Clinical Trials at Scottsdale Healthcare; CMO, CRAB-Clinical Trials Consortium
A single microRNA can impact hundreds of targets and can affect pathways controlling oncogenic processes. Data will be presented illustrating how using microRNA can impact cancer treatment decision making, validated microRNAs are associated with resistance and/or sensitivity to chemotherapy and targeted therapy, and how microRNAs could be used as therapeutics.
9:15-9:45 Bridging Early Discovery Needs and Downstream Biomarker Requirements
David von Schack, Ph.D., Senior Principal Research Scientist, Translational Immunology, Pfizer
miRNAs have been shown to be involved in a wide range of diseases and disease mechanisms and have become an integral component in building a complete understanding of molecular disease processes. Challenges remain in the interpretation of miRNA involvement in pathologic processes as they are predicted and shown to bind 10s or sometimes 100s of mRNA transcripts. While it is possible to show correlation of miRNA changes with disease processes, the understanding of these changes are hampered by our difficulties in assigning specific molecular functions to those regulated miRNAs. Target discovery efforts using miRNA profiling will be addressed and the potential for miRNA as mechanistic biomarkers of drug action or disease modulation discussed.
9:45-10:15 Integrating Contextual microRNA and Protein Signatures for Diagnostic Applications in Cancer
Lorenzo F. Sempere, Ph.D., Research Assistant Professor, Medicine, Dartmouth-Hitchcock/Norris Cotton Cancer Center
As each tumor lesion has a different ratio of cancer cells to reactive stroma and infiltrating immune cells, we implemented a multiplex tissue slide-based assay to measure microRNA expression levels at single cell resolution and to extract the contextual information embedded within these different cell types. We will present some salient examples that ascertain aberrant and dysfunctional miRNA-mediated pathways within cancer cells and other cellular compartments of the tumor microenvironment. We will also discuss a tissue slide-based strategy for integration of translational miRNA and well-established protein biomarkers to increase molecular characterization of tumors and enhance the diagnostic power of current clinical assays.
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microRNA and Stem Cells
8:40-8:45 Chairperson's Opening Remarks
8:45-9:15 Talk Title to be Announced
Richard I. Gregory, Ph.D., Assistant Professor, Children's Hospital Boston, Harvard Stem Cell Institute (HSCI), Harvard Medical School
9:15-9:45 microRNAs Play an Important Role in Cortical Neural Stem Cell Development
Tao Sun, Ph.D., Associate Professor, Cell and Developmental Biology, Weill Medical College of Cornell University
Neural stem cells can self-renew and give rise to multiple cell types in the nervous system. We have identified some microRNAs that are expressed in neural stem cells derived from the cerebral cortex. We have found that microRNAs control self-renewal and differentiation of neural stem cells by regulating distinct targets. Our results have demonstrated the importance of microRNAs in neural stem cell development.
9:45-10:15 Inducing Pluripotency by RNA
Tariq M. Rana, Ph.D., Professor and Director, Program for RNA Biology, Sanford-Burnham Medical Research Institute
Although induced pluripotent stem cells (iPSCs) hold great promise for customized regenerative medicine, the molecular basis of reprogramming is largely unknown. Overcoming barriers that maintain cell identities is a critical step in the reprogramming of differentiated cells. Since microRNAs (miRNAs) modulate target genes tissue specifically, we reasoned that distinct fibroblast-enriched miRNAs post-transcriptionally modulate proteins that function as reprogramming barriers. We will present recent findings that cellular microRNAs regulate iPSC generation. Knockdown of key microRNA pathway proteins resulted in significant decreases in reprogramming efficiency. Three microRNA clusters, miR-17~92, 106b~25 and 106a~363, were shown to be highly induced during early reprogramming stages. Several microRNAs, including miR-93 and 106b, which have very similar seed regions, greatly enhanced iPSC induction and modulated the mesenchymal-to-epithelial transition step in the initiation stage of reprogramming, and inhibiting these microRNAs significantly decreased reprogramming efficiency. Moreover, miR-iPSC clones reached the fully reprogrammed state. Further analysis revealed that Tgfbr2 and p21 are directly targeted by these microRNAs. Overall, our results demonstrate that microRNAs function in the reprogramming process and that iPSC induction efficiency can be greatly enhanced by modulating microRNA levels in cells. These studies should facilitate development of clinically-applicable reprogramming strategies.
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Technology Showcase
Sponsored by
 11:15-11:30 A Unique Biological Knowledge-Driven Method for Rapid microRNA Target Prioritization: Cancer Case Studies
Aimee Jackson, Director, Product Management, Ingenuity Systems
microRNAs are a promising source of biomarkers. However, searching databases for predicted and observed targets yields thousands of potential targets and paring these down to a manageable number is a challenge. Case studies in which the number of potential targets was quickly reduced to just a few will be presented.
Sponsored by
 11:30-12:00 Translating miRNA Discovery in Biofluids into Robust Biomarkers for Disease
Søren Jensby Nielsen, Ph.D., Head, Diagnostics, Exiqon A/SExiqon has developed a LNA™-based miRNA qPCR platform with unparalleled sensitivity and robustness to accelerate discovery and clinical development of miRNA-based biomarkers. We have profiled thousands of biofluid samples including plasma, serum and urine. To secure technical excellence and eliminate unwanted bias, an extensive QC system has been implemented including studies of pre-analytical variables and development of normal reference ranges for circulating miRNAs, which will support the development of robust biomarkers.
12:00-12:15 pm Sponsored Presentation(Opportunity available. Contact Ilana Quigley, Manager, Business Development, at 781-972-5457 or iquigley@healthtech.com.)
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Technology Showcase
11:15-12:15 pm Sponsored Presentations
(Opportunities available. Contact Ilana Quigley, Manager, Business Development, at 781-972-5457 or iquigley@healthtech.com.)
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microRNAs as Biomarkers and Diagnostics (continued)
1:45-1:50 Chairperson's Opening Remarks
1:50-2:20 microRNAs as Potential Biomarkers for Non-Small Cell Lung Cancer
Feng Jiang, M.D., Ph.D., Associate Professor, Pathology, University of Maryland School of Medicine
We proposed to develop miRNAs as sensitive and specific sputum markers for early detection of NSCLC. From paired normal and tumor lung tissues from 20 patients with NSCLC using miRNA profiling, 7 miRNAs were found to have significantly altered expressions in tumors. On the sputum samples of 36 cancer patients and 36 healthy individuals, combination of the miRNAs produced 80.6% sensitivity and 91.7% specificity for lung cancer. Validation of the marker panel in an independent set confirmed the sensitivity and specificity. The sputum markers demonstrated the potential of translation to laboratory settings for improving the early detection of NSCLC.
2:20-2:50 Blood Cell microRNAs: What Are They and What Future Do They Hold?
C.D. Atreya, Ph.D., Associate Director for Research, Office of Blood Research and Review, CBER, FDA
Storage of blood components has revolutionized health care by allowing for a managed supply of transfusion-quality blood products. During storage, blood components undergo a series of physiological changes that affect the product quality, which ultimately can interfere with the safety and efficacy of such products after transfusion. Despite continuous improvements in blood component quality and safety, it is still desirable to have in vitro standard markers of measurable characteristics that predict blood component safety and efficacy in vivo following their transfusion. There is potential for microRNAs to act as measurable characteristics (product biomarkers) for stored blood component quality and safety.
2:50-3:20 microRNAs as Effectors and Biomarkers in Cholestatic Liver Disease
Joshua Friedman, M.D., Ph.D., Assistant Professor, Pediatrics, Perelman School of Medicine, University of Pennsylvania
Consistent with their importance in many organ systems, microRNAs play critical roles in gastrointestinal development and differentiation. Using genetic tools, our laboratory has provided evidence of miRNA function in the developing and post-natal liver. Biliary atresia is a disease affecting infants in which the bile ducts are destroyed by a fibro-inflammatory process; it is the most common reason for liver transplantation in children. We have discovered that circulating miRNAs are biomarkers of the liver disease biliary atresia. In addition, we have identified a panel of miRNAs whose expression is altered in a variety of cholestatic disease models, and we have linked those miRNAs to several relevant regulatory pathways. These results will be reviewed in the context of growing evidence that microRNAs participate in the pathogenesis of digestive and liver diseases and make appealing therapeutic targets.
3:20-3:50 Circulating microRNAs as Biomarkers of Individual Sensitivity to Liver Injury in a Multistrain Panel of Inbred Mice
Igor Pogribny, M.D., Ph.D., Laboratory Director and Principal Research Investigator, Division of Biochemical Toxicology, National Center for Toxicological Research, U.S. Food and Drug Administration; Basic Science Professor, Pharmacology and Toxicology, University of Arkansas Medical Sciences
microRNAs (miRNAs) are a class of small, conserved, tissue-specific regulatory non-coding RNAs that modulate a variety of biological processes and play a fundamental role in pathogenesis of major human diseases, including nonalcoholic fatty liver disease (NAFLD), which is a major health problem and the leading cause of chronic liver disease in the United States and developed countries. In a relevant mouse model of human NAFLD, aberrant levels of several circulating miRNAs were highly correlated with a severity of NAFLD-associated pathomorphological changes in the livers. These changes correlated with an individual susceptibility to liver injury, suggesting that circulating miRNAs may be used as biomarkers of liver toxicity and, more importantly, as potential noninvasive indicators of susceptibility to NAFLD liver injury.
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microRNA Pathways in Disease
1:45-1:50 Chairperson's Opening Remarks
1:50-2:20 microRNAs, Other Small RNAs, and Schizophrenia
Clark D. Jeffries, Ph.D., Research Professor, UNC Eshelman School of Pharmacy and Renaissance Computing Institute, University of North Carolina at Chapel Hill
Judiciously chosen microRNAs have become established as consistent markers of schizophrenia in postmortem brain samples. Illumina sequencing of small RNAs has further revealed markers that are byproducts of well-known noncoding RNAs including tRNAs, rRNAs, snRNAs, and snoRNAs. There appears to be a parallel universe of small RNA agents of gene expression that inform us of schizophrenia.
2:20-2:50 NF-kB-Sensitive microRNA Signaling in Alzheimer's Disease
Walter J. Lukiw, B.S., M.S., Ph.D., Principal Investigator, LSU Neuroscience Center and Department of Ophthalmology, Louisiana State University Health Sciences Center
Human brain cells retain selective subsets of microRNAs (miRNAs), and miRNA effects on brain gene expression are mediated through their regulation of messenger RNA (mRNA) complexity. In recent studies: (a) in short postmortem interval Alzheimer's disease (AD) association neocortex, (b) in cytokine- (IL-1β,TNFα) and Aβ42 peptide-stressed human neuronal-glial (HNG), astroglial (HAG) and microglial (HMG) cells in primary culture, and (c) in several murine transgenic AD (Tg-AD) models including Tg2576 and 5xFAD, we have identified several brain-abundant miRNA species that are consistently upregulated, including miRNA-146a. This inducible, NF-kB-regulated, 22 nucleotide non-coding RNA targets the mRNA of the key, innate-immune- and inflammation-related regulatory protein, complement factor-H (CFH), resulting in significant decreases in CFH expression (p<0.01, ANOVA). HNG, HAG and HMG cells each respond differently to cytokine- and Aβ42-peptide-induced stress, in part by variation in their intrinsic miRNA-mediated responses. The complex interactive signaling of NF-kB and miRNA-146a further underscore the interplay between inducible transcription factors and potentially pathogenic miRNAs that mediate CFH expression in inflammatory neurodegeneration.
2:50-3:20 microRNA-21 in Type-1 Diabetes
Youhai H. Chen, M.D., Ph.D., Professor, Pathology and Lab Medicine, University of Pennsylvania School of Medicine
Death of pancreatic b cells is a pathological hallmark of type-1 diabetes (T1D). Recently we discovered a novel regulatory pathway of b cell death that comprises microRNA-21, its target tumor suppressor PDCD4, and its upstream transcriptional activator NF-kB. In pancreatic b cells, NF-kB activates the miR-21 gene promoter and increases miR-21 RNA levels; miR-21 in turn decreases the level of PDCD4 that is able to induce cell death. Consequently, PDCD4 deficiency in mice conferred resistance to diabetes. Thus, the NF-kB-microRNA-21-PDCD4 axis plays a crucial role in T1D and represents a novel therapeutic target for treating the disease.
3:20-3:50 Angiotensin Type 1 Receptors Regulate a Specific Set of microRNAs in Cardiovascular Tissues Including Human Arteries
Tilde V. Eskildsen, Ph.D., Postdoctoral Scientist, Clinical Biochemistry and Pharmacology, Odense University Hospital
The Angiotensin II receptor (AT1R) is a key regulator of blood pressure cardiac contractility. We asked whether miRNAs are regulated by AT1R signals in vitro using primary cardiac cells and in vivo using aortas from rats with Ang II-induced hypertension and human arteries from patients with cardiovascular disease. Ang II upregulated miRNA-7, miRNA-129-3p, miRNA-132, and miRNA-212 in the cell lines and in rat aortas after Ang II infusion. Remarkably, we found a robust decrease in miRNAs including miR-7 in human arteries treated with AT1R blockers. The perspective is that the identified miRNAs may be involved in cardiac biology and disease.
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microRNA Therapeutics: Design and Delivery
4:45-5:15 Therapeutic microRNA Mimetic and Antisense Oligonucleotides
Thomas D. Schmittgen, Ph.D., Associate Professor and Chair, Division of Pharmaceutics, Ohio State University
5:15-5:45 Chemical Strategies for Delivery of RNA Drugs
Muthiah Manoharan, Ph.D., Vice President, Drug Discovery, Alnylam
At Alnylam Pharmaceuticals we have developed and applied multiple chemistry strategies to address the challenge of cellular delivery of drugs that function through RNAi pathways. These include chemical modifications of oligonucleotides, molecular conjugates and delivery systems based on liposomal nanoparticles (LNPs). Our progress in these areas will be summarized.
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Role of microRNA in Viral Infection
4:45-5:15 Herpes Simplex Virus microRNAs and the Lytic-Latent Balance
Donald Coen, Ph.D., Professor, Biological Chemistry and Molecular Pharmacology, Harvard Medical School
Herpes simplex viruses (HSV) 1 and 2 infect much of the human population. HSV-1 and -2 establish latent infections in sensory neurons from which the viruses can reactivate to cause recurrent disease and transmit to new hosts. During latency, expression of viral genes active during productive ("lytic") infection is repressed. HSV-1 and -2 encode a score or more of microRNAs. There is increasing evidence that these microRNAs can tilt the outcome of infection either towards virus production or latency. The results have raised the possibility that latent HSV infections can be addressed by targeting HSV microRNAs.
5:15-5:45 Promoting Hepatitis C Virus Replication through miR-122 Manipulation
Matthew J. Evans, Ph.D., Assistant Professor, Microbiology, Mount Sinai School of Medicine
The hepatitis C virus (HCV) is a major cause of chronic hepatitis, affecting approximately 170 million people worldwide. HCV replication requires a direct interaction with the liver-specific microRNA miR-122, and the few cell lines that support the HCV life cycle express high levels of this microRNA. Here we show that overexpression of miR-122 in the normally HCV-nonpermissive and miR-122-deficient hepatocyte-derived HepG2 cell line renders these cells able to support the entire HCV life cycle. Thus HepG2 cells represent a novel HCV cell culture system that is particularly useful for exploring the mechanism by which this microRNA enhances HCV replication.
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