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OMB No. 0925-0001/0002 (Rev. 08/12 Approved Through 8/31/2015) BIOGRAPHICAL SKETCH Provide the following information for the Senior/key personnel and other significant contributors. Follow this format for each person. DO NOT EXCEED FIVE PAGES. NAME: Yoo, Soonmoon eRA COMMONS USER NAME (credential, e.g., agency login): soonmoonyoo POSITION TITLE: Research Scientist EDUCATION/TRAINING (Begin with baccalaureate or other initial professional education, such as nursing, include postdoctoral training and residency training if applicable. Add/delete rows as necessary.) INSTITUTION AND LOCATION DEGREE (if applicable) Completion Date MM/YYYY FIELD OF STUDY Chung-Ang University, Seoul, South Korea Chung-Ang University, Seoul, South Korea University of Texas Medical Branch, Galveston, TX B.S. M.S. Ph.D. 02/1994 08/1996 05/2003 Biology Biochemistry Physiology & Biophysics Georgetown University, Washington, DC Nemours Biomedical Research, Wilmington, DE Postdoctoral Postdoctoral 12/2006 08/2010 Neuroscience Neurobiology A. Personal Statement The long-term goal of this proposed study is: (1) to identify and characterize neuronal piRNAs in axons of sciatic nerve, (2) to profile and contrast levels of piRNAs in axonal vs. cell body compartment s at different times after injury, (3) to identify PIWI/piRNA-target genes that are transported into distal axons, and (4) examine roles of the axonal piRNAs in regenerating axons. I, a New Investigator, have worked in the field of neural regeneration for more than a decade, using in vivo and in vitro models including invertebrate giant axons, primary mammalian neurons in culture and rodent spinal cord and sciatic nerve injury, and have focused on the cellular and molecular mechanisms underlying spontaneous resealing of injured axoplasmic membrane followed by regeneration. Using adult rat peripheral injury model, we have successfully developed live cell imaging and local translation monitoring methodologies (FRAP), the qualitative and quantitative fluorescence in situ hybridization (FISH) protocols and subcellular compartment isolation technique that are necessary for studies of the axonal mRNA and miRNA localization and local protein synthesis. My laboratory is recently well-versed in a RNA-immunoprecipitation (RIP) technique and RNA-electrophoresis mobility shift assay (REMSA) to identify the cognate mRNA population. We have very recently started investigating mechanisms of mature and precursor miRNAs that are localized into axonal compartments of neurons, supported by R21 funding. The studies proposed here are built on the very exciting findings from these profiling studies using Next Generation Deep Sequencing. Although it took some time for me to start publishing my independent work from my own group, I think that we are finally producing very exciting data that the neuroscience as well as cellular/molecular biology community will accept. The studies proposed here will combine several methodologies that we have been successfully optimized and utilized in collaboration (NGS, axoplasm purification, FISH, CLIP and microfluidic device), and directly apply these systems and the unique reagents for study regulatory mechanisms of intra-axonal protein synthesis to improve regeneration in nerve injury. 1. Yudin, D., Hanz, S., Yoo, S., Iavnilovitch, E., Willis, D., Gradus, T., Vuppalanchi, D., Segal-Ruder, Y., BenYaakov, K., Hieda, M., Yoneda, Y., Twiss, J.L. & Fainzilber, M. (2008). Localized regulation of axonal RanGTPase controls retrograde injury signaling in peripheral nerve. Neuron 59(2): 241-52. PMCID: PMC2538677. 2. Yoo, S., Kim, H.H., Donnelly, C.J., Vuppalanchi, D., Kim, P., Park, M., Lee, S.J., Merianda, T.T., PerroneBizzozero, N., Twiss, J.L. (2013). A HuD-ZBP1 ribonucleoprotein complex localizes GAP-43 mRNA into axons through its 3’ untranslated region AU-rich regulatory elements. J Neurochem. 126: 792-804. PMCID: PMC3766383. 3. Kim, H.H., Kim, P., Phay, M. & Yoo, S. (2015). Identification of precursor microRNAs within distal axons of sensory neuron. J. Neurochem. 134:193-9. PMCID: PMC4490939. 4. Kim, H.H., Lee, S.J., Gardiner, A.S., Perrone-Bizzozero, N.I. & Yoo, S. (2015). Different motif requirements for the localization zipcode element of β-actin mRNA binding by HuD and ZBP1. Nucleic Acid Res. 43:7432-46. PMCID: PMC4551932. B. Positions and Honors Positions and Employment 1994-1996 Teaching Assistant, Department of Biochemistry, School of Graduate Studies, Chung-Ang University, Seoul, South Korea 1997-2002 Graduate Assistant, Department of Physiology and Biophysics, University of Texas Medical Branch, Galveston, TX 2003-2006 Postdoctoral fellow (with Dr. Jean R. Wrathall), Department of Neuroscience, Georgetown University Medical Center, Washington, DC 2006-2010 Postdoctoral fellow (with Dr. Jeffery L. Twiss), Nemours Biomedical Research, Alfred I duPont Hospital for Children, Wilmington, DE 2010-2013 Assistant Research Scientist, Human Genetics Research Laboratory, Nemours Biomedical Research, Alfred I duPont Hospital for Children, Wilmington, DE 2012Affiliated Scientist, Department of Biological Sciences, University of Delaware, Newark, DE 2012-2013 Guest Lecturer, Department of Biology, Drexel University, Philadelphia, PA 2013Research Scientist, Molecular Regeneration and Neuroimaging Laboratory, Nemours Biomedical Research, Alfred I duPont Hospital for Children, Wilmington, DE 2014Research Assistant Professor, Department of Pediatrics, Thomas Jefferson University Medical College, Philadelphia, PA Other Experience and Professional Memberships 1998Member, Society for Neuroscience 2005-2007 Member, International Society for Neurochemistry 2010-2013 Member, American Society for Cell Biology 2005Journal of Neurochemistry, ad hoc reviewer 2007Developmental Neurobiology, ad hoc reviewer 2008Journal of Neuroscience, ad hoc reviewer 2010EMBO Journal, ad hoc reviewer 2011Journal of Cell Science, ad hoc reviewer 2012Molecular & Cellular Proteomics, ad hoc reviewer 2014Editorial Board, The Journal of DNA and RNA Research, Open Access Pub 2014Cellular and Molecular Neurobiology, ad hoc reviewer Honors 2003 George Sealy Research Award in Neurology, The University of Texas Medical Branch, Galveston, Texas, USA C. Contribution to Science 1. My early interests in neurobiology/neuroscience focused on the cellular mechanisms underlying neural repair of both invertebrate and mammalian axons following mechanical injury. I demonstrated for the first time that the injury distance from the cell body is the main determinant of cell fate after injury via extracelluar calcium ions influxed and accumulated into cell body. This finding brought us to new frontiers in understanding the fundamental mechanisms for axolemmal repair after neuronal injury and multiple roles of calcium ion in neurons. I also enabled to show that axolemmal repair after injury requires some synaptic vesicle fusion proteins, such as syntaxin and synaptotagmin. These works were published in several peer- reviewed journals, in which I served as the primary or co-investigator. a. Yoo, S., Bottenstein, J.E., Bittner, G.D., and Fishman, H.M. 2004. Survival of Mammalian B104 Cells following Neurite Transection at Different Locations Depends on Somal Ca2+ Concentration. J. Neurobiol. 60:137-153. PMID: 15266646. b. Yoo, S., Nguyen, M.P., Fukuda, M., Bittner, G.D., Fishman, H.M. 2003. Plasmalemmal Sealing of Transected Mammalian Neurites is a Gradual Process Mediated by Ca2+-regulated Proteins. J. Neurosci. Res. 74:541-551. PMID: 14598298. c. Detrait, E.R., Eddleman, C.S., Yoo, S., Fukuda, M., Nguyen, M.P., Bittner, G.D., Fishman, H.M. 2000. Axonal repair requires proteins that mediate synaptic vesicle fusion. J. Neurobiol. 44:382-391. PMID: 10945894. d. Detrait, E.R., Yoo, S., Eddleman, C.S., Fukuda, M., Bittner, G.D., Fishman, H.M. 2000. Plasmalemmal repair of severed neurites of PC12 cells requires Ca2+ and synaptotamin. J. Neurosci. Res. 62: 566573. PMID: 11070500. 2. I further expanded my research agenda and explored nerve regeneration using an animal model of spinal cord injury (SCI). I was one of the first to demonstrate that SCI stimulates the proliferation of NG2+ cells with the properties of oligodendrocytes progenitor cells (OPCs) in adult rodents [13]. The method that I developed to purify OPCs from adult spinal cord enabled us to characterize the properties of precursor cells in the adult CNS and evaluate the therapeutic potential to improve recovery after SCI. My research has further shown that activated microglia and macrophages, the predominant cell types at the site of SCI, produce factors that inhibit oligodendrogenesis in vivo after SCI [14]. These studies clearly suggest that activated microglia/macrophages and factors secreted by them could contribute to an inhibitory microenvironment in the SCI lesion per se, interfering with the proliferation and differentiation of endogenous NG2+ OPCs and preventing functional recovery in SCI patients. a. Wu, X.F., Yoo, S., Wrathall, J.R. 2005. Real-time PCR analysis of temporal-spatial alterations in gene expression after spinal cord contusion. J. Neurochem. 93:943-952. PMID: 15857397. b. Zai, L.J., Yoo, S., Wrathall, J.R. 2005. Increased growth factor expression and cell proliferation after contusive spinal cord injury. Brain Res. 1052:147-155. PMID: 19005441. c. Yoo, S., Wrathall, J.R. 2007. Mixed primary culture and clonal analysis provide evidence that NG2 proteoglycan-expressing cells after spinal cord injury are glial progenitors. Dev. Neurobiol. 67:860-874. PMID: 17506499. d. * Wu, J., * Yoo, S., Wilcock. D., Lytle, J.M., Leung, P.Y., Colton, C.A., Wrathall, J.R. 2010. Interaction of NG2+ glial progenitors and microglia/macrophages from the injured spinal cord. Glia. 58:410-422. PMCID: PMC2807472 (* J. Wu and S. Yoo contributed equally to this work). 3. Neurons in CNS have little capacity for regeneration but PNS neurons show significant morphologic/ functional recovery after injury. However, it is still unclear what causes the differences between these neurons and how this must be solved for successful regeneration of CNS nerve. By focusing on molecular mechanisms that regulate RNA axonal localization and de novo protein synthesis in distal axons needed to regenerate axons of PNS neurons, I have devoted substantial effort to develop robust methods to dissect the roles of cis-elements within mRNA for localization and axonal translational control using live cell imaging approaches and quantification methods (qFISH). Although this is still an underexplored topic, I directly demonstrated that specific RNAs encoding proteins that trigger injury-induced responses in the cell body contain the localization cis-elements within the 3’UTR, which are specifically recognized by transacting RNA-binding proteins. I also enabled to show that axonally synthesized proteins participate in autonomous function of distal axons via spatial and temporal regulation of local protein synthesis. Very recently, our new studies clearly demonstrated that specific small non-coding regulatory miRNAs actively localize into distal axons and locally maturated into functional mature miRNAs to spatially regulate local protein synthesis. Current studies are expanding the mechanistic research of axonal mRNA transport and regulation of local protein synthesis medicated by miRNAs and/or piRNAs to in vivo biological relevance of these axonal mRNAs in regenerating axons. On completion of these small non-coding RNA-mediated translation regulation in distal axons, I will, for the first time, demonstrate mRNA-independent localization of precursor miRNAs and axon-specific PIWI/piRNA complexes in distal axons. The elucidation of cellular/molecular mechanisms of control of local protein synthesis within the axonal compartment at different levels of neuronal activity including regenerative processes could lead to the development of RNAbased approaches for the therapy of neurological disorders. a. Donnelly, C.J., Park, M., Spillane, M., Yoo, S., Pacheco, A., Gomes, C., Kim, H.H., Merianda, T.T., Gallo, G., Twiss, J.L. 2013. Axonally synthesized β-actin and GAP-43 proteins support distinct modes of axonal growth. J. Neurosci. 33: 3311-22. PMCID: PMC3739791. b. Yoo, S., Kim, H.H., Kim, P., Donnelly, C.J., Kalinski, A.L., Vuppalanchi, D., Park, M., Lee, S.J., Merianda, T.T., Perrone-Bizzozero, N.I., Twiss, J.L. 2013. A HuD-ZBP1 ribonucleoprotein complex localizes GAP-43 mRNA into axons through its 3’ untranslated region AU-rich regulatory element. J. Neurochem. 126: 792-804. PMCID: PMC3766383. c. Merianda, T.T., Coleman, J., Kim, H.H., Kumar Sahoo, P., Gomes, C., Brito-Vargas, P., Rauvala, H., Blesch, A., Yoo, S. & Twiss, J.L. (2015). Axonal amphoterin mRNA is regulated by translational control and enhances axon outgrowth. J Neurosci. 35(14): 5693-706. PMCID: PMC3755718 d. Phay, M., Kim, H.H., & Yoo, S. (2015). Identification of precursor microRNAs within distal axons of sensory neuron. PLoS One 10:e0137461. PMCID: PMC4557935. Complete List of Published Work in MyBibliography: http://www.ncbi.nlm.nih.gov/sites/myncbi/1NM9zAe7iHi57/bibliography/47276267/public/?sort=date&di rection=ascending D. Research Support Ongoing Research Support P20GM103464 Shaffer (PI) 09/07/10-06/30/16 NIH/NCRR/NIGMS: Centers of Biomedical Research Excellence II (COBRE II), Center for Pediatric Research “Temporal regulation of localized mRNA translation in regenerating axon” The goals of this proposal are to determine the intracellular pathways that underlay the local translational control to confer temporal specificity to local mRNA translation after axonal injury, and to examine how the axon chooses which mRNAs to translate in response to any given stimulus. Role: Target Investigator R21 NS085691 Yoo (PI) 10/01/13-09/30/16 “Profiling and characterizing axonal precursor microRNAs in regenerating nerve” The major goals of this project are to profile changes of the precursor and mature miRNA expression levels in the axonal compartments following sciatic nerve injury and to identify the cis-acting region(s) of precursor miRNAs and trans-acting factor(s) that are required for localization. Role: PI Completed Research Support Delaware INBRE Core Center Access Award Yoo (PI) 09/05/14-03/05/15 Delaware INBRE “Profiling mature microRNAs and piRNAs in regenerating peripheral nerve” The goal of this award is to determine if neuron-specific small non-coding RNAs including miRNAs and piRNAs reveal differential expression in response to injury during regeneration. Role: PI Post-doctoral Fellowship Award 124124 Yoo (PI) 07/01/2009 – 09/17/2010 Craig H. Neilsen Foundation “Temporal regulation of localized mRNA translation in regenerating axons” The objective of this grant is to determine how the axonal protein synthesis machinery is regulated to confer temporal specificity to local mRNA translation after axonal injury. Role: P.I. 2 P20 RR016472-11 Steiner (PI) 03/01/2012 – 02/28/2014 Delaware INBRE “Defining the role of Sonic hedgehog mediated regulation of Na,K-ATPase in medulloblastoma” A collaboration with Dr. Sigrid A. Langhans (a PI of the pilot grant), Yoo is helping for isolating primary granule cells from mouse medulloblastoma and providing with constructs to generate the dual reporter or Dendra2 photoconvertible systems for analyzing intracellular trafficking pathways in neurons using confocal live cell imaging. Individual Research Grants WB2-0403-2 Wrathall (PI) 01/01/2005 - 12/31/2006 Christopher Reeve Paralysis Foundation “Endogenous Precursor Cells in Chronic SCI” This project will test the hypothesis that chronically after contusion injury there remain a larger than normal population of glial precursors cells that can be stimulated to divide and differentiate to improve function after SCI. Role: Co-Investigator