Nanette Nascone-Yoder
Bio
Nanette Nascone-Yoder obtained her BS in Molecular Biology from Eckerd College, and Ph.D. in Cell and Developmental Biology at Harvard University.
Dr. Nascone-Yoder established her laboratory at NC State’s College of Veterinary Medicine in 2006 and was selected as a University Faculty Scholar in 2013. As an Associate Professor in the Department of Molecular Biomedical Sciences, Dr. Nascone-Yoder’s research is at the intersection of development, evolution and toxicology. Her lab focuses on discovering the cellular and molecular mechanisms by which organs become left-right asymmetric, as a way to understand the causes of laterality-related birth defects. The Nascone-Yoder lab recently pioneered the use of a novel model organism for the study of organogenesis, the Budgett’s frog. The lab’s research has been funded by the National Science Foundation, National Institutes of Health and American Heart Association, and recommended by Faculty of 1000.
AFFILIATIONS
Comparative Medicine Institute, Center for Human Health and the Environment, Society for Developmental Biology
CERTIFICATIONS
Ph.D., Harvard University, 1997
Assistant Professor, Eckerd College, 1997, tenured 2003
Associate Professor, Eckerd College, 2004
Area(s) of Expertise
BIOLOGICAL BARRIERS
Morphogenesis, organogenesis, digestive organs, left-right asymmetry
Publications
- Developmental regulation of cellular metabolism is required for intestinal elongation and rotation , DEVELOPMENT (2024)
- Morphoelastic models discriminate between different mechanisms of left-right asymmetric stomach morphogenesis , CELLS & DEVELOPMENT (2024)
- The people behind the papers - Julia Grzymkowski and Nanette Nascone-Yoder , DEVELOPMENT (2024)
- Rare variants in CAPN2 increase risk for isolated hypoplastic left heart syndrome , HUMAN GENETICS AND GENOMICS ADVANCES (2023)
- Normal Table of Xenopus development: a new graphical resource , DEVELOPMENT (2022)
- Single-minded 2 is required for left-right asymmetric stomach morphogenesis , DEVELOPMENT (2021)
- The twists and turns of left-right asymmetric gut morphogenesis , DEVELOPMENT (2020)
- Exome sequencing of family trios from the National Birth Defects Prevention Study: Tapping into a rich resource of genetic and environmental data , BIRTH DEFECTS RESEARCH (2019)
- Vangl2 coordinates cell rearrangements during gut elongation , DEVELOPMENTAL DYNAMICS (2019)
- The left-right asymmetry of liver lobation is generated by Pitx2c-mediated asymmetries in the hepatic diverticulum , DEVELOPMENTAL BIOLOGY (2018)
Grants
The mission of the Center for Human Health and the Environment (CHHE) is to advance understanding of environmental impacts on human health. Through a systems biology framework integrating all levels of biological organization, CHHE aims to elucidate the fundamental mechanisms through which environmental exposures/stressors interface with biomolecules, pathways, the genome, and epigenome to influence human disease. CHHE will develop three interdisciplinary research teams that represent NC State������������������s distinctive strengths. CHHE will implement specific mechanisms to promote intra- and inter-team interactions and build interdisciplinary bridges to advance basic science discovery and translational research in environmental health science along the continuum from genes to population. These teams are; - The Molecular/Cellular-Based Systems and Model Organisms Team will utilize cutting edge molecular/cellular-based systems and powerful vertebrate and invertebrate model organisms to define mechanisms, pathways, GxE interactions, and individual susceptibility to environmental agents. - The Human Population Science Team will integrate expertise on environmental exposures, epidemiology, genomics and epigenomics to identify key human pathways and link exposure and disease across populations. - Bioinformatics Team will develop novel analytics and computational tools to translate Big Data generated across high-throughput and multiscale experiments into systems-level discoveries To further increase the impact and translational capacity of these teams, CHHE will develop three new facility cores that will provide instrumentation, expertise, and training to facilitate basic mechanism- to population-based research. - The Integrative Health Sciences Facility Core will expand the ability of CHHE members to translate basic science discoveries across species and provide mechanistic insights into epidemiological studies by partnering with: a) NC State������������������s Comparative Toxicogenomics Database (CTD); b) East Carolina University Brody School of Medicine and c) NC Dept. of Health and Human Services. - The Comparative Pathobiology Core will be located at NC State������������������s top-ranked College of Veterinary Medicine and its nationally recognized veterinary pathology group to facilitate assessment of the effects of environmental stressors in the many model organisms utilized by CHHE members. - The Systems Technologies Core will introduce state-of-the-art proteomics capabilities and dedicated bioinformatics support to expand the ability of CHHE members to analyze the Next Generation Sequencing data involving the genome, transcriptome and epigenome. As a land-grant university, NC State has an extensive and active Cooperative Extension Service network throughout North Carolina. CHHE will utilize this unique network to develop a highly effective, multi-directional Community Outreach and Engagement Core to disseminate findings that will contribute to addressing disparity in exposures and health outcomes and to educate communities about environmental influences on health. A strong Career Development Core for early stage scientists that is coordinated with a robust Pilot Project Program will support cutting-edge, collaborative and multidisciplinary environmental health projects to enhance the research success and impact of our membership. Through these activities and the purposeful interfacing of different disciplines CHHE will build on NC State������������������s unique research and community outreach strengths to become a premier transformative and synergistic EHS Core Center.
Intestinal malrotation (IM) is a highly prevalent birth defect that can lead to life-threatening conditions necessitating surgical intervention and long-term supplemental nutrition. Atrazine (ATR), a ubiquitous herbicide that perturbs the photosynthetic electron transport chain (ETC), was found to cause IM at high frequency. Consistent with its ability to inhibit ETC reactions, preliminary data show that ATR decreases mitochondrial oxidative phosphorylation (oxphos) in the developing intestine, while increasing glycolytic flux. However, while early gut development can subsist on glycolysis alone, later gut development requires mitochondrial respiration; thus, ATR may block a critical metabolic switch from glycolysis to oxphos during intestine morphogenesis. During normal gut development, intestinal rotation occurs simultaneously with gut lengthening, and shorter gut lengths are often associated with IM, suggesting that gut elongation mechanisms are integral to the rotation process. Indeed, cellular analyses reveal that ATR perturbs crucial events required to drive intestinal lengthening, including early mesenchymal-to-epithelial transitions (MET) and later interkinetic nuclear migration (IKNM) -- widely used morphogenetic processes also influenced by cellular metabolic states. Together, these data suggest that a metabolic switch from glycolysis to oxphos regulates the timing and/or localization of MET and IKNM events during intestine morphogenesis, thus controlling gut elongation to drive proper rotation. This hypothesis will be tested using innovative metabolomics technologies to determine: 1) how ATR affects the normal spatiotemporal dynamics of cellular metabolism during intestine morphogenesis, and 2) how dynamic cellular metabolic states affect the timing and localization of MET and IKNM within the developing intestine. Successful completion of these aims will illuminate the poorly understood etiology of IM, with implications for the role of metabolism-altering environmental toxins, diseases and/or pregnancy conditions in the development of other structural birth defects that depend on MET- and/or IKNM-mediated morphogenesis.
Left-right (LR) differences in size, shape and/or anatomical position exist in almost every organ system. Consequently, abnormal LR asymmetry (known as heterotaxy, HTX) often leads to multiple, life threatening birth defects involving complex malformations and discordant laterality between organs. While the early embryonic events that establish global LR asymmetry have been well studied, it is the later-stage, organ-specific LR asymmetric morphogenesis events that are critical for normal anatomy; yet, for most organs, the molecular and cellular processes that sculpt their individual LR asymmetries have not been elucidated. This application will explore the novel and surprising concept that LR asymmetries in GLYCOLYSIS���the primordial metabolic pathway that breaks down glucose to generate ATP���are integrally involved in asymmetric organ morphogenesis. Published work has shown that leftward curvature of the stomach, an archetypical LR asymmetry conserved among vertebrates, depends on LR asymmetric rearrangements of mesenchymal cells into an epithelium (mesenchymal-to-epithelial transition; MET), causing thinning and expansion of the left stomach wall. Recently, left vs right transcriptome profiling subsequently revealed that glycolysis enzymes are upregulated in the left side of the stomach during curvature. As glycolysis (as opposed to mitochondrial respiration) is known to promote epithelial-mesenchymal plasticity in other contexts, this unexpected finding raises the intriguing possibility that LR asymmetric glycolysis may facilitate the LR asymmetric MET that drives curvature morphogenesis. In the proposed project, this idea will be rigorously tested by exploiting the unique attributes of two different amphibian embryos, executing metabolomics, mass spectrometry imaging, glycolytic flux analyses, pharmacological perturbations, and left- vs right-targeted tests of gene function, to determine the function of glycolysis genes (Aim 1), and glycolytic metabolism (Aim 2) in stomach curvature morphogenesis. Successful completion of this R21 will therefore substantiate the intriguing premise that organ laterality is shaped by LR asymmetries in metabolism, such that aberrant metabolic states may contribute to the development of laterality-related birth defects.
Birth defects are a leading cause of infant mortality, yet in most cases, their etiology is unknown. Some of the most common and complex malformations are found in families with abnormal left-right (LR) asymmetry, suggesting that many congenital defects result from perturbed laterality. The initial embryonic events that determine the LR body axis, including the early breaking of bilateral symmetry, and subsequent left-sided expression of determinants such as nodal and Pitx2, are now well understood. However, the genes and morphogenetic events involved in the final phases of LR development, at the organ level, remain largely unknown. The long term goal is to ascertain the mechanisms of LR asymmetric organogenesis. The objective in this application is to identify the molecular and cellular processes that generate LR asymmetry (curvature) within an individual organ, the stomach. Preliminary analyses identified LR asymmetries in radial cell rearrangements in the developing stomach as the driving force for its curvature. To identify genes likely to be proximate effectors of this novel asymmetric morphogenetic program, a new model organism (Lepidobatrachus laevis) was employed. The extra-large embryos of this species facilitated a genediscovery approach that would be infeasible in most models: transcriptome profiling (RNASeq) of tissues dissected from left vs. right halves of the embryonic stomach. Pilot datasets include genes with validated LR asymmetric expression patterns and functions during stomach curvature. The central hypothesis is that stomach curvature is determined by distinct left and right regulatory networks which differentially modulate the cellular events controlling radial cell rearrangement. The unique experimental amenability of frog embryos will be used to test this hypothesis via three specific aims: 1) Generate molecular signatures of normal and abnormal stomach curvature. Comprehensive spatiotemporal profiles of LR stomach genes will be generated and compared in the context of both normal LR asymmetry and experimentally-induced LR axis defects. 2) Determine the cellular function of stomach-specific LR genes. Select genes will be tested in loss- and gain-of-function assays to determine their influence on radial cell rearrangement in the developing stomach. 3) Determine the regulatory hierarchy that controls stomach curvature. Experimental perturbations combined with spatiotemporal profiling will reveal core gene regulatory interactions governing asymmetric morphogenesis. The overall approach is innovative because it takes advantage of distinctive attributes of a unique species to address one of the key unanswered questions in LR development: what are the proximate mechanisms by which developing organs become LR asymmetric entities? The proposed research is significant because it will immediately advance our understanding of normal laterality and laterality-related birth defects by defining the genes, morphogenetic processes and regulatory logic that govern the emergence of LR asymmetry at the organ level.
The objective of this proposal is to obtain funding from the North Carolina Biotechnology Center to purchase a unique, automated, and high throughput slide scanning imager manufactured by Olympus. The new equipment allows for rapid acquisition of highly detailed images of cells and tissues. In biological research, often the generation of large amounts of samples from experiments is desired to tackle important research questions. Yet, sample generation often is not the limiting factor. Rather the imaging and data analysis of large numbers of samples is usually prohibitive and limits the types and impact of research questions that can be asked. As described in this proposal, the Olympus imaging system would unlock a wide range of important biomedical questions in diverse systems from zebrafish to human, bone marrow to brain, by providing an over 10-fold improvement in sample imaging throughput and analysis. The instrument will be installed within an NCSU Shared Core Research Facility, the Cellular and Molecular Imaging Facility (CMIF), which lacks the proposed technology currently, and is staffed by professional staff that will manage, maintain, and operate the proposed equipment. Acquisition of the Olympus slide scanner and installation within CMIF will increase the rate of data acquisition by research groups at NCSU and, in so doing, ensure their continued competitiveness for research funding.
The causes of most human birth defects are unknown. Structural defects in the heart and digestive tract are frequently found in association with abnormal left-right asymmetries in other organ systems, suggesting that such deformities may result from perturbed laterality, yet the developmental processes that form asymmetries within individual organs remain elusive. The long term goal is to determine the morphogenetic mechanisms that control the development of anatomical left-right asymmetry. The objective of this exploratory R21 application is to gain a comprehensive view of the left-right asymmetric molecular differences within the embryonic heart and gut tubes as they undergo asymmetric ?looping?, a fundamental organogenesis event that orients the most crucial anatomical asymmetries. To accomplish this objective, the unique embryological features of a novel model amphibian, Lepidobatrachus laevis, will be developed and exploited. Lepidobatrachus has massive embryos that facilitate precise excision of the left and right halves of the early heart and gut during looping, enabling the heretofore infeasible approach of left-right transcriptome profiling during a key phase of asymmetric morphogenesis. The central hypothesis is that identifying transcripts that are differentially expressed between the contralateral halves of looping organs will identify new molecules that control left-right asymmetric morphogenesis. This hypothesis will be tested via two specific aims: 1) Identify transcripts that are differentially expressed between the left and right sides of the looping heart and gut tubes; and 2) Validate the biological relevance of left- or right-enriched transcripts for asymmetric organ morphogenesis. Under Aim 1, an RNA seq approach (supported by a draft Lepidobatrachus transcriptome already constructed by the applicant) will be used to complete genome-wide expression analyses that will reveal unilaterally-enriched transcripts associated with the formation of key anatomical asymmetries. Under Aim 2, the proven ability to precisely target exogenous reagents to the left or right side of developing organs in amphibians, and the unprecedented subcellular resolution of developing organ asymmetries provided by the sizeable Lepidobatrachus, will be used to authenticate the in vivo function of select unilaterally-enriched transcripts. The approach is innovative because it takes advantage of the distinctive attributes of a unique nonmodel organism to understand one of the key unanswered questions in the field of left-right development: what are the mechanism(s) by which developing organs acquire critical left-right asymmetric anatomical features? The proposed research is significant because it is expected to immediately accelerate our understanding of the etiology of some of the most common birth defects by identifying new classes of molecules, and new cellular processes, which shape the fundamental left-right asymmetry of the heart and gut.
The vertebrate digestive tract develops left-right asymmetric loops and chiral rotations that are essential for normal physiological function. Perturbations of these fascinating anatomical features underlie the development of life-threatening congenital defects. In the early embryo, the initial determination of ?left? versus ?right? is manifest as unique left-right asymmetric gene expression patterns, including the left-limited expression of the transcription factor, Pitx2, which is required for normal asymmetric organ morphogenesis. However, the mechanism by which this molecular asymmetry then engenders morphological asymmetries within developing organs remains poorly understood. Preliminary data indicate that the endoderm cell rearrangements that drive tissue elongation and epithelial morphogenesis in the Xenopus primitive gut tube (PGT) are governed by Wnt/Planar Cell Polarity (Wnt/PCP) signaling, and are modulated by Pitx2 expression. The objective of this proposal is to determine the individual and combined roles of Pitx2 and Wnt/PCP signaling in regulating endoderm cell shape, adhesion and/or rearrangement during the development of left-right asymmetric anatomy in the digestive tract. Loss- and gain-of-function strategies will be employed, including the use of novel photoactivable reagents that have been developed for side-, stage-, and tissue-specific spatiotemporal modulation of Wnt-PCP and Pitx2 expression in the Xenopus gut tube. Specific Aim 1 is to determine whether Wnt-PCP signaling regulates cytoskeletal architecture, adhesive remodeling and cell rearrangement in the endoderm during gut morphogenesis. Aim 2 is to determine whether asymmetric Pitx2 expression controls these parameters on the left side of the gut tube. Aim 3 is to determine whether the function of Pitx2 in asymmetric gut morphogenesis is mediated by Wnt-PCP signaling. Successful completion of the proposed research will provide unique insight into the unsolved mechanisms of asymmetric organ morphogenesis, by linking left-right asymmetric gene expression patterns to key mechanisms of asymmetric organ development. The proposed work will illuminate the currently unknown etiology of common digestive organ birth defects, such as intestinal malrotation, narrowing or occlusion of the digestive tract, and congenital short bowel. The results of this research will also have significant, broad implications for the interrelationship of Pitx2 and Wnt signaling in the morphogenesis of multiple symmetric and asymmetric organs, and in the epithelial-mesenchymal transitions that drive development, maintain gut epithelial homeostasis, and underlie metastatic progression in tumors of the digestive and other epithelia.
This is a proposal for an NCSU Individual Faculty Research and Professional Development award.
The development of complex anatomy and higher-order physiological function in organ systems is ultimately based on the properties of their component cells, but the relationship between tissue architecture and large-scale form is not well understood. During the development of the vertebrate digestive system, the initially straight cylinder of the primitive gut tube adopts a complex three-dimensional configuration generated by stereotypical elongation, looping and rotation events. The accessible, transparent embryos of the aquatic vertebrates, Xenopus laevis (frog) and Danio rerio (zebrafish), were employed to initiate a late-stage ?chemical genetic? investigation of the cellular and molecular mechanisms underlying these large-scale morphogenetic events. The preliminary results of this investigation show that late stage modulation of retinoic acid (RA) signaling induces dramatic dose-dependent, topological transformations in the normal looping, elongation and rotation of the gut tube, accompanied by alterations in gut epithelial architecture and central lumen formation that are indicative of abnormal cell adhesion, polarity and/or rearrangements. The main objective of this proposal is to determine the cellular and molecular mechanisms by which RA signaling shapes the morphogenesis of the gut tube. The first specific aim is to determine the requirement for endogenous RA signaling during gut morphogenesis. The expression patterns of RA metabolizing and signaling components will be defined in both Xenopus and zebrafish gut tubes, and the functional roles of these components in gut morphogenesis elucidated by loss of function strategies in both species. To complement this approach, existing zebrafish embryos with mutations in RA metabolizing enzymes will be evaluated for potential gut phenotypes. The second aim of this proposal is to identify the roles of specific cell properties and behaviors in RA-mediated gut morphogenesis. RA signaling will be disrupted in Xenopus and zebrafish embryos by chemical or genetic means, and the alterations in cell adhesion, polarity and intercellular rearrangement that underlie the resultant anomalous gut morphologies will be determined. The third specific aim of this proposal is to identify the molecular signaling components involved in RA-mediated gut morphogenesis. RA-treated and untreated embryos will be exposed to a library of chemical compounds that inhibit the cell signaling components known to control cell adhesion and polarity, in order to identify those compounds that mimic or rescue RA-induced defects in gut morphogenesis. Intellectual Merit: Completion of the proposed research will yield novel insight into the currently unknown relationships between molecular signaling, tissue architecture and three dimensional form during organogenesis. Knowledge of these relationships is essential for a complete understanding of the mechanisms of organ development on multiple scales and at all levels of biological organization. In addition, our comparative analysis of the morphogenetic events that shape digestive anatomy in two different species will yield insight into the evolution of anatomical spatial relationships within contiguous organ systems. Moreover, our results may advance tissue engineering strategies by providing a springboard for future qualitative and quantitative investigations of the cellular and molecular parameters underlying the construction of distinct tissue topographies. Finally, our chemical genetic results will provide a novel inroad for ascertaining the effect of environmental toxins or semiochemical cues on the plasticity and evolution of gut morphology in the vertebrate lineage. Broader Impact: The PI has an 8-year history of involving undergraduates in laboratory research and has published with 7 undergraduate co-authors. Three recent undergraduates in the PI?s lab generated much of the preliminary data for this proposal. The PI is presently continuing this trend with students in the Biotechnology and Undergraduate Honors Research programs at NCSU, and will involv
Project Summary/Abstract and Relevance This application is submitted for the purchase of a microscope that offers a unique opportunity to capture detailed images of macro as well as microscopic subjects. The Nikon AZ100 Macro/Micro Zoom Microscope was introduced to the U.S. market earlier this year and offers the advantages of a stereoscope (e.g., wide field of view with a long working distance) combined with the advantages of a compound microscope featuring high-resolution images. The AZ100 images specimens using white light as well as UV illumination over a broad range of magnifications (5-400x) using a novel vertical optical system that eliminates distortion introduced by stereoscopic inclination. This novel optical configuration facilitates the continuous switching of magnified images of a single specimen from macro to micro ranges. As a consequence of the elimination of inclination distortion, the AZ100 also facilitates real time deconvolution of macro subjects resulting in unique representations of three-dimensional subjects. We propose to purchase the AZ100 and install this device in a facility proximal to seven laboratories that will be major users. The research conducted within each of these laboratories requires imaging of macro (e.g., developing zebrafish, frog, mouse, and pig embryos, fetal mouse hearts, canine lenses) as well as micro (e.g., tissue sections) subjects using white light and UV illumination. This equipment will also be available to other interested laboratories at North Carolina State University and the broader Research Triangle research community as required. The long term objective and specific aims of this proposal are to utilize this versatile microscope to capture detailed images of a variety of macro and microscopic subjects for research and teaching purposes. We anticipate that these images will greatly enhance our capacity to analyze and document our experimental results as well as communicate with students and colleagues. The relevance of this equipment proposal is the each of the proposed major users is currently using a bevy of animal models to reveal mechanisms underlying normal human development, human developmental defects, human cancers, and virus-induced human diseases.