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Principal Investigator: Jenna Bergmann
Organization: YALE UNIVERSITY
Fiscal Year: 2024
Award: $33,958
Funding agency: Eunice Kennedy Shriver National Institute of Child Health and Human Development
Despite the high prevalence of pregnancy loss during the early post-implantation period, a detailed
understanding of cell and molecular workings at this stage of development remains elusive. During this period,
specification of the principal lineages of the future body occurs at the gastrulation stage, an evolutionary
conserved landmark event in life. While early embryonic cells are known to be sensitive to the changes in the
metabolite availability in their immediate surroundings, extremely little is known about the role of the intrauterine
metabolic environment during gastrulation and how it shapes embryo viability. The maternal metabolic
environment can be disrupted via somatic mutations in metabolic enzymes, such as the gain-of-function mutation
IDH2R140Q. This mutation leads to the conversion of the tricarboxylic acid cycle metabolite alpha-ketoglutarate
(αKG) into the epigenetically active metabolite 2-hydroxyglutarate (2-HG), which subsequently accumulates in
the bloodstream of affected patients and has downstream metabolic effects. In my preliminary work, I have
modeled maternal metabolic dysfunction by inducing this mutation in adult females, and the resulting embryos
demonstrate significant developmental delays at the time of gastrulation and failure to form distinct primary germ
layers. My work also revealed increased histone methylation, as well as differential expression of genes involved
in key developmental processes, such as cellular migration, as a response to 2-HG exposure in 2D cell culture.
Collectively, these findings suggest that maternal metabolic dysfunction driven by mutant IDH is prohibitive to
proper gastrulation. In light of these findings, I hypothesize that maternal 2-HG accumulation disrupts primary
germ layer formation via both bioenergetic and epigenetic mechanisms. My first aim is to characterize the
impact of IDH2R140Q-driven maternal metabolic dysfunction on primary germ layer formation. I will
characterize the morphological effects of maternal 2-HG accumulation using high-resolution 3D confocal
microscopy to investigate the spatiotemporal dynamics of germ layer cell specification and expansion. I further
will characterize the changes in mitochondrial activity and cell death caused by maternal 2-HG accumulation
using fluorescence-based assays. My second aim is to evaluate changes in the embryonic epigenetic
landscape caused by maternal 2-HG accumulation. I will identify variable histone modifications in exposed
embryos and the associated genomic loci using histone modification profiling followed by Cleavage Under
Targets and Tagmentation (CUT&Tag) in the embryonic portion of gastrulas. Together, this project will pave the
way toward a mechanistic and functional understanding of how maternal metabolic dysfunction modulates
embryonic development as well as adverse pregnancy outcomes. Thus, in addition to providing me with valuable
training that will further my career as a developmental biologist and reproductive medicine specialist, the
proposed research has significant potential to provide a rich source of new molecular and cellular targets for
therapeutic intervention in clinical settings where embryonic development is compromised.
Terms: <2-ketoglutarate><2-oxoglutarate><3-D><3-Dimensional><3D><Adult females><Adult women><Affect><Age><Assay><Bioassay><Bioenergetics><Biological Assay><Blood Circulation><Bloodstream><Body Tissues><Cancers><Cardiac Abnormalities><Causality><Cause of Death><Cell Body><Cell Culture Techniques><Cell Death><Cell Differentiation><Cell Differentiation process><Cell Locomotion><Cell Migration><Cell Movement><Cells><Cellular Migration><Cellular Motility><Child Health><Citric Acid Cycle><Clinical><Conceptions><Confocal Microscopy><Data><Decision Making><Defect><Derivation><Derivation procedure><Development><Developmental Biology><Developmental Delay><Developmental Delay Disorders><Developmental Process><Differential Gene Expression><Ectoderm><Embryo><Embryo Development><Embryogenesis><Embryonic><Embryonic Development><Endoderm><Environment><Enzyme Gene><Enzymes><Epigenetic><Epigenetic Change><Epigenetic Mechanism><Epigenetic Process><Etiology><Event><Exposure to><Failure><Females in adulthood><Fluorescence><Fluorescent Probes><Future><Gastrula><Gene Targeting><Genetic><Genetic Alteration><Genetic Change><Genetic Models><Genetic defect><Germ Layers><Goals><Heart Abnormalities><Hereditary Metabolic Disorder><High Prevalence><Human><Hypermethylation><Immunofluorescence><Immunofluorescence Immunologic><Inborn Errors of Metabolism><Induced DNA Alteration><Induced Mutation><Induced Sequence Alteration><Isocitrate Dehydrogenase><Krebs Cycle><Life><Light><Malignant Cell><Malignant Neoplasms><Malignant Tumor><Mediating><Mesoderm><Metabolic><Metabolic Diseases><Metabolic Disorder><Metabolic dysfunction><Methylation><Mice><Mice Mammals><Mitochondria><Modeling><Modern Man><Modification><Molecular><Molecular Target><Morphology><Murine><Mus><Mutation><Neurologic><Neurological><Oncogenesis><Organogenesis><Patients><Pattern><Photoradiation><Physicians><Pregnancy loss><Progenitor Cells><Reproductive Medicine><Research><Resolution><Role><Scientist><Shapes><Site-Directed Mutagenesis><Site-Specific Mutagenesis><Somatic Mutation><Source><Specialist><Specific Child Development Disorders><Specific qualifier value><Specified><TCA cycle><Targeted DNA Modification><Targeted Modification><Testing><Therapeutic Intervention><Thesaurismosis><Time><Tissue-Specific Differential Gene Expression><Tissue-Specific Gene Expression><Tissues><Training><Tricarboxylic Acid Cycle><Woman><Women in adulthood><Work><adverse pregnancy outcome><ages><alpha ketoglutarate><alpha-oxoglutarate><cancer cell><career><causation><cell culture><cell cultures><cell fate specification><cell motility><cellular differentiation><cellular targeting><cohort><critical period><cytotoxic><developmental><disease causation><disease causing variant><disease-causing mutation><embryo cell><epigenetically><gain of function mutation><gastrulation><gene locus><genetic locus><genome mutation><genomic location><genomic locus><histone methylation><histone modification><implantation><in vivo><inborn metabolism disorder><insight><intervention therapy><malignancy><metabolism disorder><mitochondrial><mouse development><mutant><necrocytosis><neoplasm/cancer><new approaches><novel approaches><novel strategies><novel strategy><pathogenic variant><pre-implantation><preimplantation><prevent><preventing><promoter><promotor><reproductive><reproductive fitness><reproductive success><resolutions><response><social role><somatic variant><spatiotemporal><stem cells><three dimensional><tool><tumorigenesis><α-ketoglutarate><α-oxoglutarate><αKG>