Document text
Principal Investigator: Katie Whytock
Organization: ADVENTHEALTH ORLANDO
Fiscal Year: 2024
Award: $91,500
Funding agency: National Institute of Diabetes and Digestive and Kidney Diseases
PROJECT SUMMARY
Type 2 Diabetes (T2D) is estimated to affect 30 million U.S. adults and skeletal muscle (SkM) insulin resistance
(IR) is considered to be one of the primary aberrations. Effective insulin-stimulated SkM glucose uptake is
dependent on 1) insulin-stimulated activation of terminal arteriole endothelial cells (EC) to perfuse microvascular
units, 2) transport of substrates across capillary ECs and 3) stimulation of myofiber insulin signaling, resulting in
glucose uptake. Therefore, both myofibers and ECs are critical cell types that can be dysregulated with IR in
T2D. Previous research has identified a dysregulated transcriptional profile with SkM IR in the basal state and
during insulin stimulation from a hyperinsulinemic euglycemic clamp (HE). A significant limitation of these prior
studies is the restriction to whole SkM homogenates and therefore not identifying which cells or spatial area the
dysregulated signals originate from. We have developed a pipeline for 3’ and 5’ amplification of single cells or
nuclei from SkM using full-length SMART-Seq technology with the iCELL8 platform, resulting in 2-3 fold greater
gene coverage than previous research, allowing us to investigate transcriptional aberrations at a single cell/nuclei
resolution. Recent developments in spatial transcriptomics now permit transcriptional profiling on sectioned
tissue. In this proposed research, we will leverage both full-length single cell and single nuclei RNA-Seq and
spatial transcriptomics to determine cell- and spatially-distinct transcriptional aberrations in SkM that occur with
IR in T2D. The career development training will refine my skills in; leading an independent lab, bioinformatics,
and human clinical research with a focus on assessing insulin-stimulated glucose disposal (Rd) and non-
oxidative glucose disposal (NOGD) using hyperinsulinemic euglycemic (HE) clamps with glucose tracers and
indirect calorimetry. My K99/R00 training will uniquely position me to conduct bedside-to-bench-to-bioinformatics
research to uncover the molecular profiles of SkM IR. For the first time, the transcriptional responses to insulin
in human SkM will be probed at a single cell/nuclei and spatial resolution. We will identify which cells and areas
of SkM respond to insulin and if they are spatially localized to each other and if their gene-network profiles
correlate with a greater in vivo Rd and NOGD assessed simultaneously with a HE clamp. This novel and
innovative approach will reveal which cells (and importantly, where in SkM) the aberrations occur with IR in T2D.
The Translational Research Institute at AdventHealth is an ideal environment for developing my bedside-to-
bench-to-bioinformatics research approach with cutting-edge metabolic translational research facilities and a rich
interdisciplinary environment. Completion of the proposed research and career development training will facilitate
my success as an independent translational investigator in SkM metabolism and IR. Furthermore, experiments
in this proposal will identify specific molecular targets to alleviate SkM IR that will be targeted in future grant
applications.
Terms: <21+ years old><Adipose tissue><Adult><Adult Human><Adult-Onset Diabetes Mellitus><Affect><Age><Applications Grants><Archives><Area><BMI><BMI percentile><BMI z-score><Bio-Informatics><Bioinformatics><Biopsy><Blood Vessels><Body Tissues><Body mass index><Business-Friendly Atmosphere><Capillary Endothelial Cell><Cell Body><Cell Communication and Signaling><Cell Nucleus><Cell Signaling><Cells><Clampings><Clinical Research><Clinical Study><Closure by clamp><Collaborations><Complex><D-Glucose><Data Set><Development><Development and Research><Dextrose><Dissociation><Doctor of Philosophy><Endothelial Cells><Environment><Euglycemic Clamping><Expression Signature><Fats><Fatty Tissue><Fatty acid glycerol esters><Fibroblasts><Fingerprint><Foundations><Freezing><Future><Gene Expression><Gene Expression Monitoring><Gene Expression Pattern Analysis><Gene Expression Profile><Gene Expression Profiling><Gene Transcription><Genes><Genetic Transcription><Glucose><Glucose Clamp><Grant Proposals><Human><Humulin R><Hyperinsulinemia><Hyperinsulinism><Immune><Immunes><Impairment><Indirect Calorimetry><Individual><Insulin><Insulin Resistance><Intracellular Communication and Signaling><Investigators><Ketosis-Resistant Diabetes Mellitus><Leadership><Leanness><Learning><Leiomyocyte><Length><Maturity-Onset Diabetes Mellitus><Measures><Mentors><Metabolic><Modern Man><Molecular><Molecular Fingerprinting><Molecular Profiling><Molecular Target><Mononuclear><Muscle><Muscle Fibers><Muscle Tissue><Myotubes><NIDDM><Non-Insulin Dependent Diabetes><Non-Insulin-Dependent Diabetes Mellitus><Noninsulin Dependent Diabetes><Noninsulin Dependent Diabetes Mellitus><Novolin R><Nucleus><Obesity><Organ><Participant><Perfusion><Peripheral><Ph.D.><PhD><Phase><Population><Position><Positioning Attribute><Prevention><Protocol><Protocols documentation><Quetelet index><R & D><R&D><RNA Expression><Regular Insulin><Research><Research Institute><Research Personnel><Researchers><Resolution><Respiration Calorimetry><Rhabdomyocyte><Role><Sampling><Signal Transduction><Signal Transduction Systems><Signaling><Single-Nucleus Sequencing><Skeletal Fiber><Skeletal Muscle><Skeletal Muscle Cell><Skeletal Muscle Fiber><Skeletal Myocytes><Slow-Onset Diabetes Mellitus><Smooth Muscle Cells><Smooth Muscle Myocytes><Smooth Muscle Tissue Cell><Stable Diabetes Mellitus><T2 DM><T2D><T2DM><Techniques><Technology><Therapeutic Intervention><Thinness><Time><Tissues><Tracer><Training><Transcript Expression Analyses><Transcript Expression Analysis><Transcription><Translational Research><Translational Science><Type 2 Diabetes Mellitus><Type 2 diabetes><Type II Diabetes Mellitus><Type II diabetes><Voluntary Muscle><adipose><adiposity><adult onset diabetes><adulthood><ages><analyze gene expression><arteriole><bench bed side><bench bedside><bench to bed side><bench to bedside><bench to clinic><bench to clinical practice><bench-to-bedside translation><bio-informatics pipeline><bioinformatics pipeline><biological signal transduction><business-friendly environment><career><career development><cell type><collaborative atmosphere><collaborative environment><complex data><corpulence><design><designing><developmental><experiment><experimental research><experimental study><experiments><feeding><gene expression analysis><gene expression assay><gene expression pattern><gene expression signature><gene network><global gene expression><global transcription profile><glucose disposal><glucose metabolism><glucose uptake><improved><in vivo><innovate><innovation><innovative><insulin resistant><insulin sensitivity><insulin signaling><insulin stimulated glucose disposal><insulin tolerance><interactive atmosphere><interactive environment><interdisciplinary atmosphere><interdisciplinary environment><intervention therapy><ketosis resistant diabetes><maturity onset diabetes><method development><molecular profile><molecular signature><muscular><novel><peer-group atmosphere><peer-group environment><research and development><research facility><resolutions><response><sNuc-Seq><scRNA-seq><sex><single cell RNA-seq><single cell RNAseq><single cell expression profiling><single cell transcriptomic profiling><single nucleus RNA-sequencing><single nucleus seq><single-cell RNA sequencing><single-nucleus RNA-seq><skeletal muscle metabolism><skeletal muscle protein metabolism><skills><snRNA sequencing><snRNA-seq><social role><success><transcriptional profile><transcriptional profiling><transcriptional signature><transcriptome><transcriptomics><translation research><translational investigation><translational investigator><translational researcher><translational scientist><type 2 DM><type II DM><type two diabetes><vascular><white adipose tissue><yellow adipose tissue>