Hyperglycemia is very common in patients with sepsis even if there is no history of diabetes. Insulin resistance of skeletal muscle glucose uptake (MGU) is a major cause of this hyperglycemia. Control of MGU is distributed between delivery of glucose to muscle, glucose transport into muscle, and glucose phosphorylation within muscle;insulin resistance is due to defects in one or more of them. The impact of obesity on MGU has been studied by a number of groups, but the impact of inflammation on the distribution of control of MGU is unknown. The experiments described in this proposal will examine the extent to which inflammation induced by lipopolysaccharide (LPS) redistributes the control of MGU. In this proposal the roles transport and phosphorylation play in controlling MGU will be assessed by using germline manipulation (partial knockout or over expression) of transport and phosphorylation capacity (e.g. hexokinase) to modulate a single step or multiple steps and measure the impact on MGU. We hypothesize that defects in glucose phosphorylation capacity play a central role in the inflammation induced insulin resistance. Experiments will be performed in chronically catheterized, conscious mice. This approach allows for comprehensive metabolic assessment of MGU in vivo in the absence of stress. The experimental strategy is to perturb proteins or processes involved in control of MGU and measure the effect of the perturbation on glucose influx. Whole body glucose uptake and MGU will be measured using [3-3H] glucose and [14C] 2- deoxyglucose, respectively, in combination with methods for sampling blood and tissues and measuring muscle blood flow. The relationship of MGU to long chain fatty acid (LCFA) uptake will simultaneously be measured using a radiolabeled fatty acid analog. Muscle ATP flux will be assessed using 31NMR spectroscopy. Tissues will be analyzed for glycogen synthesis, insulin signaling, oxidative stress and GLUT4 translocation.
Our specific aims are to determine: 1. The impact of LPS on the relative control transport and glucose phosphorylation have in determining MGU 2. If LPS amplifies the impact NEFA and glucose availability have in modulating MGU 3. If modulating oxidative stress (NO availability and NF-:B activation) following LPS will improve MGU by augmenting glucose phosphorylation and mitochondrial ATP flux Our long term goal is to identify the steps controlling MGU that are impacted by inflammation and assess which of those steps are more responsive to changes in oxidative stress. Future therapies can then have a more targeted approach in correcting MGU during an inflammatory stress such as sepsis.

Public Health Relevance

Hyperglycemia is very common in hospitalized patients and clinical trials suggest if the hyperglycemia can be minimized morbidity and mortality are improved. A major cause of the hyperglycemia is insulin resistance of skeletal muscle glucose uptake This proposal will determine where the defect is and address the questions is it due simply to a failure of insulin to activate it signaling pathway or are underlying defects in the mitochondria caused by the inflammation aggravating and limiting the ability of insulin to exert is beneficial effects.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK078188-05
Application #
8485594
Study Section
Integrative Physiology of Obesity and Diabetes Study Section (IPOD)
Program Officer
Laughlin, Maren R
Project Start
2009-06-01
Project End
2014-05-31
Budget Start
2013-06-01
Budget End
2014-05-31
Support Year
5
Fiscal Year
2013
Total Cost
$321,292
Indirect Cost
$113,044
Name
Vanderbilt University Medical Center
Department
Physiology
Type
Schools of Medicine
DUNS #
004413456
City
Nashville
State
TN
Country
United States
Zip Code
37212
Otero, Yolanda F; Mulligan, Kimberly X; Barnes, Tammy M et al. (2016) Enhanced Glucose Transport, but not Phosphorylation Capacity, Ameliorates Lipopolysaccharide-Induced Impairments in Insulin-Stimulated Muscle Glucose Uptake. Shock 45:677-85
Dooley, James; Garcia-Perez, Josselyn E; Sreenivasan, Jayasree et al. (2016) The microRNA-29 Family Dictates the Balance Between Homeostatic and Pathological Glucose Handling in Diabetes and Obesity. Diabetes 65:53-61
Boortz, Kayla A; Syring, Kristen E; Dai, Chunhua et al. (2016) G6PC2 Modulates Fasting Blood Glucose In Male Mice in Response to Stress. Endocrinology 157:3002-8
Boortz, Kayla A; Syring, Kristen E; Lee, Rebecca A et al. (2016) G6PC2 Modulates the Effects of Dexamethasone on Fasting Blood Glucose and Glucose Tolerance. Endocrinology 157:4133-4145
Syring, Kristen E; Boortz, Kayla A; Oeser, James K et al. (2016) Combined Deletion of Slc30a7 and Slc30a8 Unmasks a Critical Role for ZnT8 in Glucose-Stimulated Insulin Secretion. Endocrinology 157:4534-4541
House 2nd, Lawrence M; Morris, Robert T; Barnes, Tammy M et al. (2015) Tissue inflammation and nitric oxide-mediated alterations in cardiovascular function are major determinants of endotoxin-induced insulin resistance. Cardiovasc Diabetol 14:56
Cyphert, Travis J; Morris, Robert T; House, Lawrence M et al. (2015) NF-κB-dependent airway inflammation triggers systemic insulin resistance. Am J Physiol Regul Integr Comp Physiol 309:R1144-52
Chen, Sheng-Song; Otero, Yolanda F; Mulligan, Kimberly X et al. (2014) Liver, but not muscle, has an entrainable metabolic memory. PLoS One 9:e86164
Barnes, Tammy M; Otero, Yolanda F; Elliott, Amicia D et al. (2014) Interleukin-6 amplifies glucagon secretion: coordinated control via the brain and pancreas. Am J Physiol Endocrinol Metab 307:E896-905
Otero, Yolanda F; Stafford, John M; McGuinness, Owen P (2014) Pathway-selective insulin resistance and metabolic disease: the importance of nutrient flux. J Biol Chem 289:20462-9

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