Micro-RNA 132 and 212 mediated regulation of fatty acid metabolism and its effect on insulin secretion. Prelim Mock Talk by -Mufaddal S Soni Attie Lab.

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Presentation transcript:

Micro-RNA 132 and 212 mediated regulation of fatty acid metabolism and its effect on insulin secretion. Prelim Mock Talk by -Mufaddal S Soni Attie Lab

Outline Introduction Preliminary Data Specific Aims Conclusion Diabetes model Micro-RNAs Preliminary Data Specific Aims Conclusion The Economist, 12/13/03

Obesity-diabetes dichotomy Lean Lepob/ob B6 BTBR Obese Diabetic Age (weeks) 3

What are micro-RNAs? Chang, Z. C. 2005

Micro-RNA 132 and 212 enhances insulin secretion.

CACT (Slc25a20) : Most down regulated target of miRNA 132 and 212

CACT mediates translocation of FA-Carn into the mitochondria for β-oxidation.

CACT mediates translocation of FA-Carn into the mitochondria for β-oxidation.

CACT mediates translocation of FA-Carn into the mitochondria for β-oxidation.

Aim 1: Determine the molecular fate of LC-Carn when stimulating insulin secretion. Experiment 1: Decrease CPT-1 activity and determine if LC-Carn stimulates insulin secretion. Experiment 2: Determine if siRNA-mediated knockdown of CACT leads to reduced fatty acid β-oxidation. Experiment 3: Determine the molecular fate of exogenous FA-Carn under conditions when CACT activity is diminished. Experiment 4: Determine if a non-metabolized analog of fatty acyl-carnitine is capable of stimulating insulin secretion.

Aim 1: Determine the molecular fate of LC-Carn when stimulating insulin secretion. Experiment 1: Decrease CPT-1 activity and determine if LC-Carn stimulates insulin secretion. Experiment 2: Determine if siRNA-mediated knockdown of CACT leads to reduced fatty acid β-oxidation. Experiment 3: Determine the molecular fate of exogenous FA-Carn under conditions when CACT activity is diminished. Experiment 4: Determine if a non-metabolized analog of fatty acyl-carnitine is capable of stimulating insulin secretion.

Aim 1: Determine the molecular fate of LC-Carn when stimulating insulin secretion. Experiment 1: Decrease CPT-1 activity and determine if LC-Carn stimulates insulin secretion. Experiment 2: Determine if siRNA-mediated knockdown of CACT leads to reduced fatty acid β-oxidation. Experiment 3: Determine the molecular fate of exogenous FA-Carn under conditions when CACT activity is diminished. Experiment 4: Determine if a non-metabolized analog of fatty acyl-carnitine is capable of stimulating insulin secretion.

Aim 1: Determine the molecular fate of LC-Carn when stimulating insulin secretion. Experiment 1: Decrease CPT-1 activity and determine if LC-Carn stimulates insulin secretion. Experiment 2: Determine if siRNA-mediated knockdown of CACT leads to reduced fatty acid β-oxidation. Experiment 3: Determine the molecular fate of exogenous FA-Carn under conditions when CACT activity is diminished. Experiment 4: Determine if a non-metabolized analog of fatty acyl-carnitine is capable of stimulating insulin secretion.

Aim 1: Determine the molecular fate of LC-Carn when stimulating insulin secretion. Experiment 1: Decrease CPT-1 activity and determine if LC-Carn stimulates insulin secretion. Experiment 2: Determine if siRNA-mediated knockdown of CACT leads to reduced fatty acid β-oxidation. Experiment 3: Determine the molecular fate of exogenous FA-Carn under conditions when CACT activity is diminished. Experiment 4: Determine if a non-metabolized analog of fatty acyl-carnitine is capable of stimulating insulin secretion.

Aim 1: Determine the molecular fate of LC-Carn when stimulating insulin secretion. Experiment 1: Decrease CPT-1 activity and determine if LC-Carn stimulates insulin secretion. Experiment 2: Determine if siRNA-mediated knockdown of CACT leads to reduced fatty acid β-oxidation. Experiment 3: Determine the molecular fate of exogenous FA-Carn under conditions when CACT activity is diminished. Experiment 4: Determine if a non-metabolized analog of fatty acyl-carnitine is capable of stimulating insulin secretion.

Aim 1: Determine the molecular fate of LC-Carn when stimulating insulin secretion. Experiment 1: Decrease CPT-1 activity and determine if LC-Carn stimulates insulin secretion. Experiment 2: Determine if siRNA-mediated knockdown of CACT leads to reduced fatty acid β-oxidation. Experiment 3: Determine the molecular fate of exogenous FA-Carn under conditions when CACT activity is diminished. Experiment 4: Determine if a non-metabolized analog of fatty acyl-carnitine is capable of stimulating insulin secretion.

Aim 2: Identify the underlying signaling pathway for how LC-Carn enhances insulin secretion. Experiment 1: Determine if LC-Carns enhance insulin secretion through PKC activation. Experiment 2: Study the role of cAMP and PKA in LC-Carn effects on insulin secretion. Experiment 3: Determine if the insulin signaling pathway is involved in LC-Carn mediated insulin secretion. FFA

Aim 2: Identify the underlying signaling pathway for how LC-Carn enhances insulin secretion. Experiment 1: Determine if LC-Carns enhance insulin secretion through PKC activation. Experiment 2: Study the role of cAMP and PKA in LC-Carn effects on insulin secretion. Experiment 3: Determine if the insulin signaling pathway is involved in LC-Carn mediated insulin secretion. Ro 31-8220 PMA UNC-01 siRNA

Aim 2: Identify the underlying signaling pathway for how LC-Carn enhances insulin secretion. Experiment 1: Determine if LC-Carns enhance insulin secretion through PKC activation. Experiment 2: Study the role of cAMP and PKA in LC-Carn effects on insulin secretion. Experiment 3: Determine if the insulin signaling pathway is involved in LC-Carn mediated insulin secretion.

Aim 2: Identify the underlying signaling pathway for how LC-Carn enhances insulin secretion. Experiment 1: Determine if LC-Carns enhance insulin secretion through PKC activation. Experiment 2: Study the role of cAMP and PKA in LC-Carn effects on insulin secretion. Experiment 3: Determine if the insulin signaling pathway is involved in LC-Carn mediated insulin secretion. Measure increase in cAMP levels Probe PKA and Epac pathways

Aim 2: Identify the underlying signaling pathway for how LC-Carn enhances insulin secretion. Experiment 1: Determine if LC-Carns enhance insulin secretion through PKC activation. Experiment 2: Study the role of cAMP and PKA in LC-Carn effects on insulin secretion. Experiment 3: Determine if the insulin signaling pathway is involved in LC-Carn mediated insulin secretion.

Aim 2: Identify the underlying signaling pathway for how LC-Carn enhances insulin secretion. Experiment 1: Determine if LC-Carns enhance insulin secretion through PKC activation. Experiment 2: Study the role of cAMP and PKA in LC-Carn effects on insulin secretion. Experiment 3: Determine if the insulin signaling pathway is involved in LC-Carn mediated insulin secretion. Wortmannin PI3K inhibitor

Aim 3: Elucidate the role of Carnitine Acetyl Transferase (CrAT) in regulating insulin secretion. Prof. Randall Mynatt, LSU.

Aim 3: Elucidate the role of Carnitine Acetyl Transferase (CrAT) in regulating insulin secretion. Prof. Randall Mynatt, LSU.

Amplification Pathway Aim 3: Elucidate the role of Carnitine Acetyl Transferase (CrAT) in regulating insulin secretion. Experiment 2: Determine the phase of insulin secretion altered in CrAT mice. Amplification Pathway (2nd Phase) Triggering Pathway (1st Phase)

Experiment 3: Metabolic profiling of the βCrAT mouse islets. Aim 3: Elucidate the role of Carnitine Acetyl Transferase (CrAT) in regulating insulin secretion. Experiment 3: Metabolic profiling of the βCrAT mouse islets. Fat animals have elevated carnitine pools in the muscle. I will profile the carnitine metabolites in the βCrAT mouse islets to determine the correlation between insulin secretion and carnitine levels. Koves, R. T. et.al. 2007

Thank You Summary

biotin–HPDP (Biotin-HPDP-N-[6-(Biotinamido)hexyl]-3′-(2′-pyridyldithio)propionamide