Congenital malformations occur in up to 10% of babies born to diabetic women. Optimal glycemiccontrol is difficult to achieve and maintain, and even transient exposure to hyperglycemia can causemalformations. This project is formulated on the basis of our strong preliminary data. We have found 1)hyperglycemia impairs autophagy and increases the accumulation of defective mitochondria,dysfunctional proteins and swollen endoplasmic reticulum (ER) in the developing neuroepithelium; 2) thenon-toxic autophagy activator, trehalose, reverses hyperglycemia-induced autophagy impairment andneural tube defects (NTDs); 3) PKCa gene deletion, a p70S6K1 inhibitor, an ER chaperone (4-PBA) andoverexpression of sirtuin 2 (SIRT2) histone deacetylase in the neural tube, all reduce hyperglycemia-induced NTDs; 4) both p70S6K1 inhibitor and SIR2 overexpression restore levels of the autophagymarker, LC3-II. We test a novel hypothesis that maternal diabetes-induced autophagy impairmentcauses NTD formation by disrupting cellular homeostasis leading to ER stress and apoptosis, andthat maternal hyperglycemia activates p70S6K1 resulting in autophagy impairment. Restoration ofautophagy by trehalose prevents hyperglycemia-induced NTDs. In addition, reduced SIRT2 andSIRT6 expression mediates the effect of p70S6K1.
Aim 1 will determine whether trehalose preventshyperglycemia-induced NTDs by correcting autophagy impairment that causes ER stress andapoptosis. We hypothesize that maternal diabetes induces aberrant changes of Atg1 and Sqsmt1expression, which regulate autophagy, leading to autophagy impairment which induces apoptosis andNTDs, and reversal of autophagy impairment by trehalose, will restore cellular homeostasis and thusprevent diabetes-induced NTDs.
Aim 2 will investigate the activation mechanism and the role ofp70S6K1 in autophagy impairment and NTD formation in diabetic embryopathy. Our workinghypothesis is that PKCa activates p70S6K1 which causes autophagy impairment, an increase in DNA-methyltransferases (DNMTs) and a decrease in SIRT 2 and 6 gene expression leading to NTD formation.
Aim 3 will determine the underlying mechanism and the role of reduced sirt2 and sirt6 geneexpression in autophagy impairment that leads to diabetic embryopathy. We will test the hypothesisthat promoter hypermethylation and subsequent reduced transcription factor binding activities cause sirt2and sirt6 gene reduction which lead to autophagy impairment by modulating the expression of Atg1 andSqsmt1 via deacetylation of Foxo transcription factors. Our studies will provide mechanistic evidence forautophagy, ER stress, p70S6K1 and SIRT2/6 as targets for therapeutic interventions.

Public Health Relevance

Nearly two million American women at reproductive ages have diabetes and this number will be double by 2030. Maternal diabetes is a significant risk factor for structural birth defects. Diabetic women under modern preconceptional care are still three to four times more likely to have a child with birth defects than non-diabetic women. Diabetes-induced birth defects remain significant maternal-fetal health problems. In this proposal, we employ two complementary models, the type I and type II diabetic embryopathy, to delineate the mechanisms underlying diabetes-induced neural tube defects (NTDs). In a series of mechanistic studies, we will test the novel hypothesis that autophagy impairment is responsible for apoptosis leading to NTDs, and that activation of the PKCa-p70S6K1-SIRT2/6 pathway causes autophagy impairment in diabetic embryopathy. This project will provide a mechanistic basis for exploring the non-toxic autophagy activator, trehalose, p70S6K1 inhibitors, endoplasmic reticulum chemical chaperones and specific SIRT2 and SIRT6 activators as new prevention strategies against diabetes- induced NTDs.

National Institute of Health (NIH)
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
High Priority, Short Term Project Award (R56)
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Pregnancy and Neonatology Study Section (PN)
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Jones, Teresa L Z
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University of Maryland Baltimore
Obstetrics & Gynecology
Schools of Medicine
United States
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