Using a rat model of type 2 diabetes (T2D), we showed that at 2 months of T2D the decrease in bone marrow progenitor cell (BMPC) release from diabetic bone marrow (BM) is caused by BM neuropathy and that these changes precede the development of diabetic retinopathy (DR). BM neuropathy was associated with a marked reduction in clock gene expression in the BMPCs themselves which led to diminished repair by these cells and by 4 months of T2D resulted in the hallmark feature of diabetic retinopathy (DR), acellular retinal capillaries. Diabetic BMPC dysfunction was corrected by increasing levels of bioavailable nitric oxide (NO) towards normal non-diabetic levels. Since NO modulates clock gene expression, the reduced levels of bioavailable NO results in altered clock gene expression. Plaminogen activator inhibitor (PAI-1), an important modulator of hematopoetic stem cell maturation and release from the BM is an immediate downstream metabolic regulator controlled by clock genes. PAI-1 demonstrates a robust circadian pattern in health, but this pattern is altered in diabetes and elevated levels are produced by endothelial cells (EC) from diabetics. We believe this NO-modulated clock gene dysfunction leads to the loss of circadian regulation of PAI-1 and represents an underlying mechanism and therapeutic target for DR and atherosclerosis, which are the major causes of blindness and mortality in diabetics, respectively. In this application, we propose the following hypothesis: In diabetes, a reduction in bioavailable NO in ECs and BMPCs causes a diminished amplitude and frequency of the oscillatory pattern of the clock proteins, BMAL1 and PER-2, leading to a loss of circadian regulation of PAI-1 and subsequent further diminution of NO levels in EC and BMPCs. Decreasing NO bioavailability and persistent increase in PAI-1 in the vasa nervorum leads to development of BM neuropathy which results in defective EPC mobilization further exacerbating end organ complications. To test our hypothesis, we propose the following aims: 1) to determine whether NO bioavailability can affect the molecular clock and circadian rhythms in vascular ECs and BMPCs and to determine whether levels of NO will have direct and indirect effects on clock gene expression via S-nitrosylation of essential clock proteins such as BMAL1 and PER-2;2) to determine whether specific clock gene knock-out of the endothelium and BMPC recapitulates the diabetic vascular phenotype of accelerated injury and reduced repair;and 3) to determine whether dysregulation of NO and PAI-1 within the vasa nervorum leads to BM neuropathy and loss of circadian release of BMPCs into the circulation.

Public Health Relevance

This proposal represents a paradigm shift in our understanding of the pathogenesis of diabetic micro- and macro-vascular complications. We are testing whether circadian oscillators play a pivotal role in endothelial cell and bone marrow progenitor cells homeostasis. We believe that when disrupted in diabetes, they must be restored before vascular health is achieved. We specifically identify plasminogen activator inhibitor as a novel therapeutic target in diabetes to restore vascular function and circadian oscillation of nitric oxide in vascular and bone marrow cells.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
1R01DK090730-01
Application #
8026768
Study Section
Biology and Diseases of the Posterior Eye Study Section (BDPE)
Program Officer
Jones, Teresa L Z
Project Start
2010-09-30
Project End
2014-08-31
Budget Start
2010-09-30
Budget End
2011-08-31
Support Year
1
Fiscal Year
2010
Total Cost
$388,402
Indirect Cost
Name
University of Florida
Department
Pharmacology
Type
Schools of Medicine
DUNS #
969663814
City
Gainesville
State
FL
Country
United States
Zip Code
32611
Shaw, Lynn Calvin; Li Calzi, Sergio; Li, Nan et al. (2018) Enteral Arg-Gln Dipeptide Administration Increases Retinal Docosahexaenoic Acid and Neuroprotectin D1 in a Murine Model of Retinopathy of Prematurity. Invest Ophthalmol Vis Sci 59:858-869
Bhatwadekar, Ashay D; Beli, Eleni; Diao, Yanpeng et al. (2017) Conditional Deletion of Bmal1 Accentuates Microvascular and Macrovascular Injury. Am J Pathol 187:1426-1435
Bhatwadekar, Ashay D; Duan, Yaqian; Korah, Maria et al. (2017) Hematopoietic stem/progenitor involvement in retinal microvascular repair during diabetes: Implications for bone marrow rejuvenation. Vision Res 139:211-220
Basavarajappa, Halesha D; Sulaiman, Rania S; Qi, Xiaoping et al. (2017) Ferrochelatase is a therapeutic target for ocular neovascularization. EMBO Mol Med 9:786-801
Salazar, Tatiana E; Richardson, Matthew R; Beli, Eleni et al. (2017) Electroacupuncture Promotes Central Nervous System-Dependent Release of Mesenchymal Stem Cells. Stem Cells 35:1303-1315
Hu, Ping; Hunt, Nicholas H; Arfuso, Frank et al. (2017) Increased Indoleamine 2,3-Dioxygenase and Quinolinic Acid Expression in Microglia and Müller Cells of Diabetic Human and Rodent Retina. Invest Ophthalmol Vis Sci 58:5043-5055
Beli, Eleni; Dominguez 2nd, James M; Hu, Ping et al. (2016) CX3CR1 deficiency accelerates the development of retinopathy in a rodent model of type 1 diabetes. J Mol Med (Berl) 94:1255-1265
Chakravarthy, Harshini; Navitskaya, Svetlana; O'Reilly, Sandra et al. (2016) Role of Acid Sphingomyelinase in Shifting the Balance Between Proinflammatory and Reparative Bone Marrow Cells in Diabetic Retinopathy. Stem Cells 34:972-83
Dominguez 2nd, James M; Hu, Ping; Caballero, Sergio et al. (2016) Adeno-Associated Virus Overexpression of Angiotensin-Converting Enzyme-2 Reverses Diabetic Retinopathy in Type 1 Diabetes in Mice. Am J Pathol 186:1688-700
Wert, Katherine J; Mahajan, Vinit B; Zhang, Lijuan et al. (2016) Neuroretinal hypoxic signaling in a new preclinical murine model for proliferative diabetic retinopathy. Signal Transduct Target Ther 1:

Showing the most recent 10 out of 75 publications