Spatially Characterized Aortic Proteome Reveals Novel Regional Signatures and Glucocorticoid Receptor/Dipeptidase 1 Axis in Diabetic Vasculopathy

Abstract

Diabetes mellitus poses a major global health burden and is intricately linked to cardiovascular complications, yet the spatial molecular landscape of diabetic vasculopathy remains poorly defined. Since thoracic and abdominal aortas differ in embryological origin and hemodynamic microenvironments, we applied laser-capture microdissection to map their spatial proteomes in health and diabetes, using a multidimensional framework across longitudinal (region) and transverse (disease) axes. This approach uncovered region-specific protein and pathway signatures obscured by conventional bulk analyses, highlighting spatial heterogeneity in transcriptional regulators and flow-sensitive proteins. We identified dipeptidase 1 (DPEP1), a membrane-bound zinc metalloprotease, as selectively upregulated in the diabetic thoracic aorta and inducible by diabetic conditions and shear stress. Mechanistically, laminar shear stress promoted glucocorticoid receptor (GR) nuclear translocation to drive a GR/DPEP1 axis, potentially explaining region-specific DPEP1 induction and its synergy with diabetic conditions. Functionally, chronic Dpep1 inhibition by cilastatin and endothelium-specific Dpep1 knockdown attenuated neutrophilic vascular inflammation and rescued endothelial dysfunction in diabetic mice. Furthermore, the corticosteroid dexamethasone activated the shear stress-responsive GR/DPEP1 axis in vivo, yet exerted time-dependent vascular effects—acutely dampening neutrophilic inflammation, but chronically worsening hyperglycemia and aggravating vascular dysfunction. These findings reveal spatially defined biomarkers and highlight DPEP1 as a therapeutic target in diabetic vasculopathy.

Keywords

diabetes / endothelial function / inflammation / proteomics / shear stress / vasculopathy

References

[1] Over 250 Million People Worldwide Unaware They Have Diabetes, According to New Research From the International Diabetes Federation (IDF). Diabetes Research and Clinical Practice 223 (2025): 112176.

[2] Y. Li, Y. Liu, S. Liu, et al., “Diabetic Vascular Diseases: Molecular Mechanisms and Therapeutic Strategies,” Signal Transduction and Targeted Therapy 8, no. 1 (2023): 152.

[3] A. Stirban, T. Gawlowski, and M. Roden, “Vascular Effects of Advanced Glycation Endproducts: Clinical Effects and Molecular Mechanisms,” Molecular Metabolism 3, no. 2 (2014): 94–108.

[4] L. Trigueros-Motos, J. M. González-Granado, C. Cheung, et al., “Embryological-Origin–Dependent Differences in Homeobox Expression in Adult Aorta,” Arteriosclerosis, Thrombosis, and Vascular Biology 33, no. 6 (2013): 1248–1256.

[5] A. J. Boyd, D. C. S. Kuhn, R. J. Lozowy, and G. P. Kulbisky, “Low Wall Shear Stress Predominates at Sites of Abdominal Aortic Aneurysm Rupture,” Journal of Vascular Surgery 63, no. 6 (2016): 1613–1619.

[6] H. Deng, A. Eichmann, and M. A. Schwartz, “Fluid Shear Stress–Regulated Vascular Remodeling: Past, Present, and Future,” Arteriosclerosis, Thrombosis, and Vascular Biology 45, no. 6 (2025): 882–900.

[7] C. K. Cheng, N. Wang, L. Wang, and Y. Huang, “Biophysical and Biochemical Roles of Shear Stress on Endothelium: A Revisit and New Insights,” Circulation Research 136, no. 7 (2025): 752–772.

[8] K. H. Ku, N. Subramaniam, and P. A. Marsden, “Epigenetic Determinants of Flow-Mediated Vascular Endothelial Gene Expression,” Hypertension 74, no. 3 (2019): 467–476.

[9] J. J. Chiu and S. Chien, “Effects of Disturbed Flow on Vascular Endothelium: Pathophysiological Basis and Clinical Perspectives,” Physiological Reviews 91, no. 1 (2011): 327–387.

[10] F. M. Callaghan and S. M. Grieve, “Normal Patterns of Thoracic Aortic Wall Shear Stress Measured Using Four-Dimensional Flow MRI in a Large Population,” American Journal of Physiology-Heart and Circulatory Physiology 315, no. 5 (2018): H1174–H1181.

[11] C. Trenti, M. Ziegler, N. Bjarnegård, T. Ebbers, M. Lindenberger, and P. Dyverfeldt, “Wall Shear Stress and Relative Residence Time as Potential Risk Factors for Abdominal Aortic Aneurysms in Males: A 4D Flow Cardiovascular Magnetic Resonance Case-Control Study,” Journal of Cardiovascular Magnetic Resonance 24, no. 1 (2022): 18.

[12] Y. Qian, S. Xiong, L. Li, et al., “Spatial Multiomics Atlas Reveals Smooth Muscle Phenotypic Transformation and Metabolic Reprogramming in Diabetic Macroangiopathy,” Cardiovascular Diabetology 23, no. 1 (2024): 358.

[13] K. I. Matsumoto, K. Satoh, T. Maniwa, T. Tanaka, H. Okunishi, and T. Oda, “Proteomic Comparison Between Abdominal and Thoracic Aortic Aneurysms,” International Journal of Molecular Medicine 33, no. 4 (2014): 1035–1047.

[14] D. He, A. Mao, C. B. Zheng, et al., “Aortic Heterogeneity Across Segments and Under High Fat/Salt/Glucose Conditions at the Single-Cell Level,” National Science Review 7, no. 5 (2020): 881–896.

[15] A. Brückner, A. Brandtner, S. Rieck, et al., “Site-Specific Genetic and Functional Signatures of Aortic Endothelial Cells at Aneurysm Predilection Sites in Healthy and AngII ApoE-/- mice,” Angiogenesis 27, no. 4 (2024): 719–738.

[16] A. J. Fletcher, M. B. J. Syed, T. J. Aitman, D. E. Newby, and N. L. Walker, “Inherited Thoracic Aortic Disease: New Insights and Translational Targets,” Circulation 141, no. 19 (2020): 1570–1587.

[17] C. K. Cheng, J. Gao, L. Kang, and Y. Huang, “Fecal Microbiota Transfer From Young Mice Reverts Vascular Aging Hallmarks and Metabolic Impairments in Aged Mice,” Aging and Disease 16, no. 3 (2025): 1576–1585.

[18] C. K. Cheng, X. Huang, S. Meng, and Y. Huang, “The Pro-Aging and Rejuvenating Effects of Young and Aged Perivascular Adipose Tissues on Endothelial Function and Inflammation,” Biogerontology 26, no. 4 (2025): 134.

[19] M. Clément, J. Chappell, J. Raffort, et al., “Vascular Smooth Muscle Cell Plasticity and Autophagy in Dissecting Aortic Aneurysms,” Arteriosclerosis, Thrombosis, and Vascular Biology 39, no. 6 (2019): 1149–1159.

[20] M. Pacurari, R. Kafoury, P. B. Tchounwou, and K. Ndebele, “The Renin-Angiotensin-Aldosterone System in Vascular Inflammation and Remodeling,” International Journal of Inflammation 2014 (2014): 689360.

[21] H. Kidoya, M. Ueno, Y. Yamada, et al., “Spatial and Temporal Role of the Apelin/APJ System in the Caliber Size Regulation of Blood Vessels During Angiogenesis,” EMBO Journal 27, no. 3 (2008): 522–534.

[22] J. Ma, Y. Li, X. Yang, et al., “Signaling Pathways in Vascular Function and Hypertension: Molecular Mechanisms and Therapeutic Interventions,” Signal Transduction and Targeted Therapy 8, no. 1 (2023): 168.

[23] M. Nunes, M. Vlok, A. Proal, D. B. Kell, and E. Pretorius, “Data-Independent LC-MS/MS Analysis of ME/CFS Plasma Reveals a Dysregulated Coagulation System, Endothelial Dysfunction, Downregulation of Complement Machinery,” Cardiovascular Diabetology 23, no. 1 (2024): 254.

[24] J. Frösen, R. Tulamo, T. Heikura, et al., “Lipid Accumulation, Lipid Oxidation, and Low Plasma Levels of Acquired Antibodies Against Oxidized Lipids Associate With Degeneration and Rupture of the Intracranial Aneurysm Wall,” Acta Neuropathologica Communications 1 (2013): 71.

[25] H. Li, C. Burkhardt, U. R. Heinrich, I. Brausch, N. Xia, and U. Förstermann, “Histamine Upregulates Gene Expression of Endothelial Nitric Oxide Synthase in Human Vascular Endothelial Cells,” Circulation 107, no. 18 (2003): 2348–2354.

[26] A. Simon, L. Plies, A. Habermeier, U. Martiné, M. Reining, and E. I. Closs, “Role of Neutral Amino Acid Transport and Protein Breakdown for Substrate Supply of Nitric Oxide Synthase in Human Endothelial Cells,” Circulation Research 93, no. 9 (2003): 813–820.

[27] J. P. Pirruccello, M. D. Chaffin, E. L. Chou, et al., “Deep Learning Enables Genetic Analysis of the Human Thoracic Aorta,” Nature Genetics 54, no. 1 (2022): 40–51.

[28] M. He, X. Zhou, and X. Wang, “Glycosylation: Mechanisms, Biological Functions and Clinical Implications,” Signal Transduction and Targeted Therapy 9, no. 1 (2024): 194.

[29] P. Lanzer, F. M. Hannan, J. D. Lanzer, et al., “Medial Arterial Calcification: JACC State-of-the-Art Review,” Journal of the American College of Cardiology 78, no. 11 (2021): 1145–1165.

[30] A. Lau, J. J. Rahn, M. Chappellaz, et al., “Dipeptidase-1 Governs Renal Inflammation During Ischemia Reperfusion Injury,” Science Advances 8, no. 5 (2022): eabm0142.

[31] S. R. Choudhury, L. Babes, J. J. Rahn, et al., “Dipeptidase-1 Is an Adhesion Receptor for Neutrophil Recruitment in Lungs and Liver,” Cell 178, no. 5 (2019): 1205–1221.e17.

[32] D. Pandey, G. Sikka, Y. Bergman, et al., “Transcriptional Regulation of Endothelial Arginase 2 by Histone Deacetylase 2,” Arteriosclerosis, Thrombosis, and Vascular Biology 34, no. 7 (2014): 1556–1566.

[33] S. Rovira-Llopis, M. Rocha, R. Falcon, et al., “Is Myeloperoxidase a Key Component in the ROS-Induced Vascular Damage Related to Nephropathy in Type 2 Diabetes?,” Antioxidants & Redox Signaling 19, no. 13 (2013): 1452–1458.

[34] L. Wang, J. Y. Luo, B. Li, et al., “Integrin-YAP/TAZ-JNK Cascade Mediates Atheroprotective Effect of Unidirectional Shear Flow,” Nature 540, no. 7634 (2016): 579–582.

[35] A. von Mässenhausen, N. Zamora Gonzalez, F. Maremonti, et al., “Dexamethasone Sensitizes to Ferroptosis by Glucocorticoid Receptor-Induced Dipeptidase-1 Expression and Glutathione Depletion,” Science Advances 8, no. 5 (2022): eabl8920.

[36] J. Y. Ji, H. Jing, and S. L. Diamond, “Shear Stress Causes Nuclear Localization of Endothelial Glucocorticoid Receptor and Expression From the GRE Promoter,” Circulation Research 92, no. 3 (2003): 279–285.

[37] C. Bernal-Mizrachi, S. Weng, C. Feng, et al., “Dexamethasone Induction of Hypertension and Diabetes Is PPAR-Alpha Dependent in LDL Receptor-Null Mice,” Nature Medicine 9, no. 8 (2003): 1069–1075.

[38] S. C. Schäfer, T. Wallerath, E. I. Closs, et al., “Dexamethasone Suppresses eNOS and CAT-1 and Induces Oxidative Stress in Mouse Resistance Arterioles,” American Journal of Physiology-Heart and Circulatory Physiology 288, no. 1 (2005): H436–H444.

[39] C. Zeng, G. Qi, Y. Shen, et al., “DPEP1 Promotes Drug Resistance in Colon Cancer Cells by Forming a Positive Feedback Loop With ASCL2,” Cancer Medicine 12, no. 1 (2023): 412–424.

[40] M. Asada, A. Rauch, H. Shimizu, et al., “DNA Binding-Dependent Glucocorticoid Receptor Activity Promotes Adipogenesis via Krüppel-Like Factor 15 Gene Expression,” Laboratory Investigation 91, no. 2 (2011): 203–215.

RIGHTS & PERMISSIONS

2026 The Author(s). MedComm published by Sichuan International Medical Exchange & Promotion Association (SCIMEA) and John Wiley & Sons Australia, Ltd.