Research Projects

Rationale of the research: Skin repair and regeneration is regulated by pro-inflammatory M1 and pro-healing M2 macrophages in a stage-dependent manner. Prolonged inflammatory environment in diabetes impairs M2 macrophages, compromising wound healing. Differentiation and switching of M1 and M2 macrophages require deeper understanding of their metabolism. Mitochondria harbour NOS2 and Arginase II enzymes that promote M1 and M2 differentiation. We have recently demonstrated that macrophage Piezo1 channel activation mediated Ca2+ dynamics changes mitochondrial structure-function, inducing higher differentiation of macrophages to M2 phenotype under normoglycemic conditions. Scientific objectives : This project will decipher the impact of Piezo1 channel on mitochondrial dynamics and metabolism of human primary macrophages cultured in normoglycemic or hyperglycemic media. We will also use peritoneal macrophages from wild-type and diabetic ob/ob mice to better understand the role of Piezo1 channels in M1 and M2 macrophage differentiation under normoglycemic vs diabetic conditions. Piezo1 modulatory drugs will also be tested in vivo using skin wound healing assays with wild-type and diabetic mice. Hypothesis/model to be tested: We hypothesize that Piezo1 activation can promote M2 macrophage differentiation, resulting in improved non-diabetic and diabetic wound healing. Main experiments to be carried out: Primary macrophages from healthy blood donors, human THP1 cell line derived macrophages and peritoneal macrophages from wild-type and diabetic mice will be utilized in RT-PCR, Confocal microscopy, super resolution microscopy, high content imaging, western blotting, luminometry to evaluate the expression and impact of Piezo1 on macrophage differentiation under normoglycemic and diabetic conditions. In vivo intraperitoneal and skin wound healing experiments using wild-type and diabetic mice followed by immunohistochemistry and ELISA will reveal the impact of Piezo1 pharmacological modulation in phenotype swithching of macrophages and wound-healing. Significance to the field of research : This project can lead to better understanding of the role of Piezo1 channels in mitochondrial dynamics, metabolism, macrophage polarization and can provide therapeutic product for non-diabetic and diabetic wound healing.