The FDA Approved Therapeutic Lasmiditan, Enhances Primary Murine Renal Peritubular Endothelial Cells Wound Healing Capacity.

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<b>Abstract ID 16703</b> <b>Poster Board 510</b> <b>Background:</b> Acute kidney injury (AKI) is defined as an episode of sudden and rapid decline in renal function and is often accompanied by a persistent reduction in mitochondrial function, renal microvasculature dysfunction/rarefication, and tubular injury/necrosis.<b></b> AKI continues to be an immense public health concern, as there remains no effective FDA-approved treatment options. A central underlying contributor to AKI, and the subsequent development of other progressive kidney and cardiovascular related diseases, is renal microvascular endothelial cell dysfunction/rarefaction. This dysfunction/rarefaction exacerbates renal injury and increases the likelihood of AKI recurrence (∼20%). Furthermore, AKI results in a persistent reduction in mitochondrial function, number, and cellular energetics. Conversely, pharmacological stimulation of mitochondrial biogenesis (MB) has been shown to restore mitochondrial function and promote renal vascular recovery post-AKI. However, <b>minimal</b> AKI research has focused on elucidating the cellular mechanisms that govern mitochondrial biogenesis (MB) and neovascularization responses post-AKI; moreover, how stimulation of MB contributes to renal peritubular endothelial cell repair mechanisms post-AKI remains <b>unknown</b>. Therefore, we conducted in-vitro wound healing assays with primary murine renal peritubular endothelial cells (MRPEC) exposed to lipopolysaccharide (LPS) in the presence/absence of the lasmiditan, a 5-HT1F receptor agonist, and a known inducer of MB within the renal cortex of mice. <b>Methods:</b> MRPECs were isolated utilizing a refined method by Zhao et&nbsp;al. (2014). Briefly, renal enzymatic tissue digestion buffers were altered by replacing collagenase type 4 with collagenase type 1 and trypsin with accutase, respectively. Additionally, gentle MACS-C tubes with automated tissue dissociation were employed to reduce processing times and improve cell viability. Cells were then subjected to CD326+ magnetic microbead negative selection, to remove epithelial cells, followed by two positive selection cycles with CD146+ magnetic microbeads. Purified MRPECs were then seeded at a density of 70,000 cells/well into human plasma fibronectin precoated 35mm Ibidi wound healing chamber dishes and incubated overnight in microvascular specific media (EGM-MV2; Lonza). MRPECs were then treated with either LPS (1ug/mL) or normal saline in the presence/absence of lasmiditan (100nM) prior to the removal of chamber inserts. Wound healing dishes were then imaged at 0-, 6-, and 12- hours post treatment and subsequently analyzed by Ibidi FastTrackAI software. <b>Results:</b> MRPECs treated with lasmiditan displayed an increase in total wound closure percentage, 1.21 and 1.44 fold compared to normal saline and LPS treated control cells, respectively. Additionally, MRPECs treated with both LPS and lasmiditan exhibited an increase in total wound closure percentage, 1.33 fold compared to LPS treated control cells. Lastly, LPS treated MRPECs displayed a decrease in total wound closure percentage compared to normal saline controls, 1.18 fold. <b>Conclusion:</b> In conclusion, we demonstrated that lasmiditan treatment enhanced MRPEC wound healing capacity alone and following LPS induced cellular injury, in-vitro. These results suggest that lasmiditan may promote vascular recovery following kidney injury. T32 5T32ES007091-39 (National Institutes of Health; NIEHS) to ADT. BX000851 (Department of Veterans Affairs) to RGS.