We aimed to evaluate the medium-term outcomes of high uterosacral ligament suspension (HUSLS) with vaginal-assisted natural orifice transluminal endoscopic surgery (vNOTES) in patients either suffering from pelvic organ prolapse (POP) or undergoing routine prophylaxis to prevent post-hysterectomy vaginal vault prolapse. Multicenter prospective cohort study SETTING: Three tertiary care hospitals PATIENTS: Patients who received vNOTES-HUSLS between January 1, 2021 and January 1, 2023, were included in the study. vNOTES-HUSLS MEASUREMENTS AND MAIN RESULTS: The data collected included surgery duration, intraoperative and postoperative complications, postoperative Visual Analog Scores (VAS), and Female Sexual Function Index (FSFI) pain subdomain scores. The changes in prolapse levels were measured using modified POP-Q scores, including the C, Ba, and Bp scores recorded prior to surgery and during the postoperative follow-up examination. Patient satisfaction was assessed using the Patient Global Impression of Improvement (PGI-I) questionnaire. The final analysis included 55 women. Of these, vNOTES-HUSLS was performed in 43 patients (78.2%) to treat apical prolapse, while the remaining 12 patients (21.8%) underwent prophylaxis following hysterectomy to prevent vaginal vault prolapse. The median follow-up period was 23.5 months, with a range of 12-37 months. Ba,C, and Bp points significantly improved in patients who underwent surgery for prolapse (p<0.001). There was no recurrence in the apical compartment following vNOTES-HUSLS for both performed prophylactic and treatment purposes after hysterectomy. Two (3.6%) patients were diagnosed with stage 3 anterior compartment prolapse. The overall vNOTES-HUSLS success rate was 96.4% at approximately two years follow-up. Bladder injury was detected as an intraoperative complication in one patient (1.8%). The overall satisfaction rate was 98.1% at the final follow-up. vNOTES-HUSLS was effective in the treatment and prophylaxis of apical prolapse, demonstrating high anatomical success rates. The procedure demonstrated a low complication rate, with intraoperative and postoperative complications each observed in 1.8%(n=1) of patients. The Medium-Term Outcomes of vNOTES High Uterosacral Ligament Suspension.