Footprint-free generation of autologous rejuvenated skeletal myocytes for sphincter muscle repair

• Applicants: Professor Dr. Wilhelm K. Aicher; Professorin Dr. Meltem Avci-Adali
• Subject Area: Reproductive Medicine, Urology
• Term: since 2021
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 468616715

Stress urinary incontinence (SUI) is the most frequent form of urinary incontinence and affects especially a significant number of elderly women. Worldwide, one in seven women is affected by SUI. Thereby, the weakened or damaged urethral closure apparatus leads to uncontrollable leakage of urine due to pressure (stress) during physical exertion such as coughing, sneezing, or performing exercise. Medication and muscle exercise are standard treatments. If the conservative regimen fails to meet the patient’s needs, surgical interventions are an option. But the overall success of the current SUI regimen is not yet optimal. Therefore, cell-based repair of the sphincter complex of the urethra is a promising option to treat SUI. But autologous cells from the elderly may yield limited sphincter regeneration. Thus, in this project, we aim to generate rejuvenated autologous striated muscle cells (SkMCs) for individualized therapy of SUI patients. To this end, we generate iPSCs from human as well as porcine urothelial cells by exogenous delivery of self-replicating RNA (srRNA) encoding the reprogramming factors. iPSCs represent rejuvenated cells. Subsequently, we differentiate the iPSCs generated into SkMCs. The cells obtained are analyzed for their expression of SkMC specific markers, electrophysiological characteristics, and clinical safety. Procedures with human cells are conducted under conditions very close to good manufacturing practice (GMP) regulations to facilitate the translation of this method from bench to bed. In a pre-clinical animal study, porcine iPSC-derived SkMCs will be generated and injected into the sphincter muscle of female pigs with sphincter deficiency. The functional recovery of the sphincter apparatus will be analyzed for 21 up to 90 days post-surgery in an established SUI animal model. The successful footprint-free generation of autologous SkMCs using srRNAs could enable the personalized treatment of patients suffering from SUI with autologous rejuvenated cells. As the generated cells are autologous, rejection reactions are prevented. The application of rejuvenated autologous SkMCs could become an improved curative strategy for the treatment of SUI in elderly patients.

• DFG Programme: Research Grants