Year 3

Urinary incontinence (UI) is common and serious, with two-thirds of the burden borne by women. Our proposed development candidate is intended to treat UI by restoring normal function to the internal urethral sphincter via regenerative therapy modalities. The initial focus is on the treatment of UI in women refractory to other surgical interventions. The urethra is comprised of an internal urethral sphincter composed primarily of smooth muscle cells that control the flow of urine at the outlet of the bladder and an external urethral sphincter composed of voluntary skeletal muscle cells. The urethral wall also contains smooth muscle fibers and is supported by connective tissue. The total number of smooth muscle cells in the urethra decrease with aging, thus contributing to the high prevalence of incontinence amongst older individual. The discovery of genetic reprogramming methods to convert an adult cell into a cell with stem cell properties (induced pluripotent stem cells or iPSCs) has sparked strong interest in using these patient derived stem cells for regenerative therapies. The overall goal of this project is to develop an iPSC-based therapy for the treatment of stress urinary incontinence (SUI) in women. Our strategy consists of deriving iPSC lines with a non-integrative technology, differentiating lines to smooth muscle cells (SMC), assessing real-time characteristics of undifferentiated and differentiated cells, testing safety and efficacy in an animal model, and proceeding to initial discussions with the FDA.
In previous reporting periods, we derived three patient lines, characterized the lines, optimized differentiation further and established initial benchmark in vitro assays for analysis of undifferentiated cells, as well as, established methods for animal studies. We have also focused on replacement of our protocol for differentiation with a protocol that removes rodent feeder cells and is carried out in chemically-defined media. In the current reporting period, we concentrated on optimizing our differentiation protocol for our patient-specific iPSC lines, increasing production of smooth muscle precursor cells, confirming SMC precursor in vivo integration and long term survival in animals, demonstrating efficacy of our pluripotent stem cell derived therapy, and examining mechanisms of action of our target therapy. We have completed four animal efficacy studies showing reproducible efficacy of our therapy derived from a human embryonic stem cell line and different iPSC lines (from different participants). Smooth muscle cell precursors derived from these lines are all able to restore urethral function and show long term survival and safety in animals. We also filed provisional patent application on our technology.