PROJECT SUMMARY Low back pain (LBP) is a leading cause of disability and morbidity in the adult population, affecting approximately 80% of adults within their lifetime. Up to 40% of all low back pain is attributed to discogenic pain from IVD degeneration. While people of different races, ethnicity and gender suffer from chronic back pain, this disease has been shown to affect often people from underserved communities. IVD degeneration is known to affect the NP, the central part of the IVD. Despite decades of research, robust therapies targeting underlying causes rather than symptoms of IVD degeneration are still in the earliest stages of development. Conservative treatments alleviate symptoms rather than targeting the underlying disease. Stem cell therapy has been shown a great promise to regenerate IVD in animal models. Specifically, we have shown that induced pluripotent stem cells can be differentiated to notochordal cells (iNC), the original progenitors of NP cells. Different cell delivery and microencapsulation techniques will be explored to generate, iNC-microgel, preconditioned iNC-microgels and iNCs in bulk hydrogel. Cell purity, identity, viability and sterility data will be obtained. Furthermore, the material composition and properties will be investigated, and the stability of microencapsulated iNCs after preparation evaluated. Safety and efficacy of the therapeutic candidates will be evaluated in a rat model of IVD degeneration and discogenic low back pain, induced by disc puncture of the lumbar spine. After confirming successful induction of IVD degeneration, different therapeutic candidates and controls will be injected. The candidate’s regenerative potential and reproducibility of results will be evaluated by biobehavioral testing, MRI and immunohistochemical analyses. For mechanism of action studies, single cell RNA sequencing of the treated IVDs will be employed. The outcome of the study will be defined therapeutic candidate for low back pain stem cell therapy and multidisciplinary team including biologist, clinicians, translational experts and statisticians that will be set up for biomanufacturing and IND- enabling studies via U19 mechanism of the NIH HEAL initiative.

PROJECT NARRATIVE The goal of this study is to further develop an injectable, rejuvenating treatment for painful intervertebral disc degeneration using microgel-encapsulated iPSC-derived notochordal cells (iNCs).