Prolonged, Controllable Protein Production of cDNA-Encoded hGH Maintained at Therapeutic Serum Levels Following One Systemic Administration of a Non-Viral, Non-Integrating, DNA-and Liposome-Based in vivo Gene Therapy Platform in Immunocompetent Mice

Abstract Recombinant human growth hormone (rhGH), the mainstay of hGH replacement therapy, is injected daily for years to enable children achieve normal stature. Daily rhGH injections are required because its serum-T1/2 <20 minutes. Human trials testing T1/2-extending proteins promising, but even weekly administration poses potential short and long-term toxicity risks, including exacerbation diabetes or hypertension oncogenesis. Frequent produce non-physiologic IGF-I profiles, (peak serum frequently supra-normal, trough concentrations at baseline). Therefore, a safe, effective, single approach that maintains within their respective therapeutic ranges approximately one year could offer significant advantages. Accordingly, DNARx tested HEDGESTM, it’s non-viral, non-integrating, DNA-and liposome-based systemic, in vivo gene therapy levels range (1 -10 ng/ml) immunocompetent mice. Previously, systemic HEDGES-human granulocyte colony stimulating factor (hG-CSF, T1/2<3 hours) cDNA produced durable hG-CSF (Science advances, 2019, 5, 2019). Importantly, simply by modifying selected aspects HEDGES’s DNA vector liposomal components, duration protein production was controllable over broad temporal range. Long-term assessment mice assessed injecting either HEDGES followed re-dose vectors expressing A) wildtype (HEDGES-1) B) fused sequences (HEDGES-2), with these results: 1) One HEDGES-1 slowly-increasing, sub-therapeutic from day 7 99 after injection. From on, remained > 230 days; 2) Concurrently, hGH-induced elevation endogenous mouse between 1 3 fold above baseline. 3) re-injection administered 35 days initial injection, time when sub-therapeutic. By 7, further elevated cDNA-encoded into range, maintaining them >175 4) HEDGES-2 were maintained >250 days. These studies demonstrate feasibility achieving very following 1-2 injections. now focused on selectively components equivalent