👤 Deborah L Della Manna

The development of nucleic acid therapeutics using non-viral delivery systems requires efficient payload delivery to target organs for higher potency and tolerability. While lipid nanoparticle (LNP) formulations influence biodistribution, cellular uptake, and therapeutic efficacy, underlying mechanisms remain incompletely understood. This study develops potent mRNA-LNP formulations and investigates determinants of liver tropism using ornithine transcarbamylase (OTC) deficiency as a protein replacement therapy model. Systematic screening of ionizable and helper lipids, optimization of composition and process, and biophysical characterization identify a liver-tropic helper lipid-1,2-dierucoyl-sn-glycero-3-phosphoethanolamine (DEPE) that modulates LNP structure and apolipoprotein E (ApoE) binding, enhancing liver-specific delivery. Analysis of ionizable lipid chemistry reveals its role in cellular uptake mechanisms, leading to the identification of a novel ionizable lipid designed with N-(2-Hydroxyethyl)piperazine-N'-(4-butanesulfonic acid) (HEPBS) core that enables efficient delivery independent of the low-density lipoprotein receptor (LDLR) pathway. The optimized formulation achieves robust dose responsiveness, sustained therapeutic expression, and favorable tolerability in preclinical models. Therapeutic levels of OTC protein expression are observed with minimal toxicity, as indicated by stable liver function markers and cytokine levels. These findings provide mechanistic insights and establish a platform for mRNA-based protein replacement therapies, supporting broader applications in rare genetic diseases requiring hepatic gene expression. Show less

Everolimus monotherapy use for metastatic renal cell carcinoma (mRCC) has diminished due to recent approvals of immune checkpoint and VEGF inhibitors. We hypothesized that gene expression associated with everolimus benefit may provide rationale to select appropriate patients. To address this hypothesis, tumors from a phase I/II trial that compared everolimus alone or with BNC105P, a vascular disrupting agent, were profiled using Nanostring as a discovery cohort. A phase III trial (CheckMate 025) was used for validation. Clinical benefit (CB) was defined as response or stable disease for ≥6 months. A propensity score covariate adjustment was used, and model discrimination performance was assessed using the area under the ROC curve (AUC). In a discovery cohort of 82 patients, 35 (43%) were treated with everolimus alone and 47 (57%) received everolimus + BNC105P. Median PFS (mPFS) was 4.9 (95% CI, 2.8-6.2) months. A four-gene signature (ASXL1, DUSP6, ERCC2, and HSPA6) correlated with CB with everolimus ± BNC105P [AUC, 86.9% (95% CI, 79.2-94.7)]. This was validated in 130 patients from CheckMate 025 treated with everolimus [AUC, 60.2% (95% CI, 49.7-70.7)]. Among 43 patients (52.4%) with low expression of an 18-gene signature, everolimus + BNC105P was associated with significantly longer mPFS compared with everolimus alone (10.4 vs. 6.9 months; HR, 0.49; 95% CI, 0.24-1.002; Show less