Author ORCID Identifier
0009-0000-7456-2733
Defense Date
2025
Document Type
Dissertation
Degree Name
Doctor of Philosophy
Department
Pharmaceutical Sciences
First Advisor
Prof. Sandro daRocha
Second Advisor
Prof. David Edwards
Third Advisor
Dr. Thomas Roper
Fourth Advisor
Dr. Laleh Golshahi
Fifth Advisor
Dr. Fan Zhang
Abstract
Development of Lipid-based Nanoformulations of Macrophage Targeting immunotherapies for Oral Inhalation Using a Continuous Manufacturing Approach and Design of Experiments A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy at Virginia Commonwealth University By Asma Al-Terawi, B.S, M.S Director: Prof. Sandro R.P. da Rocha, Ph.D. and Prof. David Edwards, Ph.D. Osteosarcoma (OS) is the most common primary bone malignancy, with the lungs being the predominant site of metastasis. At diagnosis, approximately 80% of patients harbor undetectable pulmonary metastases, while 20% present with overt osteosarcoma lung metastases (OSLM). Despite multimodal treatment approaches, survival outcomes for OSLM have remained stagnant since the 1980s, highlighting the urgent need for novel therapeutic strategies. This dissertation investigates the development of macrophage-targeted nanoimmunotherapies for local pulmonary delivery to modulate the tumor microenvironment (TME) in OSLM. Two inhalable nanocarrier systems were developed: (1) a liposomal formulation of the CSF-1R inhibitor PLX3397 (L-PLX), and (2) lipid nanoparticles (LNPs) encapsulating mRNA encoding interferon-gamma (IFNγ), both designed for oral inhalation (OI) to reprogram tumor-associated macrophages (TAMs) toward an anti-tumorigenic phenotype. Using a continuous manufacturing platform with toroidal microfluidics and a Quality-by-Design (QbD) framework, both formulations were optimized through Design of Experiments (DOE) For L-PLX, the flow rate ratio (FRR) emerged as the most significant factor affecting hydrodynamic. 16 diameter (H.D.), with optimal conditions identified as FRR 5:1 (PBS), lipid-to-drug ratio 5:1, total flow rate of 10 mL/min, and ambient temperature. The optimized L-PLX achieved high encapsulation efficiency (>85%), drug loading (>4.5%), H.D. ≈ 155 nm, and PDI < 0.2. It remained stable over 3 months at 2–8°C and retained integrity post-nebulization, producing respirable aerosols (Dv50 ≈ 3.9 µm). In vitro, studies showed superior efficacy in reducing M2- like TAMs compared to free PLX. The second platform, mRNA-loaded LNPs encoding enhanced green fluorescent protein (eGFP), was optimized using a two-level A-optimal DOE. The final formulation, composed of SM-102, DPPC, β-sitosterol, and DMG-PEG2k, exhibited high encapsulation efficiency, H.D. < 100 nm, strong eGFP expression, and batch-to-batch reproducibility. Short-term stability studies confirmed preservation of physical properties, with a moderate decline in encapsulation and expression over time. Collectively, these findings demonstrate the feasibility and translational potential of scalable, inhaling nanocarrier systems for TAM-targeted immunotherapy in OSLM. The use of clinically relevant components including FDA-approved PLX3397 and mRNA-LNP platforms validated by vaccines underscores the promise of these systems to enhance standard of care therapies and address an unmet clinical need in metastatic osteosarcoma
Rights
© The Author
Is Part Of
VCU University Archives
Is Part Of
VCU Theses and Dissertations
Date of Submission
12-12-2025
Included in
Biotechnology Commons, Immunotherapy Commons, Medicinal and Pharmaceutical Chemistry Commons, Nanomedicine Commons, Nanoscience and Nanotechnology Commons, Pharmaceutical Preparations Commons, Pharmaceutics and Drug Design Commons