Author ORCID Identifier
Semester
Spring
Date of Graduation
2024
Document Type
Dissertation
Degree Type
PhD
College
School of Medicine
Department
Not Listed
Committee Chair
Brock Lindsey
Committee Co-Chair
J. Michael Ruppert
Committee Member
Steven Frisch
Committee Member
Edwin Wan
Committee Member
Scott Weed
Abstract
Osteosarcoma (OS) is a rare pediatric bone malignancy. Those who develop advanced disease, often in the form of metastasis to the lung, experience poor five-year survival rates near approximately 25%. Considering minimal-to-no therapeutic breakthroughs have occurred for treatment of this disease since the 1980s, efforts to develop new therapeutics are underway. The goal of this dissertation was to initiate the preclinical efforts necessary to reinvigorate the use of immunostimulatory cytokine therapy for the treatment of OS and other solid tumors. This effort would not only include the development of the immunotherapeutic known as IL12ns, but also the clinical diagnostic tools that would be necessary for safe and efficacious use of this therapy in future clinical practice. This work first details the extensive efforts made to develop IL-12-loaded poly (lactic-co-glycolic acid) (PLGA) nanospheres, IL12ns, hypothesized to limit the previous side effects and immunotoxicity observed in human clinical trials of bolus IL-12 therapy through the improved pharmacodynamics of a vector delivery platform. Through use of the double-emulsion solvent evaporation (DESE) method with homogenization, we successfully encapsulated and delivered IL12ns in a murine model, achieving systemic delivery to peripheral tissues while generating desirable peripheral immune responses associated with IL-12 therapeutic efficacy. Next, to improve the likelihood of future commercialization of IL12ns, extensive efforts were made to increase both encapsulation efficiency and fabrication yield by using both synthesis modifiers (surfactants and carrier proteins) and ultrasonication. An optimized IL12ns was then examined in a preclinical model of murine metastatic OS, exhibiting preliminary therapeutic efficacy. Furthermore, efforts to shift the treatment paradigm of OS toward the use of immunostimulatory therapies will likely necessitate the concomitant development of immune diagnostic platforms capable of monitoring disease progression and therapeutic response. Clinical tools to stratify patients by likely responder status before the initiation of alternative therapies will help best optimize patient care in a future data-oriented clinical practice driven by patient-specific immunological insights. To do so, we initiated the development of an immune diagnostic platform capable of assessing changes in the systemic immune proteome and transcriptome with IL12ns therapy through repeated blood sampling. Markers of beneficial and maladaptive immune responses to IL-12 therapy, measured by the platform, will be critical assessments utilized in future human clinical trials to prevent overt clinical toxicity while best modulating the delivery of this therapy. Additionally, both a retrospective analysis of common clinical laboratory measures (such as a complete blood count) and spatial multiplexed immunofluorescence analysis of tumor specimens revealed peripheral and local immunological patterns associated with sarcoma disease that could help identify candidates for alternative immunotherapies, as opposed to the current standard of care. Finally, through efforts to characterize the systemic proteomic profile of OS patients, apo-transcobalamin-II (APO-TCN2) was identified as a novel biomarker of overall survival in this disease. Considering its likely role in the lymphoproliferation of B and plasma cells, this circulatory protein should be considered a novel immunotherapy candidate that necessitates further preclinical examination. Ultimately, these data lay the foundation for the future of OS treatment including the deployment of alternative immunotherapeutic strategies alongside data-driven diagnostic platforms that convey patient-specific immunological insights for optimal clinical decision making, robust therapeutic responses, and better patient outcomes.
Recommended Citation
Lacinski, Ryan Austin, "Immunomodulation in Osteosarcoma: From Therapy to Diagnostics" (2024). Graduate Theses, Dissertations, and Problem Reports. 12333.
https://researchrepository.wvu.edu/etd/12333
Embargo Reason
Publication Pending