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Thesis Defense: Investigating the Ablative and Immunomodulatory Effects of High-Frequency Irreversible Electroporation on Osteosarcoma In-vitro

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M Patwardhan

Thesis Defense: Investigating the Ablative and Immunomodulatory Effects of High-Frequency Irreversible Electroporation on Osteosarcoma In-vitro

Manali Patwardhan

Graduate Student, Translational Biology, Medicine, and Health
MS Student, Tuohy Lab
April 25, 2024 at 11 a.m.,  Room 232, 1 Riverside
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About this Thesis

Osteosarcoma (OS) is the most common primary bone tumor with an annual incidence rate of 3-4 individuals per million particularly affecting children and young adults. The 5-year survival rate stands at 60-80% with the current standard of care for human OS patients who do not have metastatic disease at presentation, but this drops to 20% for patients with metastatic disease which frequently occurs in the lungs. OS is much more common in canines, with metastasis being the major contributor to mortality, same as in humans. Metastatic OS warrants novel treatment strategies to improve prognosis and survival. High-frequency irreversible electroporation (H-FIRE) is a promising, non-thermal, minimally invasive technique that induces cell death by applying pulsed electric fields in targeted regions, potentially triggering an anti-tumor immune response that could also target and prevent metastases. Such a dual functionality of H-FIRE is uniquely suited to treat pulmonary metastatic OS. The goal of this thesis was to study the ablative and immunomodulatory effects of H-FIRE on OS in-vitro with the overall hypothesis that H-FIRE completely ablates OS cells, induces the release of damage-associated molecular patterns (DAMPs), and promotes pro-inflammatory immune activating signatures in macrophages and T cells. Using an in-vitro model, my master’s thesis focused on 1) Determining the electric field strength that completely ablates OS cells 2) Evaluating the immunomodulatory effects of H-FIRE by co-culturing H-FIRE treated OS cells with macrophages and T cells separately. Our study has utilized murine, canine, and human OS and immune cells, thus demonstrating a unique cross-species approach, 3) Evaluating DAMPs (ATP, calreticulin, and HMGB1) post-H-FIRE ablation of human OS cells. Overall, our study showed that H-FIRE successfully ablated OS cells in-vitro induced the release of DAMPs from treated cells, and promoted activation signatures in immune cells. This thesis provides foundational data for future investigations developing H-FIRE as an immunomodulatory strategy for treating metastatic OS.

More About the Candidate and Project

Education

Virginia Tech, Translational Biology, Medicine, and Health, M.S. Student

University of Mumbai-St. Xavier's College, M.S., Biotechnology

University of Mumbai- V.G. Vaze College, B.S., Biotechnology        

Training

Tuohy Lab

 

Mentor

Joanne Tuohy, DVM, Ph.D., Assistant Professor, VA-MD College of Veterinay Medicine

Committee Members

  • Irving Coy Allen, Ph.D., Professor, VA-MD College of Veterinary Medicine
  • Rafael Davalos, Ph.D., Chaired Professor, Georgia Tech
  • Sheryl Coutermarsh-Ott, DVM, Ph.D., Veterinary Pathologist, VA-MD College of Veterinary Medicine
  • Hay, A., Vickers, E.R., Patwardhan, M., Allen, I.C., Gannon, J., Ruger, L., Vlaisavljevich, E., & Tuohy, J. Investigating cell death genetic signatures associated with histotripsy treatment of canine osteosarcoma: An in vitro study. Int J Hyperthermia (2023) 40(1):2279027. doi: 10.1080/02656736.2023.2279027.
  • Manali Patwardhan, Alayna Hay, Elliana Vickers, Haleigh Hixson, Lauren Ruger, Irving Allen, Eli Vlaisavljevich and Joanne Tuohy. Investigating the immunomodulatory effects of histotripsy ablation for osteosarcoma. Poster presented at Immunology2023TM, Washington D.C., United States.
  • Manali Patwardhan, Alayna Hay, Kenneth Aycock, Olivia Echols, Kailee David, Elliana Vickers, Kelsey Murphy, Irving Allen, Rafael Davalos, and Joanne Tuohy. Evaluation of ablative and immunomodulatory effects of in vitro H-FIRE for osteosarcoma. Poster presented at Virginia Tech Cancer Research Alliance 2023 Retreat, Washington D.C., The United States.
  • Manali Patwardhan, Alayna Hay, Kenneth Aycock, Olivia Echols, Kailee David, Zaid Salameh, Elliana Vickers, Ny Luong, Kelsey Murphy, Irving Allen, Rafael Davalos, Joanne Tuohy. Evaluation of ablative and immunomodulatory effects of in-vitro H-FIRE for osteosarcoma. Poster presented at TBMH Winter Symposium 2023-2024, Roanoke, Virginia, United States.
  • $1500 funding awarded for a research proposal submitted to the Graduate Research Development Program, Fall 2023, Virginia Tech. Title: Evaluation of damage-associated molecular patterns post-H-FIRE treatment of osteosarcoma
  • Manali Patwardhan, Alayna Hay Ph.D., Kenneth Aycock, Olivia Echols, Kelsey Murphy, Rafael Davalos Ph.D., Joanne Tuohy DVM, Ph.D. Evaluation of ablative and immunomodulatory effects of in-vitro H-FIRE for osteosarcoma. Poster presented and won best poster prize at TBMH Winter Symposium 2022-2023, Roanoke, Virginia, United States.
  • Grant reviewer for 3 research proposals submitted to the Graduate Research Development Program (GRDP), Spring 2024, Virginia Tech, The United States.