Full Text | PDF

Theranostics 2020; 10(8):3397-3412. doi:10.7150/thno.42243 This issue Cite

Research Paper

Novel calreticulin-nanoparticle in combination with focused ultrasound induces immunogenic cell death in melanoma to enhance antitumor immunity

Sri Nandhini Sethuraman^1,#, Mohit Pratap Singh^1,#, Girish Patil¹, Shitao Li¹, Steven Fiering², P. Jack Hoopes², Chandan Guha³, Jerry Malayer¹, Ashish Ranjan^1,✉

1. Center for Veterinary Health Sciences, Oklahoma State University, Stillwater, Oklahoma 74074
2. Geisel School of Medicine, Dartmouth, Hanover, NH03755
3. Albert Einstein College of Medicine, Bronx, New York 10461
^# Both authors contributed equally to the experimental study and drafting of the manuscript.

✉ Corresponding author: Dr. Ashish Ranjan, B.V.Sc., Ph.D., Associate Professor & Kerr Chair, Center for Veterinary Health Sciences, Oklahoma State University, Stillwater, Oklahoma 74074. Phone: 4057446292; Fax: 4057448263; Email: ashish.ranjanedu More

Abstract

Rationale: Some studies have shown that the local activation of immunogenic cell death (ICD) by upregulating calreticulin (CRT) expression in solid tumors can improve antitumor effects. Although a promising approach, a key current challenge in ICD tumor therapy is the absence of a clinically translatable method for reproducibly inducing the CRT expression. Herein, we report a novel calreticulin-nanoparticle (CRT-NP) that enhances ICD and synergizes with focused ultrasound (FUS) to achieve local and systemic antitumor effects.

Methods: Full-length clone DNA of calreticulin was encapsulated in NPs made from DOTAP and cholesterol. Three CRT-NP intratumoral injections of 20 µg each were given 2 days apart, and FUS heating (42-45°C, ~15min) was applied sequentially 24h after each injection to induce ICD. To investigate ICD specific immune effect, the splenocytes of mice vaccinated with CRT-NP (± FUS) treated B16F10 cells were evaluated ex-vivo for TRP-2 antigen specific immunity. Additionally, the long-term protection was evaluated by re-challenging with the melanoma cells in the flank regions of tumor bearing mice.

Results: CRT-NP plus FUS (CFUS) upregulated CRT expression, expanded the population of melanoma TRP-2 specific functional CD4+ and CD8+ T cells and tumor-suppressing M1 phenotype, and increased PD-1 and PD-L1 marker expression in the T cells. Therapeutically, CFUS suppressed B16 melanoma growth by >85% vs. that seen in untreated controls, and >~50% vs. CRT-NP or FUS alone, and prevented tumor growth in distal untreated sites.

Conclusions: CRT-NP amplifies the FUS and ICD therapeutic outcomes against melanoma, suggesting that the proposed combinatorial methodology may be clinically translatable.

Keywords: Immunogenic cell death, calreticulin, nanoparticle, melanoma, focused ultrasound

Citation styles

©2024 Ivyspring International Publisher. Terms of use