RNAseq transcriptome of draining lymph node (LN) and tumor of MC38 murine tumors treated with cryoablation and chitosan/IL-12

<p class="MsoNormal">Focal ablation technologies are routinely used in the clinical management of inoperable solid tumors but often result in incomplete ablations leading to high recurrence rates. Adjuvant therapies capable of safely eliminating residual tumor cells are therefore of great clinical interest. Interleukin 12 (IL-12) is a potent antitumor cytokine that can be localized intratumorally through coformulation with <span>viscous biopolymers</span> including chitosan (CS) solutions. The objective of this research was to determine if localized immunotherapy with CS/IL-12 could prevent <span>tumor recurrence after cryoablation (CA). Tumor recurrence, overall survival, and protective immunity were assessed. Systemic immunity was evaluated in spontaneously metastatic and bilateral tumor models. Temporal bulk RNA sequencing was performed on tumor and draining lymph node samples.</span> In multiple murine tumor models, the addition of CS/IL-12 to CA reduced recurrence rates by 30–55%. Altogether, this cryo-immunotherapy induced complete durable regression of large tumors in 80–100% of treated animals. <span>Mice</span> treated with CA plus adjuvant CS/IL-12 were partially or completely protected from tumor rechallenge. <span>Systemically,</span> CS/IL-12 prevented lung metastases when delivered as a neoadjuvant to CA. However, CA plus CS/IL-12 had minimal antitumor activity against established, untreated abscopal tumors. Adjuvant anti-PD-1 therapy delayed the growth of abscopal tumors. Transcriptome analyses revealed early immunological changes in <span>the dLN</span>, followed by a significant increase in gene expression associated with immune suppression and regulation. Cryo-immunotherapy with localized CS/IL-12<span> reduces recurrences and</span> enhances the elimination of large primary tumors<span>. This focal combination therapy also induces significant systemic</span> antitumor immunity <span>although further studies are necessary</span>.</p>