CRYOVATE: A Pilot Study of Lung Cancer Cryoactivation in Combination With Immunotherapy in Advanced NSCLC

Abstract

Introduction: NSCLC is the leading cause of cancer-related mortality. Although immune-checkpoint inhibitors (ICIs) have improved survival in patients with advanced NSCLC, treatment resistance remains a challenge. Cryoactivation, a technique inducing cell death by cycles of freezing and thawing, has the potential to augment tumor responses when combined with ICIs.

Methods: This single-arm phase 1 clinical trial enrolled patients with previously untreated advanced NSCLC and 50% or higher programmed cell death ligand-1 (PD-L1). Patients underwent cryoactivation followed by ICI monotherapy initiated 5 days later. The primary end point was the objective response rate. Co-secondary end points included the safety and feasibility of the procedure and overall survival. Immune cell infiltration by immunohistochemistry was performed on paired pre- and post-treatment samples, with patients dichotomized according to clinical benefit (CB) rate (CB versus no CB [NCB]).

Results: Eight patients were enrolled. Two patients achieved a partial response, yielding an objective response rate of 25%. Median progression-free survival and overall survival were 3.8 and 13.0 months, respectively. The cryoactivation procedure was well tolerated, without grade 3 to 4 adverse events. Post-hoc analysis reported a CB rate of 50%. Immunohistochemistry analysis reported a numerical difference in the cluster of differentiation 8-positive (CD8⁺) T cell infiltration in CB versus NCB in the pre- and post-treatment biopsies (p = 0.09) and an increase in CD8⁺ T cells in the post-treatment biopsies of CB versus NCB (p = 0.03).

Conclusions: Although cryoactivation combined with pembrolizumab was safe and well tolerated in patients with NSCLC, therapeutic benefits were not evident compared with historical cohorts of ICI monotherapy. Correlative analyses validated CD8⁺ T cell recruitment in patients deriving CB.

Keywords: Cryoactivation; Immune checkpoint inhibitors; Immunotherapy; Non–small cell lung cancer.

Figure 1

Swimmer’s plot of time from the start of the study to the time of the last pembrolizumab treatment, including the timing of the best response. ID, identification.

Figure 2

(A) CD8⁺ T cells immune cell infiltration in tumor samples before and after cryoactivation and first dose of pembrolizumab in a tumor sample. (B) Percentage of CD8⁺ T cells in pre- and post-treatment specimens based on clinical benefit (PD versus PR or SD) with green bars representing patients deriving clinical benefit on treatment, and black bars, those not deriving clinical benefit. (C) Percentage of CD8⁺ T cells in pre- and post-treatment specimens based on median OS, with green bars representing patients with an OS of 13 months or longer (median OS) and black bars, for those with OS less than 13 months. The p values for all comparisons are indicated on the graph. CD8, cluster of differentiation 8; OS, overall survival; PD, progressive disease; PR, partial response; SD, stable disease.