Multicenter, Single-Arm, Prospective Phase II Study of Neoadjuvant Cryo-Thermal Ablation Combined With Toripalimab and Chemotherapy for Resectable Stage IB-IIIB Non-Small Cell Lung Cancer

Pathological Complete Response (pCR) Rate

The proportion of enrolled participants who achieve pathological complete response (pCR), defined as no residual viable tumor cells in the resected primary tumor and all sampled regional lymph nodes after neoadjuvant treatment (ypT0N0). Participants who do not undergo surgical resection or for whom pathological response cannot be adequately assessed will not be considered to have achieved pCR in the primary efficacy analysis.

Objective Response Rate (ORR)

The proportion of enrolled participants who achieve a complete response (CR) or partial response (PR) as their radiographic tumor response before surgery, as assessed according to Response Evaluation Criteria in Solid Tumors (RECIST) version 1.1.

Major Pathological Response (MPR) Rate

The proportion of enrolled participants who achieve major pathological response (MPR), defined as ≤10% residual viable tumor cells within the primary tumor bed in the resected lung specimen after neoadjuvant treatment, based on pathological assessment according to IASLC recommendations. Pathological response in regional lymph nodes will be assessed and reported separately and will not be included in the primary definition of MPR.

R0 resection rate

The proportion of participants undergoing surgical resection who achieve an R0 resection, defined as complete macroscopic resection of the tumor with microscopically negative surgical margins and no residual tumor at the resection margins.

Event-Free Survival (EFS)

Event-free survival (EFS) is defined as the time from enrollment to the first occurrence of any of the following events: radiographic disease progression; local disease progression resulting in unresectability; inability to undergo the planned curative-intent surgery because of treatment-related toxicity or deterioration in clinical condition attributable to study treatment; failure to achieve tumor resection because of intraoperative unresectability; local or distant disease recurrence after surgery; or death from any cause. Participants without an EFS event will be censored at the date of the last disease assessment.

Changes in the Spatial Tumor Immune Microenvironment Assessed by Atera In Situ Spatial Transcriptomics

Atera In Situ single-cell spatial transcriptomic profiling will be performed on FFPE tumor specimens. Within-participant changes from the baseline tumor biopsy before Cryo-Thermal Ablation to the surgical tumor specimen after neoadjuvant chemoimmunotherapy will be evaluated. Spatial immune features in the surgical specimen will also be compared among residual tumor, adjacent tissue, and normal lung tissue when available. Analyses will characterize spatial distributions and transcriptional states of CD8+ T cells, regulatory T cells, myeloid populations, interferon-related signaling, antigen-presentation pathways, and T-cell activation/exhaustion programs. Features will also be compared between participants with and without MPR; pCR will be explored as a subgroup.

Changes in Peripheral and Tumor T-cell Receptor Clonality Assessed by Bulk TCR Sequencing

Bulk T-cell receptor sequencing (TCR-seq) will be performed on peripheral blood mononuclear cells and tumor tissue to characterize treatment-associated T-cell clonal dynamics. Peripheral blood will be collected at baseline before Cryo-Thermal Ablation, approximately 14 days after ablation before Cycle 1, before Cycles 2 and 3 of neoadjuvant chemoimmunotherapy, and after completion of neoadjuvant therapy before surgery. TCR repertoire diversity, clonality, and clonal expansion will be evaluated longitudinally. Tumor tissue from the baseline biopsy and surgical resection specimen will also undergo TCR-seq. Changes in intratumoral T-cell clonality and overlap between blood and tumor TCR clonotypes will be explored in relation to pathological response and treatment-related toxicity.

Tumor Genomic Alterations Assessed by a 919-Gene Next-Generation Sequencing Panel

Baseline FFPE tumor biopsy specimens will undergo next-generation sequencing using a 919-gene cancer-associated panel. Somatic genomic alterations, including relevant mutations and other detectable genomic features, will be characterized. Exploratory analyses will evaluate associations between baseline tumor genomic characteristics and pathological response to Cryo-Thermal Ablation combined with neoadjuvant chemoimmunotherapy.

Longitudinal Changes in Circulating Tumor DNA and Molecular Residual Disease

Circulating tumor DNA (ctDNA) will be longitudinally assessed to characterize changes in tumor-derived molecular signals during treatment and postoperative surveillance. During neoadjuvant treatment, ctDNA levels and detectable tumor-associated alterations will be evaluated at baseline before ablation, 14 days after ablation, and at the preoperative assessment after completion of neoadjuvant therapy. ctDNA clearance and changes in variant allele abundance will be explored according to pathological response. Serial postoperative ctDNA testing will be used to assess molecular residual disease (MRD) and its relationship with subsequent disease recurrence. Longitudinal ctDNA profiles will also be explored for changes in detectable tumor clones over time.