Lymph Node Pancreatic Cancer Vaccine Trial Results
# Methods
## Study Design and Participants
This prospective, open-label, phase I clinical trial (NCT05293168) evaluated the safety and preliminary efficacy of personalized neoantigen-specific T cell therapy in patients with advanced solid tumors. Eligible patients had histologically confirmed advanced solid tumors refractory to standard therapies,an ECOG performance status of 0 or 1,and measurable disease according to RECIST v1.1 criteria. Key exclusion criteria included prior allogeneic stem cell transplantation, active autoimmune disease, or significant cardiac, pulmonary, or renal dysfunction. All patients provided written informed consent prior to enrollment. The study was conducted in accordance with the declaration of Helsinki and approved by the Institutional Review Board (IRB) at each participating center.
## Neoantigen Identification and T Cell Production
tumor tissue was obtained from patients via biopsy or surgical resection. Whole-exome sequencing (WES) and RNA sequencing (RNA-seq) were performed on tumor and matched normal samples to identify somatic mutations. Predicted neoantigens were filtered based on high predicted immunogenicity scores, HLA binding affinity, and expression levels. A panel of up to 10 neoantigens per patient was selected for T cell targeting.
Peripheral blood mononuclear cells (PBMCs) were collected from each patient via leukapheresis. Dendritic cells (DCs) were generated from monocytes and pulsed with RNA encoding the selected neoantigens. These neoantigen-pulsed DCs were then co-cultured with patient-derived T cells to stimulate neoantigen-specific T cell expansion. Expanded T cells were cryopreserved and underwent quality control testing, including assessment of phenotype, viability, and neoantigen specificity via IFN-γ ELISpot assay.
## T Cell Therapy Administration
Prior to T cell infusion, patients received lymphodepletion with cyclophosphamide (300 mg/m2) and fludarabine (30 mg/m2) on days -5 and -4, respectively. Neoantigen-specific T cells were infused on day 0 at a target dose of 1 x 106 cells/kg. Patients were monitored closely for signs of cytokine release syndrome (CRS) and immune effector cell-associated neurotoxicity syndrome (ICANS) following T cell infusion, using established grading scales. Corticosteroids were administered as needed to manage CRS or ICANS.
## Assessments
Patient assessments included physical examinations, vital sign monitoring, and adverse event (AE) reporting.Tumor response was evaluated every 8 weeks using RECIST v1.1 criteria, including computed tomography (CT) or magnetic resonance imaging (MRI). Peripheral blood samples were collected at baseline, during T cell expansion, and post-infusion to assess T cell kinetics, neoantigen specificity, and immune-related biomarkers. Tumor biopsies were collected pre- and post-infusion, when feasible, to evaluate changes in the tumor microenvironment and T cell infiltration.
## oligonucleotide Synthesis
Mutated peptide sequences were designed (generally with the mutation centered in the middle) and then Genscript synthesized two 15-mers overlapping by 11 to cover the mutated 18-mer (18-mer sequences found in Supplementary Table 1).
## Statistical Analysis
Descriptive statistics were used to summarize demographic, medical history and safety data. Continuous variables were summarized using mean, s.d., median, minimum value and maximum value. Categorical variables were summarized using frequency counts and percentages. Clinical efficacy outcomes, such as tumor biomarker reduction or clearance, were examined for association with categorical variables, including high versus low T cell response, using the Mann-Whitney test. The Kaplan-meier method was used to estimate the survival distributions. The log-rank test was used to compare the RFS between the high and low T cell responders and the ROC analysis was performed using a logistic regression model. SAS v9.4 and R v4.4.3 were used to create Fig. 1 and Extended Data Figs. 2-4 and perform statistical analysis. GraphPad Prism v9.4 was used to create Figs. 1 and 2 and Extended Data Figs. 5 and 6 and perform statistical analysis.
## Reporting Summary
Further facts on research design is available in the nature Portfolio Reporting Summary linked to this article.