Supplementary Experimental Methods

PDF file - 104KB, Clinical features of IDH1-mutant intrahepatic cholangiocarcinoma patients in the Screening cohort.

PDF file - 67KB, Characteristics of all intrahepatic cholangiocarcinoma patients evaluated across the two cohorts.

PancSeq protocol gene list for CLIA-certified somatic and germline analysis from whole exome sequencing data.

Genes differentially expressed in ACEi alone treated group compared to control group

Complete results for GO (Supplementary Table 8) and REACTOME (supplementary Table 9) analysis on differentially expressed genes comparing lisinopril vs. control PDAC tumors

Adjusted Analysis with Multivariate Model on TCGA PDAC dataset to test for independent factors.

Selected key pathways up-regulated or down-regulated by ACEi treatment alone in Gene Set Enrichment Analysis.

PDF file - 57KB, Levels of serum 2HG relative to tumor burden in IDH1-mutant and IDH2-mutant intrahepatic cholangiocarcinoma patients in the Validation cohort.

Supplementary Figure S1: Overview of workflow and data generation. Supplementary Figure S2: Analysis of neoplastic cellularity. Supplementary Figure S3: Recurrent copy number alterations. Supplementary Figure S4: Mutational signature analysis. Supplementary Figure S5: Analysis of PDAC gene expression signatures. Supplementary Figure S6: Clinically relevant alterations in the cohort.

Treatment history for patients enrolled on the PancSeq protocol.

Importance Treatment options are limited for patients with advanced pancreatic ductal adenocarcinoma (PDAC) beyond first-line 5-fluorouracil, leucovorin, irinotecan, and oxaliplatin (FOLFIRINOX), with such individuals commonly being treated with gemcitabine and nab-paclitaxel. Objective To determine whether NPC-1C, an antibody directed against MUC5AC, might increase the efficacy of second-line gemcitabine and nab-paclitaxel in patients with advanced PDAC. Design, Setting, and Participants This multicenter, randomized phase II clinical trial enrolled patients with advanced PDAC between April 2014 and March 2017 whose disease had progressed on first-line FOLFIRINOX. Eligible patients had tumors with at least 20 MUC5AC staining by centralized immunohistochemistry review. Statistical analysis was performed from April to May 2022. Interventions Patients were randomly assigned to receive gemcitabine (1000 mg/m 2 ) and nab-paclitaxel (125 mg/m 2 ) administered intravenously on days 1, 8, and 15 of every 4-week cycle, with or without intravenous NPC-1C 1.5 mg/kg every 2 weeks. Main Outcomes and Measures The primary end point was overall survival (OS). Secondary end points were progression-free survival (PFS), objective response rate (ORR), and safety. Pretreatment clinical variables were explored with Cox proportional hazards analysis. Results A total of 78 patients (median [range] age, 62 [36-78] years; 32 [41%] women; 9 [12%] Black; 66 [85%] White) received second-line treatment with gemcitabine plus nab-paclitaxel (n = 40) or gemcitabine plus nab-paclitaxel and NPC-1C (n = 38). Median OS was 6.6 months (95% CI, 4.7-8.4 months) with gemcitabine plus nab-paclitaxel vs 5.0 months (95% CI, 3.3-6.5 months; P = .22) with gemcitabine plus nab-paclitaxel and NPC-1C. Median PFS was 2.7 months (95% CI, 1.9-4.1 months) with gemcitabine plus nab-paclitaxel vs 3.4 months (95% CI, 1.9-5.3 months; P = .80) with gemcitabine plus nab-paclitaxel and NPC-1C. The ORR was 3.1% (95% CI, 0.4%-19.7%) in the gemcitabine plus nab-paclitaxel and NPC-1C group and 2.9% (95% CI, 0.4%-18.7%) in the gemcitabine plus nab-paclitaxel group. No differences in toxicity were observed between groups, except that grade 3 or greater anemia occurred more frequently in patients treated with gemcitabine plus nab-paclitaxel and NPC-1C than gemcitabine plus nab-paclitaxel (39% [15 of 38] vs 10% [4 of 40]; P = .003). The frequency of chemotherapy dose reductions was similar in both groups (65% vs 74%; P = .47). Lower performance status, hypoalbuminemia, PDAC diagnosis less than or equal to 18 months before trial enrollment, lymphocyte-to-monocyte ratio less than 2.8, and CA19-9 greater than 2000 IU/mL were independently associated with poorer survival. Conclusions and Relevance In this randomized clinical trial of advanced PDAC, NPC-1C did not enhance the efficacy of gemcitabine/nab-paclitaxel. These data provide a benchmark for future trials investigating second-line treatment of PDAC. Trial Registration ClinicalTrials.gov Identifier: NCT01834235

Supplementary Figure 1: Clinical status of patients included in the analysis Supplementary Figure 2: Unadjusted Kaplan Meier survival curves of patients who received ACEi (green), ARB (yellow) vs. control group (blue). Supplementary Figure 3: Chronic ASI use in resected patients with hypertension. Supplementary Figure 4: Unadjusted Kaplan Meier survival curve of all resected patients stratified by BMI and chemotherapy. Supplementary Figure 5: Unadjusted Kaplan Meier curves for disease recurrence.

Supplementary Figures and Legends

Supplementary Figures, Tables, Methods

Characteristics of biopsies obtained on PancSeq protocol.

Patient characteristics, unadjusted univariate model and adjusted analysis with multivariate model analyses in metastatic patients (supplementary table 1) and locally advanced patients (supplementary table 2) Supplementary Table 3: Unadjusted Univariate Analysis of Resected Patients with Hypertension Supplementary Table 4: Propensity score analysis Supplementary Table 5: Competing risk analysis in resected patients Supplementary Table 6: Multivariate model analysis of time to recurrence.

Supplementary table and figure legends

Characteristics of patients enrolled to PancSeq protocol