American Association for Cancer Research
10780432ccr191213-sup-220540_3_supp_5682624_pvzqxd.pdf (2.21 MB)

Figure S1-S7 from Perioperative Dynamic Changes in Circulating Tumor DNA in Patients with Lung Cancer (DYNAMIC)

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journal contribution
posted on 2023-03-31, 21:05 authored by Kezhong Chen, Heng Zhao, Yanbin Shi, Fan Yang, Lien Tu Wang, Guannan Kang, Yuntao Nie, Jun Wang

Figure S1. (A) Strategy for plasma detection of oncogenic mutations. The exonic positions and nature of the hotspot mutations are shown. Two sets of bidirectional reverse primers (arrows) were positioned to robust sequences on either side of each mutation site to target and amplify each allele by inverse PCR. For detection of ALK fusions, targeting primers were progressively spaced across exon 19, intron 19, and exon 20, spanning the known ALK breakpoint region. (B) Hot Spot Gene Mutations and Diagnostic cSMART Primers. Figure S2. Comparison of the distributions of driver mutations identified in 26 matched tDNA and plasma ctDNA samples. Thirty-two mutations were detected in the plasma at time A, including 14 TP53 mutations, 8 EGFR mutations, 6 PIK3CA mutations, 3 KRAS mutations and 1 ALK mutation. Five patients had two mutations simultaneously in the plasma ctDNA: case 3 had PIK3CA and EGFR mutations; case 6 had EGFR and TP53 mutations; case 15 had two different TP53 mutations, namely, a base substitution mutation in exon 5 and a base substitution mutation in exon 8; case 17 had EGFR and TP53 mutations; and case 20 had KRAS and PIK3CA mutations simultaneously. A single mutation was detected in the other 21 cases. For the corresponding tumor tissues, a total of 29 gene mutations were detected: 14 TP53 mutations, 8 EGFR mutations, 5 PIK3CA mutations and 2 KRAS mutation. More than one mutation was detected in the tumors of 5 patients, which was completely consistent with the ctDNA results. (A) Numbers of gene mutations in tDNA vs. plasma ctDNA from matched sample pairs. (B) The inner circle shows the distribution of gene mutations, and the outer ring shows specific alterations in amino acids. Figure S3. Excluded two positive plasma A patients with CHIP validated by normal lung tissue and white blood cell. Figure S4. The plasma ctDNA concentration at each time point during the perioperative period (from time A to time P2). The X-axis represents the logarithmic value of time. Figure S5. Longitudinal ctDNA profiles of non-relapse or non-progression cases. Red curve represents the fluctuation of ctDNA concentration over time. The X-axis represents the days of post-operation. The Y-axis represents the ctDNA MAF (%). In this figure, rectangular frames represent the treatment that patients have received, as shown at the bottom, green frame: surgical treatment; yellow frame: chemotherapy; blue frame: radiotherapy and red frame: target therapy. Figure S6. The RFS (left) and OS (right)between different ctDNA statuses at time P3 (Three cases with no blood samples at this time point were excluded). Figure S7. The RFS between patients who received postoperative therapy or not based on different strategy. (A) Patients with stage II, III and above lung cancer according to TNM stage in our study. (B) Patients with positive MRD detection in our study. (C) Patient with positive ctDNA at time P2.


National Natural Science Foundation of China



No study has investigated the precise perioperative dynamic changes in circulating tumor DNA (ctDNA) in any patients with early-stage cancer. This study (DYNAMIC) investigated perioperative dynamic changes in ctDNA and determined the appropriate detection time of ctDNA-based surveillance for surgical patients with lung cancer.Experimental Design: Consecutive patients who underwent curative-intent lung resections were enrolled prospectively (NCT02965391). Plasma samples were obtained at multiple prespecified time points including before surgery (time A), during surgery after tumor resection (time B–time D), and after surgery (time P1–time P3). Next-generation sequencing–based detection platform was performed to calculate the plasma mutation allele frequency. The primary endpoint was ctDNA half-life after radical tumor resection. Thirty-six patients showed detectable mutations in time A. The plasma ctDNA concentration showed a rapid decreasing trend after radical tumor resection, with the average mutant allele fraction at times A, B, C, and D being 2.72%, 2.11%, 1.14%, and 0.17%, respectively. The median ctDNA half-life was 35.0 minutes. Patients with minimal residual disease (MRD) detection had a significant slower ctDNA half-life than those with negative MRD (103.2 minutes vs. 29.7 minutes, P = 0.001). The recurrence-free survival of patients with detectable and undetectable ctDNA concentrations at time P1 was 528 days and 543 days, respectively (P = 0.657), whereas at time P2 was 278 days and 637 days, respectively (P = 0.002). ctDNA decays rapidly after radical tumor resection. The ctDNA detection on the third day after R0 resection can be used as the baseline value for postoperative lung cancer surveillance.

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