Fig. 2. RFS in patients not treated with adjuvant chemotherapy. (A) Kaplan-Meier estimates of RFS for all patients not treated with adjuvant chemotherapy, stratified by postoperative ctDNA status. (B) Kaplan-Meier estimates of RFS in the same patients, stratified by clinicopathologic characteristics. (C) Kaplan-Meier estimates of RFS stratified by postoperative ctDNA status in patients with low-risk clinicopathologic characteristics. (D) Kaplan-Meier estimates of RFS stratified by postoperative ctDNA status in patients with high-risk clinicopathologic characteristics. The high-risk group is defined as those having mismatch repair–proficient (pMMR) tumors with at least one of the following poor prognostic features: T4, LN yield <12, LVI, and poor tumor differentiation. The low-risk group is defined as those with no poor prognostic features.
Fig. 3. ctDNA status before, during, and after adjuvant chemotherapy. (A to F) ctDNA concentrations (% mutant alleles) for the six patients with positive postoperative ctDNA who subsequently received adjuvant chemotherapy. The blue shaded box indicates the period during which adjuvant chemotherapy was administered. The dotted line indicates the time of radiologic recurrence or last follow-up (if recurrence-free). The ctDNA status of patients represented in (A) and (B) initially became negative, then became positive again at the completion of adjuvant chemotherapy; both patients subsequently suffered a radiologic recurrence. Note that CEAs were not elevated in either patient at any time point. (C to F) Four patients whose ctDNA became negative after completion of chemotherapy. Three of these patients (patients C, D, and F) remained radiologic recurrence-free at their last follow-up visit. (G) Kaplan-Meier estimates of RFS according to post-chemotherapy ctDNA status in patients treated with adjuvant chemotherapy. ND, not detected; NE, not elevated.
Fig. 4. Serial ctDNA status up to radiological recurrence. (A) Serial ctDNA measurements up to the time of radiological recurrence for the nine patients who were not treated with adjuvant chemotherapy and who were ctDNA-positive postoperatively. (B) Scatter plot of the lead time to radiological recurrence for ctDNA detection and CEA elevation, with the error bars representing the median and IQR.
- Table 1. Mutations and mutant allele fractions detected in postoperative ctDNA-positive cases.
Mutations MAF (%) TP53 p.R342X 1.631 TP53 p.G245D 0.123 APC p.L878fs 0.11 TP53 p.R248Q 0.01 TP53 p.R248Q 0.006 TP53 p.R248Q 0.017 APC p.Q1406fs 0.059 APC p.E1379X 0.235 KRAS p.G13D 0.066 TP53 p.R248Q 0.678 APC p.C1578fs 0.140 KRAS p.G12D 0.027 KRAS p.G12V 0.008 TP53 p.P151fs 0.003 APC p.S1436fs 0.046 TP53 p.R282W 1.774 APC p.D1178fs 0.007 APC p.E1408fs 0.046 TP53 p.I254fs 0.05 APC p.S1010fs 0.006 - Table 2. Patient and tumor characteristics according to postoperative ctDNA status.
Characteristics* All patients
(n = 230)Postoperative
ctDNA-positive
(n = 20)Postoperative
ctDNA-negative
(n = 210)Age (years) Median
[interquartile
range (IQR)]65 (59–73) 64 (54–68) 66 (59–74) Range 23–87 43–86 23–87 Sex, no. (%) Male 131 (57) 10 (50) 121 (58) Female 99 (43) 10 (50) 89 (42) Tumor site, no. (%)† Right colon 103 (45) 5 (25) 98 (47) Left colon 127 (55) 15 (75) 112 (53) Differentiation, no. (%) Well/moderate 193 (84) 18 (90) 175 (83) Poor 37 (16) 2 (10) 35 (17) T stage, no. (%) T3 192 (83) 17 (85) 175 (83) T4 38 (17) 3 (15) 35 (17) Lymph node yield, no. (%) <12 29 (13) 5 (25) 24 (11) ≥12 201 (87) 15 (75) 186 (89) Lymphovascular invasion, no./total no. (%) Yes 41/221 (19) 6/20 (30) 35/201 (17) No 180/221 (81) 14/20 (70) 166/201 (83) MMR status, no. (%) Proficient 189 (82) 19 (95) 170 (81) Deficient 41 (18) 1 (5) 40 (19) Clinicopathologic risk group, no. (%) Low 140/224 (63) 10 (50) 130/204 (64) High 84/224 (37) 10 (50) 74/204 (36) Adjuvant chemotherapy, no. (%)‡ Yes 52 (23) 6 (30) 46 (22) No 178 (77) 14 (70) 164 (78) *There were no significant differences between the groups in any of the characteristics listed in this table.
†Right-sided colon cancer was defined as tumors arising from the cecum, ascending, hepatic flexure, or transverse colon; left-sided colon cancer was defined as tumors arising from the splenic flexure, descending, sigmoid, or rectosigmoid colon.
‡Except for two cases where oxaliplatin-based chemotherapy was used, all other adjuvant treatment was fluorouracil-based chemotherapy.
- Table 3. Recurrence-free survival analysis by clinicopathological variables and postoperative ctDNA status.
Variable Univariate analysis Multivariate analysis HR 95% CI P HR 95% CI P Patients not treated with chemotherapy (n = 178) Age, <70 versus ≥70 0.92 0.43–2.0 0.8 Sex, male versus female 1.3 0.62–2.8 0.5 Tumor site, right versus left 1.5 0.69–3.3 0.3 Tumor differentiation, well/moderate versus poor 0.39 0.09–1.7 0.2 T stage, T3 versus T4 4.0 1.7–9.5 0.002 8.1 3.1–21 <0.001 Lymph node yield, ≥12 versus <12 3.1 1.3–7.4 0.009 Lymphovascular invasion, no versus yes 2.4 1.1–5.4 0.03 MMR status, deficient versus proficient 3.6 0.86–15 0.08 Clinicopathologic risk group, low versus high 3.2 1.5–6.9 0.002 Postoperative CEA, normal versus elevated 1.6 0.37–6.8 0.5 Postoperative ctDNA status, negative versus positive 18 7.9–40 <0.001 28 11–68 <0.001 All patients (n = 230) Age, <70 versus ≥70 1.0 0.50–2.0 1.0 Sex, male versus female 1.1 0.57–2.2 0.7 Tumor site, right versus left 1.1 0.55–2.1 0.8 Tumor differentiation, well/moderate versus poor 0.32 0.08–1.3 0.1 T stage, T3 versus T4 2.4 1.2–5.1 0.02 2.6 1.2–5.5 0.01 Lymph node yield, ≥12 versus <12 2.2 0.97–4.8 0.06 Lymphovascular invasion, no versus yes 1.9 0.92–4.1 0.08 MMR status, deficient versus proficient 3.5 0.83–14.5 0.09 Clinicopathologic risk group, low versus high 2.1 1.06–4.2 0.03 Adjuvant chemotherapy, no versus yes 0.79 0.34–1.8 0.6 Postoperative CEA, normal versus elevated 2.8 0.98–7.9 0.06 Postoperative ctDNA status, negative versus positive 13 6.6–27 <0.001 14 6.8–28 <0.001
Supplementary Materials
www.sciencetranslationalmedicine.org/cgi/content/full/8/346/346ra92/DC1
Fig. S1. Recurrence-free survival in patients not treated with adjuvant chemotherapy stratified by postoperative CEA status.
Table S1. Relationship between postoperative ctDNA, postoperative CEA, and recurrence status for patients not treated with chemotherapy.
Table S2. Time-dependent predictive accuracy of postoperative ctDNA for recurrence at 5, 6, 12, 18, 24, 36, and 40 months.
Table S3. Serial circulating biomarker status and clinical characteristics of patients not treated with adjuvant chemotherapy who experienced recurrence.
Table S4. Serial circulating biomarker status and clinical characteristics of chemotherapy-treated patients who experienced recurrence.
Table S5. Genomic and clinical summary for all patients included in the final evaluable population.
Table S6. Panel of 15 genes used for identification of somatic mutations in tumor tissue.
Additional Files
- Supplementary Material for:
Circulating tumor DNA analysis detects minimal residual disease and predicts recurrence in patients with stage II colon cancer
Jeanne Tie,* Yuxuan Wang, Cristian Tomasetti, Lu Li, Simeon Springer, Isaac Kinde, Natalie Silliman, Mark Tacey, Hui-Li Wong, Michael Christie, Suzanne Kosmider, Iain Skinner, Rachel Wong, Malcolm Steel, Ben Tran, Jayesh Desai, Ian Jones, Andrew Haydon, Theresa Hayes, Tim J. Price, Robert L. Strausberg, Luis A. Diaz Jr., Nickolas Papadopoulos, Kenneth W. Kinzler, Bert Vogelstein,* Peter Gibbs*
*Corresponding author. Email: jeanne.tie{at}mh.org.au (J.T.); vogelbe{at}jhmi.edu (B.V.); peter.gibbs{at}mh.org.au (P.G.)
Published 06 July 2016, Sci. Transl. Med. 8, 346ra92 (2016)
DOI: 10.1126/scitranslmed.aaf6219This PDF file includes:
- Fig. S1. Recurrence-free survival in patients not treated with adjuvant chemotherapy stratified by postoperative CEA status.
- Table S1. Relationship between postoperative ctDNA, postoperative CEA, and recurrence status for patients not treated with chemotherapy.
- Table S2. Time-dependent predictive accuracy of postoperative ctDNA for recurrence at 5, 6, 12, 18, 24, 36, and 40 months.
- Table S3. Serial circulating biomarker status and clinical characteristics of patients not treated with adjuvant chemotherapy who experienced recurrence.
- Table S4. Serial circulating biomarker status and clinical characteristics of chemotherapy-treated patients who experienced recurrence.
- Table S5. Genomic and clinical summary for all patients included in the final evaluable population.
- Table S6. Panel of 15 genes used for identification of somatic mutations in tumor tissue.
