Figure 1. Overview of BEECH study exploratory analysis

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Figure 1. Overview of BEECH study exploratory analysis Figure 1. Overview of BEECH study exploratory analysis. (A) Schema of plasma collection during the BEECH trial for both ... Figure 1. Overview of BEECH study exploratory analysis. (A) Schema of plasma collection during the BEECH trial for both development part A and validation part B cohorts. Red arrows indicate part A sampling only, blue part B sampling only and purple part A and B shared timepoint sampling. Tracking samples were collected on day 1 of each treatment cycle. (B) CONSORT diagrams of part A and part B exploratory plasma baseline analysis. Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) 2019. Published by Oxford University Press on behalf of the European Society for Medical Oncology.This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com Ann Oncol, mdz085, https://doi.org/10.1093/annonc/mdz085 The content of this slide may be subject to copyright: please see the slide notes for details.

Figure 2. Identification of optimal circulating tumour DNA (ctDNA) early timepoint for prediction of progression-free ... Figure 2. Identification of optimal circulating tumour DNA (ctDNA) early timepoint for prediction of progression-free survival (PFS) length in the development cohort. (A) Circulating tumour DNA ratios (CDRs) at designated timepoints in the first 4 weeks of study treatment, separated by short and long PFS in study part A. Differing numbers in the long and short groups reflect missed sample collection timepoints in some patients. Long and short PFS was determined by a scan at 12 weeks on study, the time-point of the first scan in part A. C2D1 (CDR28) was the strongest predictive timepoint P = 0.0007, P value Mann–Whitney U test. (B) PFS for development part A patients split by CDR28 suppressed (CDR28<0.25) versus CDR28 high. P value log rank test. (C) Longitudinal tracking of a PIK3CA c.1624G>A (p.E542K) mutation in a patient classified as a long PFS by ctDNA demonstrating successful suppression of ctDNA before rise before progression. (D) Longitudinal tracking of a TP53 c.815T>C (p.V272A) mutation in a patient classified as a short PFS by ctDNA demonstrating failure to suppress ctDNA in the first 4 weeks of treatment. Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) 2019. Published by Oxford University Press on behalf of the European Society for Medical Oncology.This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com Ann Oncol, mdz085, https://doi.org/10.1093/annonc/mdz085 The content of this slide may be subject to copyright: please see the slide notes for details.

Figure 3. Early ctDNA dynamics are a surrogate for progression-free survival (PFS) in the independent validation ... Figure 3. Early ctDNA dynamics are a surrogate for progression-free survival (PFS) in the independent validation cohort. (A) PFS for validation cohort part B split by CDR28 suppressed (CDR28<0.25) versus CDR28 high. P value log rank test. (B) PFS for high versus suppressed CDR28, stratified for placebo (top) and capivasertib (bottom) treatment arms. (C) Box plot of CDR28 by treatment arm for all patients and CDR28 for treatment arms subdivided by PIK3CA mutation status. Patients with undetectable ctDNA at C2 D1 were assigned an arbitrary low value for statistical analysis. P value from Mann–Whitney U test, not significant across the groups. Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) 2019. Published by Oxford University Press on behalf of the European Society for Medical Oncology.This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com Ann Oncol, mdz085, https://doi.org/10.1093/annonc/mdz085 The content of this slide may be subject to copyright: please see the slide notes for details.

Figure 4. Circulating tumour DNA (ctDNA) dynamics during treatment and lead-time over clinical progression. (A) ... Figure 4. Circulating tumour DNA (ctDNA) dynamics during treatment and lead-time over clinical progression. (A) Longitudinal tracking of a PIK3CA c.1633G>A (p.E545K) mutation in a patient who demonstrates a clear fall to a sustained nadir and a clear rise before clinical progression. (B) Longitudinal tracking of a PIK3CA c.3140 A>G (p.H1047R) mutation in a patient who demonstrates a fall in ctDNA which then fluctuates at a low level across multiple timepoints. (C) Longitudinal tracking of a PIK3R1 DelCTGAGA (p.L573_R574del) deletion in a patient who demonstrates a clear fall in ctDNA but a gradual rise over subsequent cycles before clinical progression. (D) Distribution of lead-time of calculated molecular progression before confirmed clinical progression (range 0–329 days). Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) 2019. Published by Oxford University Press on behalf of the European Society for Medical Oncology.This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com Ann Oncol, mdz085, https://doi.org/10.1093/annonc/mdz085 The content of this slide may be subject to copyright: please see the slide notes for details.

Figure 5. Clonal haematopoiesis of indeterminate potential (CHIP) is frequently detected in advanced breast cancer. (A) ... Figure 5. Clonal haematopoiesis of indeterminate potential (CHIP) is frequently detected in advanced breast cancer. (A) Clonal haematopoiesis variants detected by sequencing of baseline plasma in part B. (B) Clinical and pathological associations of CHIP detection in baseline plasma. (C) Longitudinal tracking of a PIK3CA c.3140 A>G (p.H1047R) mutation and a DNMT3A CHIP splice site donor c.25236935 C>T variant in the same patient. (D) Progression-free survival by baseline detection of CHIP for the whole study population, capivasertib and placebo treatment arms. P values from log rank test. Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) 2019. Published by Oxford University Press on behalf of the European Society for Medical Oncology.This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com Ann Oncol, mdz085, https://doi.org/10.1093/annonc/mdz085 The content of this slide may be subject to copyright: please see the slide notes for details.