eprintid: 10203239 rev_number: 13 eprint_status: archive userid: 699 dir: disk0/10/20/32/39 datestamp: 2025-03-14 13:46:45 lastmod: 2025-03-14 13:46:45 status_changed: 2025-03-14 13:46:45 type: thesis metadata_visibility: show sword_depositor: 699 creators_name: Pawlik, Piotr title: Inference of clone-specific copy numbers from bulk tumour DNA ispublished: unpub divisions: UCL divisions: B02 divisions: C10 divisions: D19 divisions: G99 note: Copyright © The Author 2025. Original content in this thesis is licensed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International (CC BY-NC 4.0) Licence (https://creativecommons.org/licenses/by-nc/4.0/). Any third-party copyright material present remains the property of its respective owner(s) and is licensed under its existing terms. Access may initially be restricted at the author’s request. abstract: Cancer is an evolutionary process. During this process, single nucleotide variants (SNVs) and somatic copy-number alterations (SCNAs) accumulate, contributing to clonal evolution. Nevertheless, making accurate inferences about their co-evolution and clone-specific copy numbers from bulk DNA sequencing poses a significant challenge. In this thesis, I present ALPACA (ALlelespecific Phylogenetic Analysis of clone Copy-number Alterations), a computational method which allows inference of clone-specific copy-numbers from bulk DNA by leveraging phylogenetic trees and SNV cellular frequencies obtained from multi-sample sequencing experiments. I validate this approach with two simulated tumour cohorts and with single-cell data, comparing the ALPACA output both with the ground truth and with the outputs of two previously published methods. I then apply ALPACA to the TRACERx421 (TRAcking non-small cell lung Cancer Evolution through therapy Frankell et al. 2023; Al Bakir et al. 2023) cohort and demonstrate that ALPACA can identify loss-ofheterozygosity events and amplifications in minor metastasis seeding clones, not detectable on bulk-sample copy-number level. By comparing seeding and non-seeding clones within the primary-metastatic matched cohort I observe evidence of increased chromosomal instability in metastasis seeding clones. Using ALPACA’s clone-specific copy-numbers I develop a novel method to quantify intra-tumour copy-number heterogeneity: clone copy-number diversity (CCD). Using this metric I show that patients who later develop a metastatic disease harbour a higher intra-tumour clone copy number diversity within their primary tumours compared to patients who do not relapse. Lastly, I show an association between CCD and patient survival. date: 2025-01-28 date_type: published full_text_type: other thesis_class: doctoral_embargoed thesis_award: Ph.D language: eng verified: verified_manual elements_id: 2352203 lyricists_name: Pawlik, Piotr lyricists_id: PPAWL31 actors_name: Pawlik, Piotr actors_id: PPAWL31 actors_role: owner full_text_status: restricted pages: 207 institution: UCL (University College London) department: UCL Cancer Institute thesis_type: Doctoral citation: Pawlik, Piotr; (2025) Inference of clone-specific copy numbers from bulk tumour DNA. Doctoral thesis (Ph.D), UCL (University College London). document_url: https://discovery.ucl.ac.uk/id/eprint/10203239/5/Thesis_PHD_Piotr_Pawlik.pdf