Summary: We devise a novel inference algorithm to effectively solve the cancer progression model reconstruction problem. Our empirical analysis of the accuracy and convergence rate of our algorithm, CAncer PRogression Inference (CAPRI), shows that it outperforms the state-of-the-art algorithms addressing similar problems. Motivation: Several cancer-related genomic data have become available (e.g. The Cancer Genome Atlas, TCGA) typically involving hundreds of patients. At present, most of these data are aggregated in a cross-sectional fashion providing all measurements at the time of diagnosis. Our goal is to infer cancer ‘progression’ models from such data. These models are represented as directed acyclic graphs (DAGs) of collections of ‘selectivity’ relations, where a mutation in a gene A ‘selects’ for a later mutation in a gene B. Gaining insight into the structure of such progressions has the potential to improve both the stratification of patients and personalized therapy choices. Results: The CAPRI algorithm relies on a scoring method based on a probabilistic theory developed by Suppes, coupled with bootstrap and maximum likelihood inference. The resulting algorithm is efficient, achieves high accuracy and has good complexity, also, in terms of convergence properties. CAPRI performs especially well in the presence of noise in the data, and with limited sample sizes. Moreover CAPRI, in contrast to other approaches, robustly reconstructs different types of confluent trajectories despite irregularities in the data. We also report on an ongoing investigation using CAPRI to study atypical Chronic Myeloid Leukemia, in which we uncovered non trivial selectivity relations and exclusivity patterns among key genomic events. Availability and implementation: CAPRI is part of the TRanslational ONCOlogy R package and is freely available on the web at: http://bimib.disco.unimib.it/index.php/Tronco Contact:daniele.ramazzotti@disco.unimib.it Supplementary information: Supplementary data are available at Bioinformatics online.

Metabolic reprogramming is a general feature of cancer cells. Regrettably, the comprehensive quantification of metabolites in biological specimens does not promptly translate into knowledge on the utilization of metabolic pathways. By estimating fluxes across metabolic pathways, computational models hold the promise to bridge this gap between data and biological functionality. These models currently portray the average behavior of cell populations however, masking the inherent heterogeneity that is part and parcel of tumorigenesis as much as drug resistance. To remove this limitation, we propose single-cell Flux Balance Analysis (scFBA) as a computational framework to translate single-cell transcriptomes into single-cell fluxomes. We show that the integration of single-cell RNA-seq profiles of cells derived from lung adenocarcinoma and breast cancer patients into a multi-scale stoichiometric model of a cancer cell population: significantly 1) reduces the space of feasible single-cell fluxomes; 2) allows to identify clusters of cells with different growth rates within the population; 3) points out the possible metabolic interactions among cells via exchange of metabolites. The scFBA suite of MATLAB functions is available at https://github.com/BIMIB-DISCo/scFBA, as well as the case study datasets. Cytotoxicity of chemotherapeutic agents and resistance to targeted treatments are the main reasons why cancer is still one of the top causes of death. As tumor cells are intrinsically resistant to therapies that target signaling pathways, targeting the metabolic hallmarks of cancer holds promise for more incisive treatments. Regrettably, the heterogeneity of cancer metabolism hinders the identification of effective treatments. To fully uncover the metabolic heterogeneity within tumors, characterization of metabolic programs (metabolic flux distributions) at the single-cell level is required. To fill the gap between current technologies for genomics and future technologies for fluxomics, both at the single-cell and the genome-wide scale, we propose to integrate cancer data from: 1) single-cell transcriptomics and 2) bulk metabolomics, into a multi-scale stoichiometric model, to deliver for the first time metabolic fluxomes at the single-cell level. To this end, we introduce a new paradigm for flux balance analysis and data integration in cancer metabolism to: 1) characterize metabolic heterogeneity, not only at the inter-, but also at the intra-tumor level 2) identify the metabolic interactions between cancer populations, whose role in resistance to metabolic treatments has been recently recognized 3) predict the collective response to drug targeting of metabolism.

Significance: A causality-based machine learning Pipeline for Cancer Inference (PiCnIc) is introduced to infer the underlying somatic evolution of ensembles of tumors from next-generation sequencing data. PiCnIc combines techniques for sample stratification, driver selection, and identification of fitness-equivalent exclusive alterations to exploit an algorithm based on Suppes’ probabilistic causation. The accuracy and translational significance of the results are studied in detail, with an application to colorectal cancer. The PiCnIc pipeline has been made publicly accessible for reproducibility, interoperability, and future enhancements.

Existing techniques to reconstruct tree models of progression for accumulative processes, such as cancer, seek to estimate causation by combining correlation and a frequentist notion of temporal priority. In this paper, we define a novel theoretical framework called CAPRESE (CAncer PRogression Extraction with Single Edges) to reconstruct such models based on the notion of probabilistic causation defined by Suppes. We consider a general reconstruction setting complicated by the presence of noise in the data due to biological variation, as well as experimental or measurement errors. To improve tolerance to noise we define and use a shrinkage-like estimator. We prove the correctness of our algorithm by showing asymptotic convergence to the correct tree under mild constraints on the level of noise. Moreover, on synthetic data, we show that our approach outperforms the state-of-the-art, that it is efficient even with a relatively small number of samples and that its performance quickly converges to its asymptote as the number of samples increases. For real cancer datasets obtained with different technologies, we highlight biologically significant differences in the progressions inferred with respect to other competing techniques and we also show how to validate conjectured biological relations with progression models.

To dissect the mechanisms underlying the inflation of variants in the SARS-CoV-2 genome, we present a large-scale analysis of intra-host genomic diversity, which reveals that most samples exhibit heterogeneous genomic architectures, due to the interplay between host-related mutational processes and transmission dynamics. The decomposition of minor variants profiles unveils three non-overlapping mutational signatures related to nucleotide substitutions and likely ruled by APOBEC, Reactive Oxygen Species and ADAR, highlighting heterogeneous host responses to SARS-CoV-2 infections. A corrected-for-signatures dN/dS analysis demonstrates that such mutational processes are affected by purifying selection, with important exceptions. In fact, several mutations appear to transit toward clonality, defining new clonal genotypes that increase the overall genomic diversity. Furthermore, the phylogenomic analysis shows the presence of homoplasies and supports the hypothesis of transmission of minor variants. This study paves the way for the integrated analysis of intra-host genomic diversity and clinical outcome of SARS-CoV-2 infections.