Search bioRxiv⌕ Search

bioRxiv · 10.1101/2020.01.21.907691

Deciphering the Signaling Network Landscape of Breast Cancer Improves Drug Sensitivity Prediction

Abstract

Although genetic and epigenetic abnormalities in breast cancer have been extensively studied, it remains difficult to identify those patients who will respond to particular therapies. This is due in part to our lack of understanding of how the variability of cellular signaling affects drug sensitivity. Here, we used mass cytometry to characterize the single-cell signaling landscapes of 62 breast cancer cell lines and five lines from healthy tissue. We quantified 34 markers in each cell line upon stimulation by the growth factor EGF in the presence or absence of five kinase inhibitors. These data - on more than 80 million single cells from 4,000 conditions - were used to fit mechanistic signaling network models that provide unprecedented insights into the biological principles of how cancer cells process information. Our dynamic single-cell-based models more accurately predicted drug sensitivity than static bulk measurements for drugs targeting the PI3K-MTOR signaling pathway. Finally, we identified genomic features associated with drug sensitivity by using signaling phenotypes as proxies, including a missense mutation in DDIT3 predictive of PI3K-inhibition sensitivity. This provides proof of principle that single-cell measurements and modeling could inform matching of patients with appropriate treatments in the future. One-linerSingle-cell proteomics coupled to perturbations improves accuracy of breast tumor drug sensitivity predictions and reveals mechanisms of sensitivity and resistance. HIGHLIGHTSO_LIMass cytometry study of signaling responses of 62 breast cancer cell lines and five lines from healthy tissue to EGF stimulation with or without perturbation with five kinase inhibitors. C_LIO_LISingle-cell signaling features and mechanistic signaling network models predicted drug sensitivity. C_LIO_LIMechanistic signaling network models deepen the understanding of drug resistance and sensitivity mechanisms. C_LIO_LIWe identify drug sensitivity-predictive genomic features via proxy signaling phenotypes. C_LI

Source connections

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Tognetti, M., Gabor, A., Yang, M., Cappelletti, V., Windhager, J., Charmpi, K., de Souza, N., Beyer, A., Picotti, P., Saez-Rodriguez, J., Bodenmiller, B.. 2020-01-21. Deciphering the Signaling Network Landscape of Breast Cancer Improves Drug Sensitivity Prediction. https://doi.org/10.1101/2020.01.21.907691

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

A three-factor model can explain symmetry breaking and the n-1 rule during X-chromosome inactivation

X-chromosome inactivation (XCI) is a mammalian gene regulatory process that via chromosome-scale transcriptional silencing, leaves active a single X chromosome per diploid genome complement. The mechanisms that enable cells to select the appropriate number of X chromosomes to inactivate while leaving others active remain unclear. We demonstrate that all known chromosomal counting scenarios during XCI can be accounted for by a three-factor mathematical model that includes (1) Xist, the long noncoding RNA (lncRNA) responsible for gene silencing during XCI in eutherians, (2) an X-linked transcriptional activator of Xist, and (3) the autosomally-encoded SPEN, an Xist RNA-binding protein that is essential for gene silencing during XCI. In our model, symmetry between active X chromosomes is broken by stochastic differences in the number of Xist molecules that associate with the X chromosomes at the onset of XCI. Subsequently, integrated positive and negative feedback loops amplify then propagate superstoichiometric assemblies of SPEN complexed with Xist on the inactive chromosome(s), and select for levels of X-linked activator that enable only one X chromosome per diploid genome complement to remain active. Thus, asymmetric propagation of the SPEN-Xist ribonucleoprotein complex may play an essential role in symmetry breaking and chromosomal counting during XCI, independently of or in concert with cis-acting networks of DNA regulatory elements. Our results also have conceptual implications for gene regulatory circuits on autosomes.

systems biology↗

stem.MAP: Identification of cell-type-specific protein complexes in mammalian embryonic stem cells.

Protein complexes carry out critical functions during development. Mapping cell-type-specific protein complexes in early embryogenesis is essential for uncovering the molecular mechanisms of embryonic development. We unfortunately currently lack a catalog of protein complexes specific for embryogenesis including embryonic stem cells. Here we develop a proteomics-based machine learning workflow to predict protein complexes present in mouse embryonic stem cells (mESC). Our method integrates data from a cell-type agnostic protein complex resource with co-fractionation mass spectrometry (CF-MS) experiments on mESC. Our resource, stem.MAP, identifies 3,986 complexes encompassing ~133,000 unique interactions, and 8,616 proteins. stem.MAP captures variant complexes critical for stem cell function including the PRC2, BAF and NuRD complexes. stem.MAP also discriminates mutually exclusive interactions specific to embryonic stem cells (e.g. Eif4ebp2 - Eif4e). Finally we observe 1,388 high confidence stem.MAP interactions found only in mESC CF-MS data, many with support from AlphaFold modeling and a gastruloid coregulation network. We make stem.MAP protein complexes available through our web resource: stem.proteincomplexes.org. We anticipate stem.MAP will be beneficial to the study of early development and we expect our proteomics-based machine learning workflow to be applicable to the creation of any context-specific protein interaction network.

systems biology↗

Coarse-graining genome-scale metabolic models for dynamic, multi-scale simulations of Pseudomonas aeruginosa in gel environments

Microbial communities are shaped by coupled interactions between microenvironment conditions and the intracellular adaptations that emerge in response external metabolic cues. For pathogenic microbes such as Pseudomonas aeruginosa, these interactions can rapidly generate heterogeneous sub-populations that exhibit adaptations for sustaining host infection and resisting antibiotic treatment. Technological strides in -omics data collection, combined with robust computational tools like genome-scale metabolic models (GEM) for predicting cellular metabolism, have yielded promising multi-scale depictions of microbial population development in simulated spatial environments. These multi-scale model simulations help establish and evaluate mechanistic contexts for how infection-enhancing behaviors might emerge in the population or be influenced by variable perturbations in the biological system. However, the number and rigorous tuning of parameters influencing the connections between intracellular-extracellular events pose barriers when shaping simulations to different in vitro systems of interest. In this work, we present a genome-scale, metabolism-informed continuum model (GEM-CM), a computational framework for coupling a reaction-diffusion model to a surrogate GEM representation that generates metabolic rate predictions informed by flux balance analysis outputs. A key feature of GEM-CM is its application of random forest regression, or ensembles, to coarse-grain the interface connecting the GEM and reaction-diffusion models down to a small set of input metabolites methodically chosen through the random forests evaluation of feature importance. To demonstrate GEM-CMs utility, we evaluated simulation predictions against P. aeruginosa PA14 populations radially propagating in low weight percentage agar gels. In addition to biomass growth, we simulated PA14 populations producing the cytotoxic secondary metabolite pyocyanin (PCN). GEM-CM could capture PA14 growth dynamics in agar gels when ensembles were provided with three carbon source inputs describing the simulated growth media. Bacterial motility parameters and the random forest ensemble allowed GEM-CM simulations to accurately predict spatial biomass distributions for PA14 populations over time and probe their PCN production dynamics. Ensembles also demonstrated robust performance for single gene knockout conditions with GEM-CM simulation accurately capturing the partial or total biomass and PCN inhibition observed in transposon mutant screenings. When evaluating transposon mutant population development, the GEM-CM framework additionally suggested that metH may be a promising target to inhibit the spatial propagation of PA14. This works proof-of-concept applications illustrated GEM-CM utility as an early stage multi-scale modeling tool for interrogating in vitro metabolic behavior in microbial populations.

systems biology↗