Inferring fitness landscapes and selection on phenotypic states from single-cell genealogical data

Recent advances in single-cell time-lapse microscopy have revealed non-genetic heterogeneity and temporal fluctuations of cellular phenotypes. While different phenotypic traits such as abundance of growth-related proteins in single cells may have differential effects on the reproductive success of cells, rigorous experimental quantification of this process has remained elusive due to the complexity of single cell physiology within the context of a proliferating population. We introduce and apply a practical empirical method to quantify the fitness landscapes of arbitrary phenotypic traits, using genealogical data in the form of population lineage trees which can include phenotypic data of various kinds. Our inference methodology for fitness landscapes determines how reproductivity is correlated to cellular phenotypes, and provides a natural generalization of bulk growth rate measures for single-cell histories. Using this technique, we quantify the strength of selection acting on different cellular phenotypic traits within populations, which allows us to determine whether a change in population growth is caused by individual cells’ response, selection within a population, or by a mixture of these two processes. By applying these methods to single-cell time-lapse data of growing bacterial populations that express a resistance-conferring protein under antibiotic stress, we show how the distributions, fitness landscapes, and selection strength of single-cell phenotypes are affected by the drug. Our work provides a unified and practical framework for quantitative measurements of fitness landscapes and selection strength for any statistical quantities definable on lineages, and thus elucidates the adaptive significance of phenotypic states in time series data. The method is applicable in diverse fields, from single cell biology to stem cell differentiation and viral evolution.Author summary: Selection is a ubiquitous process in biological populations in which individuals are endowed with heterogeneous reproductive abilities, and it occurs even among genetically homogeneous cells due to the existence of phenotypic noise. Unlike genotypes, which can remain stable for many generations, phenotypic fluctuations at the single cell level are often comparable to cellular generation times. For this reason, quantifying the contribution of specific phenotypic states to cellular fitness remains a major challenge. Here, we develop a method to measure the fitness landscape and selection strength acting on diverse cellular phenotypes by employing a novel conceptual framework in which cellular histories are regarded as a basic unit of selection. With this framework, one can tell quantitatively whether a population adapts to environmental changes by selection or through individual responses. This new analytical approach to genetics reveals the roles of heterogeneous expression patterns and dynamics without directly perturbing genes. Applications in diverse fields including stem cell differentiation and viral evolution are discussed.