1. István Bartha
  2. Jacques Fellay  Is a corresponding author
  1. École Polytechnique Fédérale de Lausanne, Switzerland

Adaptation to new environments is of fundamental importance in ecology. Infectious agents and cancer cells also relentlessly adapt to escape detection by the immune system and to avoid being killed by drugs targeted at them. The nature and extent of adaptive changes in various human pathogens have been studied under well-controlled conditions in the laboratory (Foll et al., 2014; Szamecz et al., 2014), and also in clinical samples (Mukherjee et al., 2011; Poon et al., 2012). Several mutations leading to drug resistance in Candida albicans, a species of yeast that causes oral and genital infections in humans, have been identified through targeted gene sequencing or large-scale genotyping approaches (MacCallum et al., 2010; Dhamgaye et al., 2012). Now, in eLife, Dawn Thompson and Aviv Regev of the Broad Institute of MIT and Harvard and co-workers—including Christopher Ford and Jason Funt as joint first authors—describe the genomic adaptation of C. albicans when it is exposed to a common antifungal compound (Ford et al., 2015).

C. albicans is found in a large fraction of the human population. Normally, it does not affect its host, but it can become pathogenic in people with weakened immune systems (Mayer et al., 2013). C. albicans has a diploid genome—a common feature of sexually reproducing species, featuring two sets of each chromosome—but has a predominantly asexual life cycle (Diogo et al., 2009). These genetic features are very similar to those of cancer cells (Landau et al., 2013).

To shed light on the development of drug resistant strains in vivo, Ford, Funt et al. studied samples from HIV-infected individuals diagnosed with a fungal infection called oral candidiasis and treated with fluconazole. This drug works by preventing C. albicans making a molecule called ergosterol that is incorporated into the cell membrane: without this molecule the yeast cell can't grow.

Ford, Funt et al. analyzed 43 fungal isolates collected from 11 individuals, with several samples taken from each individual over a period of several months. Using deep-sequencing technology, they obtained a whole genome sequence of C. albicans from each sample, which allowed them to produce a comprehensive catalogue of the different genetic variants of the fungus. These variants range from single nucleotide mutations in the DNA of the cells to large-scale genetic changes such as loss of heterozygosity (where one of the two copies of a chromosomal region is lost) and aneuploidies (where the cell contains either more or fewer chromosomes than normal). Of note, all sequencing data have been made publicly available, which is an unprecedented resource for the research community.

The first isolate, collected before treatment started, provided a snapshot of pre-existing genetic variation and was used to filter out non-drug-related mutations. To identify the genes that are under selection pressure during fluconazole therapy, Ford, Funt et al. searched for mutations called ‘non-synonymous coding single nucleotide polymorphisms’ that emerged and persisted in at least three of the patients under treatment. These mutations alter a single DNA nucleotide, which subsequently changes the identity of an amino acid in one of the proteins produced by the cell. Such mutations were observed in a total of 240 genes, notably including genes encoding proteins involved in fungal cell wall formation or in the regulation of the efflux pumps that move toxic substances out of the cell.

Among the larger-scale genetic variants, loss of heterozygosity events were significantly associated with higher drug resistance, whereas most aneuploidies were transient and had no detectable impact. However, aneuploidies may still indirectly help resistance to develop by increasing the likelihood that loss of heterozygosity events occur following the loss of a chromosome. Measurement of the in vitro fitness of the sequenced strains convincingly demonstrated that C. albicans had adapted to fluconazole: the measured fitness of the last isolate was indeed higher in the presence of the drug than without it.

Because a single colony was sequenced at each time point, it was not possible to distinguish between the appearance of new mutations and the selection of pre-existing minority variants in response to drug pressure. Most isolates from each individual were highly related, suggesting that samples collected in the same patient share the same common ancestor. Despite this clonal relationship, significant within-host diversity was present, as various isolates of the same patient differed by thousands of single nucleotide polymorphisms.

A remaining open question is whether pre-existing genetic variation is maintained at low levels in the C. albicans population throughout the treatment period, or if only drug-selected strains are conserved. To address this, more in-depth genomic analyses of fungal sub-populations are needed. An obvious first step would be to study the loss of heterozygosity events in greater detail, because such mutations are irreversible in clonal populations with a purely asexual life cycle. Therefore, valuable insights into the diversity of C. albicans inside a single host could be gained by investigating whether any of the resistance-inducing loss of heterozygosity events reverses after fluconazole treatment has ended.

The work by Ford, Funt et al. provides a global description of the genetic processes underlying drug resistance and adaptation in C. albicans. What else could now be learned about microbial evolution using deep sequencing technology? First, the analysis of multiple strains collected simultaneously in the same infected patient has the potential to reveal population structure, dynamics and diversity. Second, mechanistic and temporal details governing the emergence of escape mutations will certainly be gained from in vitro experiments, including the characterization of single colonies over time. Finally, the sequencing of paired host and pathogen genomes opens the door to innovative studies of host-specific adaptation.

References

Article and author information

Author details

  1. István Bartha

    School of Life Sciences, École Polytechnique Fédérale de Lausanne, Lausanne, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  2. Jacques Fellay

    School of Life Sciences, École Polytechnique Fédérale de Lausanne, Lausanne, Switzerland
    For correspondence
    jacques.fellay@epfl.ch
    Competing interests
    The authors declare that no competing interests exist.

Publication history

  1. Version of Record published:

Copyright

© 2015, Bartha and Fellay

This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 1,591
    views
  • 116
    downloads
  • 2
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. István Bartha
  2. Jacques Fellay
(2015)
Microbial Evolution: Adaptation on a genomic scale
eLife 4:e06193.
https://doi.org/10.7554/eLife.06193
  1. Further reading

Further reading

    1. Evolutionary Biology
    2. Genetics and Genomics
    Michael James Chambers, Sophia B Scobell, Meru J Sadhu
    Research Article

    Evolutionary arms races can arise at the contact surfaces between host and viral proteins, producing dynamic spaces in which genetic variants are continually pursued.  However, the sampling of genetic variation must be balanced with the need to maintain protein function. A striking case is given by protein kinase R (PKR), a member of the mammalian innate immune system. PKR detects viral replication within the host cell and halts protein synthesis to prevent viral replication by phosphorylating eIF2α, a component of the translation initiation machinery. PKR is targeted by many viral antagonists, including poxvirus pseudosubstrate antagonists that mimic the natural substrate, eIF2α, and inhibit PKR activity. Remarkably, PKR has several rapidly evolving residues at this interface, suggesting it is engaging in an evolutionary arms race, despite the surface’s critical role in phosphorylating eIF2α. To systematically explore the evolutionary opportunities available at this dynamic interface, we generated and characterized a library of 426 SNP-accessible nonsynonymous variants of human PKR for their ability to escape inhibition by the model pseudosubstrate inhibitor K3, encoded by the vaccinia virus gene K3L. We identified key sites in the PKR kinase domain that harbor K3-resistant variants, as well as critical sites where variation leads to loss of function. We find K3-resistant variants are readily available throughout the interface and are enriched at sites under positive selection. Moreover, variants beneficial against K3 were also beneficial against an enhanced variant of K3, indicating resilience to viral adaptation. Overall, we find that the eIF2α-binding surface of PKR is highly malleable, potentiating its evolutionary ability to combat viral inhibition.

    1. Evolutionary Biology
    2. Genetics and Genomics
    Giulia Ferraretti, Paolo Abondio ... Marco Sazzini
    Research Article

    It is well established that several Homo sapiens populations experienced admixture with extinct human species during their evolutionary history. Sometimes, such a gene flow could have played a role in modulating their capability to cope with a variety of selective pressures, thus resulting in archaic adaptive introgression events. A paradigmatic example of this evolutionary mechanism is offered by the EPAS1 gene, whose most frequent haplotype in Himalayan highlanders was proved to reduce their susceptibility to chronic mountain sickness and to be introduced in the gene pool of their ancestors by admixture with Denisovans. In this study, we aimed at further expanding the investigation of the impact of archaic introgression on more complex adaptive responses to hypobaric hypoxia evolved by populations of Tibetan/Sherpa ancestry, which have been plausibly mediated by soft selective sweeps and/or polygenic adaptations rather than by hard selective sweeps. For this purpose, we used a combination of composite-likelihood and gene network-based methods to detect adaptive loci in introgressed chromosomal segments from Tibetan WGS data and to shortlist those presenting Denisovan-like derived alleles that participate to the same functional pathways and are absent in populations of African ancestry, which are supposed to do not have experienced Denisovan admixture. According to this approach, we identified multiple genes putatively involved in archaic introgression events and that, especially as regards TBC1D1, RASGRF2, PRKAG2, and KRAS, have plausibly contributed to shape the adaptive modulation of angiogenesis and of certain cardiovascular traits in high-altitude Himalayan peoples. These findings provided unprecedented evidence about the complexity of the adaptive phenotype evolved by these human groups to cope with challenges imposed by hypobaric hypoxia, offering new insights into the tangled interplay of genetic determinants that mediates the physiological adjustments crucial for human adaptation to the high-altitude environment.