Category Archives: Transhuman News

A human DNA sequence is made of four types of nucleic acid called base pairs, each represented by their first letter: adenine (A), thymine (T), guanine (G) and cytosine (C). Altogether, a list 3.055-billion-letters long across 23 chromosomes makes up the human genome. Nearly two decades ago, the Human Genome Project set out to map the genetic makeup of the human species. In 2000, scientists completed the first draft of the human genome, but eight percent still remained, reports Matthew Herper for STAT.

The unsequenced remaining portion was a dizzying array of repeating letters. These missing gaps were almost impossible to decipher with the technology available at the time. Now, in a preprint published on May 27, a group of scientists describe the first "nearly" complete sequence of the human genome, reports Sarah Zhang for the Atlantic.

The feat was completed with scientists in the Telomere to Telomere (T2T) Consortium, a collaboration consisting of about 30 different institutions, reports Sara Reardon for Nature. Together, they found 115 new genes and added 200 million base pairs to a version of the human genome measured in 2013. They named the newly deciphered genome T2T-CHM13.

One of the most challenging regions to sequence in the human genome is centromeres. Each chromosome resembles an X-shaped tangle, and centromeres are located close to the pinched, knot-like center of each criss-cross. In these regions, DNA difficult to sequence because it is so densely packed and contains nearly endless repeating codes, the Atlantic reports.

But on five of the 23 total human chromosomes, the centromere is not precisely in the middle, instead favoring one end over the other, per the Atlantic. The asymmetrical point creates one long arm and one short arm on the chromosome. The previously unsequenced, repeating letters are located in these "short arms." Now, the team behind T2T-CHM13 has deciphered them.

The sequencing was made possible using new technologies developed by two private companies: Pacific Biosciences (PacBio) of Menlo Park, California, and Oxford Nanopore of Oxford Science Park in the United Kingdom.

Previously methods for genome-deciphering required cutting DNA into tiny pieces and then reassembling stretches of DNA later in a long, tedious process. Two new methods take different approaches. The Oxford Nanopore technology pulls the DNA into a small hole where longer sequences can be read. The PacBio tech uses lasers to examine 20,000 base pair sequences of DNA at a time repeatedly to create a highly accurate readout, reports STAT.

Using the Oxford Nanopore technology, the T2T-CHM13 consortium found that it can map where proteins attach to the centromere during cell division, per the Atlantic.

The sequenced DNA was derived using a cell line taken from tissue that forms when sperm fertilizes a non-viable egg that lacks a nucleus, also known as complete hydatidiform mole, reports Nature. (In other words, the sample was not taken from a person.) However, DNA is stored in the nucleus of an egg, so an egg without a nucleus does not contain gentic material from a mother. Instead, the "mole" only contains chromosomes from the father. Using a mole makes sequencing easier because researchers do not have to differentiate two sets of chromosomes from the parents.

But the T2T-CHM13 genome only represents one genome, so the researchers plan to team up with the Human Pangenome Reference Consortium to sequence over 300 genomes in the next three years from humans worldwide, using T2T-CHM13 as a reference. They also plan to sequence a Y chromosome next since the sperm used to create the hydatidiform mole only carried an X chromosome.

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Scientists Are on the Cusp of Finally Deciphering the Entire Human Genome - Smithsonian Magazine

The release of the draft human genome sequence in 2001 was a seismic moment in our understanding of the human genome, and paved the way for advances in our understanding of the genomic basis of human biology and disease.

But sections were left unsequenced, and some sequence information was incorrect. Now, two decades later, we have a much more complete version, published as a preprint (which is yet to undergo peer review) by an international consortium of researchers.

Technological limitations meant the original draft human genome sequence covered just the euchromatic portion of the genomethe 92% of our genome where most genes are found, and which is most active in making gene products such as RNA and proteins.

The newly updated sequence fills in most of the remaining gaps, providing the full 3.055 billion base pairs (letters) of our DNA code in its entirety. This data has been made publicly available, in the hope other researchers will use it to further their research.

Much of the newly sequenced material is the heterochromatic part of the genome, which is more tightly packed than the euchromatic genome and contains many highly repetitive sequences that are very challenging to read accurately.

These regions were once thought not to contain any important genetic information but they are now known to contain genes that are involved in fundamentally important processes such as the formation of organs during embryonic development. Among the 200 million newly sequenced base pairs are an estimated 115 genes predicted to be involved in producing proteins.

Two key factors made the completion of the human genome possible:

1. Choosing a very special cell type

The newly published genome sequence was created using human cells derived from a very rare type of tissue called a complete hydatidiform mole, which occurs when a fertilized egg loses all the genetic material contributed to it by the mother.

Most cells contain two copies of each chromosome, one from each parent and each parents chromosome contributing a different DNA sequence. A cell from a complete hydatidiform mole has two copies of the fathers chromosomes only, and the genetic sequence of each pair of chromosomes is identical. This makes the full genome sequence much easier to piece together.

2. Advances in sequencing technology

After decades of glacial progress, the Human Genome Project achieved its 2001 breakthrough by pioneering a method called shotgun sequencing, which involved breaking the genome into very small fragments of about 200 base pairs, cloning them inside bacteria, deciphering their sequences, and then piecing them back together like a giant jigsaw.

This was the main reason the original draft covered only the euchromatic regions of the genomeonly these regions could be reliably sequenced using this method.

The latest sequence was deduced using two complementary new DNA-sequencing technologies. One was developed by PacBio, and allows longer DNA fragments to be sequenced with very high accuracy. The second, developed by Oxford Nanopore, produces ultra-long stretches of continuous DNA sequence. These new technologies allow the jigsaw pieces to be thousands or even millions of base pairs long, making them easier to assemble.

The new information has the potential to advance our understanding of human biology including how chromosomes function and maintain their structure. It is also going to improve our understanding of genetic conditions such as Down syndrome that have an underlying chromosomal abnormality.

Well, no. An obvious omission is the Y chromosome, because the complete hydatidiform mole cells used to compile this sequence contained two identical copies of the X chromosome. However, this work is underway and the researchers anticipate their method can also accurately sequence the Y chromosome, despite it having highly repetitive sequences.

Even though sequencing the (almost) complete genome of a human cell is an extremely impressive landmark, it is just one of several crucial steps towards fully understanding humans genetic diversity.

The next job will be to study the genomes of diverse populations (the complete hydatidiform mole cells were European). Once the new technology has matured sufficiently to be used routinely to sequence many different human genomes, from different populations, it will be better positioned to make a more significant impact on our understanding of human history, biology, and health.

Both care and technological development are needed to ensure this research is conducted with a full understanding of the diversity of the human genome to prevent exacerbation of health disparities by limiting discoveries to specific populations.

This article is republished from The Conversation under a Creative Commons license. Read the original article.

Image Credit: Arek Socha /Pixabay

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Why It Took 20 Years to 'Finish' the Human Genomeand Why There's Still More to Do - Singularity Hub

NEW YORK Personal Genome Diagnosticssaid on Tuesday that it has signed an agreement to provide its Elio Tissue Complete next-generation sequencing test platform to the Duke University Health System Clinical Molecular Diagnostics Laboratory.

Elio Tissue Complete is a US Food and Drug Administration-cleared 507-gene panel kit that identifies single-nucleotide variants, small insertions and deletions, amplifications, rearrangements, microsatellite instability, and tumor molecular burden in DNA from patient tissue samples.

According to Personal Genome Diagnostics, or PGDx, the platform will be integrated into the Duke lab and be used as its primary technology for clinical oncology genomic testing initiatives. As part of the deal, the Baltimore-based company and DUHS will collaborate on a data integration system to join the health system's electronic health records with other local systems.

Financial and other terms of the collaboration were not disclosed.

"We believe strongly that the ability to provide accurate, rapid genomic profiling data and insights that can be easily accessed and incorporated into existing cancer care pathways will allow our clinicians to further optimize treatments for the benefit of our patients," Michael Datto, associate VP of DUHS Clinical Laboratories, said in a statement.

Earlier this year, PGDx raised $103 million in Series C funding that it has been usingto expand its commercial infrastructure and forge new agreements with diagnostic and pharmaceutical partners.

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Personal Genome Diagnostics Inks Cancer Genomic Profiling Deal with Duke University Health System - GenomeWeb

INTRODUCTION

Can evolutionary outcomes be predicted? Biologists have long been fascinated with this question, including Darwin and Wallaces anticipation of the existence of Morgans sphinx moth based on orchid morphology (1, 2), Vavilovs prediction of the types of morphological variants likely to occur in plants (3), and Goulds gedankenexperiment about replaying the tape of life (4). Natural examples of recurrent evolution provide a particularly favorable opportunity to study the mechanisms that influence evolutionary predictability, including molecular patterns (5, 6).

Although the predictability of evolution may appear to be in conflict with the unpredictability of historical contingency, understanding the past can yield important insights into future evolution. For example, vertebrate populations frequently harbor large reservoirs of standing genetic variation (SGV) (7) that give independent populations access to similar raw genetic material to respond to environmental challenges, as observed in diverse species including songbirds, cichlid fishes, and the threespine stickleback (Gasterosteus aculeatus) (811). SGV is often apparent in divergent species or populations where it is pretested by natural selection and then distributed by hybridization to related populations. Thus filtered and capable of leaping up fitness landscapes, SGV can also drive rapid evolution (12), helping address a very real practical challenge to testing evolutionary predictions: time.

Longitudinal studies of evolving populations have been used to estimate the tempo and strength of selection on a variety of traits in different species (1318). Rapid phenotypic evolution over contemporary time scales has enabled hypothesis testing against detailed observations at every step in the process. There is an increasing and impressive body of research examining the genomic consequences of these phenotypic changes in microbial, invertebrate, and vertebrate systems (1926).

Stickleback fish provide an outstanding system for further study of the genomic basis of recurrent evolution. At the end of the last Ice Age, threespine stickleback, including anadromous populations that migrate from the ocean to freshwater environments to breed, colonized and adapted to countless newly exposed freshwater environments created in the wake of retreating glaciers around the northern hemisphere (27, 28). This massively parallel adaptive radiation was facilitated by natural selection acting on extensive ancient SGV (8, 11). Under the transporter hypothesis, these variants are maintained at low frequencies in the marine populations by low levels of gene flow from freshwater populations (29). Reuse of ancient standing variants has enabled identification of genomewide sets of loci that are repeatedly differentiated among long-established stickleback populations (8, 3035). In addition, SGV enables new freshwater stickleback populations to evolve markedly within decades (17, 3638), including conspicuous phenotypic changes in armor plates (17) and body shape (39).

The rapidity of stickleback evolution has made it possible to begin characterizing genomic and allele frequency changes seen in very young or newly established populations under intense directional selection on multiple traits (18, 3638, 4043). Here, we identify key molecular features that underlie repeated and rapid evolution of freshwater stickleback by comparing genomes from diverse extant populations with the earliest generation-by-generation changes in a detailed genomic time series from three newly founded populations. We identify several basic genomic and genetic features that can be used to predict evolutionary outcomes in stickleback and show that they can predict genomic responses to selection in distantly related cichlids and Darwins finches.

Previous whole-genome sequencing (WGS) of threespine stickleback identified 174 loci covering 1.2 Mb with alleles shared by common descent repeatedly selected in freshwater populations around the world (8). Just as human genetic diversity is greatest in Africa, where Homo sapiens arose (44), we hypothesized that the north Pacific region where stickleback originated (27) may contain a particularly rich pool of ancient adaptive alleles. To test this hypothesis, we generated whole-genome sequence data with 76base pair (bp) paired-end Illumina reads for 38 new marine and 110 new freshwater stickleback, respectively (mean coverage of 5.5) (sections S2, S4, S6, and S7). Combined with previous stickleback sequencing (8, 41), our dataset includes 227 individual genomes: 135 genomes from 70 northeast Pacific populations in Alaska, Haida Gwaii, British Columbia, and Washington and 92 genomes from 62 populations in California, Japan, and the Atlantic coasts of North America, Iceland, and northern Europe (Fig. 1A and section S8).

(A) Marine (red) and freshwater (blue) stickleback from the locations shown were used for various analyses (table S2). (B) Detail of part of chrIV for single-nucleotide polymorphism (SNP)based analysis of differential allele distribution between marine and freshwater ecotypes in the northeast Pacific basin. SNPs within specific-threshold EcoPeaks are red. A subset of regions overlap the globally shared peaks of marine-freshwater differentiation indicated by blue-colored bars [cluster separation score (CSS), 5% false discovery rate (FDR) identified by Jones et al. (8)]. (C) As in (B), but for the whole chromosome [dashed lines from (B) to (C)]. (D) Same whole chromosome as in (C), but with genetic (not physical) distance along the x axis. (E and F) Genomewide SNP divergence between marine and freshwater ecotypes globally and in the northeastern Pacific basin, with specific-threshold EcoPeaks in red. (G) Many differentiated regions overlap the location of major quantitative trait loci (QTLs) controlling various morphological, physiological, and behavioral traits in previous genetic crosses [percent variance explained (PVE) > 20, interval < 5 Mb from Peichel and Marques (53)].

We used two methods to identify loci repeatedly differentiated in freshwater populations, both based on the expectation that variants recurrently selected from SGV will be more similar among geographically separated freshwater populations than neutral loci (section S9). First, we used a genetic distancebased approach within overlapping 2500-bp windows tiled across the genome [as in the study by Jones et al. (8)]. While statistically powerful, this approach may miss younger loci with few differences between alleles and exhibits spatial resolution dependent on window size. Second, we analyzed the distribution of variants at individual bases across the genome, which has base pairlevel resolution and less bias against younger loci, though at the cost of statistical power. After calling P valuebased peaks of ecotypic (freshwater- or marine-associated) differentiation using both methods, we accepted calls at two stringency levels, either requiring agreement between the two analyses at 1% false discovery rate (FDR) (specific) or support from either at 5% FDR (sensitive). We refer to these peaks of ecotypic differentiation as EcoPeaks. We called EcoPeaks for different geographic sets of samples to find alleles that were either shared globally, within the northeast Pacific, or within other geographic regions.

Although results of the global analysis largely matched a previous report [79 of 81 most stringent calls from Jones et al. (8) in sensitive EcoPeaks (P = 4.2 1021; table S3)], both the sensitive and specific call sets identified approximately five times as many Pacific EcoPeaks as global EcoPeaks, spanning sevenfold more of the genome (Fig. 1, E and F, and Table 1). In addition, many northeast Pacific EcoPeaks not overlapping the globally shared regions identified by Jones et al. (8) exhibit even more consistent ecotypic differentiation (assessed by P values) than others shared around the world (Fig. 1, B and C). Much smaller sets of non-global EcoPeaks were identified in the North Atlantic, subglacial Pacific, and supraglacial geographic regions (fig. S5), consistent with other reports (8, 35).

The comparisons by Jones et al. (8) are with the cluster separation score 5% FDR set (8).

As theoretical studies indicate that SGV is immediately available for evolution and may show an increased likelihood of large-effect alleles being advantageous compared to de novo mutations (12, 45), the rich genetic reservoir observed in the northeast Pacific provides a favorable system for studying the dynamics and predictability of rapid evolutionary change (section S10). Previous studies suggest that stickleback in the northeast Pacific can adapt to freshwater environments within decades (36). However, thus far, studies have lacked temporal resolution of genome evolution in the critical early years of adaptation.

To characterize the earliest stages of evolution after the establishment of new freshwater populations, we analyzed annual samples from populations that were recently founded by anadromous stickleback in three lakes in Alaska (Fig. 2A and section S1). In 1982, stickleback in Loberg Lake (LB) were exterminated to improve recreational fishing (17). Sometime between 1983 and 1988, LB was invaded by completely plated (~33 plates per side) anadromous stickleback [most likely from neighboring Rabbit Slough (RS)]. The characteristic freshwater, armor-reduced phenotype increased rapidly from ~16% in 1991 to ~50% by 1995 and to ~95% by 2017 (Fig. 2B) (17), with similarly rapid changes in overall body shape (39) and reproductive patterns (46). So as to more systematically examine even earlier generations of freshwater adaptation, Bell et al. (47) introduced ~3000 anadromous RS fish into each of two other Cook Inlet lakes without outlets that had been similarly treated to exterminate fish: Cheney Lake (CH) in 2009 and Scout Lake (SC) in 2011. Lowarmor-plated (~5 to 7 plates per side) stickleback began to appear in the second and third generation after founding in CH and SC respectively, and, by 2017, they had increased to 20 to 30% (Fig. 2B).

(A) The timing (years since founding) and approximate size of subsequent sequencing sample pools from lake populations [Loberg Lake (LB), Cheney Lake (CH), and Scout Lake (SC)] founded recently by anadromous stickleback (left) and the scenario for divergence of anadromous populations after colonizing the lakes (right). Red and blue fish represent the complete armor-plated and armor-reduced phenotypes, respectively. (B) Frequency of armor-reduced morphological phenotype across our CH, SC, and LB time series overlaid with the frequency squared for the freshwater (FW) Eda allele. LB data are based on a combination of individual genotypes and pool-seq frequencies, while CH and SC are based only on pool-seq frequencies. (C) Allele frequency trajectories for eight SNPs found within TempoPeaks on distinct chromosomes with the highest Cochran-Mantel-Haenszel (CMH) scores (except for chrIV:12823875, the Eda-plate regulatory region SNP). (D) Genomewide distribution of window-based CMH scores across chrIV for different combinations of transplant lakes discussed in the main text. Black, dark red, and teal bars above figure represent specific CH + SC + LB TempoPeaks, northeast Pacific EcoPeaks, and significant loci from Jones et al. (8) identified using CSS [5% FDR (8)], respectively.

To obtain genomewide allele frequencies across our time series, we performed pooled WGS (pool-seq) on all seven available annual samples from CH and SC since founding and eight from LB distributed between 1999 and 2017 (Fig. 2A and sections S3, S4, S7, and S13). Each freshwater pool-seq experiment consisted of 100 individuals (with three exceptions), with mean coverage of 223 per pool. In addition, we resequenced a pool of 200 anadromous RS individuals used to found the CH population in 2009 (RS2009) to 585.

We identified single-nucleotide polymorphisms (SNPs) with significant allele frequency changes, indicating directional selection, using a modified Cochran-Mantel-Haenszel (CMH) test optimized for pool-seq data (48), followed by an approach analogous to our EcoPeak analysis to define both a permissive sensitive and a stringent specific set of loci that we term TempoPeaks (sections S16 to S18). Combining all three populations into a single CMH analysis (CH + SC + LB) and using RS2009 as a proxy for the founders of LB, we identified 524 sensitive and 344 specific TempoPeaks. Despite operating over very different time spans, the visual correspondence between the Pacific EcoPeaks in long-established populations and the TempoPeaks in recently established populations is notable, particularly for the specific TempoPeaks, of which 323 of 344 (94%) overlap with the sensitive Pacific EcoPeaks (Fig. 2D and section S18). In contrast, even the most lenient set of global EcoPeaks and regions from Jones et al. (8) overlap only 96 of 344 (28%) and 47 of 344 (14%) specific TempoPeaks, respectively (tables S9 and S10), emphasizing the importance of understanding the locally available SGV. Even analyzing only CH + SC (thus focusing on <10 years of freshwater adaptation), we identified 271 sensitive and 86 specific TempoPeaks, 73% and 99% of which, respectively, overlap the sensitive Pacific EcoPeaks. This marked congruity strongly suggests that the ancient SGV represented by Pacific EcoPeaks is the primary genomic feature enabling extremely fast evolution of freshwater phenotypes in stickleback from the northeast Pacific basin.

The Eda SNP associated with armor plate variability (chrIV:12,823,875 T>G (49)) is within the second most significant specific TempoPeak on chrIV. In both CH and SC, the G allele increases rapidly from an initial frequency of <1% to over 50% within 8 years, while approaching fixation in LB by 15 years. Notably, the square of G-allele frequencies (i.e., the expected number of GG homozygotes) tracks closely with frequencies of the lowarmor plate phenotype, consistent with almost complete recessiveness (h = 0.0) for the G allele for this phenotype (Fig. 2B). Nonetheless, to fit the allele frequency trajectory of this SNP, and, in particular, the extremely rapid increase in CH and SC, it was necessary to impose a dominance coefficient (h) of 1.0 along with a very large selection coefficient (s) of 0.55, as in a recent paper focusing on this locus (18).

Like Eda, most TempoPeaks display similarly sharp left-shifted sigmoidal allele frequency trajectories, indicating very strong and dominant-positive selection (Fig. 2C and section S20). When modeling each peak SNP as independent, we find an extremely high mean s of 0.30 (5th, 95th percentile 0.08 to 0.53) and h of 0.98 (5th, 95th percentile 0.95 to 1.0) for the 344 specific TempoPeaks found in CH + SC + LB. The estimated s values for chrIV, where there are 69 TempoPeaks, are particularly high (mean s = 0.38), consistent with the accelerated evolution of this whole chromosome observed via a chromosome-wide FST analysis comparing the founding generation of CH, SC, and LB to all subsequent years (section S15).

The remarkable speed at which northeast Pacific stickleback adapt to new freshwater environments suggests that analysis of EcoPeaks may provide unique insights into optimal genomic properties for evolution. Using Gasterosteus nipponicus, Gasterosteus wheatlandi, and Pungitius pungitius for calibration, we estimated molecular divergence time between a pair of freshwater (Little Campbell upstream) and marine (Little Campbell downstream) stickleback in windows tiled across the genome (section S11). We find that EcoPeaks as a whole are significantly older than the rest of the genome [1600 thousand years (ka) versus 700 ka, P < 5 10324]. Although peaks shared globally trend older than those found just within the northeast Pacific (1800 ka versus 1600 ka, P = 0.18), the imputed ages overlap considerably (Fig. 3A). We estimate that the majority (161 of 209) are over a million years old and have cycled between freshwater and marine environments many times during this long history, likely persisting at high frequency in freshwater habitats south of the zone of glaciation during the Ice Ages and at more northerly latitudes during previous interglacials and the Holocene.

(A) Distribution of estimated molecular age for those EcoPeaks either shared worldwide (orange) or within the northeast Pacific (blue). Ma, million years. (B) EcoPeaks with older estimated molecular ages tend to be larger. (C) Estimated ages decline with distance on either side of EcoPeaks. Each dot represents mean age in 1-kb windows flanking the EcoPeak centers (gray bars, 1 SE). (D) Recombination rates tend to be lower within EcoPeaks compared to the rest of the genome, 1 SE. (E) Recombination rates and distances to nearest 20 recombination hotspots, plotted for randomly subsampled 1-kb windows tiled across the genome, with marginal histograms of all windows. Locations overlapping EcoPeaks (red) are shifted to both smaller hotspot distances and lower recombination rates compared to other genomic regions (gray). (F) Observed haploblock size in marine fish carrying freshwater EcoPeaks on the indicated chromosomes across three marine populations. For all, specific northeast Pacific EcoPeaks are used.

Contrary to our expectations that recombination would disassemble regions over time, we found that older EcoPeaks are larger than younger ones (Fig. 3B). This signature is strongest at the most significant markers within each EcoPeak, which are typically older than more distal sequences (Fig. 3C). This suggests that individual regions may grow over time, with alleles originally based on an initial beneficial mutation accumulating additional linked favorable mutations, snowballing over time to form a finely tuned haplotype with multiple adaptive changes. This is consistent with work in other species identifying examples of evolution through multiple linked mutations that together modify function of a gene (5052) and implies that progressive allelic improvement may be common.

We also observed that EcoPeaks frequently overlap major quantitative trait loci (QTLs) in stickleback [73 of 209 overlaps observed versus 32 of 209 expected, P < 1 1015; Fig. 1G (53)], suggesting that these variants underlie many mapped phenotypic traits. Just as the QTLs cluster in supergene complexes (54), so too do EcoPeaks (median observed interpeak distance 192 kb versus 795 kb expected, P = 4.88 1010). One particularly large complex (chrIV: 8 to 17 Mb) contains 22 EcoPeaks and the major QTLs controlling many aspects of both defensive armor and trophic morphology (e.g., the length of dorsal and pelvic spines, the number of armor plates through Eda, gill rakers, and teeth). Thus, clustering may have important functional effects by allowing multiple traits and underlying EcoPeaks to be selected and inherited as a single unit, especially when in tight linkage. A fine-scale recombination map of RS stickleback (generated with LDhelmet (55)) shows that EcoPeaks are highly enriched in regions of low average recombination, forming tightly linked haploblocks (Fig. 3D, compare Fig. 1, C and D; section S14). EcoPeaks are also enriched near local recombination hotspots within their neighborhood (Fig. 3E), potentially facilitating reassembly of larger haplotype blocks upon freshwater colonization (also see section S19).

To further examine the frequency and size of haploblocks in individual fish, we surveyed 1643 stickleback from three Alaskan marine populations by SNP array genotyping (sections S5 and S12). While most marine fish heterozygous for freshwater alleles carry a relatively small haploblock, some carry multi-megabase haploblocks containing multiple EcoPeaks (Fig. 3F). Thus, a proper treatment of rapid stickleback evolution needs to account for the complex linkage of EcoPeaks rather than treating them independently.

Modeling the genomic landscape of contemporary evolution. To estimate a more realistic distribution of fitness effects (DFE) that incorporates the genomes recombination landscape, we developed a deep neural network (DNN) approach that uses forward simulations (section S21). Our simulations, which are conceptually similar to those of Galloway et al. (56), attempted to replicate the dynamics of the transporter model (29), with one large (Ne = 10,000) anadromous population connected independently by gene flow to 10 smaller (Ne = 1000) established freshwater populations. After 1000 generations, we founded three new freshwater populations from the anadromous population, thus generating simulated allele frequency trajectories that reflect our annual LB, CH, and SC samples (Fig. 4A).

(A) Schematic showing evolutionary model of forward simulations under the transporter hypothesis. Red horizontal bars, anadromous (AN) ancestor; blue circles, descendant freshwater isolates; red to blue shaded circles, three adapting freshwater populations (i.e., LB, CH, and SC) founded recently by anadromous stickleback; and arrows, gene flow or founding events. (B) Genotypes across chrIV for freshwater-associated SNPs in RS (n = 750), LB in 1999 (n = 25), and LB in 2013 for (left) observed and (right) simulated data under best-fit DNN model. anadromous homozygous, red; heterozygous, yellow; and freshwater homozygous genotypes, blue; respectively. (C). Allele frequency trajectories for LB, CH, and SC in 100 simulations under the best-fit DNN model for five randomly selected SNPs. Larger points, observed data. (D) Distribution of average CMH scores in windows of 2500 bp across chrIV for (top) observed and (bottom) simulated data under best-fit DNN model. Red dotted lines, locations of SNPs under selection and used to fit DNN.

Focusing our DNN analysis on a subset of 19 specific TempoPeak SNPs separated by 0.4 cM (~100 kb) along chrIV, we closely replicated observed allele trajectories of positively selected freshwater alleles across all SNPs simultaneously using a beta distributionshaped DFE, for which the mean s across the 19 TempoPeaks was 0.063 and the standard deviation was 0.030, with reciprocal fitness costs implemented in the marine population (Fig. 4C). The estimated s from our DNN was thus substantially smaller than the mean of 0.48 when each SNP was considered independently. In addition, 18 of 19 SNPs were predicted to be fully dominant and none fully recessive under the best model.

We validated our best-fit DNN model by simulating the 19 selected TempoPeaks SNPs with the estimated DFE along with ~400k neutral SNPs distributed randomly along chrIV. Despite the neutral SNPs not being used in training the DNN, we were able to mimic the overall topology of the CMH scores across the entire genome, suggesting that our model was capturing the overall genomic architecture of freshwater adaptation (Fig. 4D). Our best-fit DNN model also appeared to recapitulate much of the haplotype structure of the array data from individuals from RS, LB1999, and LB2013 (Fig. 4B). Notably, the transition to freshwater alleles appears to be somewhat slower on the right half of chrIV, where there are fewer EcoPeaks, TempoPeaks, and QTLs, and this difference was observable in both the empirical and simulated data.

Overall, our model suggests that extremely rapid and replicable allele frequency increases on chrIV in LB, CH, and SC are mostly driven by multiple linked (primarily) dominant alleles, each with relatively smaller s values that act in concert, with recombination hotspots between them (section S19) allowing rapid reassembly of optimum freshwater haplotypes, consistent with the transporter hypothesis. The lower individual s values may allow these dominant alleles to persist in the marine environment at low frequency after being disassembled by recombination, especially if some act in epistasis.

Biological features with predictive power. Given the genomewide dynamism of the earliest stages of freshwater adaptation, we attempted to identify genomic features that predict the speed of evolution at TempoPeaks and understand why some peaks are consistently selected more rapidly than others (section S22). We used CMH scores as a proxy of evolutionary speed for each TempoPeak in CH + SC + LB and regressed these against a variety of sequence features.

The best predictor for the speed of evolution is the degree of ecotypic differentiation between marine and long-established freshwater populations (Pacific EcoPeak P value), with variants more commonly differentiated in the northeast Pacific being selected more quickly (Fig. 5A and fig. S81). Fishers geometric model indicates that alleles with large effects are usually disfavored; however, the prefiltering of ancient SGV that counters this tendency (12) largely benefits alleles that are broadly positively selected, possibly explaining this result.

(A) Variance in the speed of TempoPeak selection explained by different underlying genomic features, including colored bars: empirical recurrence of marine-freshwater differentiation (peak Pacific ecotypic P value), number of additional Pacific EcoPeaks within 10 cM, number of major QTLs overlapped, sequence divergence, and recombination rate; gray bars: genomic size of EcoPeak, total number of variable nucleotides, elevated Ka/Ks in coding regions, overlap with genic sequences, overlap with conserved noncoding sequence (PhastCons nonexonic), and carrier frequency of freshwater alleles in marine populations. (B) Precision-recall curve for predicting the locations of selected loci in CH + SC + LB lakes by either individual genomic features (dotted lines), a composite model trained with these basic predictors, or the empirical expectation of recurrence based on many extant populations. Precision is the fraction of predictions that are accurate, while recall is the fraction of true positives that are correctly predicted. No skill refers to the performance expected by random chance. (C) Performance above chance of the composite model applied to stickleback, cichlids, and two representative pairs of species of Darwins finches (ground finches: Geospiza magnirostris versus Geospiza propinqua; tree finches: Camarhynchus pauper versus Camarhynchus psittacula).

We also found that larger TempoPeaks are typically selected more rapidly. Similarly, greater TempoPeak density predicts more rapid divergence, suggesting that our simulation accurately reflects how nearby loci mutually reinforce their collective selection. Overlap with major QTLs also has a strong association with rapid evolution, while other variables such as increased sequence divergence, decreased recombination rate, increased gene overlap, increased sequence conservation, increased Ka/Ks, and decreased ancestral marine frequency have smaller contributions to predictive power for speed of selection (Fig. 5A).

We also tested whether underlying sequence characteristics could predict not only the speed of selection in CH + SC + LB but also the location of the selected regions themselves (section S23). Recombination rate, QTL overlap, allelic age, and an integrated genomic context score (section S23) that incorporate the previous features are all useful predictors (Fig. 5B). By combining these fundamental features into a logistic model trained on the survey of extant populations, the most confident predictions of selected regions in the rest of the genome achieve 85% precision. This model performs 67% as well as predictions based only on empirical repeatability in extant populations in the northeast Pacific (Fig. 5B). Thus, our understanding of underlying principles reflects an incomplete yet substantial proportion of evolutionary repeatability.

To test the generality of these predictive factors, we applied the stickleback-trained model to a dataset of 12 pairs of species of Darwins finches (section S23) (57). Darwins finches have undergone adaptive radiation in the Galpagos Islands over the last several hundred thousand years, are ~435 million years divergent from stickleback, and face very different selective pressures. As in stickleback, however, the islands of divergence of all 12 analyzed pairs of species of Darwins finches (sensu Han et al.) are enriched for ancient alleles overlapping mapped QTLs with low recombination rates. The top 100 windows predicted by the stickleback model recover a median of 28-fold more previously identified islands of divergence than expected by chance (P < 1 1010; Fig. 5C), including the Alx1 and Hmga2 loci implicated in beak morphology in multiple species pairs (even without QTL input). The model also recovers a substantial proportion of differentiated loci in a recent case of cichlid speciation (58). Thus, a handful of basic genomic properties allow strong quantitative predictions of the location of key evolutionary loci, even across widely separated branches of life.

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Predicting future from past: The genomic basis of recurrent and rapid stickleback evolution - Science Advances