Invasion Trajectories Research Articles

Two distinct epithelial-to-mesenchymal transition programs control invasion and inflammation in segregated tumor cell populations

Epithelial-to-mesenchymal transition (EMT) triggers cell plasticity in embryonic development, adult injured tissues and cancer. Combining the analysis of EMT in cell lines, embryonic neural crest and mouse models of renal fibrosis and breast cancer, we find that there is not a cancer-specific EMT program. Instead, cancer cells dedifferentiate and bifurcate into two distinct and segregated cellular trajectories after activating either embryonic-like or adult-like EMTs to drive dissemination or inflammation, respectively. We show that SNAIL1 acts as a pioneer factor in both EMT trajectories, and PRRX1 drives the progression of the embryonic-like invasive trajectory. We also find that the two trajectories are plastic and interdependent, as the abrogation of the EMT invasive trajectory by deleting Prrx1 not only prevents metastasis but also enhances inflammation, increasing the recruitment of antitumor macrophages. Our data unveil an additional role for EMT in orchestrating intratumor heterogeneity, driving the distribution of functions associated with either inflammation or metastatic dissemination.

Environmental gradients mediate dispersal evolution during biological invasions.

Rapid evolution of increased dispersal at the edge of a range expansion can accelerate invasions. However, populations expanding across environmental gradients often face challenging environments that reduce fitness of dispersing individuals. We used an eco-evolutionary model to explore how environmental gradients influence dispersal evolution and, in turn, modulate the speed and predictability of invasion. Environmental gradients opposed evolution of increased dispersal during invasion, even leading to evolution of reduced dispersal along steeper gradients. Counterintuitively, reduced dispersal could allow for faster expansion by minimizing maladaptive gene flow and facilitating adaptation. While dispersal evolution across homogenous landscapes increased both the mean and variance of expansion speed, these increases were greatly dampened by environmental gradients. We illustrate our model's potential application to prediction and management of invasions by parameterizing it with data from a recent invertebrate range expansion. Overall, we find that environmental gradients strongly modulate the effect of dispersal evolution on invasion trajectories.

Idiosyncratic invasion trajectories of human bacterial pathogens facing temperature disturbances in soil microbial communities

Current knowledge about effects of disturbance on the fate of invaders in complex microbial ecosystems is still in its infancy. In order to investigate this issue, we compared the fate of Klebsiella pneumoniae (Kp) and Listeria monocytogenes (Lm) in soil microcosms. We then used environmental disturbances (freeze–thaw or heat cycles) to compare the fate of both invaders and manipulate soil microbial diversity. Population dynamics of the two pathogens was assessed over 50 days of invasion while microbial diversity was measured at times 0, 20 and 40 days. The outcome of invasion was strain-dependent and the response of the two invaders to disturbance differed. Resistance to Kp invasion was higher under the conditions where resident microbial diversity was the highest while a significant drop of diversity was linked to a higher persistence. In contrast, Lm faced stronger resistance to invasion in heat-treated microcosms where diversity was the lowest. Our results show that diversity is not a universal proxy of resistance to microbial invasion, indicating the need to properly assess other intrinsic properties of the invader, such as its metabolic repertoire, or the array of interactions between the invader and resident communities.

Whole-mitogenome analysis unveils previously undescribed genetic diversity in cane toads across their invasion trajectory.

Invasive species offer insights into rapid adaptation to novel environments. The iconic cane toad (Rhinella marina) is an excellent model for studying rapid adaptation during invasion. Previous research using the mitochondrial NADH dehydrogenase 3 (ND3) gene in Hawai'ian and Australian invasive populations found a single haplotype, indicating an extreme genetic bottleneck following introduction. Nuclear genetic diversity also exhibited reductions across the genome in these two populations. Here, we investigated the mitochondrial genomics of cane toads across this invasion trajectory. We created the first reference mitochondrial genome for this species using long-read sequence data. We combined whole-genome resequencing data of 15 toads with published transcriptomic data of 125 individuals to construct nearly complete mitochondrial genomes from the native (French Guiana) and introduced (Hawai'i and Australia) ranges for population genomic analyses. In agreement with previous investigations of these populations, we identified genetic bottlenecks in both Hawai'ian and Australian introduced populations, alongside evidence of population expansion in the invasive ranges. Although mitochondrial genetic diversity in introduced populations was reduced, our results revealed that it had been underestimated: we identified 45 mitochondrial haplotypes in Hawai'ian and Australian samples, none of which were found in the native range. Additionally, we identified two distinct groups of haplotypes from the native range, separated by a minimum of 110 base pairs (0.6%). These findings enhance our understanding of how invasion has shaped the genetic landscape of this species.

Stiffness-Tunable Substrate Fabrication by DMD-Based Optical Projection Lithography for Cancer Cell Invasion Studies.

Cancer cell invasion is a critical cause of fatality in cancer patients. Physiologically relevant tumor models play a key role in revealing the mechanisms underlying the invasive behavior of cancer cells. However, most existing models only consider interactions between cells and extracellular matrix (ECM) components while neglecting the role of matrix stiffness in tumor invasion. Here, we propose an effective approach that can construct stiffness-tunable substrates using digital mirror device (DMD)-based optical projection lithography to explore the invasion behavior of cancer cells. The printability, mechanical properties, and cell viability of three-dimensional (3D) models can be tuned by the concentration of prepolymer and the exposure time. The invasion trajectories of gastric cancer cells in tumor models of different stiffness were automatically detected and tracked in real-time using a deep learning algorithm. The results show that tumor models of different mechanical stiffness can yield distinct regulatory effects. Moreover, owing to the biophysical characteristics of the 3D in vitro model, different cellular substructures of cancer cells were induced. The proposed tunable substrate construction method can be used to build various microstructures to achieve simulation of cancer invasion and antitumor screening, which has great potential in promoting personalized therapy.

10079-MPC-6 RADIOLOGICAL AND MOLECULAR ANALYSIS OF WHO GRADE 4 BUTTERFLY GLIOMAS WITH A TAILORED SURGICAL APPROACH

Abstract OBJECT Surgical resection for butterfly gliomas (bG) are still under debate, but recent findings have shown that surgery prolongs survival. However, clinical and molecular background of bG still remains unknown and surgical strategies based on such findings are scarce. OBJECTS AND METHODS We treated 39 bG patients between 2005 and 2023, including 32 cases of glioblastoma and 5 cases of astrocytoma, WHO grade 4, IDH-mutant. For the validation, we reviewed 968 MRI obtained from the public datasets (UPENN-GBM, n=611; TCGA-GBM, n=255; CPTAC-GBM, n=61; IvyGAP, n=41). RESULTS Molecular analyses revealed that 51.3% of TERT promoter mutations, 59.0% of EGFR amplification/gain, 38.5% of PTEN hemi/homozygous, and 51.3% of CDKN2A hemi/homozygous deletion. Sequential radiological imaging before typical bG diagnosis demonstrated that bG harbors two subtypes which are corpus callosum (CC)-type and hemispheric (H)-type. The multisampling of H-type showed that accumulating alterations in the CC lesions than hemisphere, indicating an invasive trajectory from the hemisphere to the CC region. Survival analysis presented that CC-type was significantly poorer overall survival (OS) than H-type (263 days and 691 days, respectively, P = 0.018). The validation cohort also supported the poor survival of CC-type. CONCLUSION Our detail analyses demonstrated the possibility of two subtypes in bG. Unilateral trans-cortical approach from the larger tumor size with CC removal is favored in the H-type. On the other hand, bilateral trans-sulcal and interhemispheric approaches preserving the intact cortex and cingulate gyrus is an alternative for CC-type.

Insights from the past: Invasion trajectory and niche trends of a global freshwater invader.

Freshwater ecosystems are invaded by a non-random selection of taxa, among which crayfish stand out with successful examples worldwide. Species distribution models (SDMs) have been used to detect suitable areas for invasive species and predict their potential distributions. However, these prediction exercises assume the stability of realized environmental niches, which is uncertain during invasion. Worldwide evaluations involving cosmopolitan invaders may be particularly useful but have seldom been considered. Focusing on the successful invasion history of the red swamp crayfish, Procambarus clarkii, we assessed its geographic expansion and niche trends over time. Based on global occurrences from 1854 to 2022, multiple sequential SDMs have been implemented based on a set of bioclimatic variables. The environmental suitability for each period was projected through to the next period(s) using an ensemble procedure of commonly used SDM algorithms. As the records of the species are known, it was possible to check whether the modelling projections were concordant with the observed expansion of red swamp crayfish at a global scale. This also permitted analysis of its realized niche, and its dynamics, during different expansion phases. SDM maps based on past species records showed concordance with the known crayfish distributions and yielded similar spatial patterns with outputs overperforming random combinations of cells in term of suitability. The results also reflect the stability of the species niche, which despite some expansions during the invasion process, changed little in terms of main position in functional space over time. SDMs developed in the early stages of invasion provide useful insights but also tend to underpredict the potential range compared to models that were built for later stages. Our approach can be easily transferable to other well-documented taxa and represents valuable evidence for validating the use of SDMs, considering a highly dynamic world where biogeographical barriers are often bypassed.

NIMG-61. RADIOLOGICAL, CLINICAL, AND MOLECULAR ANALYSES UNRAVEL DISTINCT SUBTYPES OF ADULT GRADE 4 BUTTERFLY GLIOMAS AFFECTING THE PROGNOSIS

Abstract Gliomas located at the corpus callosum (CC) and invading the bilateral hemispheres is known as butterfly gliomas (bG). Images of bG are frequently used as the hallmark of glioblastoma (GB); however, the molecular background and origins of bG remain unknown. We collected 37 bG patients, including 32 cases of GB and 5 cases of astrocytoma, WHO grade 4, IDH-mutant. Our dataset included extremely rare sequential radiological imaging before receiving a definitive diagnosis of bG. One case emerged from CC and subsequently became bG, whereas five cases emerged from the cerebral hemisphere and became bG, suggesting distinct radiological origins exist in bG. Reflecting this observation to further analysis, we subdivided the cohort into CC-type (n = 15) and hemispheric-type (n = 22) based on the ratio calculated by the tumor volume at CC and hemisphere. Survival analysis presented that CC-type was significantly poorer overall survival (OS) than hemispheric-type (P = 0.010), indicating that bG has clinically distinct two subtypes. Molecular analyses revealed that TERT promoter mutations and IDH1 mutations were observed in 73.3% and 0% in the CC-type, whereas 36.4% and 22.7% in the hemispheric-type, respectively. Other alterations such as MGMT promoter methylation, EGFR amplification/gain, CDKN2A hemi/homozygous deletion, and PTEN hemi/homozygous deletion remained the same between two groups. None of the patients had H3F3A, HIST1H3B, or BRAF mutations. In the hemispheric-type, the multisampling of hemispheric and CC specimens revealed accumulating alterations in the CC lesions, indicating an invasive trajectory from the hemisphere to the CC region. For the validation, we reviewed 993 MRI obtained from the public datasets (TCGA-GBM, CPTAC-GBM, IvyGap, and UPenn-GBM) and found the same trend: CC-type presented poor OS. bG is not a sole subtype with just extensive invasion from the hemisphere. Our results highlight the pathophysiology of bG and may influence its management.

The Invasion of Galinsoga quadriradiata into High Elevations Is Shaped by Variation in AMF Communities.

Mountain ranges have been previously suggested to act as natural barriers to plant invasion due to extreme environmental conditions. However, how arbuscular mycorrhizal fungi (AMF) affect invasion into these systems has been less explored. Here, we investigated how changes in AMF communities affect the performance of Galinsoga quadriradiata in mountain ranges. We performed a greenhouse experiment to study the impact of inoculations of AMF from different elevations on the performance and reproduction of invaders and how competition with native plants changes the effects of invader-AMF interactions. We found strong evidence for a nuanced role of AMF associations in the invasion trajectory of G. quadriradiata, with facilitative effects at low elevations and inhibitory effects at high elevations. Galinsoga quadriradiata performed best when grown with inoculum collected from the same elevation but performed worst when grown with inoculum collected from beyond its currently invaded range, suggesting that AMF communities can help deter invasion at high elevations. Finally, the invasive plants grown alone experienced negative effects from AMF, while those grown in competition experienced positive effects, regardless of the AMF source. This suggests that G. quadriradiata lowers its partnerships with AMF in stressful environments unless native plants are present, in which case it overpowers native plants to obtain AMF support during invasion. Finally, our results indicate that invader-AMF interactions can inhibit invasive range expansion at high elevations, and biotic interactions, in addition to harsh environmental conditions, make high-elevation mountain ranges natural barriers against continued invasion.

Tracking climate-change-induced biological invasions by metabarcoding archived natural eDNA samplers

In a time of unprecedented environmental change, understanding the response of organisms and ecosystems to change is paramount1. However, our knowledge of anthropogenic impacts on ecosystems is limited by a lack of standardized retrospective biomonitoring data2. Here, we use a four-decade time series of archived blue mussels to trace spatiotemporal biodiversity change in coastal ecosystems. The filter-feeding mussels, which were initially collected for pollution monitoring, can serve as natural eDNA samplers, carrying an imprint of the surrounding aquatic community at the time of sampling3. By sequencing the preserved DNA, we characterize highly diverse mussel-associated communities and reconstruct the invasion trajectory of an invasive species, the barnacle Austrominius modestus. We quantitatively trace population growth of the invader to the detriment of native taxa and uncover repeated population collapses and reinvasions after cold winters. By providing highly resolved temporal data on community assembly and global warming-driven invasion processes, natural eDNA sampler time series overcome a critical shortfall in our understanding of biodiversity change in the Anthropocene.

Critical role of insertion preference for invasion trajectory of transposons.

It is unclear how mobile DNA sequences (transposable elements, hereafter TEs) invade eukaryotic genomes and reach stable copy numbers, as transposition can decrease host fitness. This challenge is particularly stark early in the invasion of a TE family at which point hosts may lack the specialized machinery to repress the spread of these TEs. One possibility (in addition to the evolution of host regulation of TEs) is that TE families may evolve to preferentially insert into chromosomal regions that are less likely to impact host fitness. This may allow the mean TE copy number to grow while minimizing the risk for host population extinction. To test this, we constructed simulations to explore how the transposition probability and insertion preference of a TE family influence the evolution of mean TE copy number and host population size, allowing for extinction. We find that the effect of a TE family's insertion preference depends on a host's ability to regulate this TE family. Without host repression, a neutral insertion preference increases the frequency of and decreases the time to population extinction. With host repression, a preference for neutral insertions minimizes the cumulative deleterious load, increases population fitness, and, ultimately, avoids triggering an extinction vortex.

Multiple invasion trajectories induce niche dynamics inconsistency and increase risk uncertainty of a plant invader

AbstractOur knowledge of how niche dynamic patterns respond to invasion trajectories and influence invasion risk prediction is elusive for the majority of notorious invaders, hindering scientific understanding, biosecurity planning and practice, and management implementation of biological invasions. We used Mikania micrantha, one of the world's worst invasive alien species (IAS), to test the hypothesis that multiple invasion trajectories could induce niche dynamics inconsistency and increase risk uncertainty of IAS. We compiled a robust database of M. micrantha occurrence across its native range in Central and South America and invaded range in China. This database was used to clarify different invaded ranges and invasion trajectories of M. micrantha in China. Principal components analysis of climatic variables associated with the database was used to explore the niche dynamic patterns associated with multiple invasion trajectories of M. micrantha. Maximum entropy algorithm was used to predict the high‐risk area of M. micrantha invasion using occurrence datasets for invaded ranges where niches remained conservative, and to detect area changes with the inclusion of occurrence datasets for invaded ranges where niche shifts occurred. M. micrantha invasion occurred in three geographically distinct regions, with conservative climate niches in southern and southeastern China and climatic niche shifts in southwestern China. A high‐risk area for M. micrantha invasion spanned multiple provinces and cities and expanded considerably with the inclusion of the occurrence dataset for southwestern China. Our findings contribute to the theoretical understanding of invasion mechanisms and the practical optimization of biosecurity planning and implementation.

Why Are Invasive Plants Successful?

Plant invasions, a byproduct of globalization, are increasing worldwide. Because of their ecological and economic impacts, considerable efforts have been made to understand and predict the success of non-native plants. Numerous frameworks, hypotheses, and theories have been advanced to conceptualize the interactions of multiple drivers and context dependence of invasion success with the aim of achieving robust explanations with predictive power. We review these efforts from a community-level perspective rather than a biogeographical one, focusing on terrestrial systems, and explore the roles of intrinsic plant properties in determining species invasiveness, as well as the effects of biotic and abiotic conditions in mediating ecosystem invasibility (or resistance) and ecological and evolutionary processes. We also consider the fundamental influences of human-induced changes at scales ranging from local to global in triggering, promoting, and sustaining plant invasions and discuss how these changes could alter future invasion trajectories.

Effects of nutrient pulses on exotic species shift from positive to neutral with decreasing water availability.

Temporal fluctuation in nutrient availability generally promotes the growth of exotic plant species and has been recognized as an important driver of exotic plant invasions. However, little is known about how the impact of fluctuating nutrients on exotic species is dependent on the availability of other resources, although most ecosystems are experiencing dramatic variations in a wide variety of resources due to global change and human disturbance. Here, we explored how water availability mediates the effect of nutrient pulses on the growth of six exotic and six native plant species. We subjected individual plants of exotic and native species to well watered or water stressed conditions. For each level of water availability, we added equivalent amounts of nutrients at a constant rate, as a single large pulse, or in multiple small pulses. Under well watered conditions, nutrient pulses promoted exotic plant growth relative to nutrients supplied constantly, while they had no significant effect on natives. In contrast, under water stressed conditions, water deficiency inhibited the growth of all exotic and native species. More importantly, nutrient pulses did not increase plant growth relative to nutrients supplied constantly and these phenomena were observed for both exotic and native species. Taken together, our study shows that the impact of fluctuating nutrient availability on the growth of exotic plant species strongly depends on the variation of other resources, and that the positive effect of nutrient pulses under well watered conditions disappears under water stressed conditions. Our findings suggest that the variation in multiple resources may have complex feedback on exotic plant invasions and, therefore, it is critical to encompass multiple resources for the evaluation of fluctuating resource availability effects on exotic plant species. This will allow us to project the invasive trajectory of exotic plant species more accurately under future global change and human disturbance.

Climate Change Helps Polar Invasives Establish and Flourish: Evidence from Long-Term Monitoring of the Blowfly Calliphora vicina.

The isolated sub-Antarctic islands are of major ecological interest because of their unique species diversity and long history of limited human disturbance. However, since the presence of Europeans, these islands and their sensitive biota have been under increasing pressure due to human activity and associated biological invasions. In such delicate ecosystems, biological invasions are an exceptional threat that may be further amplified by climate change. We examined the invasion trajectory of the blowfly Calliphora vicina (Robineau-Desvoidy 1830). First introduced in the sub-Antarctic Kerguelen Islands in the 1970s, it is thought to have persisted only in sheltered microclimates for several decades. Here, we show that, in recent decades, C. vicina has been able to establish itself more widely. We combine experimental thermal developmental data with long-term ecological and meteorological monitoring to address whether warming conditions help explain its current success and dynamics in the eastern Kerguelen Islands. We found that warming temperatures and accumulated degree days could explain the species' phenological and long-term invasion dynamics, indicating that climate change has likely assisted its establishment. This study represents a unique long-term view of a polar invader and stresses the rapidly increasing susceptibility of cold regions to invasion under climate change.

Learning-Based Perimeter Invasion Design with Partial Information of Defense Model

This paper focuses on the learning-based perimeter-defense problem. Specifically, we consider a scenario where an attacker invades an area protected by a defender with only partial information about the target area and defense strategy. The attack design is challenging since the flexible and unknown defense strategy results in the highly uncertain feasible invasion space. To address the problem, we propose a learning-based method by using patrol and defense trajectories. First, we apply an ellipse fitting method to regress the perimeter of the protected area with piecewise elliptic segments. Then, we characterize the defense behaviors into different patterns and learn the conditions to activate different strategies by tentative invasions. Finally, we design a model predictive controller to solve the optimal invasion trajectory planning. Simulations are provided to illustrate the feasibility and effectiveness of the proposed method.

The COVID-19 Restrictions and Biological Invasion: A Global Terrestrial Ecosystem Perspective on Propagule Pressure and Invasion Trajectory

Biological invasions driven by climate change, transportation, and intercontinental trade, as well as land-use change and tourism, pose severe threats to biodiversity and ecosystem services worldwide. However, the COVID-19-induced shutdowns and cross-border restrictions could have significantly impacted some of these drivers. Thus, COVID-19-induced restrictions may potentially alter the invasion trajectories and propagule pressure of invasive alien species, yet very few studies have examined this possibility. Here, we provide a unique conceptual framework to examine how COVID-19-induced restrictions may influence the rate, magnitude, and trajectories of biological invasions. We also discuss the similarities between the high-hit regions of COVID-19 and the global hotspot of biological invasions. Additionally, we assessed whether previous predictions of biological invasions still hold despite the strong impact of COVID-19 on the drivers of invasions. Finally, we emphasize the possibility of harnessing such restrictive measures to manage invasive species, nature reserves, and national parks. The present study is a significant addition to the current understanding of the interplay between pandemic outbreaks and biological invasions in the context of both direct and indirect effects of global ecosystem change.

Delineating the dynamic evolution from preneoplasia to invasive lung adenocarcinoma by integrating single-cell RNA sequencing and spatial transcriptomics

The cell ecology and spatial niche implicated in the dynamic and sequential process of lung adenocarcinoma (LUAD) from adenocarcinoma in situ (AIS) to minimally invasive adenocarcinoma (MIA) and subsequent invasive adenocarcinoma (IAC) have not yet been elucidated. Here, we performed an integrative analysis of single-cell RNA sequencing (scRNA-seq) and spatial transcriptomics (ST) to characterize the cell atlas of the invasion trajectory of LUAD. We found that the UBE2C + cancer cell subpopulation constantly increased during the invasive process of LUAD with remarkable elevation in IAC, and its spatial distribution was in the peripheral cancer region of the IAC, representing a more malignant phenotype. Furthermore, analysis of the TME cell type subpopulation showed a constant decrease in mast cells, monocytes, and lymphatic endothelial cells, which were implicated in the whole process of invasive LUAD, accompanied by an increase in NK cells and MALT B cells from AIS to MIA and an increase in Tregs and secretory B cells from MIA to IAC. Notably, for AIS, cancer cells, NK cells, and mast cells were colocalized in the cancer region; however, for IAC, Tregs colocalized with cancer cells. Finally, communication and interaction between cancer cells and TME cell-induced constitutive activation of TGF-β signaling were involved in the invasion of IAC. Therefore, our results reveal the specific cellular information and spatial architecture of cancer cells and TME subpopulations, as well as the cellular interaction between them, which will facilitate the identification and development of precision medicine in the invasive process of LUAD from AIS to IAC.

Single-cell transcriptome highlights a multilayer regulatory network on an invasive trajectory within colorectal cancer progression

PurposeColorectal cancer (CRC) is one of the most common and fatal gastrointestinal malignancies, in which cancer stem cells (CSCs) were identified to enable tumor heterogeneity and initiate tumor formation. However, the process from CSCs to invasion cells is unconfirmed.MethodsSeveral bioinformatics methods, including clustering, pseudotime analysis, gene set variation analysis and gene ontology enrichment, were used to construct a path of gradual transformation of CSCs to invasive cells, called “stem-to-invasion path”. A large amount of signaling interactions were collated to build the multilayer regulatory network. Kaplan–Meier curve and time-dependent ROC method were applied to reveal prognostic values.ResultsWe validated the heterogeneity of cells in the tumor microenvironment and revealed the presence of malignant epithelial cells with high invasive potential within primary colonic carcinomas. Next, the “stem-to-invasion path” was identified through constructing a branching trajectory with cancer cells arranged in order. A multilayer regulatory network considered as the vital factor involved in acquiring invasion characteristics underlying the path was built to elucidate the interactions between tumor cell and tumor-associated microenvironment. Then we further identified a novel combinatorial biomarker that can be used to assess the prognosis for CRC patients, and validated its predictive robustness on the independent dataset.ConclusionOur work provides new insights into the acquisition of invasive potential in primary tumor cells, as well as potential therapeutic targets for CRC invasiveness, which may be useful for the cancer research and clinical treatment.Supplementary InformationThe online version contains supplementary material available at 10.1007/s00432-022-04020-2.

Divergent long-term impacts of lethally toxic cane toads (Rhinella marina) on two species of apex predators (monitor lizards, Varanus spp.).

Biological invasions can massively disrupt ecosystems, but evolutionary and ecological adjustments may modify the magnitude of that impact through time. Such post-colonisation shifts can change priorities for management. We quantified the abundance of two species of giant monitor lizards, and of the availability of their mammalian prey, across 45 sites distributed across the entire invasion trajectory of the cane toad (Rhinella marina) in Australia. One varanid species (Varanus panoptes from tropical Australia) showed dramatic population collapse with toad invasion, with no sign of recovery at most (but not all) sites that toads had occupied for up to 80 years. In contrast, abundance of the other species (Varanus varius from eastern-coastal Australia) was largely unaffected by toad invasion. That difference might reflect availability of alternative food sources in eastern-coastal areas, perhaps exacerbated by the widespread prior collapse of populations of small mammals across tropical (but not eastern) Australia. According to this hypothesis, the impact of cane toads on apex predators has been exacerbated and prolonged by a scarcity of alternative prey. More generally, multiple anthropogenically-induced changes to natural ecosystems may have synergistic effects, intensifying the impacts beyond that expected from either threat in isolation.