Dual and opposing roles for the kinesin-2 motor, KIF17, in Hedgehog-dependent cerebellar development.

While the kinesin-2 motors KIF3A and KIF3B have essential roles in ciliogenesis and Hedgehog (HH) signal transduction, potential role(s) for another kinesin-2 motor, KIF17, in HH signaling have yet to be explored. Here, we investigated the contribution of KIF17 to HH-dependent cerebellar development, where Kif17 is expressed in both HH-producing Purkinje cells and HH-responding cerebellar granule neuron progenitors (CGNPs). Germline Kif17 deletion in mice results in cerebellar hypoplasia due to reduced CGNP proliferation, a consequence of decreased HH pathway activity mediated through decreased Sonic HH (SHH) protein. Notably, Purkinje cell-specific Kif17 deletion partially phenocopies Kif17 germline mutants. Unexpectedly, CGNP-specific Kif17 deletion results in the opposite phenotype-increased CGNP proliferation and HH target gene expression due to altered GLI transcription factor processing. Together, these data identify KIF17 as a key regulator of HH-dependent cerebellar development, with dual and opposing roles in HH-producing Purkinje cells and HH-responding CGNPs.

Trouble in paradise: When two species of conservation and cultural value clash, causing a management conundrum.

Threatened species throughout the world are in decline due to various causes. In some cases, predators of conservation or cultural value are causing the decline of threatened prey, presenting a conservation conundrum for managers. We surveyed marine turtle nests on K'gari (formally known as Fraser Island), Australia, to investigate dingo predation of green and loggerhead turtle nests, where each of these species is of conservation value. Our monitoring revealed that 84% of nests were predated by dingoes. Only 16% of nests were not consumed by dingoes, and only 5.7% of nests were confirmed to have successfully hatched. Up to 94% of nests were consumed in some areas, and predation rates were similar across different dingo packs. Information on the available numbers of nests and dingoes in the area indicated that turtle nests alone are sufficient to support extant dingoes over the summer. These results indicate that marine turtle eggs represent a previously unquantified but important food source for dingoes on K'gari, and that turtle nests at this rookery site are under serious threat from dingoes. This research should highlight the importance of prioritising the protection of turtle nests from dingoes or risk losing the entire rookery forever in the near future.

Attitudes towards the Potential Use of Aversive Geofencing Devices to Manage Wild Elephant Movement.

Aversive geofencing devices (AGDs) or animal-borne satellite-linked shock collars might become a useful tool to mitigate human-elephant conflict (HEC). AGDs have the potential to condition problem elephants to avoid human-dominated landscapes by associating mild electric shocks with preceding audio warnings given as they approach virtual boundaries. We assessed the opinions of different stakeholders (experts, farmers, and others who have and have not experienced HEC; n = 611) on the potential use of AGDs on Asian elephants. Most respondents expressed positive opinions on the potential effectiveness of AGDs in managing elephant movement (62.2%). About 62.8% respondents also provided positive responses for the acceptability of AGDs if pilot studies with captive elephants have been successful in managing their movements. Some respondents perceived AGDs to be unacceptable because they are unethical or harmful and would be unsuccessful given wild elephants may respond differently to AGDs than captive elephants. Respondents identified acceptability, support and awareness of stakeholders, safety and wellbeing of elephants, logistical difficulties, durability and reliable functionality of AGDs, and uncertainties in elephants' responses to AGDs as potential challenges for implementing AGDs. These issues need attention when developing AGDs to increase support from stakeholders and to effectively reduce HEC incidents in the future.

Why humans kill animals and why we cannot avoid it.

Killing animals has been a ubiquitous human behaviour throughout history, yet it is becoming increasingly controversial and criticised in some parts of contemporary human society. Here we review 10 primary reasons why humans kill animals, discuss the necessity (or not) of these forms of killing, and describe the global ecological context for human killing of animals. Humans historically and currently kill animals either directly or indirectly for the following reasons: (1) wild harvest or food acquisition, (2) human health and safety, (3) agriculture and aquaculture, (4) urbanisation and industrialisation, (5) invasive, overabundant or nuisance wildlife control, (6) threatened species conservation, (7) recreation, sport or entertainment, (8) mercy or compassion, (9) cultural and religious practice, and (10) research, education and testing. While the necessity of some forms of animal killing is debatable and further depends on individual values, we emphasise that several of these forms of animal killing are a necessary component of our inescapable involvement in a single, functioning, finite, global food web. We conclude that humans (and all other animals) cannot live in a way that does not require animal killing either directly or indirectly, but humans can modify some of these killing behaviours in ways that improve the welfare of animals while they are alive, or to reduce animal suffering whenever they must be killed. We encourage a constructive dialogue that (1) accepts and permits human participation in one enormous global food web dependent on animal killing and (2) focuses on animal welfare and environmental sustainability. Doing so will improve the lives of both wild and domestic animals to a greater extent than efforts to avoid, prohibit or vilify human animal-killing behaviour.

Developing Future Biologists: developmental biology for undergraduates from underserved communities.

Developing Future Biologists (DFB) is an inclusive, trainee-run organization that strives to excite and engage the next generation of biologists, regardless of race, gender or socioeconomic status, in the field of developmental biology. DFB offers a week-long course consisting of active lectures, hands-on laboratory sessions, and professional development opportunities through interactions with scientists from a variety of backgrounds and careers. A major goal of DFB is to propel undergraduate students from underserved communities to pursue biomedical research opportunities and advanced degrees in science. To achieve this goal, we provide DFB participants with continuing access to a diverse network of scientists that students can utilize to secure opportunities and foster success throughout multiple stages of their research careers. Here, we describe the flourishing DFB program at the University of Michigan to encourage other institutions to create their own DFB programs.

Efficacy of Management Efforts to Reduce Food-Related Dingo-Human Interactions and Conflict on K'gari (Fraser Island), Australia.

Humans and dingoes (Canis familiaris (dingo)) share the environment of K'gari, and conflict inevitably occurs between the two species, particularly over food. Dingo attacks on humans have occurred, and some have been serious and even fatal in outcome. Wildlife feeding may cause animals to develop unnatural and potentially dangerous behaviours towards conspecifics and humans on a relatively frequent basis. Food-based attraction has been implicated in the development of human-directed aggression in the dingo population of K'gari. Supplemental feeding, whether intentional or accidental, alters wildlife foraging behaviours and may have consequences at the population and ecosystem levels. Management strategies such as education programs, prohibition of inappropriate human behaviours (compliance) and fencing of garbage dumps have each been implemented to stop the intentional or inadvertent feeding of dingoes by people. However, there has been no formal assessment of the effectiveness of these interventions at reducing food-related dingo-human incidents over time. We collated and analysed 7791 unique reports of dingo-human interactions on K'gari between 1990 and 2020, inclusive of 1307 food-related reports, including the severity of these interactions. These data showed clear seasonal peaks in the percentage of food-related dingo-human interactions, corresponding with biologically significant breeding periods in autumn and weaning and dispersing in spring. Trends in serious food-related incidents remained stable overtime. Less serious food-related incidents declined, suggesting that management efforts were successful. However, these efforts appear to have reached the limits of their effectiveness. Further innovations are required to reduce serious incidents involving the relatively few dingoes and people still experiencing conflict, and thereby provide protection to both species on K'gari.

CDON contributes to Hedgehog-dependent patterning and growth of the developing limb.

Hedgehog (HH) signaling is a major driver of tissue patterning during embryonic development through the regulation of a multitude of cell behaviors including cell fate specification, proliferation, migration, and survival. HH ligands signal through the canonical receptor PTCH1 and three co-receptors, GAS1, CDON and BOC. While previous studies demonstrated an overlapping and collective requirement for these co-receptors in early HH-dependent processes, the early embryonic lethality of Gas1;Cdon;Boc mutants precluded an assessment of their collective contribution to later HH-dependent signaling events. Specifically, a collective role for these co-receptors during limb development has yet to be explored. Here, we investigate the combined contribution of these co-receptors to digit specification, limb patterning and long bone growth through limb-specific conditional deletion of Cdon in a Gas1;Boc null background. Combined deletion of Gas1, Cdon and Boc in the limb results in digit loss as well as defects in limb outgrowth and long bone patterning. Taken together, these data demonstrate that GAS1, CDON and BOC are collectively required for HH-dependent patterning and growth of the developing limb.

Compassionate Conservation is indistinguishable from traditional forms of conservation in practice.

Animal welfare and ethics are important factors influencing wildlife conservation practice, and critics are increasingly challenging the underlying ethics and motivations supporting common conservation practices. "Compassionate Conservationists" argue that all conservationists should respect the rights of individual sentient animals and approach conservation problems from a position of compassion, and that doing so requires implementing practices that avoid direct harm to individual animals. In this way Compassionate Conservationists seek to contrast themselves with "Traditional Conservationists" who often express consequentialist decision-making processes that ostensibly aim to dispassionately minimize net animal harms, resulting in the common use of practices that directly harm or kill some animals. Conservationists and other observers might therefore conclude that the two sides of this debate are distinct and/or that their policy proscriptions produce different welfare outcomes for animals. To explore the validity of this conclusion we review the ethical philosophies underpinning two types of Compassionate Conservation-deontology and virtue ethics. Deontology focusses on animal rights or the moral duties or obligations of conservationists, whereas virtue ethics focusses on acting in ways that are virtuous or compassionate. We demonstrate that both types permit the intentional harm and killing of animals when faced with common conservation problems where animals will be harmed no matter what the conservationist does or does not do. We then describe the applied decision-making processes exhibited by Compassionate Conservationists (of both types) and Traditional Conservationists to show that they may each lead to the implementation of similar conservation practices (including lethal control) and produce similar outcomes for animals, despite the perceived differences in their ethical motivations. The widespread presence of wildlife conservation problems that cannot be resolved without causing at least some harm to some animals means that conservationists of all persuasions must routinely make trade-offs between the welfare of some animals over others. Compassionate Conservationists do this from an explicit position of animal rights and/or compassion, whereas Traditional Conservationists respect animal rights and exhibit this same compassion implicitly. These observations lead to the conclusion that Compassionate Conservation is indistinguishable from traditional forms of conservation in practice, and that the apparent disagreement among conservationists primarily concerns the effectiveness of various wildlife management practices at minimizing animal harm, and not the underlying ethics, motivations or morality of those practices.

Plexins promote Hedgehog signaling through their cytoplasmic GAP activity.

Hedgehog signaling controls tissue patterning during embryonic and postnatal development and continues to play important roles throughout life. Characterizing the full complement of Hedgehog pathway components is essential to understanding its wide-ranging functions. Previous work has identified neuropilins, established semaphorin receptors, as positive regulators of Hedgehog signaling. Neuropilins require plexin co-receptors to mediate semaphorin signaling, but the role of plexins in Hedgehog signaling has not yet been explored. Here, we provide evidence that multiple plexins promote Hedgehog signaling in NIH/3T3 mouse fibroblasts and that plexin loss of function in these cells results in significantly reduced Hedgehog pathway activity. Catalytic activity of the plexin GTPase-activating protein (GAP) domain is required for Hedgehog signal promotion, and constitutive activation of the GAP domain further amplifies Hedgehog signaling. Additionally, we demonstrate that plexins promote Hedgehog signaling at the level of GLI transcription factors and that this promotion requires intact primary cilia. Finally, we find that plexin loss of function significantly reduces the response to Hedgehog pathway activation in the mouse dentate gyrus. Together, these data identify plexins as novel components of the Hedgehog pathway and provide insight into their mechanism of action.

Combinatorial Gli activity directs immune infiltration and tumor growth in pancreatic cancer.

Proper Hedgehog (HH) signaling is essential for embryonic development, while aberrant HH signaling drives pediatric and adult cancers. HH signaling is frequently dysregulated in pancreatic cancer, yet its role remains controversial, with both tumor-promoting and tumor-restraining functions reported. Notably, the GLI family of HH transcription factors (GLI1, GLI2, GLI3), remain largely unexplored in pancreatic cancer. We therefore investigated the individual and combined contributions of GLI1-3 to pancreatic cancer progression. At pre-cancerous stages, fibroblast-specific Gli2/Gli3 deletion decreases immunosuppressive macrophage infiltration and promotes T cell infiltration. Strikingly, combined loss of Gli1/Gli2/Gli3 promotes macrophage infiltration, indicating that subtle changes in Gli expression differentially regulate immune infiltration. In invasive tumors, Gli2/Gli3 KO fibroblasts exclude immunosuppressive myeloid cells and suppress tumor growth by recruiting natural killer cells. Finally, we demonstrate that fibroblasts directly regulate macrophage and T cell migration through the expression of Gli-dependent cytokines. Thus, the coordinated activity of GLI1-3 directs the fibroinflammatory response throughout pancreatic cancer progression.

Suppression of p53 response by targeting p53-Mediator binding with a stapled peptide.

DNA-binding transcription factors (TFs) remain challenging to target with molecular probes. Many TFs function in part through interaction with Mediator, a 26-subunit complex that controls RNA polymerase II activity genome-wide. We sought to block p53 function by disrupting the p53-Mediator interaction. Through rational design and activity-based screening, we characterize a stapled peptide, with functional mimics of both p53 activation domains, that blocks p53-Mediator binding and selectively inhibits p53-dependent transcription in human cells; importantly, this "bivalent" peptide has negligible impact, genome-wide, on non-p53 target genes. Our proof-of-concept strategy circumvents the TF entirely and targets the TF-Mediator interface instead, with desired functional outcomes (i.e., selective inhibition of p53 activation). Furthermore, these results demonstrate that TF activation domains represent viable starting points for Mediator-targeting molecular probes, as an alternative to large compound libraries. Different TFs bind Mediator through different subunits, suggesting this strategy could be broadly applied to selectively alter gene expression programs.

C-X-C motif chemokine ligand 1 induced by Hedgehog signaling promotes mouse extrahepatic bile duct repair after acute injury.

BACKGROUND AND AIMS: In extrahepatic bile duct (EHBD) cholangiopathies, including primary sclerosing cholangitis, a reactive cholangiocyte phenotype is associated with inflammation and epithelial hyperproliferation. The signaling pathways involved in EHBD injury response are poorly understood. In this study, we investigated the role of Hedgehog (HH) signaling and its downstream effectors in controlling biliary proliferation and inflammation after EHBD injury. APPROACH AND RESULTS: Using mouse bile duct ligation as an acute EHBD injury model, we used inhibitory paradigms to uncover mechanisms promoting the proliferative response. HH signaling was inhibited genetically in Gli1-/- mice or by treating wild-type mice with LDE225. The role of neutrophils was tested using chemical (SB225002) and biological (lymphocyte antigen 6 complex locus G6D [Ly6G] antibodies) inhibitors of neutrophil recruitment. The cellular response was defined through morphometric quantification of proliferating cells and CD45+ and Ly6G+ immune cell populations. Key signaling component expression was measured and localized to specific EHBD cellular compartments by in situ hybridization, reporter strain analysis, and immunohistochemistry. Epithelial cell proliferation peaked 24 h after EHBD injury, preceded stromal cell proliferation, and was associated with neutrophil influx. Indian HH ligand expression in the biliary epithelium rapidly increased after injury. HH-responding cells and neutrophil chemoattractant C-X-C motif chemokine ligand 1 (CXCL1) expression mapped to EHBD stromal cells. Inhibition of HH signaling blocked CXCL1 induction, diminishing neutrophil recruitment and the biliary proliferative response to injury. Directly targeting neutrophils by inhibition of the CXCL1/C-X-C motif chemokine receptor 2/Ly6G signaling axis also decreased biliary proliferation. CONCLUSIONS: HH-regulated CXCL1 orchestrates the early inflammatory response and biliary proliferation after EHBD injury through complex cellular crosstalk.

Extrinsic KRAS Signaling Shapes the Pancreatic Microenvironment Through Fibroblast Reprogramming.

BACKGROUND & AIMS: Oncogenic Kirsten Rat Sarcoma virus (KRAS) is the hallmark mutation of human pancreatic cancer and a driver of tumorigenesis in genetically engineered mouse models of the disease. Although the tumor cell-intrinsic effects of oncogenic Kras expression have been widely studied, its role in regulating the extensive pancreatic tumor microenvironment is less understood. METHODS: Using a genetically engineered mouse model of inducible and reversible oncogenic Kras expression and a combination of approaches that include mass cytometry and single-cell RNA sequencing we studied the effect of oncogenic KRAS in the tumor microenvironment. RESULTS: We have discovered that non-cell autonomous (ie, extrinsic) oncogenic KRAS signaling reprograms pancreatic fibroblasts, activating an inflammatory gene expression program. As a result, fibroblasts become a hub of extracellular signaling, and the main source of cytokines mediating the polarization of protumorigenic macrophages while also preventing tissue repair. CONCLUSIONS: Our study provides fundamental knowledge on the mechanisms underlying the formation of the fibroinflammatory stroma in pancreatic cancer and highlights stromal pathways with the potential to be exploited therapeutically.

Identifying and prioritising climate change adaptation actions for greater one-horned rhinoceros (Rhinoceros unicornis) conservation in Nepal.

Climate change has started impacting species, ecosystems, genetic diversity within species, and ecological interactions and is thus a serious threat to conserving biodiversity globally. In the absence of adequate adaptation measures, biodiversity may continue to decline, and many species will possibly become extinct. Given that global temperature continues to increase, climate change adaptation has emerged as an overarching framework for conservation planning. We identified both ongoing and probable climate change adaptation actions for greater one-horned rhinoceros conservation in Nepal through a combination of literature review, key informant surveys (n = 53), focus group discussions (n = 37) and expert consultation (n = 9), and prioritised the identified adaptation actions through stakeholder consultation (n = 17). The majority of key informants (>80%) reported that climate change has been impacting rhinoceros, and more than 65% of them believe that rhinoceros habitat suitability in Nepal has been shifting westwards. Despite these perceived risks, climate change impacts have not been incorporated well into formal conservation planning for rhinoceros. Out of 20 identified adaptation actions under nine adaptation strategies, identifying and protecting climate refugia, restoring the existing habitats through wetland and grassland management, creating artificial highlands in floodplains to provide rhinoceros with refuge during severe floods, and translocating them to other suitable habitats received higher priority. These adaptation actions may contribute to reducing the vulnerability of rhinoceros to the likely impacts of climate change. This study is the first of its kind in Nepal and is expected to provide a guideline to align ongoing conservation measures into climate change adaptation planning for rhinoceros. Further, we emphasise the need to integrating likely climate change impacts while planning for rhinoceros conservation and initiating experimental research and monitoring programs to better inform adaptation planning in the future.

Unique lingual expression of the Hedgehog pathway antagonist Hedgehog-interacting protein in filiform papillae during homeostasis and ectopic expression in fungiform papillae during Hedgehog signaling inhibition.

BACKGROUND: Hedgehog (HH) signaling is essential for homeostasis in gustatory fungiform papillae (FP) and taste buds. However, activities of HH antagonists in these tissues remain unexplored. We investigated a potential role for HH-interacting protein (HHIP), an endogenous pathway antagonist, in regulating HH signaling during taste organ homeostasis. We found a restricted pattern of Hhip-expressing cells in the anterior epithelium of each nongustatory filiform papilla (FILIF) only. To test for roles in antagonism of HH signaling, we investigated HHIP after pathway inhibition with SMO inhibition via sonidegib and Smo deletion, Gli2 deletion/suppression, or with chorda tympani/lingual nerve cut. RESULTS: In all approaches, the HHIP expression pattern was retained in FILIF suggesting HH-independent regulation of HHIP. Remarkably, after pathway inhibition, HHIP expression was detected also in the conical, FILIF-like atypical FP. We found a close association of de novo expression of HHIP in atypical FP with loss of Gli1+, HH-responding cells. Further, we report that PTCH1 is another potential HH antagonist in FILIF that co-localizes with HHIP. CONCLUSIONS: After HH pathway inhibition the ectopic expression of HHIP correlates with a FILIF-like morphology in atypical FP and we propose that localized expression of the HH antagonist HHIP regulates pathway inhibition to maintain FILIF during tongue homeostasis.

GAS1 is required for NOTCH-dependent facilitation of SHH signaling in the ventral forebrain neuroepithelium.

Growth arrest-specific 1 (GAS1) acts as a co-receptor to patched 1, promoting sonic hedgehog (SHH) signaling in the developing nervous system. GAS1 mutations in humans and animal models result in forebrain and craniofacial malformations, defects ascribed to a function for GAS1 in SHH signaling during early neurulation. Here, we confirm loss of SHH activity in the forebrain neuroepithelium in GAS1-deficient mice and in induced pluripotent stem cell-derived cell models of human neuroepithelial differentiation. However, our studies document that this defect can be attributed, at least in part, to a novel role for GAS1 in facilitating NOTCH signaling, which is essential to sustain a persistent SHH activity domain in the forebrain neuroepithelium. GAS1 directly binds NOTCH1, enhancing ligand-induced processing of the NOTCH1 intracellular domain, which drives NOTCH pathway activity in the developing forebrain. Our findings identify a unique role for GAS1 in integrating NOTCH and SHH signal reception in neuroepithelial cells, and they suggest that loss of GAS1-dependent NOTCH1 activation contributes to forebrain malformations in individuals carrying GAS1 mutations.

Terrestrial mesopredators did not increase after top-predator removal in a large-scale experimental test of mesopredator release theory.

Removal or loss of top-predators has been predicted to cause cascading negative effects for ecosystems, including mesopredator release. However, reliable evidence for these processes in terrestrial systems has been mixed and equivocal due, in large part, to the systemic and continued use of low-inference study designs to investigate this issue. Even previous large-scale manipulative experiments of strong inferential value have been limited by experimental design features (i.e. failure to prevent migration between treatments) that constrain possible inferences about the presence or absence of mesopredator release effects. Here, we build on these previous strong-inference experiments and report the outcomes of additional large-scale manipulative experiments to eradicate Australian dingoes from two fenced areas where dingo migration was restricted and where theory would predict an increase in extant European red foxes, feral cats and goannas. We demonstrate the removal and suppression of dingoes to undetectable levels over 4-5 years with no corresponding increases in mesopredator relative abundances, which remained low and stable throughout the experiment at both sites. We further demonstrate widespread absence of negative relationships between predators, indicating that the mechanism underpinning predicted mesopredator releases was not present. Our results are consistent with all previous large-scale manipulative experiments and long-term mensurative studies which collectively demonstrate that (1) dingoes do not suppress red foxes, feral cats or goannas at the population level, (2) repeated, temporary suppression of dingoes in open systems does not create mesopredator release effects, and (3) removal and sustained suppression of dingoes to undetectable levels in closed systems does not create mesopredator release effects either. Our experiments add to similar reports from North America, Asia, Europe and southern Africa which indicate that not only is there a widespread absence of reliable evidence for these processes, but there is also a large and continually growing body of experimental evidence of absence for these processes in many terrestrial systems. We conclude that although sympatric predators may interact negatively with each other on smaller spatiotemporal scales, that these negative interactions do not always scale-up to the population level, nor are they always strong enough to create mesopredator suppression or release effects.

Animal Harms and Food Production: Informing Ethical Choices.

Ethical food choices have become an important societal theme in post-industrial countries. Many consumers are particularly interested in the animal welfare implications of the various foods they may choose to consume. However, concepts in animal welfare are rapidly evolving towards consideration of all animals (including wildlife) in contemporary approaches such as "One Welfare". This approach requires recognition that negative impacts (harms) may be intentional and obvious (e.g., slaughter of livestock) but also include the under-appreciated indirect or unintentional harms that often impact wildlife (e.g., land clearing). This is especially true in the Anthropocene, where impacts on non-human life are almost ubiquitous across all human activities. We applied the "harms" model of animal welfare assessment to several common food production systems and provide a framework for assessing the breadth (not intensity) of harms imposed. We considered all harms caused to wild as well as domestic animals, both direct effects and indirect effects. We described 21 forms of harm and considered how they applied to 16 forms of food production. Our analysis suggests that all food production systems harm animals to some degree and that the majority of these harms affect wildlife, not livestock. We conclude that the food production systems likely to impose the greatest overall breadth of harms to animals are intensive animal agriculture industries (e.g., dairy) that rely on a secondary food production system (e.g., cropping), while harvesting of locally available wild plants, mushrooms or seaweed is likely to impose the least harms. We present this conceptual analysis as a resource for those who want to begin considering the complex animal welfare trade-offs involved in their food choices.

Inhibition of Hedgehog Signaling Alters Fibroblast Composition in Pancreatic Cancer.

PURPOSE: Pancreatic ductal adenocarcinoma (PDAC) is a deadly disease characterized by an extensive fibroinflammatory stroma, which includes abundant cancer-associated fibroblast (CAF) populations. PDAC CAFs are heterogeneous, but the nature of this heterogeneity is incompletely understood. The Hedgehog pathway functions in PDAC in a paracrine manner, with ligands secreted by cancer cells signaling to stromal cells in the microenvironment. Previous reports investigating the role of Hedgehog signaling in PDAC have been contradictory, with Hedgehog signaling alternately proposed to promote or restrict tumor growth. In light of the newly discovered CAF heterogeneity, we investigated how Hedgehog pathway inhibition reprograms the PDAC microenvironment. EXPERIMENTAL DESIGN: We used a combination of pharmacologic inhibition, gain- and loss-of-function genetic experiments, cytometry by time-of-flight, and single-cell RNA sequencing to study the roles of Hedgehog signaling in PDAC. RESULTS: We found that Hedgehog signaling is uniquely activated in fibroblasts and differentially elevated in myofibroblastic CAFs (myCAF) compared with inflammatory CAFs (iCAF). Sonic Hedgehog overexpression promotes tumor growth, while Hedgehog pathway inhibition with the smoothened antagonist, LDE225, impairs tumor growth. Furthermore, Hedgehog pathway inhibition reduces myCAF numbers and increases iCAF numbers, which correlates with a decrease in cytotoxic T cells and an expansion in regulatory T cells, consistent with increased immunosuppression. CONCLUSIONS: Hedgehog pathway inhibition alters fibroblast composition and immune infiltration in the pancreatic cancer microenvironment.

Predicted declines in suitable habitat for greater one-horned rhinoceros (Rhinoceros unicornis) under future climate and land use change scenarios.

Rapidly changing climate is likely to modify the spatial distribution of both flora and fauna. Land use change continues to alter the availability and quality of habitat and further intensifies the effects of climate change on wildlife species. We used an ensemble modeling approach to predict changes in habitat suitability for an iconic wildlife species, greater one-horned rhinoceros due to the combined effects of climate and land use changes. We compiled an extensive database on current rhinoceros distribution and selected nine ecologically meaningful environmental variables for developing ensemble models of habitat suitability using ten different species distribution modeling algorithms in the BIOMOD2 R package; and we did this under current climatic conditions and then projected them onto two possible climate change scenarios (SSP1-2.6 and SSP5-8.5) and two different time frames (2050 and 2070). Out of ten algorithms, random forest performed the best, and five environmental variables-distance from grasslands, mean temperature of driest quarter, distance from wetlands, annual precipitation, and slope, contributed the most in the model. The ensemble model estimated the current suitable habitat of rhinoceros to be 2610 km2, about 1.77% of the total area of Nepal. The future habitat suitability under the lowest and highest emission scenarios was estimated to be: (1) 2325 and 1904 km2 in 2050; and (2) 2287 and 1686 km2 in 2070, respectively. Our results suggest that over one-third of the current rhinoceros habitat would become unsuitable within a period of 50 years, with the predicted declines being influenced to a greater degree by climatic changes than land use changes. We have recommended several measures to moderate these impacts, including relocation of the proposed Nijgad International Airport given that a considerable portion of potential rhinoceros habitat will be lost if the airport is constructed on the currently proposed site.