Consensus recommendations for systemic therapies in the management of relapsed Ewing sarcoma: A report from the National Ewing Sarcoma Tumor Board.

Ewing sarcoma (ES) is a malignant tumor of bone and soft tissue that most often occurs in children, adolescents, and young adults. Debate and controversy remain in the management of relapsed/refractory ES (RR-ES). The authors leveraged the expertise assembled by the National Ewing Sarcoma Tumor Board, a multidisciplinary virtual tumor board that meets monthly to discuss challenging cases of ES. In this review, they focus on select topics that apply to the management of patients with RR-ES. The specific topics covered include the initial approach of such patients and discussion of the goals of care, the role of molecular testing, chemotherapy regimens and novel agents to consider, the role of maintenance therapy, and the use of high-dose chemotherapy with autologous stem cell rescue. The data referenced are often limited to subgroup analyses and/or compiled from multiple sources. Although not intended to replace the clinical judgement of treating physicians, these guidelines are intended to support clinicians and provide some clarity and recommendations for the management of patients with RR-ES. PLAIN LANGUAGE SUMMARY: Ewing sarcoma (ES) is a bone and soft tissue cancer that most often occurs in teenagers and young adults. This article uses the experience of the National Ewing Sarcoma Tumor Board, a multi-institution, multidisciplinary virtual tumor board that meets monthly to discuss challenging cases of ES and to address questions related to the treatment of patients with relapsed ES. Although not intended to replace the clinical judgement of treating physicians and limited by available data, these consensus recommendations will support clinicians who treat patients with this challenging malignancy, made even more difficult when it recurs.

Clofarabine monotherapy in aggressive, relapsed and refractory Langerhans cell histiocytosis.

Over 50% of patients with systemic LCH are not cured with front-line therapies, and data to guide salvage options are limited. We describe 58 patients with LCH who were treated with clofarabine. Clofarabine monotherapy was active against LCH in this cohort, including heavily pretreated patients with a systemic objective response rate of 92.6%, higher in children (93.8%) than adults (83.3%). BRAFV600E+ variant allele frequency in peripheral blood is correlated with clinical responses. Prospective multicentre trials are warranted to determine optimal dosing, long-term efficacy, late toxicities, relative cost and patient-reported outcomes of clofarabine compared to alternative LCH salvage therapy strategies.

Flexible Cyclic Immunofluorescence (cyCIF) Using Oligonucleotide Barcoded Antibodies.

Advances in our understanding of the complex, multifaceted interactions between tumor epithelia, immune infiltrate, and tumor microenvironmental cells have been driven by highly multiplexed imaging technologies. These techniques are capable of labeling many more biomarkers than conventional immunostaining methods. However, multiplexed imaging techniques suffer from low detection sensitivity, cell loss-particularly in fragile samples-, and challenges with antibody labeling. Herein, we developed and optimized an oligonucleotide antibody barcoding strategy for cyclic immunofluorescence (cyCIF) that can be amplified to increase the detection efficiency of low-abundance antigens. Stained fluorescence signals can be readily removed using ultraviolet light treatment, preserving tissue and fragile cell sample integrity. We also extended the oligonucleotide barcoding strategy to secondary antibodies to enable the inclusion of difficult-to-label primary antibodies in a cyCIF panel. Using both the amplification oligonucleotides to label DNA barcoded antibodies and in situ hybridization of multiple fluorescently labeled oligonucleotides resulted in signal amplification and increased signal-to-background ratios. This procedure was optimized through the examination of staining parameters including staining oligonucleotide concentration, staining temperature, and oligonucleotide sequence design, resulting in a robust amplification technique. As a proof-of-concept, we demonstrate the flexibility of our cyCIF strategy by simultaneously imaging with the original oligonucleotide conjugated antibody (Ab-oligo) cyCIF strategy, the novel Ab-oligo cyCIF amplification strategy, as well as direct and indirect immunofluorescence to generate highly multiplexed images.

Relevance of circulating hybrid cells as a non-invasive biomarker for myriad solid tumors.

Metastatic progression defines the final stages of tumor evolution and underlies the majority of cancer-related deaths. The heterogeneity in disseminated tumor cell populations capable of seeding and growing in distant organ sites contributes to the development of treatment resistant disease. We recently reported the identification of a novel tumor-derived cell population, circulating hybrid cells (CHCs), harboring attributes from both macrophages and neoplastic cells, including functional characteristics important to metastatic spread. These disseminated hybrids outnumber conventionally defined circulating tumor cells (CTCs) in cancer patients. It is unknown if CHCs represent a generalized cancer mechanism for cell dissemination, or if this population is relevant to the metastatic cascade. Herein, we detect CHCs in the peripheral blood of patients with cancer in myriad disease sites encompassing epithelial and non-epithelial malignancies. Further, we demonstrate that in vivo-derived hybrid cells harbor tumor-initiating capacity in murine cancer models and that CHCs from human breast cancer patients express stem cell antigens, features consistent with the potential to seed and grow at metastatic sites. Finally, we reveal heterogeneity of CHC phenotypes reflect key tumor features, including oncogenic mutations and functional protein expression. Importantly, this novel population of disseminated neoplastic cells opens a new area in cancer biology and renewed opportunity for battling metastatic disease.

Measure Twice: Promise of Liquid Biopsy in Pediatric High-Grade Gliomas.

PURPOSE: To review and critique the current state of liquid biopsy in pHGG. MATERIALS AND METHODS: Published literature was reviewed for articles related to liquid biopsy in pediatric glioma and adult glioma with a focus on high-grade gliomas. RESULTS: This review discusses the current state of liquid biomarkers of pHGG and their potential applications for liquid biopsy development. CONCLUSIONS: While nascent, the progress toward identifying circulating analytes of pHGG primes the field of neuro-oncoogy for liquid biopsy development.

Wild, connected, and diverse: building a more resilient system of protected areas.

Current systems of conservation reserves may be insufficient to sustain biodiversity in the face of climate change and habitat losses. Consequently, calls have been made to protect Earth's remaining wildlands and complete the system of protected areas by establishing conservation reserves that (1) better represent ecosystems, (2) increase connectivity to facilitate biota movement in response to stressors including climate change, and (3) promote species persistence within intact landscapes. Using geospatial data, we conducted an assessment for expanding protected areas within the contiguous United States to include the least human-modified wildlands, establish a connected network, and better represent ecosystem diversity and hotspots of biodiversity. Our composite map highlights areas of high value to achieve these goals in the western United States, where existing protected areas and lands with high ecological integrity are concentrated. We also identified important areas in the East rich in species and containing ecosystems that are poorly represented in the existing protected area system. Expanding protection to these priority areas is ultimately expected to create a more resilient system for protecting the nation's biological heritage. This expectation should be subject to rigorous testing prior to implementation, and regional monitoring will ensure areas and actions are adjusted over time.

Identifying Corridors among Large Protected Areas in the United States.

Conservation scientists emphasize the importance of maintaining a connected network of protected areas to prevent ecosystems and populations from becoming isolated, reduce the risk of extinction, and ultimately sustain biodiversity. Keeping protected areas connected in a network is increasingly recognized as a conservation priority in the current era of rapid climate change. Models that identify suitable linkages between core areas have been used to prioritize potentially important corridors for maintaining functional connectivity. Here, we identify the most "natural" (i.e., least human-modified) corridors between large protected areas in the contiguous Unites States. We aggregated results from multiple connectivity models to develop a composite map of corridors reflecting agreement of models run under different assumptions about how human modification of land may influence connectivity. To identify which land units are most important for sustaining structural connectivity, we used the composite map of corridors to evaluate connectivity priorities in two ways: (1) among land units outside of our pool of large core protected areas and (2) among units administratively protected as Inventoried Roadless (IRAs) or Wilderness Study Areas (WSAs). Corridor values varied substantially among classes of "unprotected" non-core land units, and land units of high connectivity value and priority represent diverse ownerships and existing levels of protections. We provide a ranking of IRAs and WSAs that should be prioritized for additional protection to maintain minimal human modification. Our results provide a coarse-scale assessment of connectivity priorities for maintaining a connected network of protected areas.

Latent resilience in ponderosa pine forest: effects of resumed frequent fire.

Ecological systems often exhibit resilient states that are maintained through negative feedbacks. In ponderosa pine forests, fire historically represented the negative feedback mechanism that maintained ecosystem resilience; fire exclusion reduced that resilience, predisposing the transition to an alternative ecosystem state upon reintroduction of fire. We evaluated the effects of reintroduced frequent wildfire in unlogged, fire-excluded, ponderosa pine forest in the Bob Marshall Wilderness, Montana, USA. Initial reintroduction of fire in 2003 reduced tree density and consumed surface fuels, but also stimulated establishment of a dense cohort of lodgepole pine, maintaining a trajectory toward an alternative state. Resumption of a frequent fire regime by a second fire in 2011 restored a low-density forest dominated by large-diameter ponderosa pine by eliminating many regenerating lodgepole pines and by continuing to remove surface fuels and small-diameter lodgepole pine and Douglas-fir that established during the fire suppression era. Our data demonstrate that some unlogged, fire-excluded, ponderosa pine forests possess latent resilience to reintroduced fire. A passive model of simply allowing lightning-ignited fires to burn appears to be a viable approach to restoration of such forests.

Detection and classification of motor vehicle noise in a forested landscape.

Noise emanating from human activity has become a common addition to natural soundscapes and has the potential to harm wildlife and erode human enjoyment of nature. In particular, motor vehicles traveling along roads and trails produce high levels of both chronic and intermittent noise, eliciting varied responses from a wide range of animal species. Anthropogenic noise is especially conspicuous in natural areas where ambient background sound levels are low. In this article, we present an acoustic method to detect and analyze motor vehicle noise. Our approach uses inexpensive consumer products to record sound, sound analysis software to automatically detect sound events within continuous recordings and measure their acoustic properties, and statistical classification methods to categorize sound events. We describe an application of this approach to detect motor vehicle noise on paved, gravel, and natural-surface roads, and off-road vehicle trails in 36 sites distributed throughout a national forest in the Sierra Nevada, CA, USA. These low-cost, unobtrusive methods can be used by scientists and managers to detect anthropogenic noise events for many potential applications, including ecological research, transportation and recreation planning, and natural resource management.