Heterogeneous Development Research Articles

Elucidating Heterogeneous Li Insertion Using Single-Crystalline and Freestanding Layered Oxide Thin Film.

The kinetics of interfacial ion insertion govern the uniformity of electrochemical reactions, playing a crucial role in lithium-ion battery performance. In two-dimensional lithium-conducting layered-oxide battery particles, variation in insertion rates across insertion channels remains unclear due to poorly defined crystal orientation at the solid-liquid interface and solid-state-lithium-diffusion length. This ambiguity complicates understanding inhomogeneous lithium-insertion channels activation. A systematic study requires crystallographically predefined interfaces and in situ lithium-concentration mapping. Here, we fabricated a freestanding, (104)-oriented-LiNi1/3Mn1/3Co1/3O2 single-crystal thin film using dissolution-induced release and performed in situ scanning-transmission-X-ray-microscopy to spatially resolve lithium-insertion at well-defined-interfaces. We observed heterogeneous lithium-concentration evolution due to channel-by-channel insertion rate variation, despite the potential for homogeneous lithium distribution via a solid-solution-phase at equilibrium in NMC111. Increasing current density exacerbates this heterogeneity, highlighting channel-by-channel variation. Our findings provide critical insights into battery electrode utilization and lifetime management, potentially guiding the design of more efficient and durable lithium-ion batteries.

The genomic and transcriptomic landscape of metastastic urothelial cancer

Metastatic urothelial carcinoma (mUC) is a lethal cancer, with limited therapeutic options. Large-scale studies in early settings provided critical insights into the genomic and transcriptomic characteristics of non-metastatic UC. The genomic landscape of mUC remains however unclear. Using Whole Exome (WES) and mRNA sequencing (RNA-seq) performed on metastatic biopsies from 111 patients, we show that driver genomic alterations from mUC were comparable to primary UC (TCGA data). APOBEC, platin, and HRD mutational signatures are the most prevalent in mUC, identified in 56%, 14%, and 9% of mUC samples, respectively. Molecular subtyping using consensus transcriptomic classification in mUC shows enrichment in neuroendocrine subtype. Paired samples analysis reveals subtype heterogeneity and temporal evolution. We identify potential therapeutic targets in 73% of mUC patients, of which FGFR3 (26%), ERBB2 (7%), TSC1 (7%), and PIK3CA (13%) are the most common. NECTIN4 and TACSTD2 are highly expressed regardless of molecular subtypes, FGFR3 alterations and sites of metastases.

Dissolution kinetics and heterogeneous evolution of dolomite with different pore structures

Mesogenetic dissolution is a critical process in the evolution and preservation of pore spaces in carbonate reservoirs. While limestone dissolution has been extensively studied, there is a lack of research on the dissolution kinetics of dolomite reservoirs, particularly regarding differences in dissolution mechanisms and heterogeneity evolution across various pore structures. This study aims to investigate diagenesis processes and heterogeneity evolution patterns in dolomites with different pore structures by simulating reservoir dissolution using organic acids generated during kerogen cracking. Flow system dissolution experiments were conducted on four dolomites with varying pore structures in 0.2% acetic acid under high-temperature (T = 40–160 °C) and high-pressure (P = 10–50 MPa) conditions. The chemical composition of the fluid and pore structure images were analyzed using ICP-OES and X-ray computed microtomography, respectively. Pore size distribution and evolution were assessed through digital cores based on Micro-CT analysis, while fractal and multifractal analyses were employed to quantify the evolution of pore structure heterogeneity. The findings highlight the importance of an effective combination of early material base and subsequent organic acid dissolution in the formation and maintenance of deep, high-quality dolomite reservoirs.

Impact of low-carbohydrate diet on health status: an umbrella review.

The prevalence of diet-related non-communicable diseases has increased. A low-carbohydrate diet (LCDs) is one of the most popular interventions. Several systematic reviews and meta-analyses of randomised clinical trials (RCTs) and non-RCTs have linked LCDs to the management of obesity, diabetes, cardiovascular disease, epilepsy, and cancer. However, there has been limited appraisal of the strength and quality of this evidence. To systematically appraise existing meta-analyses and systematic reviews of RCTs and non-RCTs on the effects of LCDs on different health conditions. To understand their potential efficacy, we summarised the studies' findings and assessed the strength of the evidence. A search was conducted using the PubMed database from inception to October 2021 for systematic reviews and meta-analyses of RCTs and non-RCTs investigating the association between LCDs and multiple health outcomes in humans. The Academy of Nutrition and Dietetics Quality Criteria was used for the quality assessment. In addition, the evolution of heterogeneity, strength of the included studies, and effect sizes were extracted from each systematic review and meta-analysis. Ten systematic reviews and meta-analyses were included. Of the included reviews, 70% were of positive quality, 30% were neutral, and none were negative. The majority of the studies included strength in each systematic review, and the meta-analyses were of low to medium strength. The existing literature indicates that LCDs may help promote weight reduction in adults who are obese or overweight. This conclusion is supported by the findings of studies included in the analysis, which were of low to moderate strength. Furthermore, compelling data indicates a significant association between low-carbohydrate diets (LCDs) and a reduction in haemoglobin A1c levels among those diagnosed with type 2 diabetes mellitus. In contrast, there was a lack of evidence of this correlation in type 1 diabetes mellitus patients or those with cardiovascular diseases. Additionally, there was limited evidence regarding the effectiveness of LCDs in epilepsy and adult cancer patients. This review thoroughly examines the current body of information on how LCDs affect various health outcomes. Studies have presented evidence to support the idea that incorporating LCDs can positively influence weight management and HbA1c levels. However, there is a lack of information regarding the association between LCDs and individuals with Type 1 diabetes mellitus and cardiovascular diseases. Additionally, there is limited empirical evidence to substantiate the effectiveness of LCDs in the treatment of epilepsy and adult cancer patients. The long-term effects of LCDs on mortality and other chronic diseases that account for different carbohydrate subtypes is unclear. Further longitudinal cohort studies are required to reach definitive conclusions.

On the Role and Evolution of Local Grain Size Heterogeneity During Confined Compression of Boise Sandstone as Seen by X-ray Micro-CT Imaging

On the Role and Evolution of Local Grain Size Heterogeneity During Confined Compression of Boise Sandstone as Seen by X-ray Micro-CT Imaging

Coupled cellular automata-crystal plasticity modeling of microstructure-sensitive damage and fracture behaviors in deformation of α-titanium sheets affected by grain size

Concerning the micro-scale deformation of titanium metal sheets, the number of grains in the sheet thickness direction decreases, and their formability exhibits a strong grain size sensitivity. Meanwhile, the twinning-induced dynamic recrystallization (TDRX) associated with grain size significantly affects the fracture behavior in the microforming of titanium sheets. Therefore, an accurate prediction of formability to improve manufacturing reliability remains challenging in the microforming of miniaturized titanium components. To address this issue, an in-depth understanding of the grain size-dependent TDRX behavior and its role in damage and fracture development in the microforming of α-titanium sheets is critical, and a coupled cellular automata-crystal plasticity (CA-CP) modeling framework was thus developed as an approach providing efficient solutions and insightful comprehensions of the issue. For the proposed modeling framework, a kinematic model for TDRX was established and integrated into the CP model by the CA algorithm. As a result, the microstructure evolution caused by TDRX was regarded as an intrinsic part of the constitutive behavior to connect heterogeneous plastic deformation and damage evolution through data transmission between the CP model and the CA algorithm. Additionally, the coupled CA-CP modeling framework was validated with the internal defect morphologies and deformation microstructures characterized by X-ray computed tomography (X-CT) and electron backscattered diffraction (EBSD). Experiment and simulation results demonstrated that the fine recrystallized (DRXed) grains were generated after the twin fragmentation when the dislocation density at twin boundaries reached a threshold of 9.2 × 1013 /m2. After TDRX, the dislocation density and the stress concentration intensity in recrystallization regions were revealed to decrease, accounting for the ductility improvement. Nevertheless, the dislocation density at twin boundaries was determined to decrease with the increase of grain size, leading to less twin fragmentation and the absence of TDRX. The uncoordinated deformation between fine DRXed grains motivated defects to grow spherically into microvoids, thereby preventing premature intergranular cracks along twins/grain boundaries. Ultimately, the deformation microstructures resulting from TDRX with the decrease of grain size were confirmed to control the brittle to ductile fracture transition of α-titanium sheets. The presented modeling framework and simulation procedure were validated to be able to predict the material integrity affected by crystalline microstructure in the deformation of titanium metal sheets.

Synchrotron Microfocus Wide-Angle X-ray Scattering Reveals the Evolution of Radial Microstructural Heterogeneity in γ-Irradiated/Heated Polyacrylonitrile Fibers

Synchrotron Microfocus Wide-Angle X-ray Scattering Reveals the Evolution of Radial Microstructural Heterogeneity in γ-Irradiated/Heated Polyacrylonitrile Fibers

Heterogeneous evolution and driving forces of multiple hazardous air pollutants and GHGs emissions from China's primary aluminum industry

The booming of China's primary aluminum industry (PAI) brought substantial emissions of hazardous air pollutants (HAPs) and greenhouse gases (GHGs). By using life cycle assessment and bottom-up method, a comprehensive emission inventory for multiple typical HAPs and GHGs from China's PAI during 1990–2021 was developed and explored for the first time. Our results show that spatial-temporal emissions trends of HAPs and GHGs from PAI in China diverse significantly. The conventional atmospheric pollutants (including SO2, NOx and particulate matter (PM)), fluoride and per fluorinated compound (PFCs) had been effectively suppressed since 2007 due to the implementation of various environmental policies; while, emissions of CO, VOCs, CH4, heavy metals and CO2 had increased at different rates unexpectedly. From the spatial distribution perspective, Henan, Shanxi, Guizhou, Guangxi and Shandong dominated the emissions of PAI in China, but with consumption expansion and environmental constrains, PAI plants start to expand to northwest and southwest areas where are richer in sufficient and cheaper power resources, thus bring significant emission increasing there, particular for conventional atmospheric pollutants in northwest and CO and VOCs in southwest China. By underlying driving forces of PAI emissions, results show that end-of-pipe control measures at various stages have played different roles to reduce emissions of the concerned species at each period, but its reduction effect diminished gradually. Future reduction should seek underlying changes in production technology and energy system. Under constrains of environmental regulation and resource endowment, promoting circular economic development for PAI would be a key strategy to reduce HAPs and GHGs emissions simultaneously in PAI.

918P Deciphering the molecular drivers behind locoregional progression, intratumoral heterogeneity, and clonal evolution in locally advanced head and neck cancer

918P Deciphering the molecular drivers behind locoregional progression, intratumoral heterogeneity, and clonal evolution in locally advanced head and neck cancer

Loss of Heterozygosity and Mutations in the RAS-ERK Pathway Genes in Tumor Cells of Various Loci in Multiple Myeloma.

Multiple myeloma (MM) is a disease characterized by spatiotemporal heterogeneity of tumor clones. Different genetic aberrations can be observed simultaneously in tumor cells from different loci, and as the disease progresses, new subclones may appear. The role of liquid biopsy, which is based on the analysis of tumor DNA circulating in the blood plasma, continues to be explored in MM. Here, we present an analysis of the STR profiles and mutation status of the KRAS, NRAS, and BRAF genes, evaluated in plasma free circulating tumor DNA (ctDNA), CD138+ bone marrow cells, and plasmacytomas. The prospective single-center study included 97 patients, with a median age of 55 years. Of these, 94 had newly diagnosed symptomatic MM, and three had primary plasma cell leukemia. It should be noted that if mutations were detected only in ctDNA, "non-classical" codons were more often affected. A variety of adverse laboratory and clinical factors have been associated with the detection of rare KRAS or NRAS gene mutations in bone marrow or ctDNA, suggesting that these mutations may be factors of an unfavorable prognosis for MM. Liquid biopsy studies provide undeniable fundamental information about tumor heterogeneity and clonal evolution in MM. Moreover, we focus on using liquid biopsy to identify new high-risk factors for MM.

Spatio-temporal evolution law of anisotropic shear damage on rock mass joint surface affected by joint morphology

The spatial–temporal evolution law of shear damage on the joint surface is closely related to its morphological characteristics. Understanding the heterogeneous failure behavior of the joint surface is essential for revealing the controlling role of the joint morphology. The diorite from the Guanshan Tunnel is investigated through anisotropic direct shear tests in this study. Based on shear damage quantification and acoustic emission (AE) localization techniques, the heterogeneous damage evolution law of joint surfaces under different joint compression strength (JCS), normal stress (σn), and morphology parameter (M) is studied. The results indicate that the joint surface morphology, shear damage, and AE parameters exhibit similar anisotropic characteristics in different shear directions. An increase in the morphology parameter leads to an increase in shear failure, with shear fracture energy and the number of fracture events showing an increasing trend. Meanwhile, the occurrence of a large number of fracture points shifts from the pre-peak nonlinear period to the post-peak period. However, the evolution process of fracture events exhibits certain similarities, though the morphology parameters are different in two opposite shear directions. Additionally, shear damage volume and fracture energy demonstrate consistency in spatial distribution and quantitative characteristics, while the linear scale factor between cumulative shear fracture energy and damage volume decreases with increasing joint surface strength. These findings assist in a better understanding of the anti-sliding property of joint surfaces and provide crucial clues for further exploration of joint surface failure mechanisms.

Break-induced replication drives large-scale genomic amplifications in cancer cells.

DNA double-strand breaks (DSBs) are highly toxic lesions that underly the efficacy of ionizing radiation (IR) and a large number of cytotoxic chemotherapies 1-3 . Yet, abnormal repair of DSBs is associated with genomic instability and may contribute to cancer heterogeneity and tumour evolution. Here, we show that DSBs induced by IR, by DSB-inducing chemotherapeutics, or by the expression of a rare-cutting restriction endonuclease induce large-scale genomic amplification in human cancer cells. Importantly, the extent of DSB-induced genomic amplification (DIGA) in a panel of melanoma cell lines correlated with the degree of cytotoxicity elicited by IR, suggesting that DIGA contributes significantly to DSB-induced cancer cell lethality. DIGA, which is mediated through conservative DNA synthesis, does not require origin re-licensing, and is enhanced by the depletion or deletion of the methyltransferases SET8 and SUV4-20H1, which function sequentially to mono- and di-methylate histone H4 lysine 20 (H4K20) at DSBs to facilitate the recruitment of 53BP1-RIF1 and its downstream effector shieldin complex to DSBs to prevent hyper-resection 4-11 . Consistently, DIGA was enhanced in cells lacking 53BP1 or RIF1, or in cells that lacked components of the shieldin complex or of other factors that help recruit 53BP1 to DSBs. Mechanistically, DIGA requires MRE11/CtIP and EXO1, factors that promote resection and hyper-resection at DSBs, and is dependent on the catalytic activity of the RAD51 recombinase. Furthermore, deletion or depletion of POLD3, POLD4, or RAD52, proteins involved in break-induced replication (BIR), significantly inhibited DIGA, suggesting that DIGA is mediated through a RAD51-dependent BIR-like process. DIGA induction was maximal if the cells encountered DSBs in early and mid S-phase, whereas cells competent for homologous recombination (in late S and G2) exhibited less DIGA induction. We propose that unshielded, hyper-resected ends of DSBs may nucleate a replication-like intermediate that enables cytotoxic long-range genomic DNA amplification mediated through BIR.

(Invited) Thermo-Electrochemical Stability of Solid-State Batteries

Solid-state batteries (SSBs) are promising due to higher energy density and improved safety. While interface instability due to electrochemical-mechanics interaction, transport and morphological dynamics at solid-solid interfaces has been identified as a critical determinant of failure modes, the role of thermo-electrochemical effect is still elusive. A wide range of factors, such as, high reactivity of lithium, propensity for hotspot formation (e.g., at solid/solid point contacts), and cathode instability may affect thermo-electrochemical signature of solid-state architectures. In addition, spatio-temporal evolution of thermal and reaction heterogeneities and the occurrence of failure mechanisms such as lithium filament growth can be critical. In this presentation, the important role of internal thermal signature and mechanistic origin of thermo-electrochemical instability in solid-state battery architectures will be discussed.

Deep recycling of volatile elements in the mantle: Evidence from the heterogeneous B isotope in intra-plate basalts

Volatiles in the mantle are crucial for Earth’s geodynamic and geochemical evolution. Understanding the deep recycling of volatiles is key for grasping mantle chemical heterogeneity, plate tectonics, and long-term planetary evolution. While subduction transfers abundant volatile elements from the Earth’s surface into the mantle, the fate of hydrous portions within subducted slabs during intensive dehydration processes remains uncertain. Boron isotopes, only efficiently fractionating near the Earth’s surface, are valuable for tracing volatile recycling signals. In this study, we document a notably large variation in δ11B values (−14.3‰ to +8.2‰) in Cenozoic basalts from the South China Block. These basalts, associated with a high-velocity zone beneath East China, are suggested to originate from the mantle transition zone. While the majority exhibit δ11B values (−10‰ to −5‰) resembling the normal mantle, their enriched Sr-Nd-Pb isotope compositions and fluid-mobile elements imply hydrous components in their source, including altered oceanic crust and sediments. The normal δ11B values are attributed to the dehydration processes. Remarkably high δ11B values in the basalts indicate the presence of subducted serpentinites in their mantle source. A small subset of samples with low δ11B values and radiogenic isotope enrichments suggests a contribution from recycled detrital sediments, though retaining minimal volatile elements after extensive dehydration. These findings provide compelling evidence that serpentinites within subducted slabs predominantly maintain their hydrous nature during dehydration processes in subduction zones. They may transport a considerable amount of water into deep mantle reservoirs, such as the mantle transition zone.

Heterogeneous Evolution of Breast Cancer Cells-An Endogenous Molecular-Cellular Network Study.

Breast cancer heterogeneity presents a significant challenge in clinical therapy, such as over-treatment and drug resistance. These challenges are largely due to its obscure normal epithelial origins, evolutionary stability, and transitions on the cancer subtypes. This study aims to elucidate the cellular emergence and maintenance of heterogeneous breast cancer via quantitative bio-process modeling, with potential benefit to therapeutic strategies for the disease. An endogenous molecular-cellular hypothesis posits that both pathological and physiological states are phenotypes evolved from and shaped by interactions among a number of conserved modules and cellular factors within a biological network. We hereby developed a model of core endogenous network for breast cancer in accordance with the theory, quantifying its intrinsic dynamic properties with dynamic modeling. The model spontaneously generates cell states that align with molecular classifications at both the molecular and modular level, replicating four widely recognized molecular subtypes of the cancer and validating against data extracted from the TCGA database. Further analysis shows that topologically, a singular progression gateway from normal breast cells to cancerous states is identified as the Luminal A-type breast cancer. Activated positive feedback loops are found to stabilize cellular states, while negative feedback loops facilitate state transitions. Overall, more routes are revealed on the cellular transition between stable states, and a traceable count explains the origin of breast cancer heterogeneity. Ultimately, the research intended to strength the search for therapeutic targets.

Next-generation sequencing technology in cancer

Next-generation sequencing (NGS) technology has revolutionized cancer research and treatment by enabling comprehensive analysis of genetic mutations, alterations, and expression profiles. It allows for the identification of cancer-driving mutations, helping in the development of targeted therapies. NGS provides detailed insights into tumor heterogeneity, resistance mechanisms, and clonal evolution. Its high-throughput capacity facilitates large-scale studies, improving our understanding of cancer genomics. By enabling personalized treatment plans based on individual genetic profiles, NGS holds promise for more effective and tailored cancer therapies. Early reviews on cancer genomics often lacked comprehensive coverage of emerging technologies. They missed in-depth analysis of NGS advancements, their impact on cancer research, and clinical applications. The review addresses this gap by reviving a thorough examination of NGS methods, their role in identifying genetic mutations, and their potential in personalized cancer treatment, thus providing essential insights into the evolving landscape of cancer genomics. The article covers the advancements in technology and bioinformatic approaches for NGS data analysis. It delves into NGS applications in research and diagnostics, particularly for solid cancer diagnosis. The review highlights specific cancer types, including hereditary breast cancer, melanoma, prostate cancer, thyroid cancer, lung cancer, and colorectal cancer. It explores NGS contribution in understanding the genetic basis of these cancers and its potential for enhancing personalized diagnosis and treatment strategies. This review rectifies early lacunas by providing a comprehensive and updated examination of NGS technology, addressing gaps in previous analyses and emphasizes bioinformatic approaches for NGS data analysis, crucial for interpreting vast genomic data accurately. The review meets the current need for a thorough understanding of NGS’s role in personalized cancer treatment and research.

Spatial intra-tumour heterogeneity and treatment-induced genomic evolution in oesophageal adenocarcinoma: implications for prognosis and therapy

BackgroundOesophageal adenocarcinoma (OAC) is a highly heterogeneous cancer with poor survival. Standard curative treatment is chemotherapy with or without radiotherapy followed by oesophagectomy. Genomic heterogeneity is a feature of OAC and has been linked to treatment resistance.MethodsWhole-genome sequencing data from 59 treatment-naïve and 18 post-treatment samples from 29 OAC patients was analysed. Twenty-seven of these were enrolled in the DOCTOR trial, sponsored by the Australasian Gastro-Intestinal Trials Group. Two biopsies from each treatment-naïve tumour were assessed to define ‘shared’ (between both samples) and ‘private’ (present in one sample) mutations.ResultsMutational signatures SBS2/13 (APOBEC) and SBS3 (BRCA) were almost exclusively detected in private mutation populations of treatment-naïve tumours. Patients presenting these signatures had significantly worse disease specific survival. Furthermore, mutational signatures associated with platinum-based chemotherapy treatment as well as high platinum enrichment scores were only detected in post-treatment samples. Additionally, clones with high putative neoantigen binding scores were detected in some treatment-naïve samples suggesting immunoediting of clones.ConclusionsThis study demonstrates the high intra-tumour heterogeneity in OAC, as well as indicators for treatment-induced changes during tumour evolution. Intra-tumour heterogeneity remains a problem for successful treatment strategies in OAC.

Optical in situ deciphering of the surface reconstruction–assistant multielectron transfer event of single Co3O4 nanoparticles

Surface reconstruction determines the fate of catalytic sites on the near-surface during the oxygen evolution reaction. However, deciphering the conversion mechanism of various intermediate-states during surface reconstruction remains a challenge. Herein, we employed an optical imaging technique to draw the landscape of dynamic surface reconstruction on individual Co3O4 nanoparticles. By regulating the surface states of Co3O4 nanoparticles, we explored dynamic growth of the CoOx(OH)y sublayer on single Co3O4 nanoparticles and directly identified the conversion between two dynamics. Rich oxygen vacancies induced more active sites on the surface and prolonged surface reconstruction, which enhanced electrochemical redox and oxygen evolution. These results were further verified by in situ electrochemical extinction spectroscopy of single Co3O4 nanoparticles. We elucidate the heterogeneous evolution of surface reconstruction on individual Co3O4 nanoparticles and present a unique perspective to understand the fate of catalytic species on the nanosurface, which is of enduring significance for investigating the heterogeneity of multielectron-transfer events.

Chloroplast Genomes Evolution and Phylogenetic Relationships of Caragana species.

Caragana sensu lato (s.l.) includes approximately 100 species that are mainly distributed in arid and semi-arid regions. Caragana species are ecologically valuable for their roles in windbreaking and sand fixation. However, the taxonomy and phylogenetic relationships of the genus Caragana are still unclear. In this study, we sequenced and assembled the chloroplast genomes of representative species of Caragana and reconstructed robust phylogenetic relationships at the section level. The Caragana chloroplast genome has lost the inverted repeat region and wascategorized in the inverted repeat loss clade (IRLC). The chloroplast genomes of the eight species ranged from 128,458 bp to 135,401 bp and contained 110 unique genes. All the Caragana chloroplast genomes have a highly conserved structure and gene order. The number of long repeats and simple sequence repeats (SSRs) showed significant variation among the eight species, indicating heterogeneous evolution in Caragana. Selective pressure analysis of the genes revealed that most of the protein-coding genes evolved under purifying selection. The phylogenetic analyses indicated that each section forms a clade, except the section Spinosae, which was divided into two clades. This study elucidated the evolution of the chloroplast genome within the widely distributed genus Caragana. The detailed information obtained from this study can serve as a valuable resource for understanding the molecular dynamics and phylogenetic relationships within Caragana.

Characterization of transcriptional heterogeneity and novel therapeutic targets using single cell RNA-sequencing of primary and circulating Ewing sarcoma cells.

Ewing sarcoma is the second most common bone cancer in children, accounting for 2% of pediatric cancer diagnoses. Patients who present with metastatic disease at the time of diagnosis have a dismal prognosis, compared to the >70% 5-year survival of those with localized disease. Here, we utilized single cell RNA-sequencing to characterize the transcriptional landscape of primary Ewing sarcoma tumors and surrounding tumor microenvironment (TME). Copy-number analysis identified subclonal evolution within patients prior to treatment. Primary tumor samples demonstrate a heterogenous transcriptional landscape with several conserved gene expression programs, including those composed of genes related to proliferation and EWS targets. Single cell RNA-sequencing and immunofluorescence of circulating tumor cells at the time of diagnosis identified TSPAN8 as a novel therapeutic target.