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PDF file - 135K, Supplementary table 2. Donor 1 PBMC delta chain diversity.

Tables S1, S2, S3. Table S1 and S2 show the results of the Mantel test for the spatial comparison of the MRI and the histopathological biomarker for each tumors. Table S3 shows the summary of the pathological characteristics of the individual Th-MYCN tumors and their associated MRI transverse relaxation times.

Purpose: The majority of circulating human γδT lymphocytes are of the Vγ9Vδ2 lineage, and have T-cell receptor (TCR) specificity for nonpeptide phosphoantigens. Previous attempts to stimulate and expand these cells have therefore focused on stimulation using ligands of the Vγ9Vδ2 receptor, whereas relatively little is known about variant blood γδT subsets and their potential role in cancer immunotherapy.Experimental Design: To expand the full repertoire of γδT without bias toward specific TCRs, we made use of artificial antigen-presenting cells loaded with an anti γδTCR antibody that promoted unbiased expansion of the γδT repertoire. Expanded cells from adult blood donors were sorted into 3 populations expressing respectively Vδ2 TCR chains (Vδ2+), Vδ1 chains (Vδ1+), and TCR of other δ chain subtypes (Vδ1negVδ2neg).Results: Both freshly isolated and expanded cells showed heterogeneity of differentiation markers, with a less differentiated phenotype in the Vδ1 and Vδ1negVδ2neg populations. Expanded cells were largely of an effector memory phenotype, although there were higher numbers of less differentiated cells in the Vδ1+ and Vδ1negVδ2neg populations. Using neuroblastoma tumor cells and the anti-GD2 therapeutic mAb ch14.18 as a model system, all three populations showed clinically relevant cytotoxicity. Although killing by expanded Vδ2 cells was predominantly antibody dependent and proportionate to upregulated CD16, Vδ1 cells killed by antibody-independent mechanisms.Conclusions: In conclusion, we have demonstrated that polyclonal-expanded populations of γδT cells are capable of both antibody-dependent and -independent effector functions in neuroblastoma. Clin Cancer Res; 20(22); 5720–32. ©2014 AACR.

Th-MYCNCPM32 tumors and cell lines show enhanced nuclear STAT3 staining and sensitivity to CYT387.

PDF file - 504K, Supplementary figure 1. Removal of CD14+/CD11c+ cells prior to gammadeltaTCR positive selection significantly increases sample purity.

Noninvasive early indicators of treatment response are crucial to the successful delivery of precision medicine in children with cancer. Neuroblastoma is a common solid tumor of young children that arises from anomalies in neural crest development. Therapeutic approaches aiming to destabilize MYCN protein, such as small-molecule inhibitors of Aurora A and mTOR, are currently being evaluated in early phase clinical trials in children with high-risk MYCN-driven disease, with limited ability to evaluate conventional pharmacodynamic biomarkers of response. T1 mapping is an MRI scan that measures the proton spin-lattice relaxation time T1. Using a multiparametric MRI-pathologic cross-correlative approach and computational pathology methodologies including a machine learning–based algorithm for the automatic detection and classification of neuroblasts, we show here that T1 mapping is sensitive to the rich histopathologic heterogeneity of neuroblastoma in the Th-MYCN transgenic model. Regions with high native T1 corresponded to regions dense in proliferative undifferentiated neuroblasts, whereas regions characterized by low T1 were rich in apoptotic or differentiating neuroblasts. Reductions in tumor-native T1 represented a sensitive biomarker of response to treatment-induced apoptosis with two MYCN-targeted small-molecule inhibitors, Aurora A kinase inhibitor alisertib (MLN8237) and mTOR inhibitor vistusertib (AZD2014). Overall, we demonstrate the potential of T1 mapping, a scan readily available on most clinical MRI scanners, to assess response to therapy and guide clinical trials for children with neuroblastoma. The study reinforces the potential role of MRI-based functional imaging in delivering precision medicine to children with neuroblastoma.Significance:This study shows that MRI-based functional imaging can detect apoptotic responses to MYCN-targeted small-molecule inhibitors in a genetically engineered murine model of MYCN-driven neuroblastoma.

Exogenous manipulation of STAT3 levels partially mimics the effect of CYT387, Th-MYCNCPM32 cell lines show cross resistance to front line chemotherapy.

Microarray data analysis

Childhood neuroblastoma is a hypervascular tumor of neural origin, for which antiangiogenic drugs are currently being evaluated; however, predictive biomarkers of treatment response, crucial for successful delivery of precision therapeutics, are lacking. We describe an MRI-pathologic cross-correlative approach using intrinsic susceptibility (IS) and susceptibility contrast (SC) MRI to noninvasively map the vascular phenotype in neuroblastoma Th-MYCN transgenic mice treated with the vascular endothelial growth factor receptor inhibitor cediranib. We showed that the transverse MRI relaxation rate R2* (second−1) and fractional blood volume (fBV, %) were sensitive imaging biomarkers of hemorrhage and vascular density, respectively, and were also predictive biomarkers of response to cediranib. Comparison with MRI and pathology from patients with MYCN-amplified neuroblastoma confirmed the high degree to which the Th-MYCN model vascular phenotype recapitulated that of the clinical phenotype, thereby supporting further evaluation of IS- and SC-MRI in the clinic. This study reinforces the potential role of functional MRI in delivering precision medicine to children with neuroblastoma.Significance:This study shows that functional MRI predicts response to vascular-targeted therapy in a genetically engineered murine model of neuroblastoma.

PDF file - 18K, Supplementary figure 2. Anti-gammadeltaTCR clone B1 is a murine IgG1.

PML and HIF1a expression

PDF file - 89K, Supplementary figure 3. Variation in gammadeltaTCR repertoire before expansion stimulus applied.

Neuroblastoma is a pediatric cancer that is frequently metastatic and resistant to conventional treatment. In part, a lack of natively metastatic, chemoresistant in vivo models has limited our insight into the development of aggressive disease. The Th-MYCN genetically engineered mouse model develops rapidly progressive chemosensitive neuroblastoma and lacks clinically relevant metastases. To study tumor progression in a context more reflective of clinical therapy, we delivered multicycle treatment with cyclophosphamide to Th-MYCN mice, individualizing therapy using MRI, to generate the Th-MYCNCPM32 model. These mice developed chemoresistance and spontaneous bone marrow metastases. Tumors exhibited an altered immune microenvironment with increased stroma and tumor-associated fibroblasts. Analysis of copy number aberrations revealed genomic changes characteristic of human MYCN-amplified neuroblastoma, specifically copy number gains at mouse chromosome 11, syntenic with gains on human chromosome 17q. RNA sequencing revealed enriched expression of genes associated with 17q gain and upregulation of genes associated with high-risk neuroblastoma, such as the cell-cycle regulator cyclin B1-interacting protein 1 (Ccnb1ip1) and thymidine kinase (TK1). The antiapoptotic, prometastatic JAK–STAT3 pathway was activated in chemoresistant tumors, and treatment with the JAK1/JAK2 inhibitor CYT387 reduced progression of chemoresistant tumors and increased survival. Our results highlight that under treatment conditions that mimic chemotherapy in human patients, Th-MYCN mice develop genomic, microenvironmental, and clinical features reminiscent of human chemorefractory disease. The Th-MYCNCPM32 model therefore is a useful tool to dissect in detail mechanisms that drive metastasis and chemoresistance, and highlights dysregulation of signaling pathways such as JAK–STAT3 that could be targeted to improve treatment of aggressive disease.Significance:An in vivo mouse model of high-risk treatment-resistant neuroblastoma exhibits changes in the tumor microenvironment, widespread metastases, and sensitivity to JAK1/2 inhibition.

New neuroblastoma patient cohort

PDF file - 251K, Supplementary figure 6. NKG2D expression of expanded gammadeltaT cell subsets.

Purpose: Neuroblastoma is a childhood malignancy originating from the sympathetic nervous system with a complex biology, prone to metastasize and relapse. High-risk, metastatic cases are explained in part by amplification or mutation of oncogenes, such as MYCN and ALK, and loss of tumor suppressor genes in chromosome band 1p. However, it is fundamental to identify other pathways responsible for the large portion of neuroblastomas with no obvious molecular alterations.Experimental Design: Neuroblastoma cell lines were used for the assessment of tumor growth in vivo and in vitro. Protein expression in tissues and cells was assessed using immunofluorescence and IHC. The association of promyelocytic leukemia (PML) expression with neuroblastoma outcome and relapse was calculated using log-rank and Mann–Whitney tests, respectively. Gene expression was assessed using chip microarrays.Results: PML is detected in the developing and adult sympathetic nervous system, whereas it is not expressed or is low in metastatic neuroblastoma tumors. Reduced PML expression in patients with low-risk cancers, that is, localized and negative for the MYCN proto-oncogene, is strongly associated with tumor recurrence. PML-I, but not PML-IV, isoform suppresses angiogenesis via upregulation of thrombospondin-2 (TSP2), a key inhibitor of angiogenesis. Finally, PML-I and TSP2 expression inversely correlates with tumor angiogenesis and recurrence in localized neuroblastomas.Conclusions: Our work reveals a novel PML-I–TSP2 axis for the regulation of angiogenesis and cancer relapse, which could be used to identify patients with low-risk, localized tumors that might benefit from chemotherapy. Clin Cancer Res; 22(13); 3398–409. ©2016 AACR.

Pathway analysis

PDF file - 91K, Supplementary figure 5. GD2 staining of neuroblastoma cell lines.

PDF file - 85K, Supplementary figure 4. Killing of GD2+ Kelly by expanded gammadeltaT cells in the presence of ch14.18 (black) or anti-CD20 control antibody (grey).

Supplementary Figure Legends. Supplementary Figure Legends for Supp Figs 1-5 Contains Legends for: Supplementary Figure 1: Arginase II is the principal arginase isoform in neuroblastoma Supplementary Figure 2: Arginase II expression by neuroblastoma tumours creates a suppressive microenvironment Supplementary Figure 3: Neuroblastoma alters myeloid cell frequency in a murine model Supplementary Figure 4: Neuroblastoma arginase inactivates CD14+ cells and CAR T cells Supplementary Figure 5: Arginase 2 and CD14 expression are associated with a worse overall survival.

Supplemental figures 1-5. Supplementary Figure 1: Arginase II is the principal arginase isoform in neuroblastoma Supplementary Figure 2: Arginase II expression by neuroblastoma tumours creates a suppressive microenvironment Supplementary Figure 3: Neuroblastoma alters myeloid cell frequency in a murine model Supplementary Figure 4: Neuroblastoma arginase inactivates CD14+ cells and CAR T cells Supplementary Figure 5: Arginase 2 and CD14 expression are associated with a worse overall survival.

Supplementary Figure 3 from Development of Cellular Immune Responses against PAX5, a Novel Target for Cancer Immunotherapy

Supplementary Figure 2 from Migratory and Antigen Presentation Functions of IFN-Producing Killer Dendritic Cells

Supplementary Figure 1 from Development of Cellular Immune Responses against PAX5, a Novel Target for Cancer Immunotherapy

Supplementary Figure 1 from Migratory and Antigen Presentation Functions of IFN-Producing Killer Dendritic Cells

Supplementary Figure Legends 1-2 from Migratory and Antigen Presentation Functions of IFN-Producing Killer Dendritic Cells

Supplementary Figure 2 from Development of Cellular Immune Responses against PAX5, a Novel Target for Cancer Immunotherapy

Supplementary Materials and Methods. Supplementary Materials and Methods

Supp tables 1 and 2. Supplementary Table 1 Table of patient clinical characteristics for patient derived samples Supplementary Table 2 PCR Primer sequences

PAX5 is a member of the PAX family of developmental transcription factors with an important role in B-cell development. Its expression in normal adult tissue is limited to the hemopoietic system, but it is aberrantly expressed in a number of solid cancers and leukemias where it functions as an oncogene. We therefore hypothesized that anti-PAX5 immune responses could be used to target a number of malignancies without significant toxicity. We screened PAX5 peptides for the ability to bind HLA-A2 and identified a novel sequence, TLPGYPPHV (referred to as TLP). CTL lines against TLP were generated from peripheral blood of five normal HLA-A2–positive blood donors and showed specific HLA-A2–restricted killing against PAX5-expressing target cells. We generated high-avidity CTL clones from these lines capable of killing cells pulsed with

Supplementary Figure Legends 1-3 from Development of Cellular Immune Responses against PAX5, a Novel Target for Cancer Immunotherapy

Thwaites Glacier (TG) is thinning and accelerating while sitting on a landward-dipping bed, with an ice shelf that is rapidly disintegrating and losing its ability to buttress ice flow from upstream, and is in deep water that allows warm Circumpolar Deep Water (CDW) to reach its grounding zone. Significant retreat of TG would trigger loss of ice across the region. In recent decades, the mass balance of TG has become increasingly negative, suggesting that unstable retreat may have already begun. The Thwaites Offshore Research (THOR) group has just completed four field deployments aimed at understanding the recent history of TG and neighboring ice, including Pine Island Glacier (PIG). Three cruises on the RVIB N.B. Palmer, combined with sub-ice-shelf sediment coring, provide a suite of new data along the TG and PIG margins. Data include multibeam surveys, 3.5 kHz subbottom profiler, over 100 new sediment cores, and high-resolution seismic profiles. Break-up of floating ice cover in front of TG in 2019 allowed surveying of previously unmapped seafloor. Major calving of PIG in 2020 allowed marine surveying over the locations where sub-ice-shelf cores were collected in the past, allowing direct ties between ice-based and marine work. As of this writing in January 2022, we are entering into the Amundsen Sea for our third marine field season. Sediment cores record the history of grounding-zone retreat and ice interaction with the ocean over timescales from decades to several thousand years. Proxies used to reconstruct ice and ocean histories include sedimentary facies analysis, diatom and foraminiferal assemblage data, and geochemical analyses. Sedimentological analyses show a diverse array of lithofacies attributed to different environmental conditions. Many cores across the region contain laminated mud with sparse gravel and sand, suggesting deposition of meltwater deposits. Downcore 210Pb measurements are used to create age models of the past ~100 years. Combination of ages with facies models, including CT scans, reveals that the progressive detachment of Thwaites from pinning points began in the mid twentieth century, coincident with retreat of PIG (Smith et al., 2017) and with increasing advection of warm water onto the Amundsen shelf (Hillenbrand et al., 2017). Conversely, Cranton Bay, to the northeast of PIG and separated from Pine Island Bay by a shallow sill, appears to be characterized by cold deep water and high productivity, allowing it to serve as an endmember different from the records obtained proximal to the large glacial outlets where CDW is impinging. The satellite record of glacial retreat is inherently short. Observations are accumulating about forcing mechanisms that can impact the stability of ice, such as increased CDW on the Amundsen Sea shelf. However, without the time to observe the response of the ice, discovery of the forcing mechanisms is just half the story. The other half of the story is completed by using the paleo record to see how ice has responded to drivers in the past.

The coastal bathymetry of Thwaites Glacier (TG) is poorly known yet nearshore sea-floor highs have the potential to act as pinning points for floating ice shelves, or to block warm water incursions to the grounding line. In contrast, deeper areas control warm water routing. Here, we present more than 2000 km2 of new multibeam echo-sounder data (MBES) acquired offshore TG during the first cruise of the International Thwaites Glacier Collaboration (ITGC) project on the RV/IB Nathaniel B. Palmer (NBP19-02) in February-March 2019. Beyond TG, the bathymetry is dominated by a >1200 m deep, structurally-controlled trough and discontinuous ridge, on which the Eastern Ice Shelf is pinned. The geometry and composition of the ridge varies spatially with some sea-floor highs having distinctive flat-topped morphologies produced as their tops were planed-off by erosion at the base of the seaward-moving Thwaites Ice Shelf. In addition, submarine landform evidence indicates at least some unconsolidated sediment cover on the highs, as well as in the troughs that separate them. Knowing that this offshore area of ridges and troughs is a former bed for TG, we also used a novel spectral approach and existing ice-flow theory to investigate bed roughness and basal drag over the newly-revealed offshore topography. We show that the sea-floor bathymetry is a good analogue for extant bed areas of TG and that ice-sheet retreat over the sea-floor troughs and ridges would have been affected by high basal drag similar to that acting in the grounding zone today.Comparisons of the new MBES data with existing regional compilations show that high-frequency (finer than 5 km) bathymetric variability beyond Antarctic ice shelves can only be resolved by observations such as MBES and that without these data calculations of the oceanic heat flux may be significantly underestimated. This work supports the findings of recent numerical ice-sheet and ocean modelling studies that recognise the need for accurate and high-resolution bathymetry to determine warm water routing to the grounding zone and, ultimately, for predicting glacier retreat behaviour.

Two-dimensional (2D) inorganic materials have emerged as exciting platforms for (opto)electronic, thermoelectric, magnetic, and energy storage applications. However, electronic redox tuning of these materials can be difficult. Instead, 2D metal-organic frameworks (MOFs) offer the possibility of electronic tuning through stoichiometric redox changes, with several examples featuring one to two redox events per formula unit. Here we demonstrate that this principle can be extended over a far greater span with the isolation of four discrete redox states in the 2D MOFs LixFe3(THT)2 (x = 03, THT = triphenylene hexathiol). This redox modulation results in 10,000-fold greater conductivity, p- to n-type carrier switching, and modulation of antiferromagnetic coupling. Physical characterization suggests that changes in carrier density drive these trends with relatively constant charge transport activation energies and mobilities. This series illustrates that 2D MOFs are uniquely redox flexible, making them an ideal materials platform for tunable and switchable applications.

According to Army Multi-Domain Operations (MDO) doctrine, generating timely, accurate, and exploitable geospatial products from tactical platforms is a critical capability to meet threats. The US Army Corps of Engineers, Engineer Research and Development Center, Geospatial Research Laboratory (ERDC-GRL) is carrying out 6.2 research to facilitate the creation of three-dimensional (3D) products from tactical sensors to include full-motion video, framing cameras, and sensors integrated on small Unmanned Aerial Systems (sUAS). This report describes an ERDC-GRL processing pipeline comprising custom code, open-source software, and commercial off-the-shelf (COTS) tools to geospatially rectify tactical imagery to authoritative foundation sources. Four datasets from different sensors and locations were processed against National Geospatial-Intelligence Agency–supplied foundation data. Results showed that the co-registration of tactical drone data to reference foundation varied from 0.34 m to 0.75 m, exceeding the accuracy objective of 1 m described in briefings presented to Army Futures Command (AFC) and the Assistant Security of the Army for Acquisition, Logistics and Technology (ASA(ALT)). A discussion summarizes the results, describes steps to address processing gaps, and considers future efforts to optimize the pipeline for generation of geospatial data for specific end-user devices and tactical applications.

Introdução: O avanço da medicina, erradicação e controle de doenças infecciosas bem como melhorias na qualidade de vida nas últimas décadas, trouxe maior longevidade para a população, ocorrendo a transição epidemiológica das doenças, ganhando destaque as Doenças Crônicas Não Transmissíveis, como as doenças coronarianas. Esta pesquisa tem como objetivo identificar os parâmetros clínicos que a equipe de enfermagem utiliza no reconhecimento de Parada Cardiorrespiratória em Centro de Terapia Intensiva. Método: Trata-se de uma pesquisa do tipo descritiva, com abordagem qualitativa, realizada no centro de terapia intensiva de um hospital privado, localizado no município de Belém, Estado do Pará. Os participantes foram profissionais de enfermagem, que responderam a um formulário semiestruturado, para analisar as ações e conhecimento em emergência em Parada Cardiorrespiratória pelo usuário clinicamente grave. Resultados e Discussão: Pode-se observar que os participantes identificam um cenário de parada cardiorrespiratória por meio do desconforto, expressões faciais, palidez, dificuldade de respirar, sinais vitais, débito cardíaco e saturação diminuídos e ausência de pulso. No entanto, em relação à conduta no atendimento, a pesquisa mostrou-se insatisfatória em diversos aspectos abordados, identificou falta de conhecimento da equipe de enfermagem na tomada de condutas e insegurança na realização de procedimentos. Conclusão: A atuação da equipe de enfermagem em Parada Cardiorrespiratória deve continuar sendo difundida, objetivando maior segurança e autonomia desses profissionais, visando uma assistência de qualidade, a equipe de enfermagem deve estar apta a atender este tipo de emergência clínica, de modo que venha participar de treinamentos e atualizações constantes, a fim de que se capacitem e prestem atendimento com segurança e eficaz.

Introduction: The advances in medicine, eradication and control of infectious diseases, as well as improvements in quality of life in recent decades, have brought greater longevity to the population, with the epidemiological transition of diseases taking place, with emphasis on Non-communicable Chronic Diseases, such as coronary heart disease. This research aims to identify the clinical parameters that the nursing team uses in the recognition of Cardiopulmonary Arrest in an Intensive Care Unit. Method: This is a descriptive research, with a qualitative approach, carried out in the intensive care center of a private hospital, located in the city of Belém, State of Pará. The participants were nursing professionals, who answered a semi-structured form, to analyze the actions and knowledge in emergencies in Cardiorespiratory Arrest by the clinically serious user. Results and Discussion: It can be observed that the participants identify a scenario of cardiorespiratory arrest through discomfort, facial expressions, pallor, difficulty breathing, vital signs, decreased cardiac output and saturation and absence of pulse. However, in relation to the behavior in the attendance, the research proved to be unsatisfactory in several approached aspects, it identified lack of knowledge of the nursing team in the taking of conducts and insecurity in the accomplishment of procedures. Conclusion: The performance of the nursing team in Cardiorespiratory Arrest must continue to be disseminated, aiming at greater safety and autonomy of these professionals, aiming at quality care, the nursing team must be able to attend to this type of clinical emergency, so that they can participate of constant training and updates, so that they become qualified and provide safe and effective care.

Introducción: Los avances en la medicina, la erradicación y el control de las enfermedades infecciosas, así como las mejoras en la calidad de vida de las últimas décadas, han traído mayor longevidad a la población, ocurriendo la transición epidemiológica de las enfermedades, con énfasis en las Crónicas No Transmisibles. Enfermedades, como la enfermedad coronaria. Esta investigación tiene como objetivo identificar los parámetros clínicos que el equipo de enfermería utiliza en el reconocimiento de Paro Cardiopulmonar en una Unidad de Cuidados Intensivos. Método: Se trata de una investigación descriptiva, con enfoque cualitativo, realizada en el centro de cuidados intensivos de un hospital privado, ubicado en la ciudad de Belém, Estado de Pará. Los participantes fueron profesionales de enfermería, que respondieron un formulario semiestructurado, para analizar las acciones y conocimientos en emergencias en Paro Cardiorrespiratorio por parte del usuario clínicamente grave. Resultados y Discusión: Se puede observar que los participantes identifican un escenario de paro cardiorrespiratorio a través de malestar, expresiones faciales, palidez, dificultad para respirar, signos vitales, disminución del gasto cardíaco y saturación y ausencia de pulso. Sin embargo, en relación al comportamiento en la asistencia, la investigación se mostró insatisfactoria en varios aspectos abordados, identificó desconocimiento del equipo de enfermería en la toma de conductas e inseguridad en la realización de los procedimientos. Conclusión: La actuación del equipo de enfermería en Paro Cardiorrespiratorio debe continuar siendo difundida, visando mayor seguridad y autonomía de estos profesionales, visando un cuidado de calidad, el equipo de enfermería debe ser capaz de atender este tipo de emergencia clínica, de modo que puedan participar de constantes capacitaciones y actualizaciones, para que se capaciten y brinden una atención segura y eficaz.