bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract A027: Characterization of WEE1 kinase activity in myxoid liposarcoma
Authors: Heinst Lorena , Berthold Ruth , Isfort Ilka , Wosnig Svenja , Kindler Thomas , Åman Pierre , Wardelmann Eva , Scholl Claudia , Fröhling Stefan , Hartmann Wolfgang , Trautmann Marcel
Published: September 2022

Introduction: Myxoid liposarcomas (MLS), malignant soft tissue tumors of adipocyte origin, are genetically characterized by a chromosomal t(12;16)(q13;p11) translocation encoding the chimeric FUS-DDIT3 fusion gene. The resulting fusion protein drives MLS pathogenesis via (dys-)regulation of oncogenic signaling pathways. Since FUS-DDIT3 is not selectively antagonizable, counteracting the oncogenic effects of FUS-DDIT3 fusion protein represents the most promising strategy to target MLS cells. In this study, we identified cell cycle checkpoint kinase WEE1 as FUS-DDIT3 depending effector and investigated the functional requirement for WEE1 kinase activity in MLS pathogenesis. Experimental Procedures: Characterization of WEE1 expression and kinase activity was performed in multiple MLS cell lines, cell lines derived from other liposarcoma subtypes and a mesenchymal stem cell system. Modulation of WEE1 signaling was carried out by means of small-molecule inhibitor Adavosertib (MK-1775) and RNA interference (RNAi)-mediated depletion, and effects were analyzed in immunoblots, cell proliferation assays and caspase 3/7 activity-based apoptosis assays in vitro. Results: Functional genomic RNAi screening uncovered dependence of FUS-DDIT3-expressing mesenchymal stem cells and MLS cell lines on WEE1 kinase activity. Additional expression analysis revealed increased WEE1 protein levels in MLS cell lines. Moreover, pharmacologic inhibition of WEE1 demonstrated significant reduction of MLS cell viability. Accordingly, functional loss of WEE1 by inhibition or RNAi-mediated depletion was found to induce DNA damage accompanied by unscheduled mitotic entry and cell death via activation of the apoptotic program in MLS cells. Conclusions: Our results identify WEE1 kinase activity as functional liability of FUS-DDIT3 expressing MLS cells and provide first evidence that overactive WEE1 signaling represents a promising target for therapeutic intervention in MLS.

Citation Format: Lorena Heinst, Ruth Berthold, Ilka Isfort, Svenja Wosnig, Thomas Kindler, Pierre Åman, Eva Wardelmann, Claudia Scholl, Stefan Fröhling, Wolfgang Hartmann, Marcel Trautmann. Characterization of WEE1 kinase activity in myxoid liposarcoma [abstract]. In: Proceedings of the AACR Special Conference: Sarcomas; 2022 May 9-12; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2022;28(18_Suppl):Abstract nr A027.
Abstract A031: Interplay of YAP1, β-catenin and the SS18-SSX fusion protein in synovial sarcoma
Authors: Isfort Ilka , Berthold Ruth , Heinst Lorena , Wardelmann Eva , Trautmann Marcel , Hartmann Wolfgang
Published: September 2022

Introduction: Synovial sarcoma (SySa) is a rare soft-tissue malignancy characterized by a specific chromosomal translocation t(X;18). The resulting chimeric SS18-SSX fusion protein drives SySa pathogenesis by dysregulating gene transcription. As therapeutic targeting of SS18-SSX is notoriously difficult, the identification of SS18-SSX-dependent pathomechanisms may uncover novel molecular vulnerabilities. Since previous functional analyses revealed a connection between SS18-SSX and the activity of the transcriptional co-regulators YAP1/TAZ and β-catenin, this study examined a potential crosstalk between these effectors in SySa. Experimental Procedures: Nuclear abundance of YAP1/TAZ and β-catenin was analyzed by immunohistochemistry in a large cohort of SySa tissue specimens. Two SySa cell lines were employed to modulate YAP1/TAZ/β-catenin transcriptional activity by RNAi-mediated knockdown, small molecule inhibitors, and overexpression vectors in vitro. Changes in transcriptional activity were determined via luciferase reporter assays. As YAP1 and β-catenin showed highest interdependence, co-immunoprecipitation experiments were performed to investigate their interplay with the SS18-SSX fusion protein in SySa cells. Results: Immunohistochemically, a substantial subset of SySa tissue specimens showed concurrent nuclear accumulation of YAP1/TAZ and β-catenin. In vitro, SS18-SSX was found to promote YAP1/TAZ and β-catenin transcriptional activity. RNAi-mediated knockdown of YAP1, TAZ or β-catenin and small molecule inhibitor treatment mutually downregulated TEAD and TCF luciferase reporter activity, while expression of hyperactive variants led to a reciprocal luciferase reporter induction. Finally, co-immunoprecipitation assays demonstrated nuclear co-localization of SS18-SSX with YAP1 and β-catenin in SySa cell lines. Conclusions: This study reveals an interdependency between YAP1 and β-catenin activation and a complex interplay with the SySa-specific SS18-SSX fusion protein.

Citation Format: Ilka Isfort, Ruth Berthold, Lorena Heinst, Eva Wardelmann, Marcel Trautmann, Wolfgang Hartmann. Interplay of YAP1, β-catenin and the SS18-SSX fusion protein in synovial sarcoma [abstract]. In: Proceedings of the AACR Special Conference: Sarcomas; 2022 May 9-12; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2022;28(18_Suppl):Abstract nr A031.
Abstract B008: Investigating the evolution of undifferentiated soft tissue sarcomas in a genetically engineered mouse model
Authors: Chan Jason E. , Rub Jonathan , Sussman Carleigh , Grbovic-huezo Olivera , Tap William D. , Antonescu Cristina , Singer Sam , Betel Doron , Tammela Tuomas
Published: September 2022

Undifferentiated soft tissue sarcomas (USTS) are an aggressive class of sarcomas with few effective treatment options. Because of the rarity of these sarcomas and the paucity of representative disease models, little is known about how they develop. We used single cell transcriptomics to study the initiation and evolution of an USTS in an autochthonous genetically engineered mouse model. We profiled multiple stages of tumor development and identified intermediary cell states that define sarcomagenesis. Interestingly, we identify two discrete evolutionary pathways that arise in parallel in the mouse model. To determine the human relevance of our findings, we are developing machine learning approaches to compare cell states between the mouse model with single-cell transcriptomes that we generated from patient-derived xenografts. This cross-species comparison will uncover core USTS cell states and define their ancestral state transitions. Our ultimate goal is to identify essential molecular features of USTS, which may represent entry points for future therapeutic strategies.

Citation Format: Jason E. Chan, Jonathan Rub, Carleigh Sussman, Olivera Grbovic-huezo, William D. Tap, Cristina Antonescu, Sam Singer, Doron Betel, Tuomas Tammela. Investigating the evolution of undifferentiated soft tissue sarcomas in a genetically engineered mouse model [abstract]. In: Proceedings of the AACR Special Conference: Sarcomas; 2022 May 9-12; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2022;28(18_Suppl):Abstract nr B008.
Abstract B013: STEAP1 facilitates iron transport in ewing sarcoma to support mitochondrial activity
Authors: Shyp Taras , Mousavizadeh Rouhollah , Negri Gian Luca , Zhang Hai-Feng , Lizardo Michael , Sorensen Poul
Published: September 2022

Ewing sarcoma (EwS) is a highly aggressive bone-associated tumor of children and young adults with strong propensity to progress and dismal outcome for patients with advanced stages of the disease. EwS is defined by a pathognomonic recurrent potent transcription factor, in 85-90% of cases EWSR1-FLI1. One of the previously described transcriptional targets of EWSR1-FLI1 is six-transmembrane epithelial antigen of the prostate 1 (STEAP1). STEAP1 is a member of the STEAP membrane-bound protein family involved in reduction of iron and copper which is a crucial step for metal transport and overexpressed in various malignancies. However, a precise functional role of STEAP1 in cancers including EwS remains elusive. EwS cell lines with knockout (KO) or knockdown (KD) of STEAP1 were created to perform functional studies for elucidating the functions of STEAP1 in EwS, including transient metal measurements. Additionally, the measurement of functional mitochondrial activity was performed by an extracellular flux analyzer. The activity of the mitochondrial electron transport chain complexes was measured by blue native PAGE. We showed that STEAP1 functions as a metalloreductase and STEAP1 KO cells have lower intracellular iron levels. Furthermore, we discovered that STEAP1 KO cells have significantly lower levels of the mitochondrial labile iron pool. Consequently, EwS cells with STEAP1 KO demonstrated impaired mitochondrial respiration and increased glycolysis rates. Additionally, the functional activity of iron-sulfur containing proteins and the the assembly of subunits of the mitochondrial electron transport chain complexes were decreased. This study is the first to demonstrate in vitro involvement of STEAP1 in transitional metal homeostasis and mitochondrial activity. These discoveries unveil alternative therapeutic strategies for EwS patients such as using compounds that promote an iron-dependent form of cell death, ferroptosis, or drugs that decrease metabolic adaptability.

Citation Format: Taras Shyp, Rouhollah Mousavizadeh, Gian Luca Negri, Hai-Feng Zhang, Michael Lizardo, Poul Sorensen. STEAP1 facilitates iron transport in ewing sarcoma to support mitochondrial activity [abstract]. In: Proceedings of the AACR Special Conference: Sarcomas; 2022 May 9-12; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2022;28(18_Suppl):Abstract nr B013.
Abstract B026: Expression levels of chaperonin containing TCP1 in cancer are among the highest in sarcomas
Authors: Cox Amanda J. , Khaled Annette
Published: September 2022

Protein folding complexes are essential for oncogenesis. Of these, the multi-subunit complex, Chaperonin-Containing TCP-1 (CCT or TRiC), is upregulated across a spectrum of cancers and folds many of the oncoproteins that drive cancer growth. Using the UCSC Xena online database, which includes datasets from TCGA, TARGET, GTEx, and KidsFirst, we found significant increases in gene expression of the subunits (CCT1-8) that form the chaperonin complex in sarcomas, both adult and pediatric, as compared to other cancers like invasive breast carcinoma, lung carcinoma, glioblastoma multiforme, or prostate cancer. Most notably was the high expression of the second subunit (CCT2) in Rhabdomyosarcoma compared to other pediatric cancers. These findings were confirmed with other databases (e.g., Oncomine). Expression of CCT subunits, like CCT2, was decreased in normal tissues, such as muscle, kidney, and uterus, as compared to sarcoma tissues. Bioinformatic data was confirmed by finding detectable staining of CCT2 protein in sarcoma tissues. This was supported by data from the human protein atlas showing minimal staining for CCT2 in normal smooth and skeletal muscle. Cancer adjacent tissues from a metastatic breast cancer patient study had minimal staining for CCT2 in muscle, fat, and bone. These data show that CCT is increased in sarcomas, while decreased in normal tissues, and has potential for further study as a driver of sarcoma development and a target for therapeutic intervention.

Citation Format: Amanda J. Cox, Annette Khaled. Expression levels of chaperonin containing TCP1 in cancer are among the highest in sarcomas [abstract]. In: Proceedings of the AACR Special Conference: Sarcomas; 2022 May 9-12; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2022;28(18_Suppl):Abstract nr B026.
Abstract IA004: Mesenchymal progenitors and sarcomagenesis
Authors: Underhill T. Michael
Published: September 2022

Synovial sarcoma (SS) is an aggressive soft-tissue malignancy that can arise in any area of the body, and most frequently affects adolescents and young adults. The 10-year survival rate has been estimated at 50%. SS is characterized by a pathognomonic t(X;18)(p11.2;q11.2) translocation which produces a fusion oncogene named SS18-SSX. In mouse models, expression of SS18-SSX is sufficient to drive the development of SS-like tumors. Despite recent advancements in our understanding of SS biology, the cell of origin remains undefined. In most instances, it is likely mesenchymal in nature. We previously generated a mesenchymal progenitor (MP) specific Cre line that has enabled us to identify MPs within the embryo [MOU1], and characterize and define their lineage trajectories through development into the adult. This line has been used to generate a unique mouse model of SS where the human SS18-SSX fusion oncogene with an EGFP marker is conditionally expressed in adult MPs. This model shows 100% penetrant SS tumor formation with a median latency period of 16 weeks. Histologic analyses of tumors reveal characteristic cellular morphology and the expression of common markers of SS. Single-cell (sc) RNA-seq gene expression profiles of murine tumors recapitulate those observed in human SS samples and highlight genetic programs involved in sarcomagenesis. Using this model, we can identify and isolate neoplastic lesions prior to the development of grossly visible tumors and trace these cells through the complete transformation continuum. By comparing tumor cells to normal MPs, we aim to better understand the mechanisms underlying the SS18-SSX induced transformation of MPs. Together, this model identifies a cell of origin for SS and provides us with a unique opportunity to address the molecular and cellular drivers of SS.

Citation Format: T. Michael Underhill. Mesenchymal progenitors and sarcomagenesis [abstract]. In: Proceedings of the AACR Special Conference: Sarcomas; 2022 May 9-12; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2022;28(18_Suppl):Abstract nr IA004.
Abstract IA005: On the use of stem cells to generate sarcomas in vitro models
Authors: Boveé Judith V.M.G.
Published: September 2022

Sarcomas are rare cancers arising in soft tissue and bone. They are difficult to diagnose and treat, with 38-48% mortality for this often young age group. It is now generally accepted that molecular changes in cancer open avenues for personalized medicine. However, to achieve this for sarcoma there are several challenges, as representative model systems are scarce. Moreover, sarcomas are heterogeneous and >70 sarcoma types are recognized. A simple dichotomy based on molecular pathways of simple versus complex genomics is often used as a conceptual framework. My group has used mesenchymal stem cells to generate a model for complex genome sarcomas such as osteosarcoma. Murine and canine mesenchymal stem cells (MSCs) can be used to model spontaneous malignant transformation towards sarcomas with complex genomics. These MSCs have an abnormal karyotype, many structural variants and point mutations at whole genome sequencing analysis, and form sarcomas (osteosarcoma, undifferentiated pleomorphic and spindle cell sarcoma) after injection into mice. Cross-species analysis reveals that p53 loss is an early event in sarcomagenesis, and MSCs with a knockout in Trp53 transform earlier compared to wild-type MSCs. In addition, loss of p16Ink4a is shown to be a driver of osteosarcomagenesis: the majority of spontaneously transformed murine MSCs has alterations in p15Ink4b, p16Ink4a, or p19Arf and knockout of these genes in MSCs results in accelerated transformation. Of interest, osteosarcoma cells with defective p16INK4A are sensitive to the CDK4/CDK6 inhibitor Palbociclib, suggesting that a subset of osteosarcoma patients with intact Rb, but defective p16 or overexpression of CDK4 and/or CDK6 (in total 20-23%) might benefit from CDK4/CDK6 inhibition. Bone and soft tissue tumors with a simple genome often carry chromosomal translocations. We have generated a model for pseudomyogenic hemangioendothelioma (PHE), a rare locally aggressive vascular tumor. PHE has a tumor-specific t(7;19)(q22;q13) translocation resulting in a SERPINE1-FOSB fusion. We used CRISPR/Cas9 to generate this fusion in induced pluripotent stem cells (hiPSCs), which were subsequently differentiated into ECs (hiPSC-ECs) to address this. Comparison of parental with PHE hiPSC-ECs shows invasive growth in mice after transplantation, and transcriptome analysis identifies PI3K-Akt and MAPK signaling pathways as possible therapeutic targets. The modified hiPSC-ECs thus recapitulate functional features of PHE and demonstrate how these stem cell based translocation models can be used to understand tumorigenic mechanisms and identify therapeutic targets.

Citation Format: Judith V.M.G. Boveé. On the use of stem cells to generate sarcomas in vitro models [abstract]. In: Proceedings of the AACR Special Conference: Sarcomas; 2022 May 9-12; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2022;28(18_Suppl):Abstract nr IA005.
Abstract IA006: Immunobiology of sarcomas and therapeutic implications
Authors: Llosa Nicolas J.
Published: September 2022

Undoubtedly, the greatest breakthrough in cancer treatment over the past decade has been immunotherapy. However, the immunosuppressive nature of the tumor microenvironment (TME) remains the biggest obstacle to increasing the number of patients that respond to immunotherapy. Unfortunately, initial trials of immune checkpoint inhibitors in patients with soft tissue sarcoma (STS) have achieved very limited success, and the reasons for this are crudely unappreciated. Hence, it is critical to move beyond the current focus on targeting individual cell types of interest and rather adopt a more comprehensive systems-level approach in which we analyze and integrate all the TME components to identify and disable the critical nodes. Our team has characterized the immunobiology of STS via cutting–edge high-dimensional mapping platforms that we have established and standardized for analyzing specimens from patients with STS. We have discovered that tumor infiltrating myeloid cells (TIM) penetrate and dominate the TME of STS provoking the suppression of effective antitumoral T cell immune responses. Using a set of sophisticated multiscale techniques, we deconvoluted the TME of various STS creating a comprehensive map of all immune cell populations present and dissected out chemokine gradients of specific histological regions relevant for the recruitment and development of TIM in the TME of STS. STS TIM are regulated by tumor-derived cytokines to acquire a polarized immunosuppressive phenotype, which in turn de-activates the T cell compartment in the TME. In this way, we elucidated unique myeloid populations as key facilitators of STS progression and mediators of local tumor immunosuppression. Furthermore, we defined metabolic pathways utilized by TIM that contribute to their immunosuppressive properties in STS and manipulated them via glutamine antagonism in our mouse model of STS. Our intervention generated a TME that was less acidic, less hypoxic and more replete with nutrients leading to an intratumoral influx of T cells, both CD4+ and CD8+ cells, along with B cells. These modifications in the TME in combination with anti PD-1 therapy induced better endogenous anti-tumor responses and repurposed TIM markedly thereby reducing the tumor size and improving the survival of mice. Thus, we propose a metabolic strategy to therapeutically target the TME of STS and reprogram their immunosuppressive myeloid niche as a means of enhancing immunotherapy.

Citation Format: Nicolas J. Llosa. Immunobiology of sarcomas and therapeutic implications [abstract]. In: Proceedings of the AACR Special Conference: Sarcomas; 2022 May 9-12; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2022;28(18_Suppl):Abstract nr IA006.
Abstract IA007: B cell immunity generation in tertiary lymphoid structures, a major determinant of sarcoma immune response to immunotherapy
Authors: Fridman Wolf H.
Published: September 2022

The presence of tertiary lymphoid structures (TLSs) and a high intratumoral density of B lymphocytes have been associated with improved survival in several tumor types. We investigated the impact of TLS on responses to immunotherapy and, analyzing a large-scale retrospective analysis of three independent cohorts of patients with cancer, including sarcomas, who were treated with anti-PD-1 or anti-PD-L1 antibodies, we showed that the presence of mature TLSs was associated with improved objective response rates, progression-free survival and overall survival, independent of PD-L1 expression status and CD8+T cell density. To decipher the role of B cells in such responses, we performed spatial transcriptomics and examined the nature of B cell responses within TLS structure. B cells were enriched in TLS, and therein we could identify all B cell maturation stages toward plasma cell (PC) formation. B cell repertoire analysis revealed clonal diversification, selection, expansion in TLS and the presence of full mature clonotypes at distance. In TLS+ tumors, IgG producing PCs disseminated into the tumor beds along fibroblastic tracks. TLS+ tumors exhibited high frequencies of IgG-producing PCs and IgG-stained and apoptic malignant cells, suggestive of anti-tumor effector activity. We validated these findings in a prospective trial in metastatic Soft Tissue Sarcoma (STS) bearing patients. PEMBROSARC is a multi-cohort phase II study of pembrolizumab combined with low-dose cyclophosphamide in patients with advanced STS. The 6-months non-progressive rate (NPR) and the objective response rates previously reported in cohorts including all comers were 4.9% and 2.4%, respectively. In the cohort enrolling patients selected based on the presence of TLS, the 6-month NPR was 40% and the objective response rate was 30%. Exploratory analyses reveled that infiltration by IgG antibody-producing plasma cells was associated with improved clinical outcome as well as the presence of IgG antibodies on tumor cells. Altogether, these results establish that maturation, selection and amplification of B cells in intratumoral TLSs results in Plasma Cells generation which may produce anti-tumor antibodies. The latter may amplify in situ antitumor immune responses leading to better therapeutic responses to immunotherapy.

Citation Format: Wolf H. Fridman. B cell immunity generation in tertiary lymphoid structures, a major determinant of sarcoma immune response to immunotherapy [abstract]. In: Proceedings of the AACR Special Conference: Sarcomas; 2022 May 9-12; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2022;28(18_Suppl):Abstract nr IA007.
Abstract IA009: Oncogenic transcription factors drive highly tumor-specific LincRNA
Authors: Delattre Olivier , Vibert Julien , Saulnier Olivier , Waterfall Joshua
Published: September 2022

Many cancers are characterized by gene fusions encoding oncogenic chimeric transcription factors (TFs) such as EWS::FLI1 in Ewing sarcoma (EwS). EWS::FLI1 induces a complete reprogramming of mesenchymal stem cells, the putative cell of origin of Ewing sarcoma, through binding and subsequent generation of thousands of neoenhancers at GGAA microsatellite repeats. We show that EWS::FLI1 induces the robust expression of a specific set of novel spliced and polyadenylated transcripts within otherwise transcriptionally silent regions of the genome. These neo-genes (NGs) are virtually undetectable in large collections of normal tissues or non-EwS tumors and can be silenced by CRISPR interference at regulatory EWS::FLI1-bound microsatellites. Ribosome profiling and proteomics further show that some NGs are translated into highly EwS-specific peptides. More generally, hundreds of NGs can be detected in diverse cancers characterized by chimeric TFs. Altogether, we identify the transcription, processing, and translation of novel, specific, highly expressed multi-exonic transcripts from otherwise silent regions of the genome as a new activity of aberrant TFs in cancer. These neotranscripts, which can be classified as LincRNAs, or neopeptides may have specific functions in the tumor cells. The latter may also constitute ideal neoepitopes as they are tumor-specific, clonally expressed in sarcoma cells, induced by the driver genetic alterations and public for a given sarcoma.

Citation Format: Olivier Delattre, Julien Vibert, Olivier Saulnier, Joshua Waterfall. Oncogenic transcription factors drive highly tumor-specific LincRNA [abstract]. In: Proceedings of the AACR Special Conference: Sarcomas; 2022 May 9-12; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2022;28(18_Suppl):Abstract nr IA009.
Abstract IA019: How to target lung metastasis: dynamic changes in chromatin accessibility during osteosarcoma lung metastasis expose vulnerabilities
Authors: Scacheri Peter C.
Published: September 2022

The metastatic cascade describes the series of steps required for a cancer cell to successfully spread from its primary tumor to a secondary organ. Despite knowing that metastasis is a fluid process, almost all studies to date have relied on end-point assays. As a result, little is known about the continuum of molecular events required for successful growth within the secondary organ. By profiling accessible chromatin in osteosarcoma cells grown in vitro and isolated from an early and late timepoint during lung metastasis, we identify temporally distinct epigenomic reprogramming events that occur as tumors form and grow within the metastatic microenvironment. Furthermore, these data pinpoint key upstream transcription factors responsible for the dynamic chromatin changes. Through paired in vivo and in vitro CRISPR drop-out screens, we show a subset of these putative pro-metastatic transcription factors are essential for lung metastasis, but dispensable for in vitro growth. Targeting the same factors with chemical probes in vitro and ex vivo demonstrated similar context-dependent essentiality. Altogether, our study demonstrates the epigenomes of metastasizing cancer cells are tightly regulated in a dynamic and context-specific manner, and the transcription factors controlling this process may serve as novel vulnerabilities.

Citation Format: Peter C. Scacheri. How to target lung metastasis: dynamic changes in chromatin accessibility during osteosarcoma lung metastasis expose vulnerabilities [abstract]. In: Proceedings of the AACR Special Conference: Sarcomas; 2022 May 9-12; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2022;28(18_Suppl):Abstract nr IA019.
Abstract IA029: The origins of leiomyosarcoma: implications for prognosis and defining new therapeutic options
Authors: Gladdy Rebecca A.
Published: September 2022

Introduction: Leiomyosarcomas (LMS) are genetically heterogenous tumors differentiating along smooth muscle lines. Currently, LMS treatment is not informed by molecular subtyping and is associated with highly variable survival. While disease site continues to dictate clinical management, the contribution of genetic factors to LMS subtype, origins, and timing are unknown. Here we analyze 70 genomes and 130 transcriptomes of LMS, including multiple tumor regions and paired metastases, with the goal of defining the molecular origins of LMS and using both computational analysis and functional genomics to define new therapeutic arenas for metastatic disease. Results: Molecular profiling highlight the very early origins of LMS. We uncover three specific subtypes of LMS that likely develop from distinct lineages of smooth muscle cells. Of these, dedifferentiated LMS with high immune infiltration and tumors primarily of gynecological origin harbor genomic dystrophin deletions and/or loss of dystrophin expression, acquire the highest burden of genomic mutation, and are associated with worse survival. Homologous recombination defects lead to genome-wide mutational signatures, and a corresponding sensitivity to PARP trappers and other DNA damage response inhibitors, suggesting a promising therapeutic strategy for LMS. Finally, by phylogenetic reconstruction, we present evidence that clones seeding lethal metastases arise decades prior to LMS diagnosis. Conclusions: In summary, LMS, the catch all-label for any primary malignant smooth muscle neoplasm, encompasses at least three distinct entities. We report that the utility of DNA damage response inhibitors is promising and warrants further exploration. Finally, the molecular evolution and timing of LMS provide a broad window for early intervention and risk stratification for these patients.

Citation Format: Rebecca A. Gladdy. The origins of leiomyosarcoma: implications for prognosis and defining new therapeutic options [abstract]. In: Proceedings of the AACR Special Conference: Sarcomas; 2022 May 9-12; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2022;28(18_Suppl):Abstract nr IA029.
Abstract PR006: Multi-modal single-cell profiling of sarcomas from archival tissue reveals mechanisms of resistance to immune checkpoint inhibitors
Authors: Bose Sminu , Caldwell Blake , Amin Amit Dipak , Melms Johannes , Tagore Somnath , Ingham Matthew , Izar Benjamin , Schwartz Gary K.
Published: September 2022

Single-cell genomics is an enabling technology that may inform the molecular underpinnings of drug response and resistance in patient biopsies. These methods are difficult to implement in the study of rare diseases such as sarcomas due to specimen requirements and technical limitations. Here, we evolved novel methods that we recently reported in melanoma (Wang, Fan, et al., bioRxiv, 2022) which enable single-nucleus RNA, T cell receptor (snRNA/TCR-seq), and pool-matched whole-genome sequencing (WGS) from archival frozen sarcoma tissue. This enabled profiling of 75,716 cells and 788 matched TCR clonotypes from six patients with intimal sarcoma (INS) and undifferentiated pleomorphic sarcoma (UPS), including two matched pair samples from pre/post-immune checkpoint inhibitor (ICI). Our analysis revealed substantial transcriptional cancer cell heterogeneity driven by varying copy number alterations (CNAs). In one patient with INS with a complete response to ICI followed by an isolated recurrence, we identified a rare cancer cell clone defined by CNA (confirmed with WGS) and resulting transcriptional outputs that pre-existed and emerged during resistance. Furthermore, in a UPS patient with intrinsic resistance to ICI, we find adequate T cell clonal expansion, activation, and differentiation, suggesting appropriate T cell response to ICI dampened by intrinsic mechanisms of ICI resistance within the cancer cells. Non-negative matrix factorization (NMF) analysis identified cell states associated with either intrinsic or adaptive resistance to ICI that was distinct from resistance to doxorubicin. These observations are consistent with those previously reported from sequential biopsies obtained from KEYNOTE-001 in metastatic melanoma (Wang, Fan, et al., bioRxiv, 2022), which also revealed emergence of pre-existing populations of resistant clones defined by their underlying aneuploidy patterns. Together, these results demonstrate feasibility of implementing single-cell genomics from archival tissue to study sarcoma and propel our understanding of drug resistance. Conceptually, this work suggests that large-scale CNAs may allow us to identify cell sub-populations associated with ICI resistance in sarcoma and in other diseases.

Citation Format: Sminu Bose, Blake Caldwell, Amit Dipak Amin, Johannes Melms, Somnath Tagore, Matthew Ingham, Benjamin Izar, Gary K. Schwartz. Multi-modal single-cell profiling of sarcomas from archival tissue reveals mechanisms of resistance to immune checkpoint inhibitors [abstract]. In: Proceedings of the AACR Special Conference: Sarcomas; 2022 May 9-12; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2022;28(18_Suppl):Abstract nr PR006.
IgE Sensitization Profile in Patients with Netherton Syndrome
Authors: Scala Enrico , Condorelli Angelo Giuseppe , Scala Alessandro , Caprini Elisabetta , Didona Biagio , Paganelli Roberto , Castiglia Daniele
Published: September 2022

Background: Scant data are currently available on the allergen-specific immunoglobulin (Ig)E sensitization profile in primary immunodeficiencies with hyper IgE. Netherton syndrome (NS, OMIM 266500) is an extremely rare form of congenital ichthyosis characterized by congenital scaly erythroderma, hair abnormalities, and deregulated IgE reactivity associated with severe atopic manifestations. Objective: The aim of this study was to evaluate the feasibility and reliability of a multiplex proteomic approach in the detection of specific IgE in NS. Methods: Specific IgE was evaluated in 10 individuals with an established molecular diagnosis of NS using an allergenic molecules microarray (immuno-solid-phase allergen chip). Results: Polireactivity to airway allergens, mainly house dust mites and olive tree pollen, and food allergens were observed in NS. Eighty per cent of patients were responsive to LTP or profilins. A clinical history suggestive of severe egg, milk, and fish allergy was confirmed by reactivity to the thermostable molecules Gal d 1, Bod 8, and parvalbumin Gad c 1, respectively. Latex reactivity was associated with Hev b 5 and 6 reactivity. Two distinct clusters of reactivity were observed after hierarchical analysis. Extremely high IgE levels (> 10,000 kU/L) do not affect the results obtained with microarrays. Conclusion: IgE multiplex evaluation allows (i) to profile IgE polyreactivity pictures, in the presence of LTP and profilin sensitization, (ii) to verify the clinical history of food allergy to milk, egg, and seafood, (iii) to confirm the allergic events associated with latex exposure, and (iv) to disclose the presence of preclinical sensitizations in patients affected by primary immunodeficiencies with hyper IgE, such as the NS.
Method to assess the potential of photovoltaic panel based on roof design
Authors: Djafar Abdi Gunawan , Mohamad Yasin
Published: September 2022

The photovoltaic (PV) panel makes it possible for everyone to produce electricity in their own house. However, the panel is quite a costly investment and requires much consideration to maximize its potential. The roof has variables that would impact electricity generation. The roof of housing in Indonesia was built generally in a complex shape, a combination of gable and hip roofs. This research is conducted to break down factors affecting PV productivity in regard to the roof’s aspects. A computer simulation using Ladybug plugin in Rhinoceros software has been done to achieve the target. Initially, the performance of PV panels on the gable, and hip roof, is analyzed respectively. It is found that the roof’s slope, and orientation, contribute more to the amount of electricity produced than the shape itself. These factors were used to assess the PV potential in several housing models employing simple and complex roof (more than 2 surfaces) construction in the city of Gorontalo. Eventually, a comparison between the estimation of real PV production on housing, and the estimation provided by the simulation is conducted to verify the assessment method. The difference is about 4%, as proof the simulation result is reliable.
Power quality improvement of wind energy system using energy storage model and DSTATCOM
Authors: Bhadane Kishor V. , Thakre Mohan , Shriwastava Rakesh , Kadam Deepak P. , Bhadane Dnyaneshwar V. , Harne Mahesh
Published: September 2022

Nowadays power crises in different countries are observed and the main cause of the power crisis is the huge gap between the supply and demand of electricity, renewable energy sources are identified as an alternative to overcome the power crisis gap. Renewable wind energy is the most promising energy source. Increasing the integration of wind energy into the grid causes the exploitation of power quality. Hence there is a need to deal with this issue. In this case, supercapacitors and custom power devices are introduced as smart energy storage devices in grid-connected wind energy systems for power quality enhancement features. The indirect current control scheme has interfered with custom power devices based on DSTATCOM. The optimal MATLAB-based smart energy storage model and hardware results are compared and validated. power quality improvement feature of grid-connected wind energy system using DSTATCOM is highlighted. The main aim of this study is to determine and interface the optimistic energy storage device into grid connected wind energy system. So that the stability of the wind energy system is to be maintained and also able to enhance the overall efficiency of the wind energy system.
Hybrid MPPT-based predictive speed control model for variable speed PMSG wind energy conversion systems
Authors: Hashish Mai N. Abu , Daoud Ahmed Ali , Elfar Medhat Hegazy
Published: September 2022

In this research, a predictive speed control (PSC) technique based on permanent magnet synchronous generators is proposed for variable-speed wind energy conversion systems (VS-WECS) (PMSG). The control approach that has been developed makes it possible to regulate mechanical and electrical variables concurrently within the context of a single cost function. The power converter will then use the optimum switching state that will result in the lowest possible cost function when it has been chosen. The maximum power point tracking (MPPT) algorithms used in the proposed control approach are combined in order to achieve optimum efficiency. As a direct result of this, the conventional cascade structure of proportional-integral (PI) controllers has been removed, which results in an improvement in the system's dynamic responsiveness. In addition, predictive current control, also known as PCC, is implemented on the grid-side converter, also known as the GSC, in order to accomplish decoupled grid current control. Using MATLAB/SIMULINK, we analyze the performance of the suggested control methods and compare it to the performance of a traditional PI speed controller. The findings demonstrated that the MPC controller is superior than the PI controller in terms of its ability to handle system dynamics.
Inverter based implementation of maximum power point techniques
Authors: Grover Ashish , Adlakha Richa
Published: September 2022

The utilization of renewable energy sources is being pushed by both greater environmental consciousness and expanding demand. Recently, solar photovoltaic technology has found increased use for a broader range of applications. This may be ascribed to solar energy's extensive availability as well as its long-term viability and low cost. According to the global photovoltaic (PV) industry, 594 gigawatts (GW) of PV capacity were installed in 2019, with the objective of replacing conventional source-based generating facilities. The major problem in PV production, however, is identifying the maximum power point tracking (MPPT) systems that are currently in use to compute peak output. For 1240 W PV power plants, this article compares perturb & observe MPPT approaches with incremental conductance MPPT techniques. The MATLAB Simulink program was utilized to conduct the study, which was based on many factors such as voltage, current, and output power under various weather conditions. When these MPPT algorithms are applied to solar trackers, the efficiency, reaction time, and steady-state oscillations all improve.
Revision of the genus Alloplasta Förster, 1869 (Hymenoptera, Ichneumonidae, Banchinae) from Japan
Authors: WATANABE KYOHEI
Published: September 2022

Japanese species of the genus Alloplasta Förster, 1869 are revised. A total of nine species are recorded from Japan, including four new species, A. breviterebra sp. nov., A. japonica sp. nov., A. satana sp. nov. and A. septentrionalis sp. nov. and three species, A. brevipetiolaris Kang & Lee, 2020, A. kuslitzkii Kasparyan, 2007 and A. subgrisea Kasparyan, 2007, newly recorded from Japan. Amersibia longipetiolaris Uchida, 1952 is newly synonymized under A. nigripes (Meyer, 1930) (syn. nov.). A key to Japanese species of this genus is proposed.
Canada's First Experience With a Patient-Adjudicated Cancer Research Grant Competition: A Case Study
Authors: Ackerman Ruth , Desserud Don , Baudais Nathalie , Lascelle Debi , Badovinac Kimberly
Published: September 2022

Although patient partners have been part of grant review panels, they are rarely given decision-making authority and never given sole responsibility for determining what research to fund. In patient-oriented research, however, patient partners may be in the best position to determine what is needed and whether the proposed research will engage patients in meaningful ways. The objective of this case study was to demonstrate that patient partners can adjudicate patient-oriented research proposals without the inclusion of researcher/expert reviewers.
Transcriptome analysis of sweet potato responses to potassium deficiency
Authors: Wang Fang , Tan Wen-Fang , Song Wei , Yang Song-Tao , Qiao Shuai
Published: September 2022

Background

As one of three essential nutrients, potassium is regarded as a main limiting factor for growth and development in plant. Sweet potato (Ipomoea batatas L.) is one of seven major food crops grown worldwide, and is both a nutrient-rich food and a bioenergy crop. It is a typical ‘K-favoring’ crop, and the level of potassium ion (K+) supplementation directly influences its production. However, little is known about the transcriptional changes in sweet potato genes under low-K+ conditions. Here, we analyzed the transcriptomic profiles of sweet potato roots in response to K+ deficiency to determine the effect of low-K+ stress on this economically important crop.

Results

The roots of sweet potato seedlings with or without K+ treatment were harvested and used for transcriptome analyses. The results showed 559 differently expressed genes (DEGs) in low and high K+ groups. Among the DEGs, 336 were upregulated and 223 were downregulated. These DEGs were involved in transcriptional regulation, calcium binding, redox-signaling, biosynthesis, transport, and metabolic process. Further analysis revealed previously unknow genes involved in low-K+ stress, which could be investigated further to improve low K+ tolerance in plants. Confirmation of RNA-sequencing results using qRT-PCR displayed a high level of consistency between the two experiments. Analysis showed that many auxin-, ethylene- and jasmonic acid-related genes respond to K+ deficiency, suggesting that these hormones have important roles in K+ nutrient signaling in sweet potato.

Conclusions

According to the transcriptome data of sweet potato, various DEGs showed transcriptional changes in response to low-K+ stress. However, the expression level of some kinases, transporters, transcription factors (TFs), hormone-related genes, and plant defense-related genes changed significantly, suggesting that they have important roles during K+ deficiency. Thus, this study identifies potential genes for genetic improvement of responses to low-K+ stress and provides valuable insight into the molecular mechanisms regulating low K+ tolerance in sweet potato. Further research is required to clarify the function of these DEGs under low-K+ stress.
Intracranial stenting as a bail-out option for posthemorrhagic cerebral vasospasm: a single-center experience with long-term follow-up
Authors: Khanafer Ali , Cimpoca Alexandru , Bhogal Pervinder , Bäzner Hansjörg , Ganslandt Oliver , Henkes Hans
Published: September 2022

Background

Cerebral vasospasm (CVS) is a leading cause of morbidity and mortality in patients after aneurysmal subarachnoid hemorrhage (aSAH). Endovascular treatment, including intraarterial infusion of drugs with vasodilation effects, and balloon- and stentriever angioplasty, are helpful but may achieve only short-term effects. There is a clinical need for long-lasting treatment of refractory recurrent vasospasm. We report our experience in stent implantation as a treatment for recurrent severe post-SAH vasospasm.

Methods

A retrospective analysis of our institutional database of 883 patients with SAH, managed between January 2010 and December 2021, was performed. Six patients were identified as having received intracranial stenting in the context of post-SAH cerebral vasospasm. All patients were initially treated with intra-arterial infusion of nimodipine and/or milrinone. Self-expanding intracranial stents were implanted during endovascular aneurysm treatment to enable access despite impaired perfusion (Group 1) or as a bail-out strategy after failed intraarterial drug infusion or mechanical treatment (Group 2). All stented patients received dual antiplatelet therapy (DAPT) for 6 months.

Results

Nine vessels in six patients with severe post-SAH vasospasm were stented. The stents were deployed in 16 vessel segments. All attempted implantations were technically successful. All patients demonstrated radiographic and clinical improvement of the vessel narrowing. No recurrent vasospasm or permanent vessel occlusion of the stented vessels was encountered. A thrombus formation in a Group 1 patient resolved under 4 mg eptifibatide IA infusion. During long-term angiographic follow-up, neither in-stent stenosis nor stent occlusion was found.

Conclusions

Endovascular implantation of self-expanding stents is a potential ultima ratio strategy for patients with severe refractory post-SAH cerebral vasospasm. Stents with reduced thrombogenicity (avoiding DAPT) and bioabsorbable self-expanding stents might further advance this concept.
The effect of the participatory heat education and awareness tools (HEAT) intervention on agricultural worker physiological heat strain: results from a parallel, comparison, group randomized study
Authors: Chavez Santos Erica , Spector June T. , Egbert Jared , Krenz Jennifer , Sampson Paul D. , Palmández Pablo , Torres Elizabeth , Blancas Maria , Carmona Jose , Jung Jihoon , Flunker John C.
Published: September 2022

Background

Farmworkers are at risk of heat-related illness (HRI). We sought to: 1) evaluate the effectiveness of farmworker Spanish/English participatory heat education and a supervisor decision-support mobile application (HEAT intervention) on physiological heat strain; and 2) describe factors associated with HRI symptoms reporting.

Methods

We conducted a parallel, comparison group intervention study from May–September of 2019 in Central/Eastern Washington State, USA. We used convenience sampling to recruit adult outdoor farmworkers and allocated participating crews to intervention (n = 37 participants) and alternative-training comparison (n = 38 participants) groups. We measured heat strain monthly using heart rate and estimated core body temperature to compute the maximum work-shift physiological strain index (PSImax) and assessed self-reported HRI symptoms using a weekly survey. Multivariable linear mixed effects models were used to assess associations of the HEAT intervention with PSImax, and bivariate mixed models were used to describe factors associated with HRI symptoms reported (0, 1, 2+ symptoms), with random effects for workers.

Results

We observed larger decreases in PSImax in the intervention versus comparison group for higher work exertion levels (categorized as low, low/medium-low, and high effort), after adjustment for maximum work-shift ambient Heat Index (HImax), but this was not statistically significant (interaction − 0.91 for high versus low/medium-low effort, t = − 1.60, p = 0.11). We observed a higher PSImax with high versus low/medium-low effort (main effect 1.96, t = 3.81, p < 0.001) and a lower PSImax with older age (− 0.03, t = − 2.95, p = 0.004), after covariate adjustment. There was no clear relationship between PSImax and the number of HRI symptoms reported. Reporting more symptoms was associated with older age, higher HImax, 10+ years agricultural work, not being an H-2A guest worker, and walking > 3 min to get to the toilet at work.

Conclusions

Effort level should be addressed in heat management plans, for example through work/rest cycles, rotation, and pacing, in addition to education and other factors that influence heat stress. Both symptoms and indicators of physiological heat strain should be monitored, if possible, during periods of high heat stress to increase the sensitivity of early HRI detection and prevention. Structural barriers to HRI prevention must also be addressed.

Trial registration

ClinicalTrials.gov Registration Number: NCT04234802, date first posted 21/01/2020.