bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract B23: PMN-MDSCs enhance CTC metastatic properties through reciprocal interactions via ROS/Notch/Nodal Nodal signaling
Authors: Sprouse Marc , Welte Thomas , Boral Debasish , Vishnoi Monika , Liu Haowen , Glitza-Oliva Isabella , Marchetti Dario
Published: June 2020

Intratumoral infiltration of myeloid-derived suppressor cells (MDSCs) is known to promote neoplastic growth by inhibiting the tumoricidal activity of T cells. However, direct interactions between patient-derived MDSCs and circulating tumors cells (CTCs) within the microenvironment of blood remain unexplored. Dissecting interplays between CTCs and circulatory MDSCs by heterotypic CTC/MDSC clustering is critical as a key mechanism to promote CTC survival and sustain the metastatic process. We characterized CTCs and polymorphonuclear-MDSCs (PMN-MDSCs) isolated in parallel from peripheral blood of metastatic melanoma and breast cancer patients by multiparametric flow cytometry. Transplantation of both cell populations in the systemic circulation of mice revealed significantly enhanced dissemination and metastasis in mice coinjected with CTCs and PMN-MDSCs compared to mice injected with CTCs or MDSCs alone. Notably, CTC/PMN-MDSC clusters were detected in vitro and in vivo either in patients’ blood or by longitudinal monitoring of blood from animals. This was coupled with in vitro coculturing of cell populations, demonstrating that CTCs formed physical clusters with PMN-MDSCs, and induced their protumorigenic differentiation through paracrine Nodal signaling, augmenting the production of reactive oxygen species (ROS) by PMN-MDSCs. These findings were validated by detecting significantly higher Nodal and ROS levels in blood of cancer patients in the presence of naïve, heterotypic CTC/PMN-MDSC clusters. Augmented PMN-MDSC ROS upregulated Notch1 receptor expression in CTCs through the ROS-NRF2-ARE axis, thus priming CTCs to respond to ligand-mediated (Jagged1) Notch activation. Jagged1-expressing PMN-MDSCs contributed to enhanced Notch activation in CTCs by engagement of Notch1 receptor. The reciprocity of CTC/PMN-MDSC bidirectional paracrine interactions and signaling was functionally validated in inhibitor-based analyses, demonstrating that combined Nodal and ROS inhibition abrogated CTC/PMN-MDSC interactions and led to a reduction of CTC survival and proliferation. This study provides seminal evidence showing that PMN-MDSCs, additive to their immunosuppressive roles, directly interact with CTCs and promote their dissemination and metastatic potency. Targeting CTC/PMN-MDSC heterotypic clusters and associated crosstalks can therefore represent a novel therapeutic avenue for limiting hematogenous spread of metastatic disease.

Citation Format: Marc Sprouse, Thomas Welte, Debasish Boral, Monika Vishnoi, Haowen Liu, Isabella Glitza-Oliva, Dario Marchetti. PMN-MDSCs enhance CTC metastatic properties through reciprocal interactions via ROS/Notch/Nodal Nodal signaling [abstract]. In: Proceedings of the AACR Special Conference on Advances in Liquid Biopsies; Jan 13-16, 2020; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2020;26(11_Suppl):Abstract nr B23.
Abstract B51: Performance assessment of total RNA sequencing of human biofluids and extracellular vesicles
Authors: Everaert Celine , Helsmoortel Hetty , Decock Anneleen , Hulstaert Eva , Van Paemel Ruben , Verniers Kimberly , Nuytens Justine , Anckaert Jasper , Nijs Nele , Tulkens Joeri , Dhondt Bert , Hendrix An , Mestdagh Pieter , Vandesompele Jo
Published: June 2020

RNA profiling has emerged as a powerful tool to investigate the biomarker potential of human biofluids. Despite the interest in extracellular nucleic acids, RNA sequencing methods to quantify the total RNA content outside cells are rare. Therefore, we evaluated the performance of the SMARTer Stranded Total RNA-Seq method in human platelet-rich plasma, platelet-free plasma, urine, conditioned medium, and extracellular vesicles (EVs) from these biofluids. We found the method to be accurate, precise, compatible with low-input volumes, and able to quantify a few thousand genes. We picked up distinct classes of RNA molecules, including mRNA, lncRNA, circRNA, miscRNA, and pseudogenes. Notably, the read distribution and gene content drastically differed among biofluids. We also observed differences between EDTA and citrate platelet-free plasma, which are identical biofluids collected in different blood tubes and prepared with a slightly different centrifugation protocol. In conclusion, we are the first to show that the SMARTer method can be used for unbiased charting of the complete transcriptome of a wide range of biofluids and their extracellular vesicles. The advantage of the SMARTer Stranded Total RNA-Seq method is its potential to process low amounts of input material. Indeed, collecting samples is often the bottleneck in fundamental, (pre)clinical, or translational research projects, and being able to disseminate large amounts of information from only 200 μL (or less) can substantially impact research progress.

Citation Format: Celine Everaert, Hetty Helsmoortel, Anneleen Decock, Eva Hulstaert, Ruben Van Paemel, Kimberly Verniers, Justine Nuytens, Jasper Anckaert, Nele Nijs, Joeri Tulkens, Bert Dhondt, An Hendrix, Pieter Mestdagh, Jo Vandesompele. Performance assessment of total RNA sequencing of human biofluids and extracellular vesicles [abstract]. In: Proceedings of the AACR Special Conference on Advances in Liquid Biopsies; Jan 13-16, 2020; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2020;26(11_Suppl):Abstract nr B51.
Abstract B55: PIK3CA mutation enrichment and detection in clinical samples
Authors: Keraite Ieva , Alvarez-Garcia Virginia , Favero Jurgen Del , Kersaudy-Kerhoas Maiwenn , Leslie Nicholas
Published: June 2020

The PI3K/AKT/mTOR signaling pathway is one of the most frequently mutated pathways in human cancers, including breast cancer. Given the current urge for more personalized approaches, there is an increasing need for a rigorous list of biomarkers and detection methods with advanced, less-invasive, reliable, and low-cost technologies. Despite limited clinical results to date, the detection of PIK3CA mutations shows great potential to contribute in cancer patient management in the future at many different levels, including diagnosis, treatment choice and monitoring, and identification of drug resistance. However, current molecular methods still lack analytical sensitivity to detect mutations of low abundance in high wild-type DNA background. Makrigiorgos et al. recently developed a nuclease-assisted minor-allele enrichment assay with overlapping probes, which is practical and cost-effective and allows detection of very-low-abundance mutations that could have relevance in the clinic. Thus we aimed to develop PIK3CA mutation-specific nuclease-based enrichment assay and validate it with tissue biopsy and blood samples using crystal dPCR as well as combine it with our developed Sybr Green real-time qPCR detection method (Alvarez-Garcia et al. 2018), which allows a quick detection of PIK3CA mutations in clinical samples at low cost.

Citation Format: Ieva Keraite, Virginia Alvarez-Garcia, Jurgen Del Favero, Maiwenn Kersaudy-Kerhoas, Nicholas Leslie. PIK3CA mutation enrichment and detection in clinical samples [abstract]. In: Proceedings of the AACR Special Conference on Advances in Liquid Biopsies; Jan 13-16, 2020; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2020;26(11_Suppl):Abstract nr B55.
Abstract B65: Precompetitive collaboration on liquid biopsy for early cancer assessment
Authors: Sorbara Lynn , Wong David , Bob Carter , Cote Richard , Lee Hakho , Papadopoulos Nickolas , Patel Abhijit , Srivastava Sudhir
Published: June 2020

Precompetitive collaboration is a term used when competitors share early stages of research and development that are beneficial to all stakeholders. This Public-Private Partnership Consortium supports research to advance technologies and use these technologies for early cancer detection, to distinguish benign disease from cancer, and to distinguish aggressive from nonaggressive cancer. Combining the complementary expertise of both academics and industry may increase the likelihood of developing a viable diagnostic assay. In the era of personalized medicine, it is desirable to have noninvasive or minimally invasive methods to determine and follow longitudinally the molecular composition and characterization of a patient’s tumor, which will help gain a broader understanding of the disease. Liquid biopsy (CTC, ctDNA, extracellular vesicles, etc.) can be used to molecularly characterize the tumor and monitor genetic changes over time using repeat sampling of biofluids. The current technologies have shown great potential in diagnosis, predicting response to therapy, and monitoring relapse; however, the biologic and analytical challenges have rendered their usefulness for early detection/screening more difficult to establish. To address these challenges, the NCI developed this cooperative agreement program to support the work of six teams. Each team consists of a group of experts that include oncologists, bioinformaticians, researchers, and at least one industry partner. The current teams are developing a variety of unique assays/technologies that range from capture enhancements, microscope integration, nanoplasmonics, to specialized machine learning algorithms and bioinformatic analyses. Ovarian, breast, lung, and brain cancers are the model systems being studied, and there are at least two teams focused on each cancer type. The Consortium conducts work with the assistance of two working subgroups, Study Design and Biorepository, in conjunction with NCI staff. To monitor progress, there are yearly face-to-face Steering Committee meetings and monthly conference calls/Webex meetings. Progress in the first year of funding has been noteworthy. One of the teams has already achieved Clinical Laboratory Improvement Amendments (CLIA) and College of American Pathologists Certification, while another team has an insurance reimbursable extracellular vesicles test for risk assessment for prostate cancer.

Citation Format: Lynn Sorbara, David Wong, Carter Bob, Richard Cote, Hakho Lee, Nickolas Papadopoulos, Abhijit Patel, Sudhir Srivastava. Precompetitive collaboration on liquid biopsy for early cancer assessment [abstract]. In: Proceedings of the AACR Special Conference on Advances in Liquid Biopsies; Jan 13-16, 2020; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2020;26(11_Suppl):Abstract nr B65.
Abstract IA01: Plasma DNA-based molecular diagnostics: Fragments, circles and beyond
Authors: Lo Yuk Ming Dennis
Published: June 2020

My group has a longstanding interest in the diagnostic applications of cell-free DNA in plasma. For example, we have pioneered noninvasive prenatal testing (NIPT) and the use of plasma DNA for cancer screening. We believe that understanding the fundamental biology of circulating DNA is essential to further advance the field. In this regard, we have been particularly interested in the fragmentation patterns of cell-free DNA in plasma. We have shown that cell-free fetal DNA molecules in the plasma of pregnant women are shorter than those of maternal origin. The differential size profiles between circulating fetal and maternal DNA have enabled the use of plasma DNA size as an independent metric for NIPT. In the area of cancer liquid biopsy, we have used circulating Epstein-Barr virus (EBV) DNA as an example of tumor-derived DNA in plasma. Hence, circulating EBV DNA molecules are also short fragments of DNA. In another example, we have used hepatocellular carcinoma as a model system and have found that circulating tumor-derived DNA molecules have a shorter size profile than those of nontumoral origin. We are exploring enzymes that might be involved in the fragmentation of plasma DNA. We have shown that mice in which both copies of the Dnase1l3 gene have been knocked out have a larger size profile of circulating DNA. We have also developed a plasma DNA end motif metric to measure the effect of disruption of this nuclease system. In addition to the fragmentation of linear DNA in plasma, my group is also interested in exploring the presence of nonlinear DNA in plasma. Hence, we have used mitochondrial DNA (mtDNA) as a model system as mtDNA exists as a circular molecule inside mitochondria. Interestingly, we have observed the existence of both linear and circular mtDNA in plasma. Using liver and bone marrow transplantation models, we have shown that the circular form is predominantly of hematopoietic origin while the linear form is predominantly from the liver. Plasma mtDNA topologics thus holds information on the origin of circulating DNA. These studies therefore demonstrate some of the exciting findings that can be revealed through the study of plasma DNA fragmentomics and topologics. We believe that such new biologic understandings will drive new diagnostic applications.

Citation Format: Yuk Ming Dennis Lo. Plasma DNA-based molecular diagnostics: Fragments, circles and beyond [abstract]. In: Proceedings of the AACR Special Conference on Advances in Liquid Biopsies; Jan 13-16, 2020; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2020;26(11_Suppl):Abstract nr IA01.
Abstract IA05: ctDNA and MRD in colorectal cancer: Time to reinvent adjuvant clinical trials
Authors: Tie Jeanne , Papadopoulos Nickolas , Kinzler Kenneth , Vogelstein Bert , Gibbs Peter
Published: June 2020

The most substantial opportunity for improving cancer survival outcomes is in the setting of early-stage disease, where evidence indicates that adjuvant chemotherapy can reduce recurrence and prevent cancer-related death through the eradication of minimal residual disease. However, the current stage-based treatment approach lacks precision and has quite a modest survival impact. A powerful prognostic marker could revolutionize adjuvant therapy by (i) better defining the recurrence risk of individual patients and allowing delivery of a more personalized treatment approach and (ii) enriching studies of new therapies with high-risk patients, substantially reducing the size and cost of adjuvant therapy trials. Circulating tumor DNA (ctDNA) has emerged as a promising noninvasive biomarker, with the ability to detect minimal residual disease following curative intent treatment in early-stage colorectal cancer. I will discuss the potential clinical utilities of ctDNA in early-stage colorectal cancer, including ongoing biomarker-driven adjuvant randomized trials.

Citation Format: Jeanne Tie, Nickolas Papadopoulos, Kenneth Kinzler, Bert Vogelstein, Peter Gibbs. ctDNA and MRD in colorectal cancer: Time to reinvent adjuvant clinical trials [abstract]. In: Proceedings of the AACR Special Conference on Advances in Liquid Biopsies; Jan 13-16, 2020; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2020;26(11_Suppl):Abstract nr IA05.
Abstract IA13: Tracking and targeting colorectal cancer evolution
Authors: Bardelli Alberto
Published: June 2020

When metastatic cancers are challenged with targeted agents, almost invariably a subset of cells insensitive to the drug emerges. As a result, in most instances, targeted therapies are only transiently effective in patients. Strategies to prevent or overcome resistance are therefore essential to design the next generation of clinical trials. How can we overcome the near certainty of disease recurrence following treatment with targeted agents? Addressing this question means considering as a target not “only” individual oncogenes but also the evolving nature of human tumors. We used colorectal cancer (CRC) as a model system to test the hypothesis that by understanding tumors’ evolution, the emergence of drug resistance can be controlled. We find that clonal dynamics can be monitored in real time in the blood of patients, and liquid biopsies can be used to intercept the emergence of resistant clones before relapses are clinically manifest. We discovered that a multistep clonal evolution process driven by progressive increases in drug fitness underlies the development of resistance in cells and patient avatars. To have long-term efficacy, the use of targeted therapies must take into account the continuous evolution of cancer cells, that is to say, therapies must adapt to tumor evolution. One possibility is to anticipate the changes the tumors will make. For example, by knowing in advance how CRC cells overcome resistance to EGFR blockade, we devised further rounds of therapy. These findings became the bases of clinical trials aimed at targeting cancer evolution and exploiting liquid biopsies to monitor the emergence of drug resistance and to design further lines of therapies.

Citation Format: Alberto Bardelli. Tracking and targeting colorectal cancer evolution [abstract]. In: Proceedings of the AACR Special Conference on Advances in Liquid Biopsies; Jan 13-16, 2020; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2020;26(11_Suppl):Abstract nr IA13.
Abstract IA16: Enhancing detection of ctDNA in patients with early-stage or residual disease
Authors: Rosenfeld Nitzan
Published: June 2020

Liquid biopsies have become a reality as part of the diagnostic toolbox for oncology. Assays based on targeted next-generation sequencing of circulating tumor DNA (ctDNA) are now reimbursed for genotyping of advanced disease and can be used to identify resistance mechanisms. I will describe some of our work in developing and validating such tests. I will also describe methods we have developed that leverage the characteristic fragment sizes of ctDNA to aid in detecting and monitoring cancer, using shallow whole-genome sequencing and by analysis of dried blood spots. Applications of ctDNA analysis as a diagnostic tool for earlier-stage disease are making exciting progress. Recent studies have shown potential for detection of minimal residual disease and monitoring for relapse in patients who have undergone treatment with curative intent. Current tests have shown excellent specificity and positive predictive value, and most patients relapse within 6-12 months after ctDNA is detected. These findings have supported the launching of clinical trials to test the utility of ctDNA-guided adjuvant treatment. Patients for whom ctDNA has not been detected with current methods still face an uncertain prognosis. I will describe our ongoing efforts to develop tests with enhanced sensitivity, which aim to identify a larger fraction of patients destined to recur, and to provide a longer lead time prior to clinically overt recurrence, when the residual disease has had less time to grow and evolve.

Citation Format: Nitzan Rosenfeld. Enhancing detection of ctDNA in patients with early-stage or residual disease [abstract]. In: Proceedings of the AACR Special Conference on Advances in Liquid Biopsies; Jan 13-16, 2020; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2020;26(11_Suppl):Abstract nr IA16.
Abstract IA18: Molecular signatures of circulating tumor cells
Authors: Haber Daniel , Hong Xin , Ebright Richard , Micalizi Doug , Bardia Aditya , Sullivan Ryan , Toner Mehmet , Maheswaran Shyamala
Published: June 2020

Circulating tumor cells (CTCs) are shed from primary cancers, as well as from metastatic deposits, into the blood circulation, where they may travel either as single cells or as CTC clusters, facing both mechanical and oxidative stress. Only a small subset of CTCs remain sufficiently viable to initiate metastatic growth upon dissemination to distant organs; their detection in patients with localized invasive cancer may present an opportunity for early detection, while their analysis in advanced cases provides a “whole cell biomarker,” enabling single-cell DNA, RNA, and protein studies, as well as functional drug-sensitivity testing using cultured CTCs. We have applied microfluidic technologies to deplete normal hematopoietic cells from blood specimens of patients with cancer, enhancing the ability of unmanipulated CTCs to proliferate ex vivo and recapitulating features of metastatic-competent cancer cells. Cultured CTCs from patients with either breast cancer or melanoma are highly tumorigenic when injected orthotopically at low inoculum into immunosuppressed mice. However, they display sensitivity to ROS and appear to upregulate compensatory mechanisms that are not commonly observed in primary tumors. In addition, they may undergo prolonged dormancy when lodged within distant organs. By combining single-cell RNA seq of patient-derived CTCs together with CRISPR and other genome-wide screens for modulators of metastasis phenotypes, we have sought to identify novel vulnerabilities in these cellular initiators of metastasis. While CTCs transit only very briefly through the blood circulation, these exceptional conditions may present new therapeutic opportunities to suppress the metastatic recurrence of cancer.

Citation Format: Daniel Haber, Xin Hong, Richard Ebright, Doug Micalizi, Aditya Bardia, Ryan Sullivan, Mehmet Toner, Shyamala Maheswaran. Molecular signatures of circulating tumor cells [abstract]. In: Proceedings of the AACR Special Conference on Advances in Liquid Biopsies; Jan 13-16, 2020; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2020;26(11_Suppl):Abstract nr IA18.
Abstract IA22: Tumor-educated platelets for the detection of cancer
Authors: Wurdinger Thomas
Published: June 2020

Blood-based “liquid biopsies” provide a means for minimally invasive molecular diagnostics, overcoming limitations of tissue acquisition. Early detection of cancer, clinical cancer diagnostics, and companion diagnostics are regarded as important applications of liquid biopsies. It has been reported that tumor-educated platelets (TEPs) may enable blood-based cancer diagnostics. Platelets have emerged as central players in the systemic and local responses to tumor growth. Confrontation of platelets with tumor cells via transfer of tumor-associated biomolecules (“education”) is an emerging concept and can result in the sequestration of biomolecules. Moreover, external stimuli induce specific splicing of pre-mRNAs in circulating platelets. Platelets are considered to undergo queue-specific splice events in response to signals released by cancer cells and the tumor microenvironment, such as stromal and immune cells. The combination of specific splice events in response to external signals and the capacity of platelets to directly ingest (spliced) circulating mRNA can provide TEPs with a highly dynamic mRNA repertoire, with potential applicability to cancer diagnostics. We determined the diagnostic potential of TEPs by mRNA sequencing of platelet samples. Blood platelet samples of >1,000 patients with cancer covering multiple tumor types and healthy donors were analyzed. Healthy donors and early- and late-stage cancer patients were distinguished by a self-learning support vector machine (SVM) algorithm, using a set of spliced platelet transcripts with moderate to high expression. These results suggest that blood platelets may advance blood-based “liquid biopsies,” including for the detection of stage I-II tumors.

Citation Format: Thomas Wurdinger. Tumor-educated platelets for the detection of cancer [abstract]. In: Proceedings of the AACR Special Conference on Advances in Liquid Biopsies; Jan 13-16, 2020; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2020;26(11_Suppl):Abstract nr IA22.
Abstract IA26: Liquid biopsy in precision pathology: Plasma and beyond!
Authors: Netto George J.
Published: June 2020

Launched in 2006, The Cancer Genome Atlas project (TCGA) aimed at systematically exploring the entire spectrum of genomic changes involved in human cancer. This year, the colossal effort culminated with the analysis of 11,000 cases across 33 tumor types. The comprehensive multiparametric (i.e., genomic, transcriptomic, epigenomic, and proteomic) analysis has yielded staggering datasets characterizing the molecular underpinning of human cancer types. This resulted in robust molecular classifications for common malignancies. A shared functional categorization of molecular alterations across tumor types has emerged, which offers a multitude of druggable targets.

As the list of actionable genetic alterations in solid and hematologic malignancies continue to grow, molecular diagnostic applications are rapidly penetrating the daily practice of the pathologist in an unparalleled pace. “Precision pathology” is now an integral part of the practice of “precision oncology.” Despite initial response, patients receiving targeted therapy will develop resistance due to clonal evolution and the acquired survival advantage of nonresponsive tumor clones. As a result, treated patients are increasingly subjected to longitudinal surveillance for clonal evolution in search of new targets.

The need for repeated tumor sampling for surveillance is driving the search for dynamic noninvasive tumor sampling methods. Non-tissue-based sampling methods (liquid biopsy) offer easy repeated access to tumor material in cancer patient body fluids for genomic analysis. Compared to traditional tissue biopsies, liquid biopsies also offer the advantage of “global” analysis of the often heterogeneous tumor genomic landscape both within primary and metastatic sites. The promise of liquid biopsy assays has fueled billions of dollars of investment from major biotechnology players.

The current seminar will discuss the principles of liquid biopsy technology. Challenges facing the applications of plasma and urine-based liquid biopsy techniques in precision pathology will be highlighted.

Citation Format: George J. Netto. Liquid biopsy in precision pathology: Plasma and beyond! [abstract]. In: Proceedings of the AACR Special Conference on Advances in Liquid Biopsies; Jan 13-16, 2020; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2020;26(11_Suppl):Abstract nr IA26.
Epigenetic Switch–Induced Viral Mimicry Evasion in Chemotherapy-Resistant Breast Cancer
Authors: Deblois Geneviève , Tonekaboni Seyed Ali Madani , Grillo Giacomo , Martinez Constanza , Kao Yunchi Ingrid , Tai Felicia , Ettayebi Ilias , Fortier Anne-Marie , Savage Paul , Fedor Alexandra N. , Liu Xiaojing , Guilhamon Paul , Lima-Fernandes Evelyne , Murison Alex , Kuasne Hellen , Ba-alawi Wail , Cescon David W. , Arrowsmith Cheryl H. , De Carvalho Daniel D. , Haibe-Kains Benjamin , Locasale Jason W. , Park Morag , Lupien Mathieu
Published: June 2020

Tumor progression upon treatment arises from preexisting resistant cancer cells and/or adaptation of persister cancer cells committing to an expansion phase. Here, we show that evasion from viral mimicry response allows the growth of taxane-resistant triple-negative breast cancer (TNBC). This is enabled by an epigenetic state adapted to taxane-induced metabolic stress, where DNA hypomethylation over loci enriched in transposable elements (TE) is compensated by large chromatin domains of H3K27me3 to warrant TE repression. This epigenetic state creates a vulnerability to epigenetic therapy against EZH2, the H3K27me3 methyltransferase, which alleviates TE repression in taxane-resistant TNBC, leading to double-stranded RNA production and growth inhibition through viral mimicry response. Collectively, our results illustrate how epigenetic states over TEs promote cancer progression under treatment and can inform about vulnerabilities to epigenetic therapy.

Significance:

Drug-resistant cancer cells represent a major barrier to remission for patients with cancer. Here we show that drug-induced metabolic perturbation and epigenetic states enable evasion from the viral mimicry response induced by chemotherapy in TNBC. These epigenetic states define a vulnerability to epigenetic therapy using EZH2 inhibitors in taxane-resistant TNBC.

See related commentary by Janin and Esteller, p. 1258.

This article is highlighted in the In This Issue feature, p. 1241
HIF1α Deficiency in Dendritic Cells Attenuates Symptoms and Inflammatory Indicators of Allergic Rhinitis in a SIRT1-Dependent Manner
Authors: Niu Yanliu , Wang Jianquan , Li Zhen , Yao Keqing , Wang Liangliang , Song Jingjing
Published: June 2020

Background: Allergic rhinitis is the most prevalent atopic disorder worldwide. Inflammation is believed to participate in allergic rhinitis. Previous studies indicate that hypoxia-inducible factor (HIF) promotes the development of allergic rhinitis, and dendritic cells are also involved in allergic rhinitis. Methods:We explored the consequences of HIF1α deficiency in dendritic cells on allergic rhinitis. Allergic rhinitis in mice was induced by ovalbumin (OVA). The levels of IgE, leukotriene C4 (LTC4), eosinophil cationic protein (ECP), prostaglandin D2 (PGD2), IFN-γ, IL-2, IL-4, IL-5, IL-10, and IL-13 in serum or nasal lavage fluid (NLF) were detected by ELISA. Inflammatory cells in NLF were counted by hemocytometer. The protein levels of p-ERK1/2, p-p38, p-JNK2, SIRT1, p-IκBα, and p65 were determined by Western blot. Results:HIF1α deficiency in dendritic cells (HIF1αCD11c–/–) decreased sneezing and nasal rubbing, the production of OVA-specific IgE, LTC4, and ECP in serum and NLF, and the numbers of leukocytes, eosinophils, lymphocytes, and neutrophils in NLF. Th1 cytokines increased, while Th2 cytokines decreased in HIF1aCD11c–/– mice. SIRT1/NF-κB signaling was inhibited in HIF1αCD11c–/– mice, while SIRT1 inhibitor administration in HIF1αCD11c–/– mice attenuated the symptoms and inflammatory indicators of allergic rhinitis. Conclusion:HIF1α deficiency in dendritic cells attenuates symptoms and inflammatory indicators of allergic rhinitis in a SIRT1-dependent manner.
Altered Gene Expression Profiles in Peripheral Blood Mononuclear Cells in Obese Subjects
Authors: Jang Kyungho , Tong Tao , Lee Jinhui , Park Taesun , Lee Howard 
Published: June 2020

Introduction: Gene expression profiles in human peripheral blood mononuclear cells (PBMCs) may act as a useful tool to better understand obesity. We investigated gene expression levels in PMBCs for possible differences between obese and non-obese subjects (19–55 years) and evaluated correlations between gene expression in PBMCs and clinical obesity indices. Methods: Body weight, BMI, fat amount, fat percentage, waist/hip ratio, leptin, and adiponectin levels were determined in 30 obese and 20 non-obese subjects. Expression levels of 19 genes, which were differentially expressed by clinical obesity indices in the PBMCs of high fat-fed rats, were determined in their PBMCs using real-time PCR. Results: The expression of 9 of 19 previously selected genes was significantly correlated with one or more clinical obesity indices. Both TFEC and CCL2 expression were negatively correlated with BMI, fat amount, fat percentage, waist/hip ratio, and leptin concentration. Similarly, TNFAIP2, VCAN, ASSI, IRF1, and HK3 expression negatively correlated with some clinical obesity indices, such as TNFAIP2 for BMI, fat amount, fat percentage, and waist/hip ratio, VCAN for fat amount, fat percentage, and waist/hip ratio, ASS1 for BMI and fat amount, IRF1 for BMI, fat amount, and fat percentage, and HK3 for fat amount. In contrast, both TNF-α and LPL expression were positively correlated with waist/hip ratio. Conclusion: We identified 9 of 19 genes in human PBMCs that significantly correlated with one or more clinical obesity indices. Because these genes have a mechanistic basis for the development or progression of obesity and its metabolic derangements, they may help to determine possible underlying mechanisms for obesity.
Docetaxel-Induced Peripheral Neuropathy in Breast Cancer Patients Treated with Adjuvant or Neo-Adjuvant Chemotherapy
Authors: Cheng Hui Lin , Molassiotis Alex , Leung Anthony Kwun To , Wong Kam Hung
Published: June 2020

Background: Docetaxel-induced peripheral neuropathy (PN) is typically manifested as sensory and motor neuropathy. This study aimed to investigate the incidence, duration, and risk factors of sensory and motor PN and their impact on health-related quality of life (HRQOL) among breast cancer (BC) patients during the first year since starting docetaxel-based chemotherapy. Methods: We reported a secondary analysis of longitudinal data on docetaxel-induced PN and HRQOL among 127 BC patients. Results: Cumulative incidence rates of motor and sensory PN were 31.5 and 21.3%, while the median durations of motor and sensory PN were 6 and 13 weeks. A consistently significant risk factor for both PNs was a cumulative docetaxel dose of >300 mg/m2. A significant interaction between sensory PN and time was found for physical and social functioning, while a significant motor PN and time interaction effect was identified for physical functioning only. Conclusions: Motor PN was more common than sensory PN in BC patients treated with docetaxel. Both types of PN had a significant impact on physical functioning.
Confirming the Diagnosis of Amyloidosis
Authors: Wisniowski Brendan , Wechalekar Ashutosh
Published: June 2020

Amyloidosis is a general term for diseases characterised by the deposition of insoluble amyloid fibrils in organs or tissues, leading to organ dysfunction and, in many cases, death. Amyloid fibrils are derived from soluble precursor proteins, with the number of known amyloidogenic proteins increasing over time. The identity of the precursor protein often predicts the disease phenotype, although many of the amyloidoses have overlapping clinical features. Most patients with amyloidosis will require biopsy of an involved organ or tissue to confirm the diagnosis. Cardiac transthyretin amyloidosis, however, may be diagnosed without a biopsy provided stringent criteria are met. Where amyloid is confirmed histologically, the identity of the amyloidogenic protein must be determined, given several of the amyloidoses have disease-specific therapies. Laser capture microdissection and tandem mass spectrometry, LCM-MS, has revolutionised amyloid subtyping, being able to identify the amyloidogenic protein more reliably than antibody-based methods such as immunohistochemistry. Here we summarise the biopsy approach to amyloidosis, as well as the non-biopsy diagnosis of cardiac transthyretin amyloidosis. Proteomic and antibody-based methods for amyloid subtyping are reviewed. Finally, an algorithm for confirming the diagnosis of amyloidosis is presented.
SOCS2 Silencing Improves Somatic Growth without Worsening Kidney Function in CKD
Authors: Landau Daniel , Assadi Muhammad H. , Abu Hilal Rawan , Chen Yu , Rabkin Ralph , Segev Yael
Published: June 2020

Background: Growth hormone (GH) resistance in CKD is partly due to increased expression of SOCS2, a GH signaling negative regulator. In SOCS2 absence, body growth is exaggerated. However, GH overexpression in mice causes glomerulosclerosis. Accordingly, we tested whether lack of SOCS2 improves body growth, but accelerates kidney damage in CKD. Methods: Eight-week-old mutant SOCS2-deficient high growth (HG) and normal wild-type mice (N) underwent 5/6 nephrectomy (CKD) or sham operation (C) and were sacrificed after 12 weeks, generating 4 groups: C-N, C-HG, CKD-N, CKD-HG. Results: Somatic growth, inhibited in CKD-N, increased significantly in CKD-HG. Liver p-STAT5, a key intracellular signal of GH receptor (GHR) activation, was decreased in CKD-N but not in CKD-HG. Serum Cr as well as histopathological scores of renal fibrosis were similar in both CKD groups. Kidney fibrogenic (TGF-β and collagen type IV mRNA) and inflammatory precursors (IL6, STAT3, and SOCS3 mRNA) were similarly increased in C-HG, CKD-HG, and CKD-N versus C-N. Renal GHR mRNA was decreased in C-HG, CKD-HG, and CKD-N versus C-N. Kidney p-STAT5 was decreased in CKD-N but not elevated in CKD-HG. Conclusions: CKD-related growth retardation is overcome by SOCS2 silencing, in association with increased hepatic STAT5 phosphorylation. Renal insufficiency is not worsened by SOCS2 absence, as kidney GHR and STAT5 are not upregulated. This may be due to elevated kidney proinflammatory cytokines and their mediators, phospho-STAT3 and SOCS3, which may counteract for the absence in SOCS2 and explain the renal safety of prolonged GH therapy in CKD.
Analysis of Oncological Second Opinions in a Certified University Breast and Gynecological Cancer Center in Relation to Complementary and Alternative Medicine
Authors: Hack Carolin C. , Wasner Sonja , Meyer Julia , Häberle Lothar , Jud Sebastian , Hein Alexander , Wunderle Marius , Emons Julius , Gass Paul , Fasching Peter A. , Egloffstein Sainab , Beckmann Matthias W. , Lux Michael P. , Loehberg Christian R.
Published: June 2020

Introduction: Oncological second opinions are becoming increasingly important given more complex treatment strategies, simultaneously more patients use complementary and alternative medicine (CAM), and many comprehensive cancer centers initiate integrative medicine programs. The present study focuses on analyzing the effects of a second opinion in relation to attitudes toward CAM. Methods: In this prospective study patients (n = 97) with a diagnosis of breast cancer or gynecological malignancies who had requested a second opinion received a questionnaire before and after the second opinion concerning their attitudes toward CAM. Results: The majority of patients had breast cancer (72.2%, n = 70). Only 6.2% (n = 6) stated that they had been informed about CAM by the doctors who treated them first, 21.6% (n = 21) had received information about it when seeking the second opinion. After the first opinion, 42.3% (n = 41) wanted to try CAM, the same proportion trusted orthodox medicine alone. After the second opinion, 24 patients (24.7%) wanted to try CAM, while 38.1% (n = 37) relied exclusively on orthodox medicine. There was a significant correlation between an increased patients’ need for information and interest in CAM (p = 0.02). Conclusions: Today, aspects of CAM still are very often no part of oncological first and second opinions. This might hence lead to discouraging patients to try out CAM and therefore integrative medicine programs in comprehensive cancer centers might be problem-solving.
A Randomized Phase IIa Trial with Temsirolimus versus Sunitinib in Advanced Non-Clear Cell Renal Cell Carcinoma: An Intergroup Study of the CESAR Central European Society for Anticancer Drug Research-EWIV and the Interdisciplinary Working Group on Renal Cell Cancer (IAGN) of the German Cancer Society
Authors: Bergmann Lothar , Grünwald Viktor , Maute Luise , Grimm Marc-Oliver , Weikert Steffen , Schleicher Jan , Klotz Theodor , Greiner Jochen , Flörcken Anne , Hartmann Arndt , Gauler Thomas
Published: June 2020

Background: Non-clear cell renal cell cancers (nccRCC) are rare entities, and the optimal therapy in metastatic disease has still to be defined. Methods: In this small prospectively randomized phase IIa multicenter trial, we investigated temsirolimus (TEM) versus sunitinib (SUN) as first-line therapy in patients with metastatic nccRCC. The patients were randomized 1:1 to either TEM in a dose of 25 mg i.v. once a week or SUN with 50 mg p.o. daily for 4 weeks on and 2 weeks off. Primary endpoint was progression-free survival (PFS). In total, 22 patients were included with predominantly papillary RCC (16/22) followed by chromophobe RCC and others. Results: The male to female ratio was 16:6. The tumor control rate (CR + PR + SD) was 58% for TEM and 90% for SUN-treated patients. There was also a trend for improved PFS with 9.3 versus 13.2 months (HR 1.64; 95% CI 0.65–4.18) in favor of SUN. There was no trend for overall survival. Conclusions: Despite this trial had to be terminated earlier due to low recruitment, the results match the other studies published so far with the mTOR inhibitor everolimus and SUN, which show a trend in favor of SUN for ORR and PFS.
Feasibility Study of a Docosahexaenoic Acid-Optimized Nutraceutical Formulation on the Macular Levels of Lutein in a Healthy Mediterranean Population
Authors: Zanón-Moreno Vicente , Domingo Pedrol Joan C. , Sanz-González Silvia M. , Raga-Cervera Jorge , Salazar-Corral Juan , Pinazo-Durán Maria Dolores
Published: June 2020

Introduction: Macular pigment optical density (MPOD) plays a pivotal role in maintaining macular structure and functioning. Research shows that daily consumption of lutein reduces the risk of eye diseases such as age-related macular degeneration. Objective: This study analyzes the influence of a supplementation containing lutein and antioxidant vitamins either with or without docosahexaenoic acid (DHA), with the main objective of identifying MPOD changes in both eyes at the end of the follow-up using the Visucam®retinograph. The secondary end point was to determine variation in the lutein and DHA levels in plasma and red blood cell membranes (RBCMs), respectively. Methods: One hundred healthy participants (200 eyes) aged 40–70 years (mean age 49.3 years, SEM = 13.7) were randomized in a 1:1 ratio to receive daily one of the following supplements for 3 months: lutein group (LT-G, n = 49) and lutein plus DHA group (LT/DHA-G, n = 51). The MPOD was measured at baseline and end of the follow-up by retinography (Visucam®retinograph). Lutein in plasma was determined by HPLC, and DHA in RBC membranes was analyzed by gas chromatograph/mass spectrometer. Results: From baseline, MPOD showed significantly higher values in the LT/DHA-G than in the LT-G at the end of the study (p < 0.0001). Significantly higher lutein in plasma (p < 0.0001) and DHA (p < 0.0001) levels in the RBC membrane were seen in the LT/DHA-G than in the LT-G at the 3-month follow-up. Conclusion: Lutein supplementation improves MPOD in healthy subjects from a Mediterranean population being significantly increased in the presence of DHA. Therefore, our findings highlight the relevance of the adjunctive role of DHA for better lutein availability.
Abstract 100: HIV Protein Tat Induces Macrophage Dysfunction and Atherosclerosis Development in LDLR-deficient Mice
Authors: Meng Zhaojie , Lu Weiwei , Gwag Taesik , Zhou Changcheng
Published: June 2020

Introduction:
As the average lifespan of HIV-infected patients receiving anti-retroviral therapy lengthens, morbidity and mortality from cardiovascular disease pose considerable challenges. HIV infection is consistently associated with increased risk of atherosclerosis development, but the underlying mechanisms remain elusive. HIV-1 Tat protein, a transcriptional activator of HIV virus, has been shown to activate NF-κB signaling and promote inflammation in vitro. However, the atherogenic effects of HIV-1 Tat have not been investigated in vivo. Macrophage is one of the major cell types involved in the initiation and progression of atherosclerosis. We and others have previously demonstrated that NF-κB signaling functions in macrophages to regulate atherogenesis. This study aims to investigate the impact of HIV-1 Tat exposure on macrophage functions and atherogenesis.

Hypothesis:
HIV-1 Tat activates IκB kinase β (IKKβ), a central coordinator in inflammation through activation of NF-κB, to induce macrophage dysfunction and atherosclerosis development.

Methods:
To investigate the effects of HIV-1 Tat on macrophage IKKβ signaling and atherosclerosis development in vivo, myeloid-specific IKKβ-deficient LDLR-deficient (IKKβ
ΔMye
LDLR
-/-
) mice and their control littermates (IKKβ
F/F
LDLR
-/-
) were exposed to recombinant HIV-1 Tat for 12 weeks. The effects of HIV-1 Tat on macrophage functions including inflammatory responses, adhesion and migration properties were also studied.

Results and Conclusions:
HIV-1 Tat significantly increased atherosclerotic lesion size in aortic root and brachiocephalic artery of IKKβ
F/F
LDLR
-/-
but not IKKβ
ΔMye
LDLR
-/-
mice. Deficiency of myeloid IKKβ attenuated macrophage inflammatory responses and decreased atherosclerotic lesional inflammation in IKKβ
ΔMye
LDLR
-/-
mice. In addition, HIV-1 Tat stimulated adhesion and migration properties of control macrophages but had no effects on IKKβ-deficient macrophages. In conclusion, our findings reveal the atherogenic effects of HIV-1 Tat in vivo and demonstrate a pivot role of myeloid IKKβ in HIV-1 Tat-driven atherogenesis.
Variability of mandible shape in the freshwater glacial relict Eurytemora lacustris (Poppe, 1887) (Copepoda, Calanoida, Temoridae)
Authors: Sługocki Łukasz
Published: June 2020

The mandibles of copepods are used to classify the feeding modes of certain species, because these appendages have specific shapes. The number, shape, location, and structure of the mandibular teeth are traits that are related to the feeding preferences of copepods. Therefore, the mandibles have been used as parameters in order to determine the role of the corresponding species in the trophic network. In this study, we aimed to determine the morphological variability of the mandible shape in a rare calanoid, Eurytemora lacustris (Poppe, 1887). The size of the mandibles of E. lacustris is closely related to its body size; however, considering the proportions of the width of the mandibles to that body size, young individuals have relatively larger mandibles. Despite the large body size of males, the females are characterized by larger mandibles than those of males. The teeth of males were found to be smaller, because of intensive abrasion. Moreover, the season and habitat affected the size of the mandibles and also the values of the edge index (EI). The food conditions in the studied lakes fluctuated within seasons, which led to changes in the shape of the dental crowns. Using this index (EI), E. lacustris was classified as an omnivorous species; however, the range of calculated values varied considerably. We note that the values of the EI decrease along with maturation, which suggests that with increase in age, the copepod becomes more herbivorous; however, this contradicts the paradigm that copepods become omnivorous during maturation. All those morphological differences, furthermore, indicate the high phenotypic plasticity of the species, which was found to be related to maturity, sex, phenology, and the environmental conditions of the habitat.
A multi-year investigation of the Temoridae (Copepoda, Calanoida) assemblage succession within the interplay waters of the northern South China Sea
Authors: Tseng Li-Chun , Wang Yan-Guo , Lian Guang-Shan , Hwang Jiang-Shiou
Published: June 2020

The South China Sea is the largest marginal sea in the world. The northern South China Sea has a complex hydrographic system affected by interplay waters of the Kuroshio Branch Current and the South China Sea waters. To understand the changes in the community structure of the planktonic calanoid copepod family Temoridae in relation to the interplay waters, we collected zooplankton samples in the northern South China Sea from surface waters (0 to 2 m) from November 2001 to January 2007 during 24 research cruises using a standard North Pacific zooplankton net (with a mesh size of 200 μm, a length of 1.8 m, and a mouth diameter of 0.45 m). Among a total of 253 samples, 5 species belonging to 3 genera of Temoridae were identified: Eurytemora pacifica Sato, 1913, Temora discaudata Giesbrecht, 1889, Temora stylifera (Dana, 1849), Temora turbinata (Dana, 1849), and Temoropia mayumbaensis Scott T., 1894. The 3 most abundant species were: T. turbinata (relative abundance, RA: 53.28%; mean density ± standard deviation, MD: 29.33 ± 89.53 ind. m−3), T. stylifera (RA: 36.02%, MD: 19.83 ± 145.22 ind. m−3), and T. discaudata (RA: 7.70%, MD: 4.24 ± 11.82 ind. m−3). The 3 most frequently occurring species were: T. turbinata (occurrence ratio, OR: 52.17%), T. discaudata (OR: 35.18%), and T. stylifera (OR: 9.88%). A noteworthy discovery in our samples was E. pacifica, a marine and brackish water species, collected from 3 stations located near the Kuroshio Branch Current in April 2003. This is the only month when this species was recorded during the investigation period. Variations of dominant species of Temoridae show a clear pattern of seasonal succession. The total abundance was significantly higher in the third quarter (September-November, 116.98 ± 314.49 ind. m−3) than in the fourth quarter (December-February, 24.26 ± 47.72 ind. m−3) (, one-way ANOVA). In general, the present results demonstrate that the assemblages of the Temoridae are very much structured by the water masses of the Kuroshio Current and the South China Sea.