Prognosis Monitoring Research Articles

Immunological significance of prognostic alternative splicing signature in hepatocellular carcinoma

BackgroundHepatocellular carcinoma (HCC) ranks the sixth prevalent tumors with high mortality globally. Alternative splicing (AS) drives protein diversity, the imbalance of which might act an important factor in tumorigenesis. This study aimed to construct of AS-based prognostic signature and elucidate the role in tumor immune microenvironment (TIME) and immunotherapy in HCC.MethodsUnivariate Cox regression analysis was performed to determine the prognosis-related AS events and gene set enrichment analysis (GSEA) was employed for functional annotation, followed by the development of prognostic signatures using univariate Cox, LASSO and multivariate Cox regression. K-M survival analysis, proportional hazards model, and ROC curves were conducted to validate prognostic value. ESTIMATE R package, ssGSEA algorithm and CIBERSORT method and TIMER database exploration were performed to uncover the context of TIME in HCC. Quantitative real-time polymerase chain reaction was implemented to detect ZDHHC16 mRNA expression. Cytoscape software 3.8.0 were employed to visualize AS-splicing factors (SFs) regulatory networks.ResultsA total of 3294 AS events associated with survival of HCC patients were screened. Based on splicing subtypes, eight AS prognostic signature with robust prognostic predictive accuracy were constructed. Furthermore, quantitative prognostic nomogram was developed and exhibited robust validity in prognostic prediction. Besides, the consolidated signature was significantly correlated with TIME diversity and ICB-related genes. ZDHHC16 presented promising prospect as prognostic factor in HCC. Finally, the splicing regulatory network uncovered the potential functions of splicing factors (SFs).ConclusionHerein, exploration of AS patterns may provide novel and robust indicators (i.e., risk signature, prognostic nomogram, etc.,) for prognostic prediction of HCC. The AS-SF networks could open up new approach for investigation of potential regulatory mechanisms. And pivotal players of AS events in context of TIME and immunotherapy efficiency were revealed, contributing to clinical decision-making and personalized prognosis monitoring of HCC.

Microfluidic Chip-Based Cancer Diagnosis and Prediction of Relapse by Detecting Circulating Tumor Cells and Circulating Cancer Stem Cells.

Simple SummaryMetastasis is the main cause of cancer-related death. Circulating cancer stem cells have recently attracted attention because they have higher tumorigenicity than non-stem-like circulating tumor cells. Despite the strong scientific evidence for circulating cancer stem cells and secondary tumor formation, the exact mechanisms behind the generation and characteristics of circulating cancer stem cells are not yet fully understood because of their extreme scarcity. This review aims to introduce the recent advances in the detection and analysis of circulating tumor cells and circulating cancer stem cells.Detecting circulating tumor cells (CTCs) has been considered one of the best biomarkers in liquid biopsy for early diagnosis and prognosis monitoring in cancer. A major challenge of using CTCs is detecting extremely low-concentrated targets in the presence of high noise factors such as serum and hematopoietic cells. This review provides a selective overview of the recent progress in the design of microfluidic devices with optical sensing tools and their application in the detection and analysis of CTCs and their small malignant subset, circulating cancer stem cells (CCSCs). Moreover, discussion of novel strategies to analyze the differentiation of circulating cancer stem cells will contribute to an understanding of metastatic cancer, which can help clinicians to make a better assessment. We believe that the topic discussed in this review can provide brief guideline for the development of microfluidic-based optical biosensors in cancer prognosis monitoring and clinical applications.

Effect of cell-nanostructured substrate interactions on the capture efficiency of HeLa cells

Circulating tumor cells (CTCs) are regarded as an effective biomarker for cancer detection, diagnosis and prognosis monitoring. CTCs capture based on nanostructured substrates is a powerful technique. Some specific adhesion molecule antibody coated on the surface of nanostructured substrates, such as EpCAM, is commonly used to enhance the CTCs capture efficiency. Substrate nanotopographies regulate the interaction between the substrates and captured cells, further influencing cell capture efficiency. However, the relationship between cell capture efficiency and cell–substrate interaction remains poorly understood. Here, we explored the relationship between cell capture efficiency and cell–substrate interaction based on two sets of nanostructures with different nanotopographies without antibody conjugation. Given the urgent demand for improving the capture efficiency of EpCAM-negative cells, we used HeLa (EpCAM-negative) cells as the main targets. We demonstrated that HeLa cells could be more effectively captured by two nanostructural substrates, especially by double-layer composite nanoforests. Therefore, the morphological and migrating interaction between HeLa cells and distinct substrates was associated with cell capture efficiency. Our findings demonstrated the potential mechanism for optimizing the nanotopography for higher capture efficiency, and provide a potential foundation for cancer detection, diagnosis and treatment.

PTX3 gene polymorphism associated with cryptococcosis in HIV-uninfected Chinese patients.

For Chinese Han populations, cryptococcosis are more likely to occur in HIV-uninfected patients instead of HIV-infected patients compared with other countries and regions, implying that there may be genetic predisposing factors for cryptococcosis in the Chinese Han populations. However, the retail mechanism has not been clarified. We aimed to conduct an association analysis between the single nucleotide polymorphisms (SNPs) of pattern recognition receptors (PRR) genes and the susceptibility to cryptococcosis in HIV-uninfected Chinese patients, which may provide new genetic predisposing factors for early-risk predictionof disease, individualised treatment and prognosis monitoring. Using the SNaPshot SNP typing technique, eight SNPs of PRR genes (Dectin-2, Dectin-1, PTX3, CXCL8, IL12B, IFIH1, TLR1 and CD209) were typed on 97 HIV-uninfected cryptococcosis patients and 120 healthy controls who admitted to West China Hospital, Sichuan University, China, from 1 March 2018 to 30 December 2018. The results were analysed by the SHEsis software and SPSS 20.0 software. It was found that that PTX3 rs2305619 polymorphism was associated with cryptococcosis in HIV-uninfected patients. Compared with the GG genotype, AA genotype increased the risk of cryptococcosis in HIV-uninfected patients (p=.015, OR, 2.579; 95% CI, 1.202-5.535). In the immunocompetent patients, the AA genotype had a higher risk (p=.002, OR, 4.399; 95% CI, 1.745-11.088). Further verification found that the plasma PTX3 level of the AA genotype was significantly higher than the GA or GG genotype (60.28±16.12 vs 7.32±0.79, p<.001). PTX3 rs2305619 polymorphism was associated with cryptococcosis in HIV-uninfected Chinese patients. The AA genotype increased the risk of cryptococcosis, and its plasma PTX3 level was significantly higher than that of GA or GG genotype.

Long Noncoding RNAs in the Metastasis of Oral Squamous Cell Carcinoma.

Oral squamous cell carcinoma (OSCC) is a common malignant tumor worldwide. Metastasis is the main cause of the death of OSCC patients. Long noncoding RNAs (lncRNAs), one of the key factors affecting OSCC metastasis, are a subtype of RNA with a length of more than 200 nucleotides that has little or no coding potential. In recent years, the important role played by lncRNAs in biological processes, such as chromatin modification, transcription regulation, RNA stability regulation, and mRNA translation, has been gradually revealed. More and more studies have shown that lncRNAs can regulate the metastasis of various tumors including OSCC at epigenetic, transcriptional, and post-transcriptional levels. In this review, we mainly discussed the role and possible mechanisms of lncRNAs in OSCC metastasis. Most lncRNAs act as oncogenes and only a few lncRNAs have been shown to inhibit OSCC metastasis. Besides, we briefly introduced the research status of cancer-associated fibroblasts-related lncRNAs in OSCC metastasis. Finally, we discussed the research prospects of lncRNAs-mediated crosstalk between OSCC cells and the tumor microenvironment in OSCC metastasis, especially the potential research value of exosomes and lymphangiogenesis. In general, lncRNAs are expected to be used for screening, treatment, and prognosis monitoring of OSCC metastasis, but more work is still required to better understand the biological function of lncRNAs.

OGDHL closely associates with tumor microenvironment and can serve as a prognostic biomarker for papillary thyroid cancer.

BackgroundPapillary thyroid cancer (PTC) is the most common type of thyroid cancer. However, due to the lack of reliable prognostic biomarkers for PTC, overtreatment has been on the rise. Therefore, our research aims to identify new and promising prognostic biomarkers and provide fresh perspectives for clinical decision making.MethodsThe RNA‐seq data and clinical data of PTC samples were obtained from The Cancer Genome Atlas data portal. GSE64912 and GSE83520 datasets were downloaded through the GEOquery R package. The difference in the expression of oxoglutarate dehydrogenase like (OGDHL) between PTC and normal tissues was explored by the Wilcoxon test. Kaplan–Meier (KM) and Cox regression analyses were used to further explore the prognostic value of OGDHL. The tumor microenvironments of PTC patients were explored based on ssGSEA and Tumor Immune Estimation Resource online database. Gene Set Enrichment Analysis (GSEA) was performed to explore the biological processes associated with OGDHL.ResultsThe expression level of OGDHL in PTC was significantly altered compared to that in normal tissues (p < 0.05). Various biological processes associated with OGDHL were also explored through GSEA. KM analysis suggested that the low‐OGDHL group had a better overall survival [OS, p = 3.49e‐03, hazard ratio (HR) = 4.567]. The receiver operating characteristic curve also indicated the favorable prognostic potential of OGDHL. Moreover, OGDHL was proved to be an independent prognostic indicator in Cox analysis (p = 1.33e‐02, HR = 0.152). In the analysis of the tumor microenvironment, the low‐OGDHL group showed a lower immune score and stromal score, while tumor purity was higher. The expression of OGDHL was also closely correlated with the infiltration of immune cells.ConclusionOur study elucidated the influence of OGDHL on the prognosis of PTC and demonstrated its potential as a novel biomarker, which would provide new insights into the prognosis monitoring and clinical decision making in PTC patients.

Mechanism of KLF4 in the development and progression of hepatocellular carcinoma

The pathogenesis of hepatocellular carcinoma is complex and has not been fully clarified. KLF4, also known as gut-enriched Kruppel-like factor (GKLF), plays the dual regulatory role of tumor suppressor and tumor promoter. KLF4 plays an anti-cancer role in liver cancer. This article introduces the biological function and regulatory mechanism of KLF4 in the progression of liver cancer, and research on the mechanism of action of KLF4 in liver cancer is expected to provide new ideas for targeted therapy and prognosis monitoring of liver cancer.

Mechanisms and Functions of MiR-200 Family in Hepatocellular Carcinoma.

Hepatocellular carcinoma (HCC) is one of the most common clinically malignant tumors of the digestive system. It ranks the sixth most common malignant tumor in the world and ranks fourth among cancer-related death worldwide. At present, early diagnosis and prognosis monitoring of hepatocellular carcinoma mainly use alpha-fetoprotein combined with ultrasonography, which leads to clinical frequently missed diagnosis or even misdiagnosis. Therefore, seeking specific diagnostic and monitoring molecules of hepatocellular carcinoma are still hot topics in contemporary medical practice. MicroRNA is an endogenous non-coding small RNA that regulates the expression of the target molecule and participates in various biological processes in vivo. The miR-200 family, the most common celebrity family of microRNAs, is commonly lower expression in a variety of cancers and is closely associated with tumorigenesis and outcome, especially hepatocellular carcinoma. This review mainly discusses the expression changes, specific molecular mechanisms, biological functions and clinical values of miR-200 family in hepatocellular carcinoma. Moreover, we highlighted utilization of miR-200 family as molecular biomarkers for early diagnosis, prognostic monitoring and appropriate therapy in hepatocellular carcinoma.

Simple and fast isolation of circulating exosomes with a chitosan modified shuttle flow microchip for breast cancer diagnosis.

Tumor-derived exosomes have been recognized as promising biomarkers for early-stage cancer diagnosis, tumor prognosis monitoring and individual medical treatment. However, it is a huge challenge to separate exosomes from trace biological samples in clinics for disease diagnosis. Herein, we propose a simple, quick, and label-free method for isolating circulating exosomes from serum of patients. The strategy synergistically integrates chitosan electrostatic-adsorption, micro-patterned substrates, and microfluidic shuttle flow control to enable the capture/release of circulating exosomes in a simple manner. Using this microchip, we can isolate exosomes from trace samples (10 μl) with relative purity over 90% and high RNA recovery ratio over 84% within 15 minutes, which is impossible for traditional ultracentrifugation methods. We then validate the application of the microchip using 24 serum samples from clinical breast cancer and breast fibroma patients. The isolated exosomes are subjected to miRNA sequencing and RT-PCR, followed by pathway prediction analysis. The results showed that exosomes were relevant to the invasion and metastasis of breast cancer cells and hsa-miR-18a-3p might have the potential to become a new biomarker for distinguishing breast cancer from breast fibroma (AUC = 0.83, P value = 0.019). This established method is simple, quick and easy to operate with integration. And it may pave a new way for clinical research on exosomes and tumor relevant diagnosis.

Colorimetric detection of exosomal microRNA through switching the visible-light-induced oxidase mimic activity of acridone derivate

Exosomal microRNAs (miRNAs) are vital biomarkers for early diagnosis and prognosis monitoring of cancer. Yet, convenient and controllable detection of exosomal miRNA still remains challenges because of lacking of adequately simple and robust assay platforms. In this paper, it is first time to study the visible-light-induced oxidase mimic activity of 10-methyl-2-amino-acridone (MAA) being able to be switched by Cu2+ and DNA. Based on this phenomenon, a series of visual molecular logic gates are constructed, and a colorimetric strategy has been developed to achieve exosomal microRNA-21 (miR-21) detection with a signal amplification approach. The visible-light-induced oxidase mimic activity of MAA can be inhibited by Cu2+. In presence of target, a large amount of capture probes partly complementary with miR-21 are hydrolyzed with the assist of duplex-strand specific nuclease (DSN), releasing guanine-rich oligodeoxynucleotides that can chelate Cu2+, resulting in catalytic activity of MAA being recovered under irradiation. This strategy allows the detection of miR-21 with a light modulating temporal controllable manner, and the linear range is from 50 fM to 3000 fM with the limit of detection (LOD) being 44.76 fM. More importantly, the proposed method can achieve quantitative measurement of exosomal miR-21 that is derived from three-dimensional multicellular tumor spheroids with different size, which is able to monitor the growth of tumor spheroids. This work is potential to provide a feasible tool for application in exosomal miRNAs-based cancer diagnosis. Ultimately, MAA is expected to be a signal probe in biomedical field by virtue of its fascinating visible-light-induced oxidase mimic activity.

Nanotechnology-Assisted Isolation and Analysis of Circulating Tumor Cells on Microfluidic Devices.

Circulating tumor cells (CTCs), a type of cancer cell that spreads from primary tumors into human peripheral blood and are considered as a new biomarker of cancer liquid biopsy. It provides the direction for understanding the biology of cancer metastasis and progression. Isolation and analysis of CTCs offer the possibility for early cancer detection and dynamic prognosis monitoring. The extremely low quantity and high heterogeneity of CTCs are the major challenges for the application of CTCs in liquid biopsy. There have been significant research endeavors to develop efficient and reliable approaches to CTC isolation and analysis in the past few decades. With the advancement of microfabrication and nanomaterials, a variety of approaches have now emerged for CTC isolation and analysis on microfluidic platforms combined with nanotechnology. These new approaches show advantages in terms of cell capture efficiency, purity, detection sensitivity and specificity. This review focuses on recent progress in the field of nanotechnology-assisted microfluidics for CTC isolation and detection. Firstly, CTC isolation approaches using nanomaterial-based microfluidic devices are summarized and discussed. The different strategies for CTC release from the devices are specifically outlined. In addition, existing nanotechnology-assisted methods for CTC downstream analysis are summarized. Some perspectives are discussed on the challenges of current methods for CTC studies and promising research directions.

Abstract 3130: Evaluation of TK1 protein changes using the AroCell TK 210 ELISA in an acute myeloid leukemia (AML) xenograft model during 5-fluoruracil treatment

Abstract Background Thymidine kinase 1 (TK1) is a cytosolic enzyme involved in DNA precursor synthesis and it is up-regulated in proliferating cells. Serum TK1 is an established biomarker for prognosis and therapy monitoring of malignancies. However, commercial TK1 enzyme activity assays have limitations for their use in xenograft models as they cross-react with TK1 activity from the host. The AroCell TK 210 ELISA is specific for human TK1 and offers the opportunity to measure the release of TK1 from human xenografts free from the background of TK1 release from the host (mouse). Here we studied the changes in serum TK1 protein levels using the AroCell TK 210 ELISA in a xenograft model during 5-fluorouracil (5FU) treatment of Acute Myeloid Leukemia (AML). Experimental Procedures Human MV-4-11 leukemic cells from ATCC were cultured with 95% O2 and 5% CO2 at 37°C and in IMD medium (Sigma) supplemented with 10% FBS (HI media) containing 4mmol/L L-glutamine, 5 ng/mL GM-CSF and 1% penicillin and streptomycin. The cells were harvested during exponential growth and injected subcutaneously into the left flank of athymic BALB/c nude mice 8-10 weeks old. The mice were divided into controls that received no therapy (n=8) and an active group receiving 5FU (n=8). The treatment was initiated when the average tumor volume reached 50-100 mm3. Tumor bearing mice were treated with PBS (control) or 5-FU (active group 10 mg/kg, once daily) for three weeks. Serum samples (70-80 µL) were collected before treatment initiation, during treatment (once weekly for 3 weeks) and a final termination. Serum samples were stored at -80°C and TK1 protein levels were determined with the AroCell TK 210 ELISA. All the animal experiments were performed at TheraIndx life sciences Pvt Ltd, India. Results The tumor volumes were measured post-implantation of cells every 3 days throughout the study. There was a significant difference (p&amp;lt;0.05) in the mean tumor volume between 5-FU treated (5-FU) (108.75± 36.3 mm3) and untreated controls (294.63± 50.17 mm3) at the end of treatment. There was a significant reduction in the tumor volume from 2nd week onwards in 5-FU treated mice up to termination (4th week) compared to control mice. TK1 protein concentrations increased significantly from the 2nd week onwards in 5-FU treated mice (0.42± 0.19 µg/L) in comparison to controls (0.1± 0.02 µg/L) in spite of the decrease in tumor volume in response to treatment. No cross-reactivity of TK 210 ELISA with mouse TK1 was observed. Conclusions This study is the first evaluation of TK 210 ELISA as a biomarker for monitoring therapy with anticancer drugs in an AML xenograft model. The changes in TK1 protein levels were significant enough to monitor the response of the xenograft to 5FU therapy in this animal model. Furthermore, the use of the AroCell TK 210 ELISA in xenograft models may enable more accurate measurement of the anti-cancer drug effects on cell proliferation and these results encourage further applications of TK 210 ELISA in drug development projects. Citation Format: Kiran Kumar Jagarlamudi, Ramesh Jayaraman, Sundaresh Babu, Martin Shaw, Staffan Eriksson. Evaluation of TK1 protein changes using the AroCell TK 210 ELISA in an acute myeloid leukemia (AML) xenograft model during 5-fluoruracil treatment [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3130.

Abstract 1989: Enhanced whole exome profiling of tumor circulating cell-free DNA enables sensitive assessment of tumor mutations

Abstract An increasing number of studies have demonstrated the potential use of circulating cell-free DNA (cfDNA) for diagnosis, prognosis, and disease progression monitoring. However, many of these studies utilize assays covering a limited set of genes, typically tens to a few hundred genes, and therefore can miss biologically and clinically important genetic alterations such as DNA repair pathways, immuno-modulatory pathways, mechanisms of resistance and changes in neoantigen status. To address this, we have developed a whole-exome scale cfDNA platform, NeXT Liquid Biopsy, that enables sensitive detection and tracking of mutations in approximately 20000 genes. To enable sensitive detection across the exome, we developed an enhanced exome assay and chemistry that augments hard to sequence genomic regions such as regions of high GC content, to enable more uniform coverage across the exome. Additionally, we achieve a high average depth of approximately 2000X for the entire exome, with additional boosted depth for 248 clinically relevant oncogenic and tumor suppressor genes to further enhance sensitivity. For analysis, we developed a computational pipeline for our NeXT Liquid Biopsy assay optimized to lower the noise floor for variant detection, enabling sensitive monitoring and de novo detection of variants over multiple time points. We have evaluated the sensitivity of our NeXT Liquid Biopsy platform using three approaches. First, we evaluated the sensitivity using both Horizon and Seracare reference materials at multiple allele frequency (AF) dilutions. Our platform identified all 25 Seracare SNV events at 1% AF, all 25 events at 2% AF, and 24 out of 25 events at 0.5% AF. We detected no variants from the negative control. Next, we expanded our sensitivity evaluation using a much larger reference panel of 555 SNVs from the Acrometric Oncology HotspotControl. Similarly, we were able to achieve high sensitivity at our detection limit. Further, to enable sensitivity analysis at the whole exome scale, we developed a cell culture media system that models the shed and degraded tumor DNA fragments seen in human plasma samples. Finally, we demonstrated our ability to capture mutations across the exome using a head and neck cancer cohort on checkpoint therapy, observing a large fraction of mutations in genes not covered by commercially available targeted panels. In conclusion, we have developed a whole-exome scale NeXT Liquid Biopsy platform that enables sensitive monitoring and detection of somatic SNVs from cfDNA across approximately 20000 genes. The NeXT Liquid Biopsy platform generates a much broader view of the tumor mutational landscape from the plasma than typical liquid biopsy platforms that are focused on a much smaller set of genes. The platform enables broader monitoring of changes in response to cancer therapy, acquired mechanisms of resistance, mechanisms of drug resistance, and intra- and inter-tumor heterogeneity. Citation Format: Simo V. Zhang, Mengyao Tan, Josette M. Northcott, Shuyuan Ma, Christopher S. Nelson, L. Gordon Bentley, Manju Chinnappa, Devayani P. Bhave, Dan Norton, Jason Harris, Sean M. Boyle, John West, Richard Chen. Enhanced whole exome profiling of tumor circulating cell-free DNA enables sensitive assessment of tumor mutations [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1989.

Positive result of Sars-Cov-2 in faeces and sputum from discharged patients with COVID-19 in Yiwu, China.

BackgroundWith the effective prevention and control of COVID‐19 in China, the number of cured cases has increased significantly. Further monitoring of the disease prognosis and effective control of the “relapse” of the epidemic has become the next focus of work. This study analysed the clinical prognosis of discharged COVID‐19 patients by monitoring their SAR‐CoV‐2 nucleic acid status, which provided a theoretical basis for medical institutions to formulate discharge standards and follow‐up management for COVID‐19 patients.MethodsWe included 13 discharged COVID‐19 patients who were quarantined for 4 weeks at home. The patient's daily clinical signs were recorded and sputum and faecal specimens were regularly sent for detection of SARS‐CoV‐2 nucleic acid.ResultsThe time between initial symptoms and meeting discharge criteria was 18 to 44 days with an average of 25 ± 6 days. The faecal samples of two patients still tested positive after meeting the discharge criteria and the sputum samples of four patients returned positive 5 to 14 days after discharge. The rate of the recurring positive test result in samples from the respiratory system was 31% (4/13).ConclusionUnder the present discharge criteria, the high presence of SARS‐CoV‐2 nucleic acid in faecal and respiratory samples of discharged COVID‐19 patients indicates potential infectivity. Therefore, we suggest that faecal virus nucleic acid should be tested as a routine monitoring index for COVID‐19 and a negative result be added to the criteria. Simultaneously, we should strengthen the regular follow‐up of discharged patients with continuous monitoring of the recurrence of viral nucleic acid.

The role of peripheral blood eosinophil counts in COVID-19 patients.

BackgroundCoronavirus disease 2019 (COVID‐19) emerged in Wuhan city and rapidly spread globally outside China. We aimed to investigate the role of peripheral blood eosinophil (EOS) as a marker in the course of the virus infection to improve the efficiency of diagnosis and evaluation of COVID‐19 patients.Methods227 pneumonia patients who visited the fever clinics in Shanghai General Hospital and 97 hospitalized COVID‐19 patients admitted to Shanghai Public Health Clinical Center were involved in a retrospective research study. Clinical, laboratory, and radiologic data were collected. The trend of EOS level in COVID‐19 patients and comparison among patients with different severity were summarized.ResultsThe majority of COVID‐19 patients (71.7%) had a decrease in circulating EOS counts, which was significantly more frequent than other types of pneumonia patients. EOS counts had good value for COVID‐19 prediction, even higher when combined with neutrophil‐to‐lymphocyte ratio. Patients with low EOS counts at admission were more likely to have fever, fatigue, and shortness of breath, with more lesions in chest CT and radiographic aggravation, and longer length of hospital stay and course of disease than those with normal EOS counts. Circulating EOS level gradually increased over the time, and was synchronous with the improvement in chest CT (12 days vs 13 days, P = .07), later than that of body temperature (12 days vs 10 days, P = .014), but earlier than that of the negative conversion of nucleic acid assays (12 days vs 17 days, P = .001).ConclusionPeripheral blood EOS counts may be an effective and efficient indicator in diagnosis, Evaluation, and prognosis monitoring of COVID‐19 patients.

Abstract B21: Circulating tumor cell by microfluidic magnetophoresis-based gene expression changes for monitoring of prognosis in advanced prostate cancer patient: A pilot study

Abstract Introduction: By using the circulating tumor cells (CTCs), we can get useful genomic information that enables monitoring of patients’ prognosis or selecting appropriate treatment over time by repeat biopsy. The aim of this study is to separate CTCs from prostate cancer patients using new microfluidic technique that is based on disposable lateral magnetophoretic microseparator and to assess the gene expression that is specific for prostate cancer using mRNA from isolated CTCs measured by droplet digital PCR (ddPCR) method. We also evaluated the treatment-dependent gene-profile changes in four metastatic castration-resistant prostate cancer patients (mCRPC). Patients and Methods: Total 55 samples of 49 patients who underwent treatment for prostate cancer between 06/2018 and 05/2019 were prospectively enrolled. Approximately 10 mL of whole blood was drawn from prostate cancer patients; Group A, 14 samples (25.4%) with localized stage II-III cancer; Group B, 5 samples (9.1%) with stage IV; Group C, 8 samples (14.5%) with metastatic hormone-sensitive prostate cancer (mHSPC); and Group D, 29 samples (52.7%) with mCRPC. In particular, more than two repeated blood collections were performed for 4 mCRPC. CTCs were positively isolated by using assembly-disposable CTC-μChip. CTCs from 5 mL of blood were used for counting to identify isolated CTC numbers with immunofluorescence detection procedure. CTCs from the remaining 5 mL of blood were used for gene analysis by ddPCR for 6 prostate cancer-related genes, such as androgen receptor (AR), androgen receptor splice variant 7 (AR-V7), prostate specific membrane antigen (PSMA), prostate specific antigen (PSA), cytokeratin-19 (CK-19), and epithelial cell adhesion molecule (EpCAM). Results: We identified the average of 3, 16, 14, and 26 CTCs per 1ml in group A, B, C, and D, respectively (P&amp;lt; 0.01). The gene positive rates of AR, AR-V7, KRT-19, EpCAM, PSA, and PSMA were 85.4% (47/55), 16.4% (9/55), 56.4% (31/55), 92.7% (51/55), 41.8% (23/55), and 50.9% (28/55), respectively. The copy number of PSA, PSAM, and KRT-19 increased in proportion as the clinical stage increased. 92.7% of samples showed EpCAM positive, whereas AR-V7 was a rare event, with only in 9 out 28 samples positive with mCRPC, but not in other stages. In particular, of the four mCRPC patients who repeated blood sampling, the number of CTCs and the expression of AR, PSA, PSMA, and KRT-19 decreased in patients who responded to drug (enzalutamide and docetaxel). On the other hand, patients who were resistant to drug showed increased expression of AR-V7 and KRT-19 with the increased number of CTC. Conclusions: This study demonstrated that CTC-based multigene profiling could be analyzed using ddPCR. We have also identified several genes associated with progression in mCRPC patients initiating enzalutamide or chemotherapy. While this is a pilot study and prospective validation is needed, these findings highlight the potential role for this approach in real-time monitoring of advanced prostate cancer patients. Citation Format: Jae-Seung Chung, Hyungseok Cho, Ki-Ho Han. Circulating tumor cell by microfluidic magnetophoresis-based gene expression changes for monitoring of prognosis in advanced prostate cancer patient: A pilot study [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 B21.

Application of 18F-fluorodeoxyglucose positron emission tomography/computerized tomography in mantle cell lymphoma.

The study is to investigate the application of F-fluorodeoxyglucose (F-FDG) PET/computerized tomography (CT) for the evaluation of mantle cell lymphoma (MCL). We retrospectively analyzed 39 patients who were pathologically diagnosed with MCL and underwent F-FDG PET/CT before treatment between August 2007 and August 2018. We compared the clinical information and PET/CT imaging characteristics in different groups based on bone marrow invasion, spleen invasion or International Prognostic Index (IPI) score. We also assessed the efficacy of PET/CT evaluation basing on the follow-up PET CT findings of 21 MCL patients and their biopsies. Thirty-five patients were stage IV according to the Revised Ann Arbor Staging System. Lymph node involvement was observed in all 39 cases. The maximum diameter of the affected lymph nodes (4.33 ± 3.09 cm) and maximum standardized uptake value (SUVmax) (8.38 ± 4.99) was positively correlated (r = 0.486, P = 0.002). Extranodal invasion was identified in 38 patients with MCL, and the SUVmax of extranodal invasion was 7.34 ± 3.31. Extranodal invasion was most common in the spleen (25/38) and bone marrow (18/38). The group with bone marrow invasion was more prone to nasopharyngeal, lung and renal invasions (all P < 0.05). The groups with bone marrow invasion or spleen invasion were more likely to have decreased hemoglobin (Hgb) and platelets (all P < 0.01). The IPI high-risk group was more prone to lung involvement, elevated LDH and CRP, and decreased Hgb (all P < 0.05). Among the follow-up of 30 MCL patients, the 2-year progression-free survival and overall survival rates were 73.33 and 87.50%, respectively. PET/CT reexaminations of 21 MCL patients after treatment showed that the sensitivity, specificity, negative predictive value, positive predictive value and accuracy of the efficacy evaluation were 80, 90.91, 88.89, 83.33 and 85.71%, respectively. F-FDG PET/CT imaging has important application value in the diagnosis, staging, treatment efficacy assessment and prognosis monitoring of MCL, especially in the systemic assessment of advanced MCL.

Plasma Exosomal Long Noncoding RNA lnc-SLC2A12-10:1 as a Novel Diagnostic Biomarker for Gastric Cancer.

PurposeExosomes participate in cellular communications by transmitting active molecules, including long noncoding RNAs (lncRNAs) and are regarded as suitable candidates for disease diagnosis. This study aimed to identify gastric cancer (GC)-specific exosomal lncRNA and investigate the potential diagnostic value of plasma exosomal lncRNA in GC.Patients and MethodsExosomes from the culture media (CM) of four GC cells (GCCs) and human gastric epithelial cells were isolated. Exosomal RNA was extracted, and lncRNA microarray assay was performed to identify GC-specific exosomal lncRNAs. The expression levels of the candidate exosomal lncRNAs were validated in 120 subjects via quantitative reverse transcription PCR (qRT-PCR). The receiver operating characteristic (ROC) curve and area under curve were used to estimate the diagnostic capacity. We investigated the potential relationship between plasma exosomal lncRNA expression and the clinicopathological parameters of GC.ResultsA total of 199 exosomal lncRNAs were expressed at considerable higher levels in GCCs than those in normal controls, among which the top 10 upregulated lncRNAs were selected for further validation in cell, CM, and plasma. qRT-PCR revealed that lnc-SLC2A12-10:1 was remarkably upregulated in exosomes derived from patients with GC and GCCs. The area under the ROC curve was 0.776, which was higher than the diagnostic accuracies of CEA, CA 19–9, and CA72–4. The expression level of exosomal lnc-SLC2A12-10:1 was also significantly correlated with tumor size, TNM stage, lymph node metastasis, and degree of differentiation. The postoperative expression levels of exosomal lnc-SLC2A12-10:1 were lower compared with those of preoperative levels.ConclusionOur study suggested that exosomal lnc-SLC2A12-10:1 may be a potential noninvasive biomarker for the diagnosis and prognosis monitoring of GC. Further large-scale studies are necessary to validate its performance in GC progression.

Endoscopic decompression of acute intestinal distension is associated with reduced mortality in critically ill patients

BackgroundEndoscopic placement of intestinal decompression tubes is a feasible technique for treatment of acute intestinal dilation. Given the heterogeneity of the underlying diseases leading to intestinal obstruction data on the significance of endoscopic procedures for treatment of these conditions are sparse.MethodsIn the study period from 2008 to 2019 all patients receiving a decompression tube were identified by retrospective chart review and analyzed.ResultsA total of 59 decompression tubes were placed in 50 patients. Technical success was achieved in 98% (58/59 tubes). As major complication one small bowel perforation occurred (1/59; 1.7%). Causes for impaired intestinal transit comprised tumor stenoses 22% (11/50), infections 18% (9/50), post-operative paralysis 14% (7/50), neurological diseases 8% (4/50), trauma 2% (1/50) and others 36% (18/50). Most patients (74%; 37/50) were critically ill and treated on intensive care unit. Treatment response after tube insertion was documented in 76% of patients (38/50) whereas 24% (12/50) did not fulfill response criteria. Patients with treatment response showed a significantly better outcome compared to non-responders. Responders had a median survival of 113 days (95% CI 41–186) compared to 15 days (95% CI 6–24) in non-responders (p = 0.002). Analysis of laboratory parameters after stratification in responders and non-responders to endoscopic therapy showed that non-responders had significantly higher levels of CRP and lower platelet count at baseline (CRP 262 mg/L (IQR 101–307) vs. 94 mg/L (IQR 26–153): p = 0.027; platelets 69 thsd/μL (IQR 33–161) vs. 199 thsd/μL (IQR 138–289): p = 0.009).ConclusionsEndoscopic decompression is a safe procedure for acute management of impaired intestinal transit even in critically ill patients. Response to therapy is associated with improved outcome and markers of inflammation and organ function such as CRP, platelet count and serum lactate have to be taken into account for therapy monitoring and evaluation of prognosis.

Characterizing the gut microbiota in patients with chronic kidney disease

ABSTRACT Objectives: Emerging evidence suggests that gut microbiota dysbiosis plays a critical role in chronic kidney disease (CKD). However, the relationship between altered gut microbiome profiles and disease severity remains unclear. In this study, we sought to characterize the gut microbiota in CKD patients compared to healthy controls, and to explore potential relationships between gut microbiota composition and disease severity. Methods: Fecal samples were collected from 95 patients at different stages of CKD (non-dialysis patients from stage 1 to 5) and 20 healthy controls. Bacterial DNA was extracted for 16S ribosomal DNA sequencing targeting the V3-V4 region. The diversity and relative abundance of gut microbiota were analyzed as outcome indicators. Results: Differences were observed in the microbial composition and diversity of fecal samples from CKD patients and healthy controls. Specifically, disease severity was found to alter gut microbiota composition. Compared to that in healthy controls, CKD patients showed an increased abundance of Proteobacteria and decreased Synergistetes, most notably in disease stage 5. Lower levels of butyrate-producing bacteria and higher levels of potential pathogens were also detected in CKD patients. Further, Pyramidobacter and Prevotellaceae_UCG-001 were significantly decreased in the CKD1 group compared with healthy controls. Notably, nine microbial genera, including Escherichia-Shigella, Parabacteroides, Roseburia, rectale_group, Ruminococcaceae_NK4A214_group, Prevotellaceae_UCG.001, Hungatella, Intestinimonas, and Pyramidobacter, identified using a random forest model, distinguished between patients with CKD and healthy controls with high accuracy. Functional analysis also revealed that fatty acid and inositol phosphate metabolism were enriched in the CKD group, while aminoacyl-tRNA biosynthesis, oxidative phosphorylation, phenylalanine, tyrosine, and tryptophan biosynthesis, thiamine metabolism, pantothenate, and CoA biosynthesis, as well as valine, leucine, and isoleucine biosynthesis were enriched in healthy controls. Conclusion: Gut microbiota composition and function are associated with CKD severity. And, specific gut microbes are potentially helpful for CKD early diagnosis and prognosis monitoring.