Fecal microbial biomarkers combined with multi-target stool DNA test improve diagnostic accuracy for colorectal cancer

BACKGROUND Colorectal cancer(CRC)is a major global health burden.The current diagnostic tests have shortcomings of being invasive and low accuracy.AIM To explore the combination of intestinal microbiome composition and multi-target stool DNA(MT-sDNA)test in the diagnosis of CRC.METHODS We assessed the performance of the MT-sDNA test based on a hospital clinical trial.The intestinal microbiota was tested using 16S rRNA gene sequencing.This case-control study enrolled 54 CRC patients and 51 healthy controls.We identified biomarkers of bacterial structure,analyzed the relationship between different tumor markers and the relative abundance of related flora components,and distinguished CRC patients from healthy subjects by the linear discriminant analysis effect size,redundancy analysis,and random forest analysis.RESULTS MT-sDNA was associated with Bacteroides.MT-sDNA and carcinoembryonic antigen(CEA)were positively correlated with the existence of Parabacteroides,and alpha-fetoprotein(AFP)was positively associated with Faecalibacterium and Megamonas.In the random forest model,the existence of Streptococcus,Escherichia,Chitinophaga,Parasutterella,Lachnospira,and Romboutsia can distinguish CRC from health controls.The diagnostic accuracy of MT-sDNA combined with the six genera and CEA in the diagnosis of CRC was 97.1%,with a sensitivity and specificity of 98.1%and 92.3%,respectively.CONCLUSION There is a positive correlation of MT-sDNA,CEA,and AFP with intestinal microbiome.Eight biomarkers including six genera of gut microbiota,MT-sDNA,and CEA showed a prominent sensitivity and specificity for CRC prediction,which could be used as a non-invasive method for improving the diagnostic accuracy for this malignancy.

A novel multimodal prediction model based on DNA methylation biomarkers and low-dose computed tomography images for identifying early-stage lung cancer

Objective:DNA methylation alterations are early events in carcinogenesis and immune signalling in lung cancer.This study aimed to develop a model based on short stature homeobox 2 gene (SHOX2)/prostaglandin E receptor 4gene (PTGER4) DNA methylation in plasma,appearance subtype of pulmonary nodules (PNs) and low-dose computed tomography (LDCT) images to distinguish early-stage lung cancers.Methods:We developed a multimodal prediction model with a training set of 257 individuals.The performance of the multimodal prediction model was further validated in an independent validation set of 42 subjects.In addition,we explored the association between SHOX2/PTGER4 DNA methylation and driver gene mutations in lung cancer based on data from The Cancer Genome Atlas (TCGA) portal.Results:There were significant differences between the early-stage lung cancers and benign groups in the methylation levels.The area under a receiver operator characteristic curve (AUC) of SHOX2 in patients with solid nodules,mixed ground-glass opacity nodules and pure ground-glass opacity nodules were 0.693,0.497 and 0.864,respectively,while the AUCs of PTGER4 were 0.559,0.739 and 0.619,respectively.With the highest AUC of0.894,the novel multimodal prediction model outperformed the Mayo Clinic model (0.519) and LDCT-based deep learning model (0.842) in the independent validation set.Database analysis demonstrated that patients with SHOX2/PTGER4 DNA hypermethylation were enriched in TP53 mutations.Conclusions:The present multimodal prediction model could more efficiently distinguish early-stage lung cancer from benign PNs.A prognostic index based on DNA methylation and lung cancer driver gene alterations may separate the patients into groups with good or poor prognosis.

Genetic susceptibility loci of lung cancer are associated with malignant risk of pulmonary nodules and improve malignancy diagnosis based on CEA levels

Objective:The heightened prevalence of pulmonary nodules(PN)has escalated its significance as a public health concern.While the precise identification of high-risk PN carriers for malignancy remains an ongoing challenge,genetic variants hold potentials as determinants of disease susceptibility that can aid in diagnosis.Yet,current understanding of the genetic loci associated with malignant PN(MPN)risk is limited.Methods:A frequency-matched case-control study was performed,comprising 247 MPN cases and 412 benign NP(BNP)controls.We genotyped 11 established susceptibility loci for lung cancer in a Chinese cohort.Loci associated with MPN risk were utilized to compute a polygenic risk score(PRS).This PRS was subsequently incorporated into the diagnostic evaluation of MPNs,with emphasis on serum tumor biomarkers.Results:Loci rs10429489G>A,rs17038564A>G,and rs12265047A>G were identified as being associated with an increased risk of MPNs.The PRS,formulated from the cumulative risk effects of these loci,correlated with the malignant risk of PNs in a dose-dependent fashion.A high PRS was found to amplify the MPN risk by 156%in comparison to a low PRS[odds ratio(OR)=2.56,95%confidence interval(95%CI),1.40−4.67].Notably,the PRS was observed to enhance the diagnostic accuracy of serum carcinoembryonic antigen(CEA)in distinguishing MPNs from BPNs,with diagnostic values rising from 0.716 to 0.861 across low-to high-PRS categories.Further bioinformatics investigations pinpointed rs10429489G>A as an expression quantitative trait locus.Conclusions:Loci rs10429489G>A,rs17038564A>G,and rs12265047A>G contribute to MPN risk and augment the diagnostic precision for MPNs based on serum CEA concentrations.

The biogenesis,function and clinical significance of circular RNAs in breast cancer

Circular RNAs(circ RNAs)are noncoding RNAs that form covalently closed loop structures.Circ RNAs are dysregulated in cancer and play key roles in tumorigenesis,diagnosis,and tumor therapy.Circ RNAs function as competing endogenous RNAs or micro RNA sponges that regulate transcription and splicing,binding to proteins,and translation.Circ RNAs may serve as novel biomarkers for cancer diagnosis,and they show potential as therapeutic targets in cancers including breast cancer(BC).In women,BC is the most common malignant tumor worldwide and the second leading cause of cancer death.Although evidence indicates that circ RNAs play a critical role in BC,the mechanisms regulating the function of circ RNAs in BC remain poorly understood.Here,we provide literature review aiming to clarify the role of circ RNAs in BC and summarize the latest research.We provide a systematic overview of the biogenesis and biological functions of circ RNAs,elaborate on the functional roles of circ RNAs in BC,and highlight the value of circ RNAs as diagnostic and therapeutic targets in BC.

Involvement of parathyroid hormone-related peptide in the aggressive phenotype of colorectal cancer cells

Colorectal cancer(CRC)remains one of the leading causes of mortality from malignant diseases worldwide.In general terms,CRC presents high heterogeneity due to the influence of different genetic and environmental factors;also,the neoplastic cells are strongly influenced by the extracellular matrix and several surrounding cells,known together as the tumor microenvironment(TME).Bidirectional communication takes place between the tumor and the TME through the release of autocrine and paracrine factors.Parathyroid hormone-related peptide(PTHrP)is a cytokine secreted by a wide variety of tissues and is able to regulate several cellular functions both in physiological as well as in pathological processes.It exerts its effects as a paracrine/autocrine factor,although its mode of action is mainly paracrine.It has been shown that this peptide is expressed by several tumors and that the tumor secretion of PTHrP is responsible for the malignant humoral hypercalcemia.Eight years ago,when our research group started studying PTHrP effects in the experimental models derived from intestinal tumors,the literature available at the time addressing the effects of PTHrP on colorectal tumors was limited,and no articles had been published regarding to the paracrine action of PTHrP in CRC cells.Based on this and on our previous findings regarding the role of PTH in CRC cells,our purpose in recent years has been to explore the role of PTHrP in CRC.We analyzed the behavior of CRC cells treated with exogenous PTHrP,focalizing in the study of the following events:Survival,cell cycle progression and proliferation,migration,chemoresistance,tumor-associated angiogenesis,epithelial to mesenchymal transition program and other events also associated with invasion,such us the induction of cancer stem cells features.This work summarizes the major findings obtained by our investigation group using in vitro and in vivo CRC models that evidence the participation of PTHrP in the acquisition of an aggressive phenotype of CRC cells and the molecular mechanisms involved in these processes.Recently,we found that this cytokine induces this malignant behavior not only by its direct action on these intestinal cells but also through its influence on cells derived from TME,promoting a communication between CRC cells and surrounding cells that contributes to the molecular and morphological changes observed in CRC cells.These investigations establish the basis for our next studies in order to address the clinical applicability of our findings.Recognizing the factors and mechanisms that promote invasion in CRC cells,evasion to the cytotoxic effects of current CRC therapies and thus metastasis is decisive for the identification of new markers with the potential to improve early diagnosis and/or to predict prognosis,to predetermine drug resistance and to provide treatment guidelines that include targeted therapies for this disease.

HER2 changes to positive after neoadjuvant chemotherapy in breast cancer: A case report and literature review

BACKGROUND As the most common cancer in women,breast cancer is the leading cause of death.Most patients are initially diagnosed as stage I-III.Among those without distant metastases,64%are local tumors and 27%are regional tumors.Patients in stage IIA-IIIC and those who meet the breast-conserving criterion with the exception of tumor size can consider neoadjuvant chemotherapy(NACT).It is worth noting that the status of tumor cell biomarkers is not consistently static.Endocrine-related estrogen receptor(ER),progesterone receptor(PR)and human epidermal growth factor receptor 2(HER2)encoded by erythroblastic leukemia viral oncogene homolog 2 gene can all alter from positive to negative or vice versa,especially in luminal B subtype after NACT.In addition,determination of HER2 status currently mainly relies on immunohistochemistry(IHC)and fluorescence in situ hybridization(FISH),but FISH is commonly used when the result of IHC is uncertain.HER2 is regarded as negative when the IHC result is 0/1+without the addition of FISH.To the best of our knowledge,this is the first report of a case harboring HER2 status transformation and IHC1+with positive amplification by FISH after NACT.CASE SUMMARY A 49-year-old woman discovered a mass in her right breast and underwent diagnostic workup.Biopsies of the right breast lesion and axillary lymph nodes were obtained.The results pointed to invasive ductal carcinoma with the IHC result for ER(80%),PR(60%),Ki-67(20%)and ambiguous expression of HER2(IHC 2+)with negative amplification by FISH(HER2/CEP17 ratio of 1.13).She underwent surgery after NACT.The pathological findings of the surgically resected sample supported invasive ductal carcinoma with the tumor measuring 1.1 cm×0.8 cm×0.5 cm and had spread to one of fifteen dissected lymph nodes.Retesting of the specimen showed that the tumor was positive for ER(2+,85%)and PR(2+,10%)but negative for HER2 by IHC(1+).Also Ki-67 had dropped to 2%.The patient was regularly monitored every 3 mo without evidence of recurrence.CONCLUSION Biomarker status should be reassessed after NACT especially in luminal subtypes.

Novel biomarkers and the future of targeted therapies in cholangiocarcinoma:a narrative review

Background and Objectives:Cholangiocarcinoma is a highly aggressive and heterogenous group of biliary malignancies arising from any site in the biliary tree,comprising 15%of all primary liver cancers.The nature of the disease and nonspecific presentation leads to late diagnosis and ultimately poor outcomes for patients.Combination gemcitabine and cisplatin has been the standard of care for cholangiocarcinoma(CCA)since 2010,with a median overall survival of 11.7 months.The five-year survival for CCA remains 5-10%,revealing a clear need for improved treatment options.Methods:This targeted review highlights the role of next generation sequencing in CCA and the clinically relevant tumor biomarkers that have become the focus of therapeutic development.Key Content and Findings:These tumor biomarkers or actionable mutations hold the potential to enable earlier diagnosis,provide prognostic information,and guide treatment decisions for patients with CCA.Specifically,the FGFR2 fusion and IDH1 mutation have shown considerable promise in development of targeted therapies.Clinical trials with inhibitors targeting FGFR2 fusion and IDH1 mutation have created expectations that these drugs will soon enter clinical practice.Other biomarkers including KRAS and B-raf protooncogenes,Her2/neu genes,and BRCA1 and 2 tumor-suppressor genes have also been touted as potential targets for future therapies,with early data showing promise for new drug development.Conclusions:The discovery of these actionable mutations and identification of targeted therapies have challenged the notion of a“one-size fits all”for treatment of CCA,and generated optimism that these novel treatments will soon be available for patients with CCA.

In vitro cultures of circulating tumor cells:a potential tool to unravel drug sensitivity

Since taking part as leading actors in driving the metastatic process,circulating tumor cells(CTCs)have displayed a wide range of potential applications in the cancer-related research field.Besides their well-proved prognostic value,the role of CTCs in both predictive and diagnostics terms might be extremely informative about cancer properties and therefore highly helpful in the clinical decision-making process.Unfortunately,CTCs are scarcely released in the blood circulation and their counts vary a lot among different types of cancer,therefore CTC detection and consequent characterization are still highly challenging.In this context,in vitro CTC cultures could potentially offer a great opportunity to expand the number of tumor cells isolated at different stages of the disease and thus simplify the analysis of their biological and molecular features,allowing a deeper comprehension of the nature of neoplastic diseases.The aim of this review is to highlight the main attempts to establish in vitro CTC cultures from patients harboring different tumor types in order to highlight how powerful this practice could be,especially in optimizing the therapeutic strategies available in clinical practice and potentially preventing or contrasting the development of treatment resistance.

Mass Spectrometry and Mass Spectrometry Imaging-based Thyroid Cancer Analysis

Thyroid carcinoma is one of the most common endocrine malignant diseases worldwide.With the rapid development of medical technology,early and effective diagnostic methods could be able to improve the survival rate and quality of life of patients suffering from the disease.Considering the complexity of cancer,some specific detection method is desired for diagnosis and treatment.Mass spectrometry imaging(MSI)is an emerging technique for acquiring molecular information from biological tissues without staining and labeling,including qualitative,quantitative and spatial distribution information.Over the past several decades,MSI has been widely used for pharmacological monitoring,biomolecular imaging of cells and tissues.In this review,we introduce the tumor progression and histological characteristics of thyroid cancer,and focus mainly on the preparation of biological specimens for MSI and mass spectrometry(MS)analysis,as well as the recent progress in MS and MSI-based thyroid cancer research.This review thoroughly discusses the importance of MS and MSI for clinical diagnosis,identification and prognosis of thyroid cancer,and provides some new clues for molecular mechanisms research and tumor metastasis.

Cholangiocarcinoma:early detection and screening in high-risk population

Cholangiocarcinoma(CCA)is a highly lethal malignancy that comprises approximately 15%of all the primary liver tumors and 3% of gastrointestinal cancers.Diagnosis is often done when the disease is already at advanced stages,resulting in poor outcomes.Prevention of risk factors and early diagnosis are the cornerstones for improving survival.Early diagnosis is feasible in the setting of surveillance programs in patients at high risk of CCA such as patients with primary sclerosing cholangitis.Regrettably,surveillance of CCA in this population is hampered by the low diagnostic accuracy of current tumor markers at earlier stages,the difficulties of imaging techniques for the differential diagnosis between benign and malignant biliary strictures,and the need for invasive procedures for diagnostic confirmation.In this review we discuss the rationale for surveillance of CCA in high-risk populations,particularly patients with primary sclerosing cholangitis,the recommended tools for surveillance and diagnostic work-up,and future perspectives.

Monoclonal antibodies to the exon 18 encoded moiety of NCAM

Aim: Exon 18 expression of NCAM has been recognized as a biomarker for small cell lung cancer (SCLC). To use this finding for an improved diagnosis of SCLC and personalized treatment of patients, techniques to identify and quantitate E18, the exon 18 encoded protein moiety of NCAM, are needed. We developed three monoclonal antibodies for this purpose. Methods: The his-tagged E18 antigen was expressed in E. coli and, after purification, used to immunize mice. Hybridoma's were isolated by standard procedures and tested for their reaction with E18. Results: Three monoclonal antibodies, MUM-1, MUM-4 and MUM-6 were obtained. They reacted with E18 in western blots, with SCLC cell line NCI-H82, but not with unrelated his-tagged proteins. Only permeabilized NCI-H82 cells stained with the antibodies, confirming the intracellular position of E18. Next an enzyme-linked immunosorbent assay was developed using the earlier isolated monoclonal antibody MUMi-21B2, coated on the surface of microtiter wells as capture antibody and biotinylated MUM-6 as second antibody. Using streptavidin conjugated to horse radish peroxidase a linear dose response curve to his-tagged E18 antigen was obtained between 0 and 5 μg/mL with a sensitivity of at least 0.5 μg/mL or 50 ng/well. Conclusion: Four monoclonal antibodies are available to be used in assays for the identification and quantification of SCLC biomarker E18. This will enable the development of liquid biopsies to follow the tumor load in patients.