Expression Of Molecular Biomarkers Research Articles

Integrative Clinical, Hormonal and Molecular Data associate with Invasiveness in Acromegaly: REMAH Study.

Growth-hormone (GH)-secreting pituitary-tumors (GHomas) are the most common acromegaly cause. At diagnosis, most of them are macroadenomas, and up to 56% display cavernous sinus invasion. Biomarkers assessment associated with tumor-growth and invasion are important to optimize their management. To identify clinical/hormonal/molecular-biomarkers associated with tumor-size and invasiveness in GHomas, and to analyze the influence of pre-treatment with somatostatin-analogs or dopamine-agonists in key molecular biomarkers expression. Clinical/analytical/radiological-variables were evaluated in 192 patients from the REMAH-study (ambispective multicenter post-surgery study of the Spanish Society of Endocrinology and Nutrition). Expression of somatostatin/ghrelin/dopamine-systems components, and key pituitary/proliferation-markers were evaluated in GHomas after the first surgery. Univariate/multivariate regression studies were performed to identify association between variables. 80% of patients harbor macroadenomas (63.8% with extrasellar-growth). Associations between larger and more invasive GHomas with younger age, visual-abnormalities, higher IGF1-levels, extrasellar/suprasellar-growth and/or cavernous-sinus invasion were found. Higher GH1 and lower PRL/POMC/CGA/AVPR1B/DRD2T/DRD2L expression levels (p<0.05) were associated to tumor invasiveness. LASSO´s penalized regression identified combinations of clinical and molecular features with AUCs between 0.67-0.82. Preoperative therapy with dopamine-agonist or somatostatin-analogs did not alter the expression of any of the markers analyzed except for DRD1/AVPR1B (up-regulated with dopamine-agonist), and FSHB/CRHR1 (down-regulated with somatostatin-analogs). A specific combination of clinical/analytical/molecular variables was found to be associated with tumor invasiveness and growth capacity in GHomas. Pre-treatment with first-line drugs for acromegaly did not significantly modify the expression of the most relevant biomarkers in our association model. These findings provide valuable insights for risk stratification and personalized management of GHomas.

Prognostic Molecular Biomarkers in Breast Cancer Lesions with Non-Mass Enhancement on MR.

Clustered ring enhancement (CRE) is a new lexicon for non-mass enhancement (NME) of breast MR in the 5th BIRADS, indicating a high suspicion of malignancy. We wonder if the presence of CRE correlates with expression of prognostic molecular biomarkers of breast cancer. A total of 58 breast lesions, which MRI reported with NME, were collected between July 2013 and December 2018. The patterns of enhancement including CRE were reviewed and the pathological results with expression of molecular biomarkers were collected. The association between MRI NME, pathological, and IHC stain findings were investigated under univariate analysis. A total of 58 breast lesions were pathologically proven to have breast cancer, comprising 31 lesions with CRE and 27 lesions without CRE on breast MRI. The expression of the estrogen receptor (ER) (p = 0.017) and the progesterone receptor (PR) (p = 0.017) was significantly lower in lesions with CRE as compared with those without CRE. The expression of Ki-67 (≥25%) was significantly higher in lesions with CRE (p = 0.046). The lesions with CRE had a lower expression ratio of ER (50.71 ± 45.39% vs. 74.26 ± 33.59%, p = 0.028). Our study indicated that lesions with CRE may possess different features from those without CRE in molecular expression, bearing a more aggressive behavior.

Differential molecular biomarker expression in corals over a gradient of water quality stressors in Maunalua Bay, Hawaii.

Coral reefs globally face unprecedented challenges from anthropogenic stressors, necessitating innovative approaches for effective assessment and management. Molecular biomarkers, particularly those related to protein expressions, provide a promising avenue for diagnosing coral health at the cellular level. This study employed enzyme-linked immunosorbent assays to evaluate stress responses in the coral Porites lobata along an environmental gradient in Maunalua Bay, Hawaii. The results revealed distinct protein expression patterns correlating with anthropogenic stressor levels across the bay. Some proteins, such as ubiquitin and Hsp70, emerged as sensitive biomarkers, displaying a linear decrease in response along the environmental gradient, emphasizing their potential as indicators of stress. Our findings highlighted the feasibility of using protein biomarkers for real-time assessment of coral health and the identification of stressors. The identified biomarkers can aid in establishing stress thresholds and evaluating the efficacy of management interventions. Additionally, we assessed sediment and water quality from the inshore areas in the bay and identified organic contaminants, including polycyclic aromatic hydrocarbons and pesticides, in bay sediments and waters.

PDL1 and molecular biomarkers expression in non-small cell lung cancer in Tunisian patients.

In cancer treatment, programmed death-1 (PD-1) and programmed death ligand-1 (PD-L1) inhibitors are thriving. Activated T lymphocytes expressed PD-1, it works with its ligand PD-L1 to limit T lymphocyte activation and prevent autoimmune disease. The expression of molecular biomarkers and PD-L1 in lung cancer determines the appropriate treatment strategy for patients with lung cancer. The purpose of this study was to look at the prevalence of molecular biomarkers and PD-L1 expression in a large group of Tunisian patients with advanced non-small cell lung cancer. We conducted an observational retrospective study in which medical/treatment history data were extracted retrospectively from medical records and archived tissue samples between January 1st 2019 and December 31st 2021. We gathered 157 patients who had recently been diagnosed with non-small cell lung carcinoma. In 36.9%of the cases, there was no molecular genotyping. EGFR (28.6%), KRAS (5.73%), and ALK gene rearrangement were the most common genotyping mutations (3.8%). ROS1 rearrangement was not present. There was a link between EGFR and gender, HER and age, and KRAS and biopsy tissue origin. Six of the tested cases with PD-L1 met the cut-off (³50%). PD-L1 positivity was more common in solid type adenocarcinoma (1.9%) than in acinar or papillary adenocarcinoma. There were no significant differences in PD-L1 expression across clinical and demographic parameters. High PD-L1 expression and molecular abnormalities were found in 1 case of EGFR, 1 case of BRAF, and 1 case of KRAS (3 cases). All of the other specimens with abnormalities had a PD-L1 <50%. ALK, ROS1, BRAF, KRAS, and MET were found to be significantly associated with PD-L1 expression. Our study is one of the country's largest, describing a large panel of biomarkers and their clinicopathologic/histopathologic associations in Tunisian lung cancer patients. We have the same molecular profile as European patients with an EGFR mutation, which is not the most common genotype abnormality in Tunisian patients. There is only one mutation at any given time. The expression of PD-L1 is determined by the histologic type and the origin of the biopsy tissue.

Ultrasound-based radiomics model for predicting molecular biomarkers in breast cancer.

Breast cancer (BC) is the most common cancer in women and is highly heterogeneous. BC can be classified into four molecular subtypes based on the status of estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2) and proliferation marker protein Ki-67. However, they can only be obtained by biopsy or surgery, which is invasive. Radiomics can noninvasively predict molecular expression via extracting the image features. Nevertheless, there is a scarcity of data available regarding the prediction of molecular biomarker expression using ultrasound (US) images in BC. To investigate the prediction performance of US radiomics for the assessment of molecular profiling in BC. A total of 342 patients with BC who underwent preoperative US examination between January 2013 and December 2021 were retrospectively included. They were confirmed by pathology and molecular subtype analysis of ER, PR, HER2 and Ki-67. The radiomics features were extracted and four molecular models were constructed through support vector machine (SVM). Pearson correlation coefficient heatmaps are employed to analyze the relationship between selected features and their predictive power on molecular expression. The receiver operating characteristic curve was used for the prediction performance of US radiomics in the assessment of molecular profiling. 359 lesions with 129 ER- and 230 ER+, 163 PR- and 196 PR+, 265 HER2- and 94 HER2+, 114 Ki-67- and 245 Ki-67+ expression were included. 1314 features were extracted from each ultrasound image. And there was a significant difference of some specific radiomics features between the molecule positive and negative groups. Multiple features demonstrated significant association with molecular biomarkers. The area under curves (AUCs) were 0.917, 0.835, 0.771, and 0.896 in the training set, while 0.868, 0.811, 0.722, and 0.706 in the validation set to predict ER, PR, HER2, and Ki-67 expression respectively. Ultrasound-based radiomics provides a promising method for predicting molecular biomarker expression of ER, PR, HER2, and Ki-67 in BC.

Conspectus on nanodiagnostics as an incipient platform for detection of oral potentially malignant disorders and oral squamous cell carcinoma.

One of the most common causes of death around the globe is cancer. One of the most prevalent forms of cancer is the cancer of the oral cavity and oropharynx known as oral squamous cell carcinoma (OSCC). The susceptivity to various viruses, smoking, overindulgence in tobacco, alcohol consumption, occupational exposure, increased sun exposure, and nutritional deficiencies resulting from low socioeconomic status are the preponderant risk factors which lead to genetic and epigenetic transmutation1,2. OSCC potentially spread quickly and the initial stage is referred to as the oral potentially malignant disorder (OPMD). The OPMD symptoms are white or red lesions in the mouth, paresthesia, trismus or jaw hypomobility gradually pronounce into severe vocal changes and oral ulcers which are basically nontreatable3. This illness has the potential to spread quickly, and it can frequently invade nearby tissues4. Since the OPMD indications were usually staved off, the OSCC becomes life-threatening and is thus associated with high morbidity rates. Hence, early detection of the very early OPMD stage is very critical in order to improve the health benefits of the patient and curb the prognosis of the disease. This article provides a brief summary of diagnostic strategies in the detection of oral cancer especially using nanotechnology, and the current trends and future prospects in nanodiagnostics. There are numerous tests available to diagnose oral and oropharyngeal cancer. Common techniques for diagnosing OSCC are endoscopy, oral brush biopsy, human papillomavirus testing, positron emission tomography scan, ultrasound, MRI, computed tomography, and radiography. Emerging detection techniques like nanodiagnostics provide advantages over the contrast agents used in modern imaging. Nanoparticles (NPs) will target specific surface molecules, are simpler to synthesize, and are more biocompatible than larger particles. Nanodiagnostics have exceptional characteristics like their size, high mechanical strength, shape, and surface. Nanomaterial-based sensors are significantly more sensitive than normal sensors. They also offer real-time tracking5, surface-enhanced Raman spectroscopy, MRI, optical coherence tomography, nano-based ultrasensitive biomarker detection, Quantum Dots (QDs) imaging, photoacoustic imaging, diffusion reflection imaging, and surface plasmon resonance scattering are some of the implementations of nano-based diagnostics6. The NPs currently used in oral cancer diagnosis are listed in Table 1. Table 1 - Nanoparticles for diagnosis of oral cancer Nanoparticles Description References Liposomes Liposomes are extensive in diagnostic research. Compared to 18F-FDG, 64Cu liposomes are more capable to recognize tumors in the early stage 7 Dendrimers The detection of oral malignancies can be done by an attractive device, Dendrimers. Wei et al. constructed DNA-dendrimer and poly-pyrrole (DDPpy) sensors for the detection of oral cancer biomarkers such as interleukin-1 protein, interleukin-8 RNA and interleukin-8 protein, which have improved specificity and greater bioaffinity 8 Gold NPs A noninvasive, efficient method for diagnosing OSCC is SERS 9 According to Xue and Wang’s study, the SERS approach could be utilized to examine and distinguish between OSCC and even the various tumor stages. Gold NPs can greatly increase the SERS spectrum’s intensities by adding to serum samples 10 For the electrochemical detection of oral cancer as an immunosensing platform, a customized AuNPs-rGO (gold-NP-reduced grapheneoxide) based bioelectrode has been designed 11 Magnetic NPs (MNPs) For MRI contrast agents, MNPs have been researched. They also assist in refining proton relaxation, then progressively developing as a contrast in both fields of biological and medical diagnostics with useful probes 12 MNPs can also be administered to the location of the tumor with precision, avoiding adjacent organs 13 Polymeric NPs Fluorescent endoscopic detection of oral cancer has been described by Yang et al. using Highly performing NPs. The folate receptors on oral cancer cells are targeted for nanoparticle endocytosis that is improved by Folic acid conjugated chitosan NPs. By reducing the interface between chitosan and the drug, the release of 5-aminolevulinic acid (5-ALA) in oral cancer cells can be boosted by the N-succinyl chitosan (SCHI) polymer with a negative charge 14 NP, nanoparticle; OSCC, oral squamous cell carcinoma; SERS, surface-enhanced Raman spectroscopy. Numerous developments have been made in the field of diagnostics as an outcome of the urgent need for sensitive, precise, strong, and quick diagnosis of oral cancer15,16. Recent developments in the field of nanotechnology offer the necessary incentive for nanobiosensor technology development17. Nanobiosensing techniques for OSCC detection are substrate selection, surface cleaning, surface enhancement by incorporation of NPs, surface enhancement by polymers, surface functionalization, biorecognition probe, and its immobilization (Fig. 1)18. In order to diagnose oral cancer, artificial intelligence is applied, such as the detection of precancerous as benign and postcancerous as malignant regions using automated deep learning CNN (Convolutional Neural Network) technique19. CNN is a deep learning algorithm. The major purpose of CNN is that it automatically identifies desired features and provide image recognition, with no human assistance. For the screening of oral cancer, deep CNNs can be a practical way to create low-cost embedded vision systems with constrained memory and processing power20. A significant advancement in recent years is the biosensor based on QDs that help with the diagnosis of some oral cancer biomarkers21. By conjugating QDs with complements of molecular biomarkers, multifunctional NP probes have been developed. QD probes can be often used to determine the molecular biomarker expression in oral cancer cells.Figure 1: Nanomaterial promising structures for the management of oral cancer.It is now evident that the NPs have demonstrated great promise and success in fundamentally altering the strategy for detecting oral cancer. Also, we sought to discuss key aspects of NP synthesis such as design criteria, cutting-edge experimental techniques, and NP platforms for OSCC diagnosis. The use of unique molecular markers will aid in the future development of active targeting magnetic NPs to significantly enhance tumor localization and diagnosis. Ethical approval Not applicable. Sources of funding None. Author contribution D.K.S.: investigation, writing – original draft preparation. S.C.A.: data collection, visualization, and original draft preparation. R.A.N.: investigation and data curation. M.S.: conceptualization, writing – reviewing and editing and supervision. J.A.: investigation, writing – reviewing and editing and supervision. M.A.B.: conceptualization, writing – reviewing and editing and supervision. Conflicts of interest disclosure The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Research registration unique identifying number (UIN) None. Guarantor Melaku Ashagrie Belete. Data statement All data are available in the manuscript.

Type IV collagen-derived angiogenesis inhibitor: canstatin low expressing in brain-invasive meningiomas using liquid chromatography–mass spectrometry (LC-MS/MS)

PurposeBrain invasion in meningiomas is considered an indicator of more aggressive behavior and worse prognosis. But the precise definition and the prognostic role of brain invasion remains unsolved duo to lacking a standardized workflow of surgical sampling and the histopathological detection. Searching for molecular biomarker expression correlating with brain invasion, could contribute to establish a molecular pathological diagnosis without problems of subjective interobserver variation and deeply understand the mechanism of brain invasion and develop innovative therapeutic strategies.MethodsWe utilized liquid chromatography tandem mass spectrometry to quantify protein abundances between non-invasive meningiomas (n = 21) and brain-invasive meningiomas (n = 21) spanning World Health Organization grades I and III. After proteomic discrepancies were analyzed, the 14 most up-regulated or down-regulated proteins were recorded. Immunohistochemical staining for glial fibrillary acidic protein and most likely brain invasion-related proteins was performed in both groups.ResultsA total of 6498 unique proteins were identified in non-invasive and brain-invasive meningiomas. Canstatin expression in the non-invasive group was 2.1-fold that of the brain-invasive group. The immunohistochemical staining showed canstatin expressed in both groups, and the non-invasive group showed stronger staining for canstatin in the tumor mass (p = 0.0132) than the brain-invasive group, which showed moderate intensity.ConclusionThis study demonstrated the low expression of canstatin in meningiomas with brain invasion, a finding that provide a basis for understanding the mechanism of brain invasion of meningiomas and may contribute to establish molecular pathological diagnosis and identify novel therapeutic targets for personalized care.

Colon tumor CD31 expression is associated with higher disease-free survival in patients with metabolic syndrome.

Metabolic syndrome (MS) is recognized as a risk factor for colon cancer (CC). However, how does the interplay between metabolic dysfunction caused by MS and its individual components affect CC microenvironment and prognosis remains unexplored. Angiogenesis and lymphangiogenesis are fundamental processes for tumor progression and dissemination, ensuring oxygen and nutrient delivery and supporting one of the most important pathways of tumor dissemination, contributing to metastasis. Thus, our aim was to evaluate whether the expression of molecular biomarkers involved in angiogenic and lymphangiogenic processes influenced CC clinicopathological features and prognosis in patients with MS. Clinical and pathological data of 300 patients submitted to CC surgical resection at a single tertiary hospital were retrospectively retrieved from hospital records. Tumor tissue microarrays of archived paraffin-embedded blocks were used to assess CD31, VEGF-A and D2-40 tissue expression by immunohistochemistry. The percentage of stained area was quantified by computerized morphometric analysis. No association between tissue expression of angiogenesis and lymphangiogenesis biomarkers and tumor clinical and pathological characteristics was found. However, in subgroup analysis of patients with MS, dysglycemia was associated with lower D2-40 expression (p=0.007) and high waist-circumference was associated with higher D2-40 (p=0.0029) and VEGF-A expression (p=0.026). In an adjusted Cox proportional hazard model CD31 expression was significantly associated with greater disease-free survival (HR=0.62; 95% CI: 0.41-0.95, p=0.028). No association was found between D2-40 and VEGF-A expression and CC prognosis. Our data reinforces previous reports that suggest the potential use of CD31 as a CC prognostic biomarker. Additionally, our data further supports the evidence for an interplay between metabolic dysfunction, tumor microenvironment, and vascularization pathways.

Prediction of Initial Risk Group, B/T Subtype, and ETV6-RUNX1 Translocation in Pediatric Acute Lymphoblastic Leukemia By Deep Convolutional Neural Network Analysis of Giemsa-Stained Whole Slide Images

Background: Acute lymphoblastic leukemia (ALL) is the most common malignancy in children. Diagnosis of ALL relies on subjective morphologic criteria by analyzing Giemsa-stained bone marrow (BM) smears under a microscope. Such examination, however, does not provide information about the ALL subtype and molecular profile, which is essential for diagnosis and guidance of treatment. To this end, clinicians resort to advanced techniques, such as immunophenotyping and genomic assays. Unlike Giemsa staining, these additional assays are expensive, time consuming, require special labs and trained clinicians, and are inaccessible in countries with limited resources. Convolutional neural networks (CNNs) are machine learning methods, which currently provide state-of-the-art performance for image analysis. It has been recently shown that CNNs can reveal information from the tumor morphology that is unseen by the human eye, such as molecular biomarker expression from hematoxylin and eosin-stained histological images. Nevertheless, it is yet unclear if BM smears, which lack tissue architecture and structured layout of cells, could also provide such information. We sought to evaluate whether image analysis of Giemsa-stained BM samples by CNNs could predict B and T ALL subtypes, ETV6-RUNX1 translocation, and initial risk group stratification. Methods: We collected a total of 276 Giemsa-stained BM slides from 163 pediatric patients diagnosed with ALL (n = 69), acute myeloid leukemia (AML) (n = 35), and non-leukemic BM (n = 59), between 2009 and 2022, at the Pediatric Hemato-Oncology Department, Souraski medical center, Israel. Information regarding patients included: age, white blood cell (WBC) count on presentation, B/T subtype, central nervous system (CNS) involvement and the presence of ETV6-RUNX1 translocation (Table 1). For each patient, we collected 1-4 Geimsa stained smears from the diagnostic BM, and scanned them at 0.25 micron/pixel, 10× magnification. The patients were split to train (80%) and test (20%) sets, and the images were automatically segmented to remove the background. Each slide was split to 256x256 non-overlapping tiles, resulting in overall 138,000 tile images containing cells (Figure 1). Importantly, we skipped the common step of cell segmentation. This end-to-end approach allows the computational model to exploit the entire smear information, including the morphology of all cells and the global diversity and arrangement of the cells in the smear. Using only the training cases, a deep CNN was trained to classify each Giemsa tile to control, ALL, and AML. The model was further trained to classify ALL tiles to either B or T subtype and to predict the presence of ETV6-RUNX1 translocation in B-ALL. The model was also trained to stratify patients to initial low risk (LR) or high risk (HR) groups, where LR is defined as age<10, WBC<50K, CNS=1 or 2, and B-ALL. The model was then applied to the held-out test set, and tile scores were aggregated to produce per-slide and per-patient prediction scores. The AUC for each prediction task was calculated. To obtain robust statistical analysis, the entire process was repeated 5 times, such that in each time a different test group was selected, and the AUC results were averaged. Results: The AUC for classification of ALL versus control and ALL versus AML were above 0.99 at the patient level (Table 1), demonstrating the CNN's ability to distinguish ALL cases from the rest almost perfectly. The AUC for classification of B/T subtypes, initial HR/LR subroups and ETV6-RUNX1 translocation were 0.79, 0.68 and 0.68, respectively, at a patient level, all having significant p-values. Notably, the system managed to make its predictions using slides scanned at a magnification significantly lower than routine magnification used for diagnosis by clinicians. Conclusions: These results show, for the first time, that machine learning computational models can not only reliably identify pediatric ALL, but can also predict the initial risk group, B/T subtype, and presence of ETV6-RUNX1 translocation from Giemsa stained BM samples - features that cannot be concluded from morphological analysis by a human observer. Utilizing such a system may enable physicians in countries that lack immunophenotyping and molecular analysis capabilities to refine ALL diagnosis and initial risk stratification based on Giemsa-stained BM smears alone. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Molecular Biomarker Expression in Window of Opportunity Studies for Oestrogen Receptor Positive Breast Cancer-A Systematic Review of the Literature.

Simple SummaryWindow of opportunity (WoO) trials allow the opportunity to assess the use of drugs in breast cancer research before treatment has commenced. The aim of this review of the literature is to review WoO trials in patients with oestrogen receptor-positive (ER+) breast cancer to help guide treatment. This will be useful for patients who receive drug treatment before surgery, or as an alternative to surgery in older, more frail adults.Window of opportunity (WoO) trials create the opportunity to demonstrate pharmacodynamic parameters of a drug in vivo and have increasing use in breast cancer research. Most breast cancer tumours are oestrogen receptor-positive (ER+), leading to the development of multiple treatment options tailored towards this particular tumour subtype. The aim of this literature review is to review WoO trials pertaining to the pharmacodynamic activity of drugs available for use in ER+ breast cancer in order to help guide treatment for patients receiving neoadjuvant and primary endocrine therapy. Five databases (EMBASE, Cochrane, MEDLINE, PubMed, Web of Science) were searched for eligible studies. Studies performed in treatment-naïve patients with histologically confirmed ER+ breast cancer were included if they acquired pre- and post-treatment biopsies, compared measurement of a proteomic biomarker between these two biopsies and delivered treatment for a maximum mean duration of 31 days. Fifteen studies were eligible for inclusion and covered six different drug classes: three endocrine therapies (ETs) including aromatase inhibitors (AIs), selective oestrogen receptor modulators (SERMs), selective oestrogen receptor degraders (SERDs) and three non-ETs including mTOR inhibitors, AKT inhibitors and synthetic oestrogens. Ki67 was the most frequently measured marker, appearing in all studies. Progesterone receptor (PR) and ER were the next most frequently measured markers, appearing five and four studies, respectively. All three of these markers were significantly downregulated in both AIs and SERDs; Ki67 alone was downregulated in SERMs. Less commonly assessed markers including pS6, pGSH3B, FSH and IGF1 were downregulated while CD34, pAKT and SHBG were significantly upregulated. There were no significant changes in the other biomarkers measured such as phosphate and tensin homolog (PTEN), Bax and Bcl-2.WoO studies have been widely utilised within the ER+ breast cancer subtype, demonstrating their worth in pharmacodynamic research. However, research remains focused upon routinely measured biomarkers such ER PR and Ki67, with an array of less common markers sporadically used.

Molecular biomarker expression within window of opportunity studies for oestrogen receptor-positive breast cancer

Introduction: Window of opportunity trials allow the opportunity to demonstrate pharmacodynamic parameters of a drug in vivo and are increasingly used in the context of breast cancer. Most breast cancer tumours are oestrogen receptor-positive (ER+), and multiple treatment options exist for this tumour subtype. The aim of this systematic review was to collate window of opportunity trials pertaining to the pharmacodynamic activity of drugs available for use in oestrogen receptor-positive breast cancer.

Molecular Hallmarks of Ischemia with Non-Obstructive Coronary Arteries: The "INOCA versus Obstructive CCS" Challenge.

Up to 4 million patients with signs of myocardial ischemia have no obstructive coronary artery disease (CAD). The absence of precise guidelines for diagnosis and treatment in non-obstructive CAD encourages the scientific community to fill the gap knowledge, to provide non-invasive and less expensive diagnostic tools. The aim of our study was to explore the biological profile of Ischemia with Non-Obstructive Coronary Arteries (INOCA) patients with microvascular dysfunction compared to patients presenting with obstructive chronic coronary syndrome (ObCCS) in order to find specific hallmarks of each clinical condition. We performed a gene expression array from peripheral blood mononuclear cells (PBMCs) isolated from INOCA (n = 18) and ObCCS (n = 20) patients. Our results showed a significantly reduced gene expression of molecules involved in cell adhesion, signaling, vascular motion, and inflammation in INOCA as compared to the ObCCS group. In detail, we found lower expression of Platelet and Endothelial Cell Adhesion Molecule 1 (CD31, p < 0.0001), Intercellular Adhesion Molecule-1 (ICAM1, p = 0.0004), Tumor Necrosis Factor (TNF p = 0.0003), Transferrin Receptor (TFRC, p = 0.002), and Vascular Endothelial Growth Factor A (VEGFA, p = 0.0006) in the INOCA group compared with ObCCS. Meanwhile, we observed an increased expression of Hyaluronidase (HYAL2, p < 0.0001) in INOCA patients in comparison to ObCCS. The distinct expression of molecular biomarkers might allow an early and non-invasive differential diagnosis between ObCCS and INOCA, improving clinical management and treatment options, in the era of personalized medicine.

Preoperative ultrasound radiomics analysis for expression of multiple molecular biomarkers in mass type of breast ductal carcinoma in situ

BackgroundThe molecular biomarkers of breast ductal carcinoma in situ (DCIS) have important guiding significance for individualized precision treatment. This study was intended to explore the significance of radiomics based on ultrasound images to predict the expression of molecular biomarkers of mass type of DCIS.Methods116 patients with mass type of DCIS were included in this retrospective study. The radiomics features were extracted based on ultrasound images. According to the ratio of 7:3, the data sets of molecular biomarkers were split into training set and test set. The radiomics models were developed to predict the expression of estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2), Ki67, p16, and p53 by using combination of multiple feature selection and classifiers. The predictive performance of the models were evaluated using the area under the curve (AUC) of the receiver operating curve.ResultsThe investigators extracted 5234 radiomics features from ultrasound images. 12, 23, 41, 51, 31 and 23 features were important for constructing the models. The radiomics scores were significantly (P < 0.05) in each molecular marker expression of mass type of DCIS. The radiomics models showed predictive performance with AUC greater than 0.7 in the training set and test set: ER (0.94 and 0.84), PR (0.90 and 0.78), HER2 (0.94 and 0.74), Ki67 (0.95 and 0.86), p16 (0.96 and 0.78), and p53 (0.95 and 0.74), respectively.ConclusionUltrasonic-based radiomics analysis provided a noninvasive preoperative method for predicting the expression of molecular markers of mass type of DCIS with good accuracy.

Morphological and molecular alterations of reactive astrocytes without proliferation in cerebral cortex of an APP/PS1 transgenic mouse model and Alzheimer's patients

Astrocytes are fundamental for maintaining brain homeostasis and are commonly involved in the progression of neurodegenerative diseases including Alzheimer's disease (AD). In response to injury or toxic material, astrocytes undergo activation that results in hypertrophy and process ramification. Although numerous studies have shown that reactive astrocytes are intimately related to the pathogenesis of AD, their characteristic features including morphological and molecular alterations that occur during different stages of AD progression remain to be elucidated. Here, we crossed astrocyte-specific reporter mice hGFAP-CreERT2;Rosa-tdTomato with APP/PS1 mice, and then used genetic tracing to characterize the morphological profiles and expression of molecular biomarkers associated with progressive β-amyloid deposits in the cortical region of AD mice. Expression of glutamine synthetase (GS) was lower in cortical reactive astrocytes, in contrast to the higher expression of glial fibrillary acidic protein, of APP/PS1 mice and AD patients relative to that in cortical astrocytes of wild-type mice and age-matched controls, respectively. GS activity was also decreased obviously in the cortex of APP/PS1 mice at 6 and 12 months of age relative to that in the wild-type mice of the same ages. Furthermore, cortical reactive astrocytes in APP/PS1 mice and AD patients did not undergo proliferation. Finally, based on RNA-sequencing analysis, we identified differentially expressed transcripts of signal transduction molecules involved in early induction of reactive astrocytes in the cortex of APP/PS1 mice. These findings provide a morphological and molecular basis with which to understand the function and mechanism of reactive astrocytes in the progression of AD.

Biomarkers in wound drainage fluids of head and neck squamous cell carcinoma patients receiving neck dissection: A pilot study

AimIn a pilot prospective study, we aimed to test the feasibility and report on the preliminary results on the expression of molecular biomarkers in wound drainage fluids (WDFs) of operated head and neck squamous cell carcinoma (HNSCC) patients. Material and methodsNineteen patients undergoing primary tumor resection with en-block neck dissection were enrolled. In postoperative days 1–3, the expression of several biomarkers in WDFs was measured using enzyme-linked immunosorbent assay (ELISA) kits and correlated with clinical and histopathologic features. ResultsThe expression of stromal cell-derived factor 1 (CXCL-12) was significantly increased in WDFs in presence of lymph node metastases, extranodal extension (ENE), and in case of close resection margins. In addition, Osteopontin expression was significantly increased in presence of ENE, whereas transforming growth factor beta (TGF-β) detection was significantly reduced. At multivariate analysis, CXCL-2 levels in both day 1 and 3 post-surgery were the only factor which retained significance in the prediction of close surgical margins (p = 0.028 and 0.025 for day 1 and day 3, respectively). Both CXCL-2 and Ostepontin assays were significantly correlated with ENE (p = 0.018 and 0.035 for day 1; 0.052 and 0.025 for day 3, respectively) whereas TGF- β expression was significant at day 1 only (p = 0.038) ConclusionsOur pilot study showed that WDFs could qualify as a potential source of relevant postoperative information. Further studies are needed to confirm the prognostic impact of CXCL-12, Osteopontin and TGF-β expressed in WDFs on the personalized management of HNSCC.

Dietary Tomato, but Not Lycopene Supplementation, Impacts Molecular Outcomes of Castration-resistant Prostate Cancer in the TRAMP Model (P05-015-19)

ObjectivesThe objective was to determine whether dietary tomato or lycopene supplementation impacted the expression of biomarkers associated with aggressive tumors and poorer prognostic outcomes in castration-resistant prostate cancer (CRPC). MethodsTransgenic adenocarcinoma of the mouse prostate (TRAMP) mice were castrated at 12 weeks of age to mimic the effects of androgen deprivation therapy (ADT) leading to CRPC. Mice were separated into three dietary groups (n = 30/group) and fed powdered, modified AIN-93 G diets containing 10% tomato powder (TP), lycopene beadlets matched for lycopene content of TP (LYCO), or placebo beadlets (PLAC). Tumors were detected and monitored for growth by ultrasound scan. After 5 weeks of CRPC growth, mice were euthanized, and prostate and tumor tissues were collected. Histological analysis was used to identify presence of tumor neuroendocrine (NE) phenotype (by synaptophysin [syp] expression) and androgen receptor (AR) expression. ELISA and Western blot were used to quantify interleukin-6 (IL-6) and STAT3, and RT-PCR was used to measure downstream targets of both AR and STAT3. ResultsNo differences were observed between AR expression or presence of the NE phenotype between dietary groups. However, lower tumor weight was associated with both AR and syp expression in mice fed TP. A positive correlation (P = 0.03) was observed between reduced tumor IL-6 (P = 0.012) and reduced phosphorylation (activation) of STAT3 (P = 0.017) in tumors of mice fed TP when compared to PLAC or LYCO. RNA analysis showed reduced expression of genes related to invasion (mmp-2; P = 0.006), cell proliferation (cdk1; P = 0.01), NE phenotype (ngfr; P = 0.001), and androgen metabolism (srd5a2; P = 0.03) in tumors of TP-fed mice. ConclusionsPhysiological levels of dietary tomato, but not lycopene supplementation, reduced the expression of molecular biomarkers typically upregulated in CRPC tissues that are associated with tumor growth and metastasis. Funding SourcesThis research is supported by the National Institute of Biomedical Imaging and Bioengineering of the National Institutes of Health.

Early Alterations in the Expressions of MCTS and HIF‐1α in Experimental Diabetes Mellitus

AimTo evaluate alterations due to the establishment of experimental diabetes mellitus (DM) alloxan‐induced in biochemical parameters and the expression of genes MCT1, MCT4, CD147 and HIF‐1α in different tissues and in the blood profile.MethodsWistar rats (120–180g) were conditioned at the FMABC bioterium in conditions of 12 hours light/dark, climate temperature (21 ± 2 °C) and ad libitum supply of water and food. Animals of the DM group were treated with Alloxan monohydrate (120 mg/kg) intraperitoneal (ip); the control group sham (NDS) received injections of sodium chloride 0,9% (ip). The glycemic values were measured weekly during the period of treatment through a commercial glycosimeter by puncturing of the caudal vein of the animal; animals with glycemia above ≥200 mg / dL were determined as diabetic. Evaluated the biochemical parameters: glycemia, blood creatinine and urea, and the expression of molecular biomarkers in the heart, brain, kidney and blood by the RT‐qPCR technique. Data was analyzed by GraphPad Prism® 6.0, statistical differences were observed by the Mann‐Whitney test and Student T test with an established significance level in 5% (descriptive value of p &lt;0.05).ResultsThe biochemical parameters found were: glycemia (DM7 619.1 ± 140.7 * p &lt;0.005, n = 10 vs NDS 192.5 ± 43.64, n = 4), blood creatinine (DM7 0.46 ± 0.05 * p &lt;0.005, n = 10 vs NDS 0.40 ± 0.07, n = 4) and urea (DM7 96.7 ± 26.6 * p &lt;0.005, n = 10 vs. NDS 46.5 ± 7.0, n = 4) these values follow the established standards for the diabetic profile in the present study. In the molecular analyzes an increase of CD147 in the brain was observed (DM7 26.72 ± 27.45, * p &lt;0.005, n = 9 vs NDS 1,489 ± 1,648, n = 4). In the blood profile an enhancement in the expressions of all the genes studied in the animals with DM were found; MCT1 (DM7 3,739 ± 3,057 * p &lt;0.005, n = 10 vs NDS 0.009745 ± 0.007489, n = 4); MCT4 (DM7 7.364 ± 7.3, * p &lt;0.005 n = 10 vs NDS 0.06399 ± 0.04844, n = 3); CD147 (DM7 4,295 ± 4,424 * p &lt;0.005, n = 10 vs NDS 0.2212 ± 0.01653, n = 4) and HIF‐1α (DM7 1,413 ± 1,087 * p &lt;0.005, n = 10 vs NDS 0.003833 ± 0.003741, n = 4). A positive correlation between glucose and MCT1 in the blood profile (r = 0.8061, * p &lt;0.05, n = 10) was also noted, Spearman test.ConclusionsPreliminary data demonstrate that even early DM promotes serious biochemical changes accompanied by modifications in the expression of genes involved with glycolytic metabolism and in the cellular hypoxia mechanism. Apparently, the brain is the first tissue affected by these changes.Support or Funding InformationCAPES and CNPqThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Histo-genomics: digital pathology at the forefront of precision medicine.

The toughest challenge OMICs face is that they provide extremely high molecular resolution but poor spatial information. Understanding the cellular/histological context of the overwhelming genetic data is critical for a full understanding of the clinical behavior of a malignant tumor. Digital pathology can add an extra layer of information to help visualize in a spatial and microenvironmental context the molecular information of cancer. Thus, histo-genomics provide a unique chance for data integration. In the era of a precision medicine, a four-dimensional (4D) (temporal/spatial) analysis of cancer aided by digital pathology can be a critical step to understand the evolution/progression of different cancers and consequently tailor individual treatment plans. For instance, the integration of molecular biomarkers expression into a three-dimensional (3D) image of a digitally scanned tumor can offer a better understanding of its subtype, behavior, host immune response and prognosis. Using advanced digital image analysis, a larger spectrum of parameters can be analyzed as potential predictors of clinical behavior. Correlation between morphological features and host immune response can be also performed with therapeutic implications. Radio-histomics, or the interface of radiological images and histology is another emerging exciting field which encompasses the integration of radiological imaging with digital pathological images, genomics, and clinical data to portray a more holistic approach to understating and treating disease. These advances in digital slide scanning are not without technical challenges, which will be addressed carefully in this review with quick peek at its future.

Modulation of Molecular Biomarker Expression in Response to Chemotherapy in Invasive Ductal Carcinoma.

Breast cancer (BC) has varied morphological and biological features and is classified based on molecular and morphological examinations. Molecular classification of BC is based on biological gene-expression profiling. In this study, biomarker modulation was assessed during BC treatment in 30 previously untreated patients. Heterogeneity among patients was pathologically diagnosed and classified into luminal and basal-like immunohistochemical profiles based on estrogen, progesterone, and human epidermal growth factor receptor (ER/PR/HER2) status. Marker heterogeneity was compared with mRNA biomarker expression in patients with BC before and after therapy. Reverse transcription-polymerase chain reaction was performed for molecular characterization. Expression and modulation of biological markers, CK19, hMAM, CEA, MUC, Myc, Ki-67, HER2/neu, ErbB2, and ER, were assessed after treatment, where the expression of the biomarkers CK19, Ki-67, Myc, and CEA was noted to be significantly decreased. Marker expression modulation was determined according to different stages and pathological characteristics of patients; coexpression of three markers (CK19, Ki-67, and Myc) was specifically modulated after therapy. In the histopathologically classified basal-like group, two markers (CK19 and Ki-67) were downregulated and could be considered as diagnostic biomarkers. In conclusion, pathological characteristics and marker variation levels can be evaluated to decide a personalized treatment for patients.

[Clinical significance of targeting drug-based molecular biomarkers expression in ovarian clear cell carcinoma].

Objective: To assess the expression level of targeting drug-based molecular biomarkers in ovarian clear cell carcinoma (OCCC) tissues and its clinical significance. Methods: A total of 63 OCCC patients included 40 primary OCCC and 23 recurrent OCCC for secondary cytoreductive surgery (SCS), who had received primary surgeries at Fudan University Shanghai Cancer Center between January, 2008 and December, 2015 were enrolled, and immunohistochemistry SP method was used to test human epidermal growth factor receptor (EGFR), human epidermal growth factor receptor-2 (HER2), aurora kinase A (AURKA), breast cancer susceptibility gene 1 (BRCA1), BRCA2 and programmed death-ligand 1 (PD-L1)protein expression in paraffin-embedded tissues. Results: The positive rates of EGFR, HER2, AURKA,BRCA1, BRCA2 and PD-L1 in primary and recurrent tumor tissues were respectively 20% (8/40) vs 30% (7/23) , 22% (9/40) vs 35% (8/23) , 38% (15/40) vs 35% (8/23) , 42% (17/40) vs 39% (9/23) , 20% (8/40) vs 22% (5/23) , 25% (10/40) vs 17% (4/23) , and there were no significant differences between primary and recurrent OCCC (all P>0.05). χ(2)-test or Fisher exact analysis revealed that HER2 expression in recurrent tumor tissues had a relationship with chemoresistance (P<0.05), while the expression of other biomarkers showed no significant relationship with chemoresistance (all P>0.05). Further, Kaplan-Meier survival analysis showed that patients with HER2 and AURKA-positive expression had a significantly shorter progression-free survival time in primary OCCC (4 months vs 10 months, log-rank test, P<0.05 for HER2; and 4 months vs 10 months, P<0.05 for AURKA); and a shorter overall survival time after SCS in recurrent OCCC (10 months vs 44 months, P<0.05 for HER2; and 13 months vs 43 months, P<0.05 for AURKA). However, multivariate Cox proportional hazards regression analysis indicated that none of these 6 biomarkers was independent risk factor of progression-free survival time of primary OCCC or overall survival time after SCS for recurrent OCCC (P>0.05). Conclusion: HER2 and AURKA could serve as prognostic factors in ovarian clear cell carcinoma.