Abstract Diagnostic delays in lung cancer compromise patient survival. This study aims to develop a deep learning-based framework that integrates rapid on-site evaluation (ROSE) cytomorphology and serum tumor markers to enable accurate pre-pathological subtyping, thereby facilitating earlier treatment initiation. A dataset of 156 matched cases with both ROSE cytology images and five serum biomarkers (squamous cell carcinoma antigen (SCCA), pro-gastrin-releasing peptide (ProGRP), carcinoembryonic antigen (CEA), neuron-specific enolase (NSE), cytokeratin-19 fragment (CYFRA21-1)) was retrospectively analyzed. Two deep learning models were developed: (1) a ROSE Image-Only Model (RIOM) using ResNet-50 with spatial attention, and (2) a ROSE-Serum Marker Model (RSMM) incorporating cross-modal feature alignment between ROSE images and serum biomarkers. A consensus strategy was implemented to stratify patients based on prediction agreement between the two models. Manual ROSE assessment achieved 84.0% overall diagnostic accuracy. The RIOM matched this performance (84.5% accuracy), while the multimodal RSMM significantly surpassed both, achieving 91.6% accuracy in five-class classification (Benign, lung adenocarcinoma (LUAD), lung squamous cell carcinoma (LUSC), large cell lung carcinoma (LCLC), small cell lung cancer (SCLC)). The consensus strategy identified 80.6% of cases as prediction-consistent, for which the accuracy for malignancy determination, non-small cell lung cancer (NSCLC) discrimination, and final subtype classification reached 98.4%. The proposed multimodal decision-support framework provides a rapid and reliable tool for pre-pathological lung cancer subtyping. By enabling high-confidence diagnoses immediately after biopsy, this approach could rationalize clinical workflows through risk-stratified management and significantly accelerate reflex molecular testing and therapy initiation, particularly for the majority of patients with prediction-consistent results.
Abstract Immune checkpoint blockade (ICB) therapy only induces durable responses in a subset of cancer patients. The underlying mechanisms of such selective efficacy remain largely unknown. By analyzing the expression profiles of immune checkpoint molecules in different statuses of murine tumors, we found that tumor progression generally randomly upregulated multiple immune checkpoints, thus increased the Heterogeneity of Immune checkpoint Signature (HIS) and resulted in immunotherapeutic resistance. Interestingly, overexpressing one pivotal immune checkpoint in a tumor hindered the upregulation of a majority of other immune checkpoint genes during tumor progression via suppressing interferon γ, resulting in HIS-low. Indeed, PD-L1 high-expression sensitized baseline large tumors to anti-PD1 therapy without altering the sensitivity of baseline small tumors. In line with these preclinical results, a retrospective analysis of a phase III study involving patients with non-small cell lung cancer (NSCLC) revealed that PD-L1 tumor proportion score (TPS) ≥ 50% more reliably predicted therapeutic response in NSCLC patients with baseline tumor volume (BTV)-large compared to patients with BTV-small. Notably, TPS combined with BTV significantly improved the predictive accuracy. Collectively, the data suggest that HIS reflects the dynamic features of tumor immune evasion and dictates the selective efficacy of ICB in a tumor size-dependent manner, providing a potential novel strategy to improve precision ICB. These findings highlight the application of ICB to earlier stages of cancer patients. The integration of PD-L1 with BTV may immediately improve patient stratification and prediction performance in the clinic.
Karen Diaz-Palacios, Pilar López Navajas, Bárbara Rodrigo Martín
et al.
Abstract Focal Adhesion Kinase (FAK) is a key signaling molecule in focal adhesions (FAs) orchestrating the formation, maturation and turnover of the FA complex. A controlled FA lifecycle is essential for various cellular processes requiring mesenchymal cell migration and is harnessed by advanced cancers to initiate cancer invasion and metastasis. The mechanical force for migration is transmitted from actin stress fibers to FAs via specialized force transduction components in FAs. These forces are known to activate FA signaling, suggesting a communication between FA force transduction and FA signaling components, yet how this occurs mechanistically is unknown. Here we demonstrate that paxillin can act as an adaptor protein to connect FAK with the force transduction component vinculin. Our data show that this connection forms inefficient in the basal state but suggest Y925 phosphorylation in FAK as a key mechanism for optimal formation of the FAK:paxillin:vinculin linkage. This is achieved by switching binding of the paxillin LD2 motif from FAK to vinculin while keeping paxillin LD4 tethered to FAK. We further provide the first high-resolution crystal structure of LD2 bound to the vinculin tail domain, which importantly shows that vinculin can simultaneously link to actin. This therefore ensures an intact force transduction role of vinculin while tethered via paxillin to the signaling apparatus in FAs. With this data, all interactions of the force transmitting tether to FAK are structurally defined and we provide an atomic model for FAK force activation. In summary, we propose a phospho-regulated connection between signaling and force transduction components in FAs allowing for force induced activation of FA signaling.
Rana Al-Awadhi, Moody AlRoomy, Hawraa Dashti
et al.
Context:
Persistent infection with high-risk human papillomavirus (hr-HPV) types is associated with high-grade cervical abnormalities.
Aims:
The aim of the study was to find most hr-HPV types causing persistent infection in abnormal cytological samples using Aptima HPV testing and discuss the compatibility of the Gardasil 9 vaccine in targeting most types.
Settings:
The study was conducted in a cytology laboratory in a tertiary hospital.
Design:
This is a retrospective observational study.
Methods and Materials:
Cytology and HPV Aptima test reports were obtained for abnormal cervical samples for a 6-year period.
Statistical Analysis Used:
Pearson Chi-square test.
Results:
Reports of 2834 abnormal cervical samples were analyzed. Aptima testing was positive in 21% of samples, including 92% of squamous cell carcinoma (SCC), 76.4% of the high-grade squamous intraepithelial lesion (HSIL), 52% of low-grade squamous intraepithelial lesion (LSIL), 40% of adenocarcinoma (ADC), and 21% of atypical squamous cells that cannot exclude HSIL (ASC-H). The Aptima other hr-HPV group was most common (60%), HPV16 was 26%, HPV18/45 was 9.6%, and double HPV infection was 4.3%. HPV16 was the most common infection in HSIL+ cases. HPV infection was most common in age groups (30–39) and (40–49), and a shift to age groups (50–59) and ≥60 was seen in HSIL+ cases.
Conclusions:
This study is the first of its kind in correlating age with hr-HPV and cytology findings in the Middle East and adds to previous knowledge related to the prevalence and Aptima testing of HPV. The outcome could be used as a baseline for the Gardasil 9 vaccine and for the assessment of its effectiveness after three or five years from initiation.
Abstract DNA damage resulting from genotoxic injury can initiate cellular senescence, a state characterized by alterations in cellular metabolism, lysosomal activity, and the secretion of factors collectively known as the senescence-associated secretory phenotype (SASP). Senescence can have beneficial effects on our bodies, such as anti-cancer properties, wound healing, and tissue development, which are attributed to the SASP produced by senescent cells in their intermediate stages. However, senescence can also promote cancer and aging, primarily due to the pro-inflammatory activity of SASP. Studying senescence is complex due to various factors involved. Genotoxic stimuli cause random damage to cellular macromolecules, leading to variations in the senescent phenotype from cell to cell, despite a shared program. Furthermore, senescence is a dynamic process that cannot be analyzed as a static endpoint, adding further complexity. Investigating SASP is particularly intriguing as it reveals how a senescence process triggered in a few cells can spread to many others, resulting in either positive or negative consequences for health. In our study, we conducted a meta-analysis of the protein content of SASP obtained from different research groups, including our own. We categorized the collected omic data based on: i) cell type, ii) harmful agent, and iii) senescence stage (early and late senescence). By employing Gene Ontology and Network analysis on the omic data, we identified common and specific features of different senescent phenotypes. This research has the potential to pave the way for the development of new senotherapeutic drugs aimed at combating the negative consequences associated with the senescence process. Video Abstract
Cancer, while a multifactorial chronic disease with an increasing prevalence, has been the subject of intense investigation, not only because of the growing need to find the main triggers that motivate its onset but essentially because of the need to discover increasingly safer and effective therapeutic options that have fewer adverse effects and associated toxicity [...]
Mirco Masi, Alessandro Attanzio, Marco Racchi
et al.
The role of protein misfolding, deposition, and clearance has been the dominant topic in the last decades of investigation in the field of neurodegeneration. The impairment of protein synthesis, along with RNA metabolism and RNA granules, however, are significantly emerging as novel potential targets for the comprehension of the molecular events leading to neuronal deficits. Indeed, defects in ribosome activity, ribosome stalling, and PQC—all ribosome-related processes required for proteostasis regulation—can contribute to triggering stress conditions and promoting the formation of stress granules (SGs) that could evolve in the formation of pathological granules, usually occurring during neurodegenerating effects. In this review, the interplay between proteostasis, mRNA metabolism, and SGs has been explored in a neurodegenerative context with a focus on Alzheimer’s disease (AD), although some defects in these same mechanisms can also be found in frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS), which are discussed here. Finally, we highlight the role of the receptor for activated C kinase 1 (RACK1) in these pathologies and note that, besides its well characterized function as a scaffold protein, it has an important role in translation and can associate to stress granules (SGs) determining cell fate in response to diverse stress stimuli.
Abstract Background Numerous studies have revealed that long noncoding RNAs (lncRNAs) are closely related to the development of many diseases and carcinogenesis. However, their specific biological function and molecular mechanism in oesophageal squamous cell carcinoma (ESCC) remains unclear. Methods RNA-Seq was performed to determine the differential expressions of lncRNAs in ESCC, and the level of SNHG16 expression was detected in ESCC and intraepithelial neoplasia (IEN) samples. In vitro and in vivo experiments were performed to explore the role of SNHG16 and the interaction of EIF4A3 and Ras homologue family member U (RhoU) signalling. Results One hundred and seventy-five upregulated and 134 downregulated lncRNAs were identified by RNA-Seq. SNHG16 was highly expressed in ESCC and intraepithelial neoplasia (IEN) samples, and its expression level was correlated with tumour differentiation and T stage. Overexpression of SNHG16 can facilitate ESCC cell proliferation and metastasis. Mechanistically, we noticed that SNHG16 could bind RNA binding protein (RBP)-eukaryotic translation initiation factor (EIF4A3) and interact with it to form a complex. Importantly, the coalition of SNHG16 and EIF4A3 ultimately regulated Ras homologue family member U (RhoU). SNHG16 modulated RhoU expression by recruiting EIF4A3 to regulate the stability of RhoU mRNA. Knockdown of RhoU further alleviated the effect of the SNHG16 oncogene in ESCC cells. Conclusions The newly identified SNHG16–EIF4A3–RhoU signalling pathway directly coordinates the response in ESCC pathogenesis and suggests that SNHG16 is a promising target for potential ESCC treatment.
Michela Terlizzi, Chiara Colarusso, Pasquale Somma
et al.
Sphingosine-1-phosphate (S1P) is involved in inflammatory signaling/s associated with the development of respiratory disorders, including cancer. However, the underlying mechanism/s are still elusive. The aim of this study was to investigate the role of S1P on circulating blood cells obtained from healthy volunteers and non-small cell lung cancer (NSCLC) patients. To pursue our goal, peripheral blood mononuclear cells (PBMCs) were isolated and stimulated with S1P. We found that the administration of S1P did not induce healthy PBMCs to release pro-inflammatory cytokines. In sharp contrast, S1P significantly increased the levels of TNF-α and IL-6 from lung cancer-derived PBMCs. This effect was S1P receptor 3 (S1PR3)-dependent. The pharmacological blockade of ceramidase and sphingosine kinases (SPHKs), key enzymes for S1P synthesis, completely reduced the release of both TNF-α and IL-6 after S1P addition on lung cancer-derived PBMCs. Interestingly, S1P-induced IL-6, but not TNF-α, release from lung cancer-derived PBMCs was mTOR- and K-Ras-dependent, while NF-κB was not involved. These data identify S1P as a bioactive lipid mediator in a chronic inflammation-driven diseases such as NSCLC. In particular, the higher presence of S1P could orchestrate the cytokine milieu in NSCLC, highlighting S1P as a pro-tumor driver.
Valentina A. Babenko, Denis N. Silachev, Tatyana I. Danilina
et al.
The use of stem cells is part of a strategy for the treatment of a large number of diseases. However, the source of the original stem cells for use is extremely important and determines their therapeutic potential. Mesenchymal stromal cells (MSC) have proven their therapeutic effectiveness when used in a number of pathological models. However, it remains an open question whether the chronological age of the donor organism affects the effectiveness of the use of MSC. The asymmetric division of stem cells, the result of which is some residential stem cells acquiring a non-senile phenotype, means that stem cells possess an intrinsic ability to preserve juvenile characteristics, implying an absence or at least remarkable retardation of senescence in stem cells. To test whether residential MSC senesce, we evaluated the physiological changes in the MSC from old rats, with a further comparison of the neuroprotective properties of MSC from young and old animals in a model of traumatic brain injury. We found that, while the effect of administration of MSC on lesion volume was minimal, functional recovery was remarkable, with the highest effect assigned to fetal cells; the lowest effect was recorded for cells isolated from adult rats and postnatal cells, having intermediate potency. MSC from the young rats were characterized by a faster growth than adult MSC, correlating with levels of proliferating cell nuclear antigen (PCNA). However, there were no differences in respiratory activity of MSC from young and old rats, but young cells showed much higher glucose utilization than old ones. Autophagy flux was almost the same in both types of cells, but there were remarkable ultrastructural differences in old and young cells.
Donato Cappetta, Oxana Bereshchenko, Eleonora Cianflone
et al.
Glucocorticoids (GCs) are essential in regulating functions and homeostasis in many biological systems and are extensively used to treat a variety of conditions associated with immune/inflammatory processes. GCs are among the most powerful drugs for the treatment of autoimmune and inflammatory diseases, but their long-term usage is limited by severe adverse effects. For this reason, to envision new therapies devoid of typical GC side effects, research has focused on expanding the knowledge of cellular and molecular effects of GCs. GC-induced leucine zipper (GILZ) is a GC-target protein shown to mediate several actions of GCs, including inhibition of the NF-κB and MAPK pathways. GILZ expression is not restricted to immune cells, and it has been shown to play a regulatory role in many organs and tissues, including the cardiovascular system. Research on the role of GILZ on endothelial cells has demonstrated its ability to modulate the inflammatory cascade, resulting in a downregulation of cytokines, chemokines, and cellular adhesion molecules. GILZ also has the capacity to protect myocardial cells, as its deletion makes the heart, after a deleterious stimulus, more susceptible to apoptosis, immune cell infiltration, hypertrophy, and impaired function. Despite these advances, we have only just begun to appreciate the relevance of GILZ in cardiovascular homeostasis and dysfunction. This review summarizes the current understanding of the role of GILZ in modulating biological processes relevant to cardiovascular biology.
Abstract Tumor progression requires the communication between tumor cells and tumor microenvironment (TME). Cancer-associated fibroblasts (CAFs) are major components of stromal cells. CAFs contribute to metastasis process through direct or indirect interaction with tumor cells; however, the underlying mechanism is largely unknown. Here, we reported that autophagy was upregulated in lung cancer-associated CAFs compared to normal fibroblasts (NFs), and autophagy was responsible for the promoting effect of CAFs on non-small cell lung cancer (NSCLC) cell migration and invasion. Inhibition of CAFs autophagy attenuated their regulation on epithelial–mesenchymal transition (EMT) and metastasis-related genes of NSCLC cells. High mobility group box 1 (HMGB1) secreted by CAFs mediated CAFs’ effect on lung cancer cell invasion, demonstrated by using recombinant HMGB1, HMGB1 neutralizing antibody, and HMGB1 inhibitor glycyrrhizin (GA). Importantly, the autophagy blockade of CAFs revealed that HMGB1 release was dependent on autophagy. We also found HMGB1 was responsible, at least in part, for autophagy activation of CAFs, suggesting CAFs remain active through an autocrine HMGB1 loop. Further study demonstrated that HMGB1 facilitated lung cancer cell invasion by activating the NFκB pathway. In a mouse xenograft model, the autophagy specific inhibitor chloroquine abolished the stimulating effect of CAFs on tumor growth. These results elucidated an oncogenic function for secretory autophagy in lung cancer-associated CAFs that promotes metastasis potential, and suggested HMGB1 as a novel therapeutic target.
Małgorzata Kucała, Michał Saładyga, Ariel Kaminski
Cyanotoxins are harmful to aquatic and water-related organisms. In this study, Lemna trisulca was tested as a phytoremediation agent for three common cyanotoxins produced by bloom-forming cyanobacteria. Cocultivation of <i>L. trisulca</i> with <i>Dolichospermum flos-aquae</i> in BG11 medium caused a release of the intracellular pool of anatoxin-a into the medium and the adsorption of 92% of the toxin by the plant—after 14 days, the total amount of toxin decreased 3.17 times. Cocultivation with <i>Raphidopsis raciborskii</i> caused a 2.77-time reduction in the concentration of cylindrospermopsin (CYN) in comparison to the control (62% of the total pool of CYN was associated with the plant). The greatest toxin limitation was noted for cocultivation with <i>Microcystis aeruginosa</i>. After two weeks, the microcystin-LR (MC-LR) concentration decreased more than 310 times. The macrophyte also influenced the growth and development of cyanobacteria cells. Overall, 14 days of cocultivation reduced the biomass of <i>D. flos-aquae</i>, <i>M. aeruginosa</i>, and <i>R. raciborskii</i> by 8, 12, and 3 times, and chlorophyll a concentration in comparison to the control decreased by 17.5, 4.3, and 32.6 times, respectively. Additionally, the macrophyte stabilized the electrical conductivity (EC) and pH values of the water and affected the even uptake of cations and anions from the medium. The obtained results indicate the biotechnological potential of <i>L. trisulca</i> for limiting the development of harmful cyanobacterial blooms and their toxicity.
Nardtiwa Chaivoravitsakul, Katriya Chankow, Kongthit Horoongruang
et al.
Background and Aim: Ultrasound-guided fine-needle sample collection for cytology with manual restraint is frequently used for the primary assessment of diffuse liver disease in veterinary patients in Thailand. For better diagnosis, repeated collection of samples ensures the collection of adequate, representative samples, which increase diagnostic accuracy. However, in those that are unable to receive general anesthesia, it is difficult to collect the samples from several liver locations in manually restrained dogs and cats. The study aimed to compare the cytologic diagnosis of the ultrasound-guided fine-needle non-aspiration technique between the left and right liver lobes in dogs and cats with neoplastic and non-neoplastic diffuse liver disease.
Materials and Methods: This prospective study included 25 client-owned dogs and cats with diffuse liver diseases. Two liver samples were randomly collected from the left and right liver lobes under ultrasound guidance for cytologic examination. All slides were subsequently examined blindly by experienced pathologists for cytologic analysis with cytologic agreement scores (CASs).
Results: Among all 50 samples obtained from ultrasound-guided fine-needle sample collection of the left and right liver, 78% were diagnostic and 22% were non-diagnostic. In the diagnostic group, 73.3% of fine-needle samples had concordant results between the left and right liver, which exhibited 100% cytologic agreement in lymphoma and 63.6% in non-neoplastic groups. Samples collected from the left liver had slightly higher CAS and higher cytologic quality than had those from the right liver lobe (p=0.053).
Conclusion: The location and number of sample collections did not have a significant difference in the cytologic diagnosis of diffuse liver disease, especially in patients with lymphoma. For manually restrained patients, one time ultrasound-guided non-aspiration cytology procedure from the left liver lobe not only decreased restraint duration and minimized tissue trauma but also allowed for an adequate cytologic diagnosis in diffuse liver disease compared to multiple collections.
Introduction: Palpable breast lumps are common problem in
females, which can be benign or malignant. The most common
cancer after cervical cancer is breast cancer. Fine Needle
Aspiration Cytology (FNAC) is a simple, rapid, reliable and costbeneficial procedure to make early diagnosis and planning of
treatment modalities.
Aim: To study the spectrum of various breast lesions on
cytomorphology in third decade females and correlation of
cytomorphological and clinical features.
Materials and Methods: This was a two-year prospective study
from January 2018 to January 2020 in Rama Medical College
Kanpur, including 200 FNAC samples aspirated from palpable
breast lumps. Physical examination of breast lumps was done by
palpation. Diagnosis was made based on the cytomorphological
criteria and clinical assessment for diagnosis of diseases.
Result: On cytomorphological study of 200 breast lump
aspirates in females, 52 (26%) were inflammatory lesions, 123
(61.5%) benign, 13 (6.5%) were atypical/probably benign, 02
(1%) malignant and 10 (5%) were unsatisfactory. The most
common benign lesion in the present study was fibroadenoma
60 (48.8%), and the most common malignant lesion was
medullary carcinoma 02 (100%) cases.
Conclusion: FNAC is a rapid and reliable tool to provide
effective diagnosis in palpable breast lumps. It should be used
as routine diagnostic procedure to provide early diagnosis and
treatment to the patients with breast lesions.
Roman Tikhomirov, Benedict Reilly-O’ Donnell, Francesco Catapano
et al.
Fibrosis is a significant global health problem associated with many inflammatory and degenerative diseases affecting multiple organs, individually or simultaneously. Fibrosis develops when extracellular matrix (ECM) remodeling becomes excessive or uncontrolled and is associated with nearly all forms of heart disease. Cardiac fibroblasts and myofibroblasts are the main effectors of ECM deposition and scar formation. The heart is a complex multicellular organ, where the various resident cell types communicate between themselves and with cells of the blood and immune systems. Exosomes, which are small extracellular vesicles, (EVs), contribute to cell-to-cell communication and their pathophysiological relevance and therapeutic potential is emerging. Here, we will critically review the role of endogenous exosomes as possible fibrosis mediators and discuss the possibility of using stem cell-derived and/or engineered exosomes as anti-fibrotic agents.