Frontiers in bioscience (Landmark edition)最新文献

Background: Accurate biodosimetry is essential for effective radiological triage, precise clinical monitoring, and assessment of risks from diagnostic exposures. Immunofluorescent detection of phosphorylated histone H2AX (γH2AX) and p53-binding protein 1 (53BP1) DNA double-strand break repair foci provides high sensitivity for radiation dose assessment within the first hours after exposure. However, inter-laboratory reproducibility of γH2AX/53BP1-based biodosimetry remains limited, and the contribution of pharmacological modifiers is unresolved.

Methods: Here, we conducted an inter-laboratory comparison of in vitro radiation dose-response relationships of foci yields measured in cryopreserved umbilical cord blood lymphocytes (UCBLs) and freshly isolated peripheral blood lymphocytes (PBLs) from healthy donors using fluorescent microscopy with emphasis on workflow harmonization and reproducibility across laboratories.

Results: Under low-dose γ irradiation, both UCBLs and PBLs exhibited a strong linear, dose-dependent induction of γH2AX, 53BP1, and co-localized foci, with co-localization emerging as the most sensitive endpoint. γH2AX pan-nuclear staining was observed exclusively in UCBLs and functioned as a distinct endpoint under the examined conditions. Under harmonized low-dose conditions, calyculin A at a non-toxic concentration of 1 nM did not provide measurable stabilization or enhancement of ionizing radiation-induced foci (IRIF) yields. Although IRIF yields differed between the two laboratories, dose-response slopes were highly concordant, demonstrating reproducibility under harmonized experimental conditions.

Conclusions: These findings demonstrate that inter-laboratory reproducibility of γH2AX/53BP1-based biodosimetry is achieved primarily through disciplined workflow harmonization rather than through pharmacological enhancement. By reinforcing assay reproducibility and biological consistency, this work supports the translational applicability of IRIF-based biodosimetry for broader application in radiation exposure assessment.

Colorectal cancer (CRC) is a globally prevalent malignancy with rising incidence and mortality rates over the past decades. N6-methyladenosine (m6A) is the most abundant internal RNA modification in eukaryotes, and plays a pivotal role in post-transcriptional regulation. m6A is dynamically modulated by three core components, namely methyltransferases (writers), demethylases (erasers), and binding proteins (readers), which together govern the transcription, processing, translation, decay, and stability of mRNA. There has been accumulating evidence for the association of dysregulated m6A modification with CRC pathogenesis, metastasis, and therapeutic resistance. This review summarizes the biogenesis of m6A modification and its regulatory mechanisms, and discusses the dysregulation of m6A-related factors in CRC and the functional impacts. Most importantly, the review highlights the key roles of m6A modification in mediating CRC resistance to chemotherapy, targeted therapy, and immunotherapy. These insights may facilitate the development of novel therapeutic strategies for CRC.

Background: Acute myeloid leukemia (AML) is a hematological malignancy of the myeloid lineage with poor clinical outcomes due to limited targeted therapies. This study elucidates the role of inositol 1,4,5-trisphosphate receptor type 2 (ITPR2) in regulating cell apoptosis by modulating mitochondrial calcium (Ca²⁺) levels and underscores the clinical significance of ITPR2.

Methods: ITPR2 expression in patients with AML compared with that of healthy controls using the quantitative real-time PCR (RT-qPCR) method. The role of ITPR2 in AML and its association with immune infiltration levels were investigated through bioinformatics analyses. ITPR2 knockdown in Tohoku Hospital Pediatrics-1 (THP-1) cells were achieved using small interfering RNA (siRNA) and 2-aminoethyl diphenylborinate (2-APB), followed by comprehensive molecular characterization employing RT-qPCR, western blotting (WB), cell counting kit-8 (CCK-8) assays, and flow cytometry.

Results: ITPR2 was validated as being highly expressed in patients with AML, and this expression correlated with risk stratification and poor prognosis. Functional enrichment analysis revealed that ITPR2 is involved in Ca²⁺ signaling pathways and mitochondrial-related biological processes, and its expression level was negatively correlated with immune infiltration levels. Knockdown of ITPR2 or inhibition of its activity with 2-APB reduced Ca²⁺ ion concentrations in both the cytoplasm and mitochondria, leading to mitochondrial dysfunction (characterized by elevated intracellular reactive oxygen species (ROS) levels, reduced mitochondrial membrane potential (MMP) levels, and reduced mitochondrial DNA copy number) and eventually AML cell apoptosis.

Conclusion: ITPR2 facilitates AML progression via the Ca²⁺-mitochondrial axis and may serve as a prognostic factor and potential therapeutic target.

Background: Malignant gliomas remain largely refractory to current therapies, in part because abnormal tumor vasculature and a disrupted blood-brain barrier limit intratumoral drug delivery and cause severe tumor-associated edema. The oncolytic adenovirus KD01 is a conditionally replicating adenovirus type 5 with a 27-bp deletion in Early Region 1A (E1A) and a truncated BH3-Interacting Domain Death Agonist (tBID) expression cassette inserted into the E3 region. This study investigated whether bevacizumab-induced vascular normalization would enhance the delivery and efficacy of KD01 in gliomas.

Methods: The oncolytic activity and mitochondrial effects of KD01 were evaluated in human glioma cell lines using cell viability assays, JC-1 staining, quantitative real-time-polymerase chain reaction (qRT-PCR), and western blotting for BID/tBID. An orthotopic LN229 nude mouse model was used to assess a sequential bevacizumab→KD01 regimen. Mice were randomized to receive PBS, bevacizumab, KD01, or combination treatment. Body weight and survival were recorded. Tumor cell proliferation (Ki-67), tumor vasculature (CD31), brain water content (ΔWater%), serum biochemistry, coagulation parameters, and organ weights were analyzed to evaluate antitumor activity, edema, and systemic safety.

Results: KD01 induced robust dose- and time-dependent cytotoxicity in glioma cells and caused marked mitochondrial depolarization, accompanied by increased BID mRNA expression, loss of full-length BID, and accumulation of tBID. In the orthotopic LN229 model, bevacizumab administered 48 h before KD01 significantly improved overall outcomes compared with either monotherapy. The bevacizumab→KD01 group showed improved preservation of body weight, pronounced prolongation of survival, and the lowest Ki-67 labeling index. This group also exhibited reduced brain water content (ΔWater%), consistent with sparser CD31-positive vessels resulting from vascular normalization and oncolysis, indicating effective attenuation of tumor-associated edema. Serum liver and kidney function tests, platelet counts, coagulation indices, and major organ weights were comparable across treatment groups, suggesting no additional systemic toxicity associated with combination treatment.

Conclusions: KD01 exerts potent tBID-mediated mitochondrial oncolytic activity against glioma cells. When used as a priming strategy, transient vascular normalization induced by bevacizumab enhanced the intratumoral efficacy of KD01 in an orthotopic glioma model while maintaining a favorable safety profile. These findings support a simple, sequence-dependent combination approach integrating anti-VEGF therapy with oncolytic virotherapy for the treatment of malignant gliomas.

Background: The metabolic profile of cancer includes changes in energy metabolism and biosynthetic (plastic) metabolism, and redox balance of tumor cells. This study aimed to identify clinically significant salivary metabolic features associated with breast cancer phenotypes.

Methods: This study included 660 patients with breast cancer (age 54.6 ± 12.7 years) and 127 healthy volunteers (49.3 ± 14.2 years). Saliva samples were collected from all participants, strictly before the initiation of treatment, and the biochemical composition of saliva was determined, including indicators of antioxidant system activity, lipid profile, cytokines, and free amino acids.

Results: Salivary metabolic features correlated with the breast cancer phenotype. In particular, for luminal A breast cancer, which has the most favorable prognosis, the presence of an active inflammatory process in saliva (C-reactive protein +136.6%, p < 0.0001; IL-1β +317.7%, p = 0.0004) and a pronounced immune anti-inflammatory response (INF-γ +79.1%, p = 0.0004) were shown. In contrast, for triple-negative breast cancer, a low anti-inflammatory response (INF-γ -4.1%) and active cell proliferation (glutamine +45.0%, p = 0.0342) were shown, which correlated with the disease severity, low immunogenicity, and the least favorable prognosis for this subtype of breast cancer.

Conclusions: Overall, salivary composition reflects systemic metabolic changes in breast cancer, which makes it possible to construct a metabolic portrait of breast cancer across distinct phenotypes.

Hepatocellular carcinoma (HCC) is a leading cause of cancer-related mortality worldwide, owing to its molecular complexity and limited therapeutic options. Three-dimensional (3D) in vitro models more accurately recapitulate in vivo conditions than traditional two-dimensional (2D) models, due to their ability to accurately reproduce the tumor microenvironment (TME). Among these models, HepG2 cell-derived spheroids have become an important tool for drug screening, toxicity assessment, and liver cancer research. This review highlights the advantages and limitations of currently available 3D culture systems. In particular, special attention is given to the multifaceted role of leucine-rich repeat kinase 2 (LRRK2), a gene traditionally associated with neurological disorders but increasingly implicated in cancer, a kinase, which emerges as a promising therapeutic target in HCC since it regulates oxidative stress, lipid metabolism, and treatment responses, all of which contribute to tumor progression. Finally, we explore future directions, including organ-on-chip technologies and co-culture systems, which hold considerable promise for improving precision medicine and translational research in HCC.

The pathogenesis of multiple sclerosis (MS) is a complex topic that involves many individual aspects, several MS risk factors and various alterations in the immune system. In addition to the aspects still being researched in MS today, such as the activation of pro-inflammatory lymphocytes and macrophages, a particular scientific interest refers to the dysregulation of iron and to ferroptosis as an important form of programmed cell death (PCD), which is closely associated with this dysregulation. These two aspects are particularly significant because the new McDonald criteria include the diagnostic determination of so-called "paramagnetic rim lesions" (PRLs) in MS, which are areas of abnormal iron accumulation visible on magnetic resonance imaging (MRI) scans. Along with other diagnostic criteria, these PRLs are an important criterion for the diagnosis of MS. This review begins with an overall description of pathogenic risk factors and the dysregulation of the immune system. However, the priority of the review is the dysregulation of the cerebral iron balance and ferroptosis, as well as other forms of programmed cell death, which have also been demonstrated in MS: apoptosis, pyroptosis, necroptosis and NETosis. One of the main current findings on the forms of PCD in MS is their non-physiological occurrence in the brain's own cells such as oligodendrocytes and neurons, and the extensive protection of pro-inflammatory cells from the occurrence of PCD. Emerging evidence indicates that ferroptosis occurs at an early stage in MS and appears to play a role in its pathogenesis. This review will present the distinct characteristics of the various PCDs in MS, alongside other pathological mechanisms, genetic factors, and current experimental approaches of therapeutic concepts that affect especially ferroptosis and pyroptosis. The aim is to provide an overview on iron dysregulation and forms of PCD as key aspects for understanding the pathogenesis of MS.

The dynamic evolution of signaling pathway remodeling and molecular regulation within the tumor microenvironment (TME) of hepatocellular carcinoma (HCC) play a critical role in the onset and progression of this malignancy. As chronic hepatitis progresses to cirrhosis and ultimately to HCC, the signaling pathways and TME show stage-specific characteristics that provide important insights into the therapeutic challenges and opportunities. In this review, we profiled the principal components of the HCC TME, along with pivotal signaling pathways, including the receptor tyrosine kinase (RTK) and extracellular signal-regulated kinase (ERK) pathways. Furthermore, we characterized the dynamic transformation of the TME from an inflammatory state in the hepatitis phase to a fibrotic state in the cirrhosis phase. Ultimately, we assessed the therapeutic potential of current HCC targets emphasizing emerging strategies for precision and personalized treatment.

Background: ​ Altered homeostasis of the ocular surface microenvironment is a hallmark of dry eye disease (DED). The alpha-7 nicotinic acetylcholine receptor (α7nAChR) plays a key role in DED pathophysiology. In this study, we established a rabbit model of DED using scopolamine hydrobromide (Scop) to determine the effect of electroacupuncture (EA) on ocular surface damage in DED and to explore its underlying mechanisms.​ Methods: New Zealand White rabbits (1.5-2.0 kg) were subcutaneously administered Scop for 21 days prior to EA treatment. After 35 days, the homeostasis of the ocular surface microenvironment was evaluated using the Schirmer I test (SIT), tear break-up time (BUT), corneal fluorescein (FL) staining, and measurement of tear osmolarity. The expression levels of ACh, α7nAChR, and high mobility group box 1 (HMGB1) were detected via histopathological examination of the cornea, lacrimal glands, and conjunctiva, combined with immunohistochemistry and western blotting. Additionally, protein chip technology was used to determine the expression levels of downstream factors.​ Results: EA stimulation significantly improved the homeostasis of the ocular surface microenvironment, as evidenced by increased SIT values and BUT, reduced corneal FL intensity, and decreased tear osmolarity. It also alleviated pathological damage to the cornea, conjunctiva, and lacrimal glands; upregulated the expression of ACh and α7nAChR; and downregulated the expression of HMGB1 and related inflammatory factors. However, these changes were reversed following administration of α-Bungarotoxin.​ Conclusion: EA stimulation improves ocular surface homeostasis and reduces inflammation in DED, potentially via activation of the α7nAChR signaling pathway, which in turn inhibits the expression of HMGB1 and inflammatory factors.​.