Integrative multi-omics analyses identify molecular subtypes of head and neck squamous cell carcinoma with distinct therapeutic vulnerabilities.

Substantial heterogeneity in molecular features, patient prognoses, and therapeutic responses in head and neck squamous cell carcinomas (HNSCC) highlights the urgent need to develop molecular classifications that reliably and accurately reflect tumor behavior and inform personalized therapy. Here, we leveraged the similarity network fusion bioinformatics approach to jointly analyze multi-omics datasets spanning copy number variations, somatic mutations, DNA methylation, and transcriptomic profiling and derived a prognostic classification system for HNSCC. The integrative model consistently identified three subgroups (IMC1-3) with specific genomic features, biological characteristics, and clinical outcomes across multiple independent cohorts. The IMC1 subgroup included proliferative, immune-activated tumors and exhibited a more favorable prognosis. The IMC2 subtype harbored activated EGFR signaling and an inflamed tumor microenvironment with cancer-associated fibroblast/vascular infiltrations. Alternatively, the IMC3 group featured highly aberrant metabolic activities and impaired immune infiltration and recruiting. Pharmacogenomics analyses from in silico predictions and from patient-derived xenograft model data unveiled subtype-specific therapeutic vulnerabilities including sensitivity to cisplatin and immunotherapy in IMC1 and EGFR inhibitors (EGFRi) in IMC2, which was experimentally validated in patient-derived organoid models. Two signatures for prognosis and EGFRi sensitivity were developed via machine learning. Together, this integrative multi-omics clustering for HNSCC improves current understanding of tumor heterogeneity and facilitates patient stratification and therapeutic development tailored to molecular vulnerabilities.

Comprehensive 3-dimensional Positional and Morphological Analyses of Condyle and Glenoid Fossa in Patients with Skeletal Class II Malocclusion following Bimaxillary Orthognathic Surgery.

This study aimed to comprehensively and quantitatively characterize 3-dimensional (3D) positional and morphological changes of the condyle and glenoid fossa in patients with skeletal Class II malocclusion treated with bimaxillary orthognathic surgery. Twenty eligible patients treated at our institution from January 2016 to December 2021 with more than 12 months of postoperative follow-up were retrospectively enrolled. Radiographic data of cone-beam computed tomography (CBCT) for each patient were collected at 3 stages: 1 week preoperatively (T0), immediately after surgery (T1), and at least 12 months postoperatively (T2). Positional changes, surface and volumetric alterations of condyle, and bone remodeling in glenoid fossa were measured and compared based on voxel- and surface registrations in visual 3D methods. Most patients exhibited a tendency for condyles to shift posteriorly, laterally, superiorly, and rotated outward, downward, and forward immediately after surgery. Posterior, medial, superior movement and outward, upward, and backward rotation of condyles were observed during follow-up (T1-T2). Bone resorption frequently occurred in the posterior area of condylar surfaces, while bone remodeling was more common in the anterior region of the glenoid fossa. Reduced volume of the condyle was found in most cases, which was not associated with the amount of mandibular advancement. Overall, the condyle and its corresponding glenoid fossa remained relatively stable during the follow-up. Our results reveal positional and morphological alterations in the condyle and the glenoid fossa after bimaxillary orthognathic surgery in patients with skeletal class II malocclusion. These changes predominantly fall within the spectrum of physical adaption.

Oral pathogen aggravates atherosclerosis by inducing smooth muscle cell apoptosis and repressing macrophage efferocytosis.

Periodontitis imparting the increased risk of atherosclerotic cardiovascular diseases is partially due to the immune subversion of the oral pathogen, particularly the Porphyromonas gingivalis (P. gingivalis), by inducing apoptosis. However, it remains obscure whether accumulated apoptotic cells in P. gingivalis-accelerated plaque formation are associated with impaired macrophage clearance. Here, we show that smooth muscle cells (SMCs) have a greater susceptibility to P. gingivalis-induced apoptosis than endothelial cells through TLR2 pathway activation. Meanwhile, large amounts of miR-143/145 in P.gingivalis-infected SMCs are extracellularly released and captured by macrophages. Then, these miR-143/145 are translocated into the nucleus to promote Siglec-G transcription, which represses macrophage efferocytosis. By constructing three genetic mouse models, we further confirm the in vivo roles of TLR2 and miR-143/145 in P. gingivalis-accelerated atherosclerosis. Therapeutically, we develop P.gingivalis-pretreated macrophage membranes to coat metronidazole and anti-Siglec-G antibodies for treating atherosclerosis and periodontitis simultaneously. Our findings extend the knowledge of the mechanism and therapeutic strategy in oral pathogen-associated systemic diseases.

Integrative Single-Cell and Bulk Transcriptomes Analyses Identify Intrinsic HNSCC Subtypes with Distinct Prognoses and Therapeutic Vulnerabilities.

PURPOSE: Tumor heterogeneity in head and neck squamous cell carcinoma (HNSCC) profoundly compromises patient stratification, personalized treatment planning, and prognostic prediction, which underscores the urgent need for more effective molecular subtyping for this malignancy. Here, we sought to define the intrinsic epithelial subtypes for HNSCC by integrative analyses of single-cell and bulk RNA sequencing datasets from multiple cohorts and assess their molecular features and clinical significance. EXPERIMENTAL DESIGN: Malignant epithelial cells were identified from single-cell RNA sequencing (scRNA-seq) datasets and subtyped on the basis of differentially expressed genes. Subtype-specific genomic/epigenetic abnormalities, molecular signaling, genetic regulatory network, immune landscape, and patient survival were characterized. Therapeutic vulnerabilities were further predicted on the basis of drug sensitivity datasets from cell lines, patient-derived xenograft models, and real-world clinical outcomes. Novel signatures for prognostication and therapeutic prediction were developed by machine learning and independently validated. RESULTS: Three intrinsic consensus molecular subtypes (iCMS1-3) for HNSCC were proposed from scRNA-seq analyses and recapitulated in 1,325 patients from independent cohorts using bulk-sequencing datasets. iCMS1 was characterized by EGFR amplification/activation, stromal-enriched environment, epithelial-to-mesenchymal transition, worst survival, and sensitivities to EGFR inhibitor. iCMS2 was featured by human papillomavirus-positive oropharyngeal predilection, immune-hot, susceptibilities to anti-PD-1, and best prognosis. Moreover, iCMS3 displayed immune-desert and sensitivities to 5-FU and MEK, STAT3 inhibitors. Three novel, robust signatures derived from iCMS subtype-specific transcriptomics features were developed by machine learning for patient prognostication and cetuximab and anti-PD-1 response predictions. CONCLUSIONS: These findings reiterate molecular heterogeneity of HNSCC and advantages of scRNA-seq in pinpointing cellular diversities in complex cancer ecosystems. Our HNSCC iCMS regime might facilitate accurate patient stratification and individualized precise treatment.

Three-dimensional quantitative changes of condyle in patients with skeletal class III malocclusion after bimaxillary orthognathic surgery with 5-year follow-up.

OBJECTIVE: The present study aimed to characterize three-dimensional (3D) long-term quantitative condyle change including positional, surface, and volumetric alterations in patients with skeletal class III malocclusion treated with bimaxillary orthognathic surgery. MATERIAL AND METHODS: Twenty-three eligible patients (9 males, 14 females, mean age: 28.28 years old) treated from Jan. 2013 to Dec. 2016 with postoperative follow-up over 5 years were retrospectively enrolled. Cone-beam computed tomography scan for each patient was conducted at 4 stages: 1 week preoperatively (T0), immediately after surgery (T1), 12 months postoperatively (T2), and 5-year postoperatively (T3). Positional changes, surface, and volumetric remodeling of condyle were measured in segmented visual 3D models and statistically compared between stages. RESULTS: Our 3D quantitative calibrations revealed that the condylar center shifted in anterior (0.23 ± 1.50 mm), medial (0.34 ± 0.99), and superior (1.11 ± 1.10 mm) directions and rotated outward (1.58 ± 3.11°), superior (1.83 ± 5.08°), and backward (4.79 ± 13.75°) from T1 to T3. With regard to condylar surface remodeling, bone formation was frequently observed in the anteromedial areas, while bone resorption was commonly detected in the anterolateral area. Moreover, condylar volume remained largely stable with a minimal reduction during the follow-up. CONCLUSION: Collectively, although condyle undergoes positional changes and bone remodeling after bimaxillary surgery in patients with mandibular prognathism, these changes largely fall in the range of physical adaptations in the long run. CLINICAL RELEVANCE: These findings advance the current understanding of long-term condylar remodeling after bimaxillary orthognathic surgery in skeletal class III patients.

circFNDC3B Accelerates Vasculature Formation and Metastasis in Oral Squamous Cell Carcinoma.

Emerging evidence has demonstrated that circular RNAs (circRNA) are involved in cancer metastasis. Further elucidation of the role of circRNAs in oral squamous cell carcinoma (OSCC) could provide insights into mechanisms driving metastasis and potential therapeutic targets. Here, we identify a circRNA, circFNDC3B, that is significantly upregulated in OSCC and is positively associated with lymph node (LN) metastasis. In vitro and in vivo functional assays showed that circFNDC3B accelerated the migration and invasion of OSCC cells and the tube-forming capacity of human umbilical vein endothelial cells and human lymphatic endothelial cells. Mechanistically, circFNDC3B regulated ubiquitylation of the RNA-binding protein FUS and the deubiquitylation of HIF1A through the E3 ligase MDM2 to promote VEGFA transcription, thereby enhancing angiogenesis. Meanwhile, circFNDC3B sequestered miR-181c-5p to upregulate SERPINE1 and PROX1, which drove epithelial-mesenchymal transition (EMT) or partial-EMT (p-EMT) in OSCC cells and promoted lymphangiogenesis to accelerate LN metastasis. Overall, these findings uncovered the mechanistic role of circFNDC3B in orchestrating cancer cell metastatic properties and vasculature formation, suggesting circFNDC3B could be a potential target to reduce OSCC metastasis. SIGNIFICANCE: Dual functions of circFNDC3B in enhancing the metastatic ability of cancer cells and promoting vasculature formation through regulation of multiple pro-oncogenic signaling pathways drive lymph node metastasis of OSCC.

Evaluation of a respiratory motion-corrected image reconstruction algorithm in 2-[18F]FDG and [68Ga]Ga-DOTA-NOC PET/CT: impacts on image quality and tumor quantification.

Background: Respiratory motions may cause artifacts on positron emission tomography (PET) images that degrade image quality and quantification accuracy. This study aimed to evaluate the effect of a respiratory motion-corrected image reconstruction (MCIR) algorithm on image quality and tumor quantification compared with nongated/nonmotion-corrected reconstruction. Methods: We used a phantom consisting of 5 motion spheres immersed in a chamber driven by a motor. The spheres and the background chamber were filled with 18F solution at a sphere-to-background ratio of 5:1. We enrolled 42 and 16 patients undergoing 2-deoxy-2-[18F]fluoro-D-glucose {2-[18F]FDG} and 68Ga-labeled [1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid]-1-Nal3-octreotide {[68Ga]Ga-DOTA-NOC} PET/computed tomography (CT) from whom 74 and 30 lesions were segmented, respectively. Three reconstructions were performed: data-driven gating-based motion correction (DDGMC), external vital signal module-based motion correction (VSMMC), and noncorrection reconstruction. The standardized uptake values (SUVs) and the volume of the spheres and the lesions were measured and compared among the 3 reconstruction groups. The image noise in the liver was measured, and the visual image quality of motion artifacts was scored by radiologists in the patient study. Results: In the phantom study, the spheres' SUVs increased by 26-36%, and the volumes decreased by 35-38% in DDGMC and VSMMC compared with the noncorrection group. In the 2-[18F]FDG PET patient study, the lesions' SUVs had a median increase of 10.87-12.65% while the volumes had a median decrease of 14.88-15.18% in DDGMC and VSMMC compared with those of noncorrection. In the [68Ga]Ga-DOTA-NOC PET patient study, the lesions' SUVs increased by 14.23-15.45%, and the volumes decreased by 19.11-20.94% in DDGMC and VSMMC. The image noise in the liver was equal between the DDGMC, VSMMC, and noncorrection groups. Radiologists found improved image quality in more than 45% of the cases in DDGMC and VSMMC compared with the noncorrection group. There was no statistically significant difference in SUVs, volumes, or visual image quality scores between DDGMC and VSMMC. Conclusions: MCIR improves tumor quantification accuracy and visual image quality by reducing respiratory motion artifacts without compromised image noise performance or elongated acquisition time in 2-[18F]FDG and [68Ga]Ga-DOTA-NOC PET/CT tumor imaging. The performance of DDG-driven MCIR is as good as that of the external device-driven solution.

SIRT6-regulated macrophage efferocytosis epigenetically controls inflammation resolution of diabetic periodontitis.

Rationale: Diabetes exacerbates the prevalence and severity of periodontitis, leading to severe periodontal destruction and ultimately tooth loss. Delayed resolution of inflammation is a major contributor to diabetic periodontitis (DP) pathogenesis, but the underlying mechanisms of this imbalanced immune homeostasis remain unclear. Methods: We collected periodontium from periodontitis with or without diabetes to confirm the dysfunctional neutrophils and macrophages in aggravated inflammatory damage and impaired inflammation resolution. Our in vitro experiments confirmed that SIRT6 inhibited macrophage efferocytosis by restraining miR-216a-5p-216b-5p-217 cluster maturation through ''non-canonical'' microprocessor complex (RNA pulldown, RIP, immunostaining, CHIP, Luciferase assays, and FISH). Moreover, we constructed m6SKO mice that underwent LIP-induced periodontitis to explore the in vitro and in vivo effect of SIRT6 on macrophage efferocytosis. Finally, antagomiR-217, a miRNA antagonism, was delivered into the periodontium to treat LIP-induced diabetic periodontitis. Results: We discovered that insufficient SIRT6 as a histone deacetylase in macrophages led to unresolved inflammation and aggravated periodontitis in both human and mouse DP with accumulated apoptotic neutrophil (AN) and higher generation of neutrophil extracellular traps. Mechanistically, we validated that macrophage underwent high glucose stimulation resulting in disturbance of the SIRT6-miR-216/217 axis that triggered impeded efferocytosis of AN through targeting the DEL-1/CD36 axis directly. Furthermore, we demonstrated the inhibitory role of SIRT6 for MIR217HG transcription and identified a non-canonical action of microprocessor that SIRT6 epigenetically hindered the splicing of the primary miR-216/217 via the complex of hnRNPA2B1, DGCR8, and Drosha. Notably, by constructing myeloid-specific deletion of SIRT6 mice and locally delivering antagomir-217 in DP models, we strengthened the in vivo effect of this axis in regulating macrophage efferocytosis and inflammation resolution in DP. Conclusions: Our findings delineated the emerging role of SIRT6 in mediating metabolic dysfunction-associated inflammation, and therapeutically targeting this regulatory axis might be a promising strategy for treating diabetes-associated inflammatory diseases.

The deubiquitinase USP7 promotes HNSCC progression via deubiquitinating and stabilizing TAZ.

Dysregulated abundance, location and transcriptional output of Hippo signaling effector TAZ have been increasingly linked to human cancers including head neck squamous cell carcinoma (HNSCC). TAZ is subjected to ubiquitination and degradation mediated by E3 ligase ß-TRCP. However, the deubiquitinating enzymes and mechanisms responsible for its protein stability remain underexplored. Here, we exploited customized deubiquitinases siRNA and cDNA library screen strategies and identified USP7 as a bona fide TAZ deubiquitinase in HNSCC. USP7 promoted cell proliferation, migration, invasion in vitro and tumor growth by stabilizing TAZ. Mechanistically, USP7 interacted with, deubiquitinated and stabilized TAZ by selectively removing its K48-linked ubiquitination chain independent of canonical Hippo kinase cascade. USP7 potently antagonized ß-TRCP-mediated ubiquitin-proteasomal degradation of TAZ and enhanced its nuclear retention and transcriptional output. Importantly, overexpression of USP7 correlated with TAZ upregulation, tumor aggressiveness and unfavorable prognosis in HNSCC patients. Pharmacological inhibition of USP7 significantly suppressed tumor growth in both xenograft and PDX models. Collectively, these findings identify USP7 as an essential regulator of TAZ and define USP7-TAZ signaling axis as a novel biomarker and potential therapeutic target for HNSCC.

Macrophage miR-149-5p induction is a key driver and therapeutic target for BRONJ.

Bisphosphonate-related (BP-related) osteonecrosis of the jaw (BRONJ) is one of the severe side effects of administration of BPs, such as zoledronic acid (ZA), which can disrupt the patient's quality of life. Although the direct target of skeletal vasculature and bone resorption activity by BPs has been phenomenally observed, the underlying mechanism in BRONJ remains largely elusive. Thus, it is urgently necessary to discover effective therapeutic targets based on the multifaceted underlying mechanisms in the development of BRONJ. Here, we determined the inhibitory role of ZA-treated macrophages on osteoclast differentiation and type H vessel formation during tooth extraction socket (TES) healing. Mechanistically, ZA activated the NF-κB signaling pathway and then induced p65 nuclear translocation in macrophages to promote miR-149-5p transcription, resulting in impaired osteoclast differentiation via directly binding to the Traf6 3'-UTR region. Moreover, we identified that miR-149-5p-loaded extracellular vesicles derived from ZA-treated bone marrow-derived macrophages could regulate biological functions of endothelial cells via the Rap1a/Rap1b/VEGFR2 pathway. Furthermore, local administration of chemically modified antagomiR-149-5p was proven to be therapeutically effective in BRONJ mice. In conclusion, our findings illuminate the dual effects of miR-149-5p on skeletal angiogenesis and bone remolding, suggesting it as a promising preventive and therapeutic target for BRONJ.

Differential expression profiles and function prediction of tRNA-derived fragments in fibrous dysplasia.

OBJECTIVE: Fibrous dysplasia (FD) is a bone disease featuring that normal bone matrix is replaced by fibrous tissue and immature bone tissue. Transfer RNA-derived RNA fragments (tRFs) and tRNA halves (tiRNAs) are types of small non-coding RNA that produced by specific shearing of mature tRNA. Here, we conducted a comparative analysis of the expression of tRFs/tiRNAs in BMSCs and FD BMSCs. DESIGNS: The differential expression of tRFs/tiRNAs was detected by high-throughput sequencing technique, and validated by qRT-PCR. Bioinformatics analyses including prediction of target genes, gene ontology (GO) enrichment and KEGG pathway analysis and protein-protein interaction (PPI) network analysis were performed. RESULTS: The results detected 1 significantly upregulated tDR (tRNA-derived small RNA) and 3 downregulated tDRs in FD BMSCs. Predictions of target genes, GO and KEGG pathway analysis indicated that these tRFs were mainly involved in regulation of immune response, osteoclast differentiation, calcium ion transport, apoptotic signaling pathway, cell proliferation and endocrine system development. The PPI network analysis showed that PPP2R5A, ADAMTS1, PPARA, and POLR2C were the most frequently interacted proteins in target genes. CONCLUSIONS: Our findings provided a comprehensive analysis of the expression of tRFs/tiRNAs in FD BMSCs and BMSCs, and they could serve as potential therapeutic targets.

Quantitative sensory testing of mandibular somatosensory function following orthognathic surgery-A pilot study in Chinese with class III malocclusion.

BACKGROUND: Somatosensory changes after sagittal split ramus osteotomy (SSRO) have not been fully studied in Chinese patients by the latest technologies. OBJECTIVE: To provide a comprehensive analysis of somatosensory function at the lower lip and chin at different time points following SSRO in a Chinese population. METHODS: A total of 22 patients (18-27 years; nine men) with skeletal III malocclusion and scheduled for SSRO were recruited. Quantitative sensory testing (QST) was performed at pre-operation (baseline), 1 week (1W), 1, 3 and 6 months (1M, 3M, 6M) post-operatively. Cold detection threshold (CDT), warm detection threshold (WDT), cold pain threshold (CPT), heat pain threshold (HPT), mechanical detection threshold (MDT), mechanical pain threshold (MPT), pressure pain threshold (PPT) and two-point discrimination threshold (2PD) were tested at the lower lip and chin. RESULTS: Except for PPT at both test sides at 1W and 1M, all QST values indicated a significantly reduced sensitivity (p < .05). All values had returned to baseline values at 3M with exception of HPT at the right chin which, however, had recovered at 6M (p > .05). CONCLUSIONS: Somatosensory function at the lower lip and chin appears to be fully recovered in the majority of young Chinese adults 6 months after SSRO for skeletal class III malocclusion.

LncRNA NEAT1 controls the lineage fates of BMSCs during skeletal aging by impairing mitochondrial function and pluripotency maintenance.

Aged bone marrow mesenchymal stem cells (BMSCs) exhibit aberrant self-renewal and lineage specification, which contribute to imbalanced bone-fat and progressive bone loss. In addition to known master regulators of lineage commitment, it is crucial to identify pivotal switches governing the specific differentiation fate of aged BMSCs. Here, we profiled differences in epigenetic regulation between adipogenesis and osteogenesis and identified super-enhancer associated lncRNA nuclear-enriched abundant transcript 1 (NEAT1) as a key bone-fat switch in aged BMSCs. We validated that NEAT1 with high enhancer activity was transcriptionally activated by ATF2 and directed aged BMSCs to a greater propensity to differentiate toward adipocytes than osteoblasts by mediating mitochondrial function. Furthermore, we confirmed NEAT1 as a protein-binding scaffold in which phosphorylation modification of SOX2 Ser249/250 by CDK2 impaired SOX2/OCT4 complex stability and dysregulated downstream transcription networks of pluripotency maintenance. In addition, by sponging miR-27b-3p, NEAT1 upregulated BNIP3L, BMP2K, and PPARG expression to shape mitochondrial function and osteogenic/adipogenic differentiation commitment, respectively. In extracellular communication, NEAT1 promoted CSF1 secretion from aged BMSCs and then strengthened osteoclastic differentiation by extracellular vesicle delivery. Notably, Neat1 small interfering RNA delivery induced increased bone mass in aged mice and decreased fat accumulation in the bone marrow. These findings suggest that NEAT1 regulates the lineage fates of BMSCs by orchestrating mitochondrial function and pluripotency maintenance, and might be a potential therapeutic target for skeletal aging.

miR-34a-5p functions as a tumor suppressor in head and neck squamous cell cancer progression by targeting Flotillin-2.

While a number of therapeutic advances have been made in recent years, the overall survival of patients with head and neck squamous cell cancer (HNSCC) remains poor. MicroRNAs (miRNAs) are key drivers of oncogenic progression, with miR-34a-5p downregulation having been observed in many different tumor types. Here, we assessed the link between miR-34a-5p and HNSCC progression and the mechanistic basis for this relationship. Levels of miR-34a-5p in HNSCC tumors and cell lines were assessed via qPCR, after which we explored the functional importance of this miRNA in this oncogenic setting. Through luciferase reporter assays, the ability of miR-34a-5p to regulate flotillin-2 (FLOT-2) was further clarified. Overall, these analyses revealed that HNSCC tumors and cells exhibited marked miR-34a-5p downregulation that was linked to the progression of this tumor type. At a functional level, miR-34a-5p constrained the proliferation, migratory/invasive activity, and epithelial-mesenchymal transition induction in HNSCC cells. At the mechanistic level, miR-34a-5p was found to suppress FLOT-2 expression and to activate the MEK/ERK1/2 pathway. Overall, these results suggest that miR-34a-5p can function as a tumor suppressor miRNA in HNSCC owing to its ability to target FLOT-2, highlighting the promise of targeting this regulatory axis to treat HNSCC.

Identification of diagnostic and prognostic signatures derived from preoperative blood parameters for oral squamous cell carcinoma.

BACKGROUND: We aimed to develop novel diagnostic and prognostic signatures based on preoperative inflammatory, immunological, and nutritional parameters in blood (PIINPBs) by machine learning algorithms for patients with oral squamous cell carcinoma (OSCC). METHODS: A total of 486 OSCC patients and 200 age and gender-matched non-OSCC patients who were diagnosed and treated at our institution for noninfectious, nontumor diseases were retrospectively enrolled and divided into training and validation cohorts. Based on PIINPB, 6 machine learning classifiers including random forest, support vector machine, extreme gradient boosting, naive Bayes, neural network, and logistic regression were used to derive diagnostic models, while least absolute shrinkage and selection operator (LASSO) analyses were employed to construct prognostic signatures. A novel prognostic nomogram integrating a PIINPB-derived prognostic signature and selected clinicopathological parameters was further developed. Performances of these signatures were assessed by receiver operating characteristic (ROC) curves, calibrating curves, and decision tree. RESULTS: Diagnostic models developed by machine learning algorithms from 13 PIINPBs, which included counts of white blood cells (WBC), neutrophils (N), monocytes (M), lymphocytes (L), platelets (P), albumin (ALB), and hemoglobin (Hb), along with albumin-globulin ratio (A/G), neutrophil-lymphocyte ratio (NLR), platelet-lymphocyte ratio (PLR), lymphocyte-monocyte ratio (LMR), systemic immune-inflammation index (SII), and prognostic nutritional index (PNI), displayed satisfactory discriminating capabilities in patients with or without OSCC, and among OSCC patients with diverse pathological grades and clinical stages. A prognostic signature based on 6 survival-associated PIINPBs (L, P, PNI, LMR, SII, A/G) served as an independent factor to predict patient survival. Moreover, a novel nomogram integrating prognostic signature and tumor size, pathological grade, cervical node metastasis, and clinical stage significantly enhanced prognostic power [3-year area under the curve (AUC) =0.825; 5-year AUC =0.845]. CONCLUSIONS: Our results generated novel and robust diagnostic and prognostic signatures derived from PIINPBs by machine learning for OSCC. Performance of these signatures suggest the potential for PIINPBs to supplement current regimens and provide better patient stratification and prognostic prediction.

A CREB1-miR-181a-5p loop regulates the pathophysiologic features of bone marrow stromal cells in fibrous dysplasia of bone.

BACKGROUND: Fibrous dysplasia (FD) is a bone marrow stromal cell (BMSC) disease caused by activating mutations of guanine nucleotide-binding protein alpha-stimulating activity polypeptide (GNAS) and is characterized by increased proliferative activity and disrupted osteogenesis of BMSCs. However, the molecular mechanisms regulating the pathophysiologic features of BMSCs in FD remain unknown. This study aimed to identify and verify the roles of the CREB1-miR-181a-5p regulatory loop in FD pathophysiology. METHODS: MicroRNA (miRNA) sequencing analysis was used to identify the possible miRNAs implicated in FD. The proliferation, apoptosis, and osteogenic differentiation of BMSCs, as well as the osteoclast-induced phenotype, were measured and compared after exogenous miR-181a-5p transfection into FD BMSCs or miR-181a-5p inhibitor transfection into normal BMSCs. Chromatin immunoprecipitation and luciferase reporter assays were performed to verify the interactions between CREB1 and miR-181a-5p and their effects on the FD pathological phenotype. RESULTS: Compared to normal BMSCs, FD BMSCs showed decreased miR-181a-5p levels and exhibited increased proliferative activity, decreased apoptotic capacity, and impaired osteogenesis. FD BMSCs also showed a stronger osteoclast activation effect. miR-181a-5p overexpression reversed the pathophysiologic features of FD BMSCs, whereas miR-181a-5p suppression induced an FD-like phenotype in normal BMSCs. Mechanistically, miR-181a-5p was the downstream target of CREB1, and CREB1 was posttranscriptionally regulated by miR-181a-5p. CONCLUSIONS: Our study identifies that the interaction loop between CREB1 and miR-181a-5p plays a crucial role in regulating the pathophysiologic features of FD BMSCs. MiR-181a-5p may be a potential therapeutic target for the treatment of FD.

Secretory carcinoma of the breast with multiple distant metastases in the brain and unfavorable prognosis: a case report and literature review.

BACKGROUND: Secretory carcinoma of the breast is one of the rarest entities, accounting for less than 0.15 % of all infiltrating breast carcinomas. It has characteristic histopathological and molecular features and, in general, a more favorable prognosis. In this case report, we describe a local, advanced secretory carcinoma of the breast with aggressive course and an unfavorable outcome. CASE PRESENTATION: A hard, painless, and palpably bossed mass approximately 12.0 cm in diameter occupied most of the left breast of a 39-year-old woman with fixation to the overlying skin. Breast ultrasonography and magnetic resonance imaging (MRI) scans gave the same grading as BI-RADS IV. A needle biopsy was performed, and the pathological diagnosis was secretory carcinoma. Neoadjuvant chemotherapy (NAC) was then performed, after which ultrasonography and MRI scans revealed chemo-resistance of the tumor to NAC. Left breast mastectomy and axillary lymphadenectomy were subsequently performed. Tumor cells were triple-negative and positive for S-100 and periodic acid-Schiff (PAS) staining. Fluorescence in-situ hybridization (FISH) analysis indicated a fusion arrangement of the ETV6-NTRK3 gene. The patient developed multiple distant metastases in the brain and died of these metastases 19 months after initial diagnosis. CONCLUSIONS: Secretory carcinomas of the breast have been described as a low-grade histologic subtype with a favorable prognosis. This case showed chemo-resistance to neoadjuvant chemotherapy, multiple distant metastases, and a final unfavorable outcome. Further research is needed to better understand the behavior and treatment of this rare tumor.

Immune landscape and subtypes in primary resectable oral squamous cell carcinoma: prognostic significance and predictive of therapeutic response.

BACKGROUND: Immune landscape of cancer has been increasingly recognized as a key feature affecting disease progression, prognosis and therapeutic response. Here, we sought to comprehensively characterize the patterns of tumor-infiltrating immune cells (TIIs) in primary oral squamous cell carcinoma (OSCC) and develop immune features-derived models for prognostication and therapeutic prediction. METHODS: A total number of 392 patients with OSCC receiving ablative surgery at three independent centers were retrospectively enrolled and defined as training, testing and validation cohorts. Detailed features of 12 types of TIIs at center of tumor and invasive margin were assessed by immunohistochemistry coupled with digital quantification. TIIs abundance in OSCC was also estimated by bioinformatics approaches using multiple publicly available data sets. Prognostic models based on selected immune features were trained via machine learning approach, validated in independent cohorts and evaluated by time-dependent area under the curves and concordance index (C-index). Immune types of OSCC were further identified by consensus clustering and their associations with genetic, molecular features and patient survival were clarified. RESULTS: Patterns of TIIs infiltration varied among patients and dynamically evolved along with tumor progression. Prognostic models based on selected TIIs were identified as efficient and sensitive biomarkers to stratify patients into subgroups with favorable or inferior survival as well as responders or non-responders to postoperative radiotherapy or immunotherapy. These models outperformed multiple conventional biomarkers and immune-related scores in prognostic prediction. Furthermore, we identified two main immune subtypes of OSCC (immune-hot and immune-cold) which harbored characteristic TIIs infiltrations and genomic and molecular features, and associated with patient survival. CONCLUSIONS: Our results delineated immune landscape and subtypes in OSCC, consolidated their clinical values as robust biomarkers to predict patient survival and therapeutic benefits and reinforced key roles of TIIs and tumor-immune interactions underlying oral tumorigenesis, ultimately facilitating development of tailed immunotherapeutic strategies.

Identification of the canonical and noncanonical role of miR-143/145 in estrogen-deficient bone loss.

Rationale: Postmenopausal-induced bone loss is mainly caused by declining core transcription factors (TFs) of bone mesenchymal stem cells (BMSCs), but little is known about how miRNAs regulate chromatin structure remodeling of TFs gene to maintain BMSCs function in bone homeostasis. Methods: We examined the serum, salivary and bone samples from Pre- and Post-menopause women by paired analysis and confirmed canonical ceRNA role of MIR143HG and miR-143/145 complexes in cytoplasm and noncanonical role for SOX2 transcription in nucleus (FISH, qRT-PCR, immunostaining, Luciferase assays and ChIP). Moreover, we took advantage of transgenic mice under OVX-induced osteoporosis, studying the in vitro and in vivo effect of miR-143/145 deletion on BMSCs function and bone homeostasis. Last, using miRNA antagonism, antagomiR-143/145 were delivered into bone marrow to treat estrogen-deficient bone loss. Results: Here, we identified miR-143/145 as potential diagnostic candidates for postmenopausal osteoporosis, and miR-143/145 overexpression impaired BMSCs self-renewing and differentiation function. Mechanistically, we confirmed that cytoplasmic miR-143/145 and LncRNA MIR143HG, that controlled by ERß, cooperatively regulated pluripotency genes translation via canonical ceRNA pathway, and MIR143HG cooperates with miR­143 to nuclear translocation for co-activation of SOX2 transcription via opening promoter chromatin. Meanwhile, miR­143/145 were shuttled into osteoclasts in extracellular vesicles and triggered osteoclastic activity by targeting Cd226 and Srgap2. Furthermore, miR-143/145-/- mice or using chemically­modified antagomiR-143/145 significantly alleviated estrogen-deficient osteoporosis. Conclusions: Our findings reveal a canonical and noncanonical role of miR-143/145 in controlling BMSCs pluripotency and unfold their dual effect on bone formation and bone resorption, suggesting miR-143/145 as promising therapeutic targets for treating estrogen-deficient bone loss.

Blood and Salivary MicroRNAs for Diagnosis of Oral Squamous Cell Carcinoma: A Systematic Review and Meta-Analysis.

PURPOSE: This meta-analysis aimed to compare and evaluate the diagnostic accuracy of blood and salivary microRNAs (miRNAs) in discriminating oral squamous cell carcinoma (OSCC). METHODS: The PubMed, Embase, Web of Science, and Cochrane Library were searched (updated to February 2020) to identify all articles describing the diagnostic value of blood and salivary miRNAs for OSCC. The pooled parameters were calculated using Revman (v.5.3) and STATA (v.14.0). RESULTS: Twenty articles involving 1,106 patients and 732 controls were included in this meta-analysis. The pooled sensitivity and specificity of salivary miRNAs were 0.70 (95% CI: 0.63-0.77) and 0.82 (95% CI: 0.72-0.90). For blood miRNAs, they were 0.79 (95% CI: 0.73-0.84) and 0.82 (95% CI: 0.77-0.86). The areas under receiver operating characteristic curve in saliva, blood, and body fluid miRNAs were 0.80 (95% CI: 0.77-0.84), 0.88 (95% CI: 0.84-0.90), and 0.87 (95% CI: 0.84-0.90), respectively. CONCLUSIONS: The results of this meta-analysis indicate a moderate diagnostic accuracy of blood and salivary miRNAs presented for OSCC. These findings may provide less invasive and relatively reliable diagnostic tools for OSCC detection.