Iphigenia Gintoni

Background: Tall stature is a multifactorial phenotypic trait encountered in the general pediatric population; however, it could constitute a manifestation of an underlying genetic and/or endocrine disorder. Mutations in genes involved in gene transcription, genomic imprinting and intracellular vesicle trafficking may lead to dysregulation of certain cellular pathways associated with different genetic syndromes with overgrowth and tall stature. Objective: Considering the facial changes that growth hormone surplus induces, it would be reasonable to hypothesize that the maxillofacial phenotype is also affected in certain genetic conditions causing excessive linear growth. The aim of this review is to assess the cephalometric data for certain genetic and/or endocrine diseases causing tall stature and overgrowth in children and discuss the potential role of cephalometric analysis in diagnosis of such phenotypes and better understanding of the underlying causing molecular mechanisms. Design: A literature search was conducted in the PubMed electronic database. The scientific articles that provided cephalometric data for genetic syndromes and conditions associated with tall stature or overgrowth were included. The following keywords were utilized: tall stature cephalometry, overgrowth syndromes cephalometry, gigantism cephalometry, hemihyperplasia cephalometric, hemihypertrophy cephalometric, Simpson–Golabi–Behmel cephalometric, Prader–Willi cephalometry, and Marfan and Klinefelter cephalometry. Results: Although the number of studies found remains limited (n = 37), the consensus findings indicate that the growth of the craniofacial complex of patients with genetic syndromes causing tall stature is affected. The cephalometric measurements assessing the cranial base, the jaws and the facial soft tissue differ from the norm and are suggestive of irregularities in both the sagittal and vertical planes. Conclusions: As the literature in this field is particularly rare, more data are required to establish relations between excessive linear growth and the maxillofacial phenotype exhibited in certain genetic and/or endocrine syndromes. Nevertheless, lateral cephalometric radiography is an extensively utilized imaging method that could potentially serve as a promising tool in the early diagnosis, phenotype–genotype comparative research and better understanding of the underlying causing molecular mechanisms of conditions associated with tall stature.

Background: Pediatric thrombosis (PT) is a rare clinical entity manifesting from neonatal life to adolescence with possibly severe clinical complications and increasing prevalence in the last decade. Its pathogenesis has been attributed to a multitude of factors, all of which share stress and inflammation as a common denominator. Although stress-induced inflammation, marked by the upregulation of proinflammatory interleukins, such as IL-1β, creates a prothrombotic environment, its relation with thrombosis remains incompletely defined, especially in the context of PT, which differs substantially from adult thrombosis. Here, we aimed to epigenetically explore the potential link between PT and stress-induced inflammation through a combinatorial approach. Methods: Initially, we bioinformatically identified candidate stress-specific miRNAs in childhood, followed by subsequent experimental assessment in serum samples of PT patients and healthy controls in relation to subjectively perceived stress levels and the functional +3953C/T genetic polymorphism of the IL1B gene, which affects the circulating levels of the proinflammatory and stress-induced interleukin 1beta (IL-1β). Results: The bioinformatic analysis identified miR-34a-5p as the most stress-specific candidate microRNA in childhood, exhibiting the highest gene-target coverage (80.6%) of the developed panel of adversity-related genes, but is also known to regulate IL1B expression. The expression of serum miR-34a-5p was not significantly influenced by subjectively perceived stress; however, its expression was significantly reduced in the presence of the T allele of the functional +3953C/T polymorphism of its target gene, IL1B, associated with increased levels of the stress-induced proinflammatory and simultaneously prothrombotic IL-1β cytokine (FC:0.57; p = 0.024). Conclusions: The observed downregulation of miR-34a-5p in the presence of the IL-1β-increasing T allele possibly indicates a dynamic regulatory relationship, which, alongside its previously established long-term downregulation in PT, renders miR-34a-5p a promising candidate molecule for bridging the biological processes of stress-associated inflammation and thrombosis. In addition, its bioinformatically yielded candidate stress-specific role holds great potential for the non-invasive evaluation of adversity-related stress in childhood.

Oral squamous cell carcinoma (OSCC) is characterized by consistently high mortality rates (≤60%) despite therapeutic advances. This is attributable to diagnostic delays arising from the asymptomatic early stages and time-consuming protocols. Hence, the establishment of reliable biomarkers for the routine assessment of the oral mucosa is imperative. MicroRNAs (miRNAs), key epigenetic regulators of gene expression, represent ideal candidates given their characteristic dysregulation across different pathologies. Here, we aimed to identify novel OSCC-specific miRNAs for the saliva-based detection of OSCC from the presymptomatic stage of early invasion. Through a multistep bioinformatic workflow, four miRNAs (miR-20b-5p, miR-484, miR-185-5p and miR-181d-5p) were identified as disease-specific since they simultaneously regulated >65% of a panel encompassing the 15 primarily overexpressed oncogenes in OSCC and a stage-specific panel including the six upregulated genes that genetically define the malignant stages of sequential oral carcinogenesis. The salivary expression of the identified miRNAs was studied in 31 OSCC patients and 31 healthy controls, using quantitative real-time PCR, followed by statistical analysis and an evaluation of the diagnostic accuracy. All studied miRNAs were significantly downregulated in the saliva of OSCC patients compared to controls (miR-484, p < 0.001; miR-181d-5p, p < 0.001; miR-185, p = 0.008; miR-20b, p = 0.026) and exhibited combinatory diagnostic performance of 95.4% (p < 0.001) for OSCC detection. Their expression remained uninfluenced by lifestyle and clinicopathological parameters, including smoking/alcohol, tumor site, grade and disease stage. The proposed 4-miRNA panel exhibits high diagnostic performance for the early, saliva-based detection of OSCC, irrespective of histopathological and lifestyle confounders, highlighting its potential as a robust non-invasive screening tool.

Background: Oral squamous cell carcinoma (OSCC) is a highly aggressive neoplasm characterized by grim survival outcomes, despite significant therapeutic advances. Mortality rates (up to 70%) have remained unaltered for decades, predominantly due to profound diagnostic delays. These derive from the asymptomatic nature of the early stages of oral carcinogenesis and the emergence of dysplastic areas in previously benign lesions, acting as the bridge to malignant transformation. Hence, the establishment of reliable salivary biomarkers is crucial for non-invasive OSCC detection, even from the premalignant stage of dysplasia. Based on our previous bioinformatic research identifying stage-specific miRNAs throughout OSCC progression, which yielded miR-34a-5p as the most significant, we aimed to experimentally investigate its role in oral oncogenesis and explore its stage-reflecting biomarker potential for liquid biopsy. Methods: The expression of miR-34a was evaluated using quantitative real-time PCR in saliva samples from 9 patients with oral premalignant dysplastic lesions, 10 patients with OSCC and 10 healthy controls. The diagnostic accuracy of miR-34a expression profiles was assessed using ROC-curve analyses. Results: The expression of salivary miR-34a differed significantly across the studied groups, demonstrating a steep decrease in the presence of epithelial premalignant dysplasia, significant upregulation in OSCC and intermediate levels in normal oral mucosa (p < 0.001). The ROC results indicate strong diagnostic performance for the detection of oral dysplasia (AUC = 0.93; p < 0.001), OSCC (AUC = 0.77; p = 0.01) and excellent accuracy for the discrimination between premalignant and OSCC lesions (AUC = 0.98; p < 0.001). Conclusions: Our findings reveal a state-dependent dysregulation of miR-34a in oral carcinogenesis, suggesting its complex role as a pathogenetic agent that allows for malignant transformation through its diminished expression, and as a secondary reactive mechanism attempting to suppress tumor development. Salivary miR-34a holds great, stage-specific diagnostic potential, thereby reflecting the health state of oral mucosa in real time.

Pediatric thrombosis (PT) represents a rare condition that can manifest from neonatal life to adolescence, encompassing life-threatening complications. Its pathogenesis is attributed to immature hemostasis in conjunction with environmental and genetic factors, predominantly including those resulting in increased levels of plasminogen activator inhibitor 1 (PAI-1), the principal inhibitor of fibrinolysis, which is subject to upstream regulation by angiotensin-converting enzyme (ACE). Although the implication of microRNAs (miRNAs), epigenetic modulators of gene expression, has been demonstrated in adult thrombosis, evidence is lacking in the pediatric setting. Here, we investigated the involvement of two miRNA regulators of PAI-1 (SERPINE1 gene) in PT, in relation to clinical and genetic parameters that induce PAI-1 fluctuations. Following bioinformatic target-prediction, miRNA expression was assessed by quantitative real-time PCR in serum-samples of 19 pediatric patients with thrombosis (1–18 months post-incident), and 19 healthy controls. Patients were genotyped for the SERPINE1-4G/5G and ACE-I/D polymorphisms by PCR-based assays. Genotypic and thrombosis-related clinical data were analyzed in relation to miRNA-expression. Two miRNAs (miR-145-5p, miR-34a-5p) were identified to target SERPINE1 mRNA, with miR-34a additionally targeting the mRNA of ACE. The expression of miR-34a was significantly decreased in patients compared to controls (p = 0.029), while no difference was observed in miR-145 expression. Within patients, miR-34a expression demonstrated a peak 1–3 months post-thrombosis and was diminished upon treatment completion (p = 0.031), followed by a slight long-term increase. MiR-34a levels differed significantly by thrombosis site (p = 0.019), while a significant synergistic interaction between site and onset type (provoked/unprovoked) was detected (p = 0.016). Finally, an epistatic modification was observed in cerebral cases, since double homozygosity (4G/4G + D/D) led to a miR-34 decrease, with D/D carriership reversing the 4G/4G-induced upregulation of miR-34a (p = 0.006). Our findings suggest that in pediatric thrombosis, downregulation of miR-34a is indicative of impaired fibrinolytic capacity, attributed to deficient regulation of the inhibitory ACE/PAI-1 axis.

Background: Pediatric thrombosis (PT) represents a rare condition that can manifest from neonatal life to adolescence, encompassing life-threatening complications. Its pathogenesis is attributed to immature hemostasis in conjunction with environmental and genetic factors, predominantly including those resulting in increased levels of plasminogen activator inhibitor 1 (PAI-1), the principal inhibitor of fibrinolysis, subject to upstream regulation by ACE (angiotensin-converting enzyme). Although the implication of microRNAs (miRNAs), epigenetic modulators of gene-expression, is demonstrated in adult-thrombosis, evidence is lacking in the pediatric setting. Here, we investigated the involvement of two miRNA-regulators of PAI-1 in PT, in relation to clinical and genetic parameters that induce PAI-1 fluctuations. Methods: Following bioinformatic target-prediction, miRNA expression was assessed by quantitative real-time PCR, in serum-samples of 19 pediatric patients with thrombosis (1-18 months post-incident), and 19 controls. Patients were genotyped for the PAI-1-4G/5G and ACE-I/D polymorphisms by PCR-based assays. Genotypic and thrombosis-related clinical data were analyzed in relation to miRNA-expression. Results: Two miRNAs (miR-145-5p, miR-34a-5p) were identified to target PAI-1 mRNA, with miR-34a additionally targeting ACE mRNA. The expression of miR-34a was significantly decreased in patients compared to controls (p=0.029), while no difference was observed for miR-145. Within patients, miR-34a demonstrated a peak 1-3 months post-thrombosis and was diminished upon treatment-completion (p=0.031), followed by a slight long-term increase. MiR-34a expression differed significantly by thrombosis site (p=0.019), while a significant synergistic interaction between site and onset-type (provoked/unprovoked) was detected (p=0.016). Finally, an epistatic modification was observed in cerebral cases, since double homozygosity (4G/4G+D/D) led to miR-34a diminishing, with DD carriership reversing the 4G4G-induced upregulation of miR-34a (p=0.006). Conclusions: Our findings suggest that in pediatric thrombosis, downregulation of miR-34a is indicative of impaired fibrinolytic capacity, attributed to deficient regulation of the inhibitory ACE/PAI-1 axis.

Background: Osteodysplastic syndromes comprise a very diverse group of clinically and genetically heterogeneous disorders characterized by defects in bone and connective tissue development, as well as in bone density. Here, we report the case of a 48-year-old female with a complex medical history characterized by bone dysplasia, hyperostosis, and partial tooth agenesis. Methods: Genetic testing was performed using WES analysis and Sanger sequencing. Molecular modeling analysis and dynamics simulation explored the impact of detected pathogenic variants. Results: The genetic analysis detected multiple pathogenic variants in genes CREB3L1, SLCO2A1, SFRP4, LRP5, and LRP6, each of which has been associated with rare osteodysplastic syndromes. The patient was homozygous for the same rare alleles associated with three of the identified autosomal recessive disorders osteogenesis imperfecta type XVI, primary hypertrophic osteoarthropathy, and metaphyseal dysplasia Pyle type. She also had a variant linked to autosomal dominant endosteal hyperostosis and a variant previously associated with increased risk of osteoporosis and bone fractures. Two of the detected variants are predicted to cause abnormal splicing, while molecular modeling and dynamics simulations analysis suggest that the other three variants probably confer altered local secondary structure and flexibility that may have functionally devastating consequences. Conclusions: Our case highlights the rare coexistence of multiple osteodysplastic syndromes in a single patient that may complicate differential diagnosis. Furthermore, this case emphasizes the necessity for early genetic investigation of such complex cases with overlying phenotypic traits, followed by genetic counseling, facilitating orchestration of clinical interventions and allowing prevention and/or prompt management of manifestations.

Background: Osteodysplastic syndromes compose a very diverse group of clinically and genetically heterogeneous disorders characterized by defects in bone and connective tissue development, as well as in bone density. Here, we report the case of a 48-year-old female with a complex medical history characterized by bone dysplasia, hyperostosis, and partial tooth agenesis. Methods: Genetic testing was performed using WES analysis and Sanger sequencing. Molecular modeling analysis and dynamics simulation explored the impact of detected pathogenic variants. Results: The genetic analysis detected multiple pathogenic variants in genes CREB3L1, SLCO2A1, SFRP4, LRP5 and LRP6, which are associated with rare osteodysplastic syndromes. The patient was homozygous for the same rare alleles associated with three of the identified autosomal recessive disorders, such as osteogenesis imperfecta type XVI, primary hypertrophic osteoarthropathy, and metaphyseal dysplasia Pyle type. She also had a variant linked to autosomal dominant endosteal hyperostosis and a variant previously associated with increased risk of osteoporosis and fractures. Two of the detected variants are predicted to cause abnormal splicing, while molecular modelling and dynamics simulations analysis suggest that the other three variants probably confer altered local secondary structure and flexibility that may be functionally devastating consequences. Conclusions: Our case highlights the rare coexistence of multiple osteodysplastic syndromes in a single patient that may complicate differential diagnosis. Furthermore, this case emphasizes the necessity for early genetic investigation of such complex cases with overlying phenotypic traits, followed by genetic counseling, in order to orchestrate clinical interventions and prevent or promptly manage manifestations.

Oral squamous cell carcinoma (OSCC) is a highly prevalent and aggressive malignancy, with mortality rates reaching 60%, mainly due to its excessive diagnostic delay. MiRNAs, a class of crucial epigenetic gene-expression regulators, have emerged as potential diagnostic biomarkers, with >200 molecules exhibiting expressional dysregulation in OSCC. We had previously established an in silico methodology for the identification of the most disease-specific molecules by bridging genetics and epigenetics. Here, we identified the stage-specific miRNAs that govern the asymptomatic early stages of oral tumorigenesis by exploiting seed-matching and the reverse interplay between miRNA levels and their target genes’ expression. Incorporating gene-expression data from our group’s experimental hamster model of sequential oral oncogenesis, we bioinformatically detected the miRNAs that simultaneously target/regulate >75% of the genes that are characteristically upregulated or downregulated in the consecutive stages of hyperplasia, dysplasia, and early invasion, while exhibiting the opposite expressional dysregulation in OSCC-derived tissue and/or saliva specimens. We found that all stages share the downregulation of miR-34a-5p, miR124-3p, and miR-125b-5p, while miR-1-3p is under-expressed in dysplasia and early invasion. The malignant early-invasion stage is distinguished by the downregulation of miR-147a and the overexpression of miR-155-5p, miR-423-3p, and miR-34a-5p. The identification of stage-specific miRNAs may facilitate their utilization as biomarkers for presymptomatic OSCC diagnosis.

Oral squamous cell carcinoma (OSCC) is a highly prevalent and aggressive malignancy, with mortality rates that reach 60%, mainly due to an excessive diagnostic delay. MiRNAs, a class of crucial epigenetic gene-expression regulators, have emerged as potential diagnostic biomarkers, with >200 molecules reported to be dysregulated in OSCC. We had previously established an in silico methodology for the identification of the most disease-specific molecules by bridging genetics and epigenetics. Here, we identified the stage-specific miRNAs that govern the asymptomatic early stages of oral tumorigenesis by exploiting the reverse interplay between miRNA-levels and their target-genes’ expression. Incorporating gene expression data from our group’s experimental hamster model of sequential oral oncogenesis, we bioinformatically detected the miRNAs that simultaneously target/regulate >75% of the genes that are characteristically upregulated or downregulated in the consecutive stages of hyperplasia, dysplasia and early-invasion OSCC, while exhibiting the opposite expressional dysregulation in OSCC-derived tissue and/or saliva specimens. We found that all stages share the downregulation of miR-34a-5p, miR124-3p, and miR-125b-5p, while miR-1-3p is underexpressed in dysplasia and early-invasion. Malignant early-invasion stage is distinguished by the downregulation of miR-147a and the overexpression of miR-155-5p, miR-423-3p and miR-34a-5p. The identification of stage-specific miRNA molecules may facilitate their utilization as biomarkers for presymptomatic OSCC diagnosis.