PDK1- and PDK2-mediated metabolic reprogramming contributes to the TGFß1-promoted stem-like properties in head and neck cancer.

BACKGROUND: Resistance to chemotherapeutic drugs is a key factor for cancer recurrence and metastases in head and neck cancer (HNC). Cancer stem cells (CSCs) in tumors have self-renewal, differentiation, and higher drug resistance capabilities, resulting in a poor prognosis for patients. In glucose metabolism, pyruvate dehydrogenase kinase (PDK) inhibits pyruvate dehydrogenase and impedes pyruvate from being metabolized into acetyl-CoA and entering the tricarboxylic acid cycle to generate energy. Studies have reported that PDK1 and PDK2 inhibition suppresses the growth, motility, and drug resistance of cancer cells. Furthermore, while TGFß1 levels are persistently elevated in HNC patients with poor prognosis, the role of PDK isoforms in the TGFß1-promoted progression and stem-like properties of HNC is unclear. METHODS: Levels of PDK1 and PDK2 were evaluated in HNC tissue microarrays by immunohistochemistry to explore potential clinical relevance. PDK1 and PDK2 were knocked down by the lentivirus shRNA system to investigate their role in TGFß1-promoted tumor progression in vitro. RESULTS: We found that PDK2 levels were increased in the later stage of HNC tissues compared to constant PDK1 expression. After PDK1 and PDK2 knockdown, we discovered increased ATP production and decreased lactate production in TGFß1-treated and untreated HNC cells. However, only PDK2 silencing significantly inhibited the clonogenic ability of HNC cells. We subsequently found that TGFß1-promoted migration and invasion capabilities were decreased in PDK1 and PDK2 knockdown cells. The tumor spheroid-forming capability, motility, CSC genes, and multidrug-resistant genes were downregulated in PDK1 and PDK2 silencing CSCs. PDK1 and PDK2 inhibition reversed cisplatin and gemcitabine resistance of CSCs, but not paclitaxel resistance. CONCLUSION: The results demonstrated that the PDK1- and PDK2-mediated Warburg effect contributes to the TGFß1-enhanced stemness properties of HNC. Therefore, PDK1 and PDK2 may serve as molecular targets for the combination therapy of HNC.

The Efficacy of DW and T1-W MRI Combined with CT in the Preoperative Evaluation of Cholesteatoma.

OBJECTIVE: This study aims to assess the efficacy of diffusion-weighted (DW) and T1-weighted (T1W) magnetic resonance imaging (MRI) combined with high-resolution computed tomography (HRCT) (together as DW-T1W-CT) in the preoperative evaluation of the presence and extent of cholesteatoma, which helps determine whether a patient is suitable for transcanal endoscopic ear surgery (TEES). METHODS: This retrospective study included 35 patients (18 male and 17 female) aged from 2 to 81 years diagnosed with chronic otitis media with or without cholesteatoma, who had received surgical treatment and a preoperative MRI and HRCT during the period of December 2015 to December 2020 at Cathay General Hospital. We compared the preoperative DW-T1W-CT findings with the intraoperative findings and final pathologic diagnosis. The accurate predictive value was evaluated using the presence of cholesteatoma and its extent. RESULTS: Regarding the efficacy of detecting cholesteatoma, we found a sensitivity of 92% (23/25 cases with cholesteatoma), a specificity of 90% (9/10 cases without cholesteatoma), and an overall accurate predictive value of 91.4% (32/35) by using combined DW-T1W-CT imaging. With regard to evaluating the extent of cholesteatoma, the combined DW-T1W-CT images obtained an accurate predictive value of 84% (21/25 cases of cholesteatoma). CONCLUSION: Combined DW-T1W-CT has been proven to be a reliable tool in detecting the presence of cholesteatoma. It is also useful in preoperatively depicting the extent of cholesteatoma, which is crucial for determining whether a patient is suitable for TEES, aiding in surgical planning and patient consultation.

CXCR3 isoform A promotes head and neck cancer progression by enhancing stem-like property and chemoresistance.

BACKGROUND: The chemokine network orchestrates the cancer stem-like property and consequently participates in cancer progression. CXCR3 contributes cancer progressive property and immunomodulation in the tumor microenvironment. The two major isoforms of CXCR3 are scrutinized and the divergence is showed that CXCR3A promotes cancer cell growth and motility while CXCR3B functions contrarily in many studies. However, rare studies illustrate the role of CXCR3 isoforms in cancer stem-like property and chemoresistance, especially in head and neck cancer (HNC). METHODS: Levels of CXCR3, CXCR3B, and Sox2 were determined in HNC tissue microarray by immunohistochemistry staining to explore potential clinical relevance. Lentivirus-mediated CXCR3-isoform overexpression with MTS assay, clonogenic assay, transwell migration, sphere formation, and chemo-drug susceptibility were implemented to investigate the role of CXCR3-isofoms in HNC. RESULTS: High levels of CXCR3 were significantly associated with advanced stage (p < 0.01), regional lymph node metastasis (p < 0.05), and poor differentiation (p < 0.005) and further correlated with worse survival rate in oral cancer patients (p = 0.036). Higher levels of CXCR3B were found in regional lymphatic invasion of HNC and progressive stage of squamous cell carcinoma. Elevated Sox2 expression was significantly associated with the advanced stage of HNC in the oral cavity, and demonstrated a co-expression pattern with CXCR3B. Furthermore, lentivirus-mediated overexpression of CXCR3A and CXCR3B in SAS human oral cancer cells promoted cell mobility. CXCR3A overexpression enhanced sphere-forming ability and chemoresistance of CSCs by upregulating stemness-related genes. CONCLUSION: This study first provides a novel insight of CXCR3 isoform A in HNC cancer progression via regulating cancer stem-like properties and chemoresistance, suggesting that CXCR3A may be a prognostic marker and novel target for HNC therapy.

The anatomic applicability of transcanal endoscopic ear surgery in children.

OBJECTIVE: The feasibility of TEES in dealing with pediatric middle ear disease is still under investigation. The goal of this study was to compare anatomical dimensions of the EAC between children and adults, and to analyze the anatomic applicability of TEES in children. METHODS: Forty pediatric (18 years old and younger) and 20 adult patients who received TB-HRCT to evaluate middle ear conditions were enrolled from December 2010 to December 2015. Dimensions including the diameters of the EAC orifice, isthmus and tympanic membrane annulus, the length of EAC, the height of the middle ear, and the angle between tympanic membrane and EAC axis were determined according to the TB-HRCT data. RESULTS: The diameters of the EAC orifice and isthmus and length of EAC in the pediatric patients were slightly smaller than those in the adult patients. The anatomical dimensions of middle ear were similar in both groups. Simple regression analysis indicated that the diameters of the EAC orifice and isthmus and the length of the EAC were positively correlated with the age of the pediatric patients. In the pediatric patients, 67 ears (83.75%) had an EAC isthmus diameter larger than 4 mm and are sufficient with a 3-mm endoscope manipulation. CONCLUSION: TEES is applicable for most pediatric patients using an endoscope with a diameter of 3 mm or smaller. With an appropriate endoscope and instruments, TEES is a safe and effective alternative to treat pediatric middle ear disease.

Characterization of the structure-function relationship at the ligament-to-bone interface.

Soft tissues such as ligaments and tendons integrate with bone through a fibrocartilaginous interface divided into noncalcified and calcified regions. This junction between distinct tissue types is frequently injured and not reestablished after surgical repair. Its regeneration is also limited by a lack of understanding of the structure-function relationship inherent at this complex interface. Therefore, focusing on the insertion site between the anterior cruciate ligament (ACL) and bone, the objectives of this study are: (i) to determine interface compressive mechanical properties, (ii) to characterize interface mineral presence and distribution, and (iii) to evaluate insertion site-dependent changes in mechanical properties and matrix mineral content. Interface mechanical properties were determined by coupling microcompression with optimized digital image correlation analysis, whereas mineral presence and distribution were characterized by energy dispersive x-ray analysis and backscattered scanning electron microscopy. Both region- and insertion-dependent changes in mechanical properties were found, with the calcified interface region exhibiting significantly greater compressive mechanical properties than the noncalcified region. Mineral presence was only detectable within the calcified interface and bone regions, and its distribution corresponds to region-dependent mechanical inhomogeneity. Additionally, the compressive mechanical properties of the tibial insertion were greater than those of the femoral. The interface structure-function relationship elucidated in this study provides critical insight for interface regeneration and the formation of complex tissue systems.

Characterization of the mechanical properties and mineral distribution of the anterior cruciate ligament-to-bone insertion site.

The anterior cruciate ligament (ACL) connects the femur to the tibia through direct insertion sites and functions as the primary restraint to anterior tibial translation. The ACL-to-bone insertion sites exhibit a complex structure consisting of four zones of varied cellular and matrix components, consisting of ligament, non-mineralized fibrocartilage, mineralized fibrocartilage and bone, which allow for the effective load transfer from ligament to bone, thereby minimizing stress concentrations and preventing failure. The mineral content and distribution within the fibrocartilage region may be an important structural component of the insertion site which may influence the mechanical properties. The goals of this study are to characterize the compressive mechanical properties of the fibrocartilage region of the ACL-to-bone insertion site and evaluate how the mineral distribution at the interface relates to these compressive properties. In order to determine the compressive mechanical properties we have utilized a novel microscopic mechanical testing method combined with digital image correlation and employed energy dispersive X-ray analysis (EDAX) in order to evaluate the mineral content and distribution across the femoral and tibial insertion sites. The results reveal that a regional mineral gradient is observed across the fibrocartilage which corresponds to depth-dependent variations in compressive mechanical properties. This depth- dependent mechanical inhomogeneity strongly correlates to the increase in mineral content of the mineralized fibrocartilage (MFC) region compared to the non-mineralized fibrocartilage (NFC). Additionally, the tibial NFC and MFC mechanical properties are greater than those of the femoral NFC and MFC which corresponds to a greater mineral content in the NFC and MFC regions of the tibial insertion. The findings of this study suggest that a structure-function relationship exists at the ACL-to-bone interface.